Pharmaceutical compounds

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula (I) or pharmaceutically acceptable salts, solvates or tautomers thereof, where substitute M is selected from groups D1 and D2, having structural formulae given below, and R1, E, A and X are as described in the formula of invention. Disclosed also are pharmaceutical compositions which contain these compounds, methods for synthesis of these compounds, intermediate compounds and synthesis methods thereof, as well as use of compounds of formula (I) in preventing or treating diseases mediated by CDK kinases, GSK-3 kinases or Aurora kinases.

EFFECT: high effectiveness of the compounds.

40 cl, 8 dwg, 18 tbl, 84 ex

 

The invention relates to the derivatives of pyrazole, which inhibit or modulate the activity of cyclin-dependent kinases (CDK), kinases glycogen synthase (GSK) and Aurora kinases, the use of these compounds in the treatment or prevention mediated by kinases diseases or conditions, as well as to new compounds with inhibitory or modulating activity against kinases. In addition, the pharmaceutical compositions containing these compounds, as well as new chemical intermediate compounds.

BACKGROUND of the INVENTION

Protein kinases constitute a large family of structurally related enzymes that are responsible for managing a variety of processes transmission signals in the cell (Hardie, G. and Hanks, S. (1995) The Protein Kinase Facts Book. I and II, Academic Press, San Diego, CA). Kinases can be classified into families on the substrates they phosphorylate (e.g., protein-tyrosine, protein-serine/threonine, lipids, etc). Were identified characteristic fragments of the sequences, which generally correspond to each of these families of kinases (e.g., Hanks, S.K., Hunter, T., FASEB J. 9:576-596 (1995); Knighton, et al., Science, 253: 407-414 (1991); Hiles, et al., Cell, 70: 419-429 (1992); Kunz, et al., Cell, 73: 585-596 (1993); Garcia-Bustos, et al., EMBO J. 13: 2352-2361 (1994)).

Protein kinases can be characterized by the mechanisms of their regulatory actions. These mechanisms on the hunger for example, autophosphorylation, transphosphorylation other kinases, protein interaction protein interaction protein-lipid and protein interaction-polynucleotide. Individual protein kinases can be regulated by more than one mechanism.

Kinases regulate many different cellular processes, including, but not limited to, proliferation, differentiation, apoptosis, motility, transcription, translation and other processes of the transmission signal, by attaching phosphate groups to target proteins. These acts of phosphorylation act as molecular switches on/off, which can modulate or regulate the biological function of the target protein. Phosphorylation of target proteins occurs in response to various extracellular signals (hormones, neurotransmitters, growth factors and differentiation etc), events of the cell cycle, stress, environmental or food etc. of the protein kinases function in signal transduction pathways are to activate or inactivate (either directly or indirectly), for example, a metabolic enzyme regulatory protein, a receptor, a protein of cytoskeleton, ion channel or pump, or a transcription factor. Unmanaged signal transmission due to impaired control phosphorylation of proteins including both the who and a number of diseases, including, for example, inflammation, cancer, Allergy/asthma, disease and immune system diseases and conditions of the Central nervous system and angiogenesis.

Cyclin-dependent kinase

The process of dividing eukaryotic cells can be broadly divided into a series of successive phases called G1, S, G2 and M. it Was shown that the correct passing through different phases of the cell cycle depends critically on the spatial and temporal regulation of a family of proteins known as cyclin-dependent kinase (CDK) and a diverse group related protein partner called cyclename. CDK are CDC2 (also known as CDK1)-homologous protein serine-threonine kinases that are able to use ATP as a substrate in the phosphorylation of various polypeptides depending on the surrounding sequence. Cycline represent a family of proteins characterized by a homologous region containing approximately 100 amino acids and is called "cyclenbuy Boxing", which is used to bind with specific CDK protein partners and determines the selectivity towards them.

Modulation of expression levels, speeds decomposition and activation levels of different CDK and tsiklonov during the cell cycle leads to the formation of cyclename number of complexes of CDK/cyclin, the cat is ryh CDK enzyme are active. The formation of these complexes controls the passage through a separate checkpoint of the cell cycle and, thus, makes possible the continuation of the process of cell division. Failure to meet pre-established biochemical criteria in this checkpoint of the cell cycle, i.e. the impossibility of the formation of the desired complex of CDK/cyclin, can lead to cell cycle arrest and/or cell apoptosis. Abnormal cell proliferation, which occurs when cancer can often be attributed to the loss of proper control of the cell cycle. Therefore, inhibition of the enzymatic activity of the CDK provides the means by which you can stop the division of abnormally dividing cells or kill them. The diversity of CDK and CDK complexes and their crucial role in mediating cell cycle provide a wide range of potential therapeutic targets selected on the basis of the specific biochemical study.

The movement from phase G1 to the S phase of the cell cycle is mainly regulated by CDK2, CDK3, CDK4 and CDK6 by binding with cyclename types D and E. Collini type D, apparently, make it possible to pass through the limiting point G1, whereas the complex CDK2/cyclin E is the key to the transition from phase G1 to phase s Further passage through the S phase and entry into G2, it is to be considered, requires complex CDK2/cyclin A. mitosis, and the transition from the G2 phase to the M phase, which runs it, are regulated by CDK1 complexes and tsiklonov types A and B.

During the G1 phase protein retinoblastoma (Rb) and the related pocket proteins ("pocket" proteins), such as, for example, p130, are substrates for CDK complexes(2, 4 and 6)/cyclin. The passage through the G1 phase partially facilitated by hyperphosphorylation and, thus, inactivation of Rb and p130 complexes of CDK(4/6)/cyclin-D. Hyperphosphorylated Rb and p130 triggers the release of transcription factors such as E2F, and, thus, gene expression, such as gene cycline E required for passage through G1 and entry into S-phase. Expression cycline E facilitates the formation of a complex of CDK2/cyclin E, which increases or maintains the levels of E2F by additional phosphorylation of Rb. The complex CDK2/cyclin E also phosphorylates other proteins required for DNA replication, such as NPAT, which is involved in the biosynthesis of histone. The passage G1 and the transition G1/S are also regulated mitogen-stimulated signalling by Myc, which is included in the signal path of the complex CDK2/cyclin E. in Addition, CDK2 associated with mediated p53 by response to DNA damage by regulating the levels of p21 under the action of p53. p21 is a protein inhibitor complex CDK2/cyclin E and due to this he ways the Yong to block or delay the transition G1/S. The complex CDK2/cyclin E may, therefore, be a point at which, to some extent, the United biochemical incentives for signaling pathways Rb, Myc and p53. Therefore, CDK2 and/or a complex of CDK2/cyclin E are good targets for therapeutic methods designed to stop the cell cycle or resume its control in abnormally dividing cells.

The exact role CDK3 in the cell cycle is unclear. Still not been identified related cyclenbuy partner, but a dominant negative form of CDK3-delayed G1 cells is, thus, to assume that CDK3 plays a role in managing the transition G1/S.

Though most of the cdks involved in cell cycle regulation, there is evidence that some members of the CDK family involved in other biochemical processes. An example of such CDK CDK5 is required for proper development of neurons and, in addition, involved in the phosphorylation of several neural proteins, such as Tau, NUDE-1, synapsin, DARPP32 and complex Munc18/syntaxin. Neuronal CDK5 is activated by binding with proteins p35/p39. However, the activity of CDK5 may be rosregulirovanie binding p25, i.e. a truncated version of the p35. Conversion of p35 to p25 and disrupting the regulation of CDK5 activity may be caused by ischemia, ek is the cytotoxicity and β-amyloid peptide. Therefore, p25 involved in the pathogenesis of neurodegenerative diseases such as Alzheimer's disease and, therefore, is of interest as a target for therapeutic methods directed against these diseases.

CDK7 is a nuclear protein that has CDC2 CAK activity, and is associated with cyclin H. CDK7 was identified as a component of the TFIIH transcription complex, which had activity against C-terminal domain (CTD) of RNA polymerase II. This was associated with the regulation of transcription of HIV-1 by Tat-mediated biochemical signaling pathways. CDK8 binds cyclin C and is involved in the phosphorylation of CTD of RNA polymerase II. Similarly, complex CDK9/cyclin T1 (complex P-TEFb) is involved in the control of elongation of RNA polymerase II. In addition, P-TEFb is required for activation of transcription of the genome of HIV-1 viral transactivator Tat due to its interaction with cyclin T1. Therefore, CDK7, CDK8, CDK9 and complex P-TEFb are potential targets for ways antiviral therapy.

The mediating activity of the complex of CDK/cyclin at the molecular level requires a series of acts of stimulating and inhibitory phosphorylation or dephosphorylation. The CDK phosphorylation was carried out by a CDK-activating kinase (CAK) and/or kinases such as wee1, Myt1 and Mik1. D. the phosphorylation is carried out such phosphatase, as cdc25(a and c), pp2a or KAP.

Further, the activity of the complex of CDK/cyclin may be regulated by two families of endogenous cellular protein inhibitors: a family of Kip/Cip or family INK. Proteins INK-specific bind CDK4 and CDK6. p16ink4(also known as MTS1) is a genome, the vast potential of the tumor, which is subjected to mutation or deleted in a large number of primary cancers. The family of Kip/Cip includes such proteins as, for example, p21Cip,Waf1, p27Kip1and p57Kip2. As discussed earlier, p21 is induced by p53, and he is able to inactivate the complexes of CDK2/cyclin E/A) and CDK4/cyclin D1/D2/D3). Atipicheskie low levels of p27 expression was observed in cancer of the breast, colon and prostate. In contrast, overexpression of cycline E in solid tumors, have been shown to correlate with poor patient prognosis. Excessive expression cycline D1 was associated with carcinoma of the esophagus, breast, squamous carcinoma and non-small cell carcinoma of the lung.

The key role of cdks and their associated proteins in coordinating and promoting cell cycle in proliferating cells was highlighted above. In addition, they described some of the biochemical signaling pathways that cdks play a key role. Therefore, development of methods monotherapy for the treatment of proliferative disorders, to the to, for example, various types of cancer, using a variety of therapies aimed at CDK in General, or on specific CDK, is potentially highly desirable. The CDK inhibitors, presumably, could also be used to treat other conditions, as, for example, among other things, viral infections, autoimmune diseases and neurodegenerative diseases. In addition, therapeutic methods aimed at CDK, can provide clinical benefits in the treatment of the above diseases when used in the framework of combination therapy in conjunction with either existing or new therapy. Aimed at CDK ways anticancer therapy could potentially have advantages over many existing anticancer remedies because they do not interact directly with DNA and, therefore, would reduce the risk of development of secondary tumors.

Diffuse B-both lymphoma (DLBCL)

The development of the cell cycle is regulated by the combined action of tsiklonov, cyclin-dependent kinases (CDK) and CDK inhibitors (CDKi), which are negative regulators of the cell cycle. p27KIP1 is a key CDKi in cell cycle regulation, and its disintegration necessary for the transition G1/S. Despite the absence of p27KIP1 expression in proliferating is imposita, it was reported that some aggressive B-cell lymphoma demonstrate abnormal staining of p27KIP1. Abnormally high expression of p27KIP1 was detected in lymphomas of this type. The analysis of the clinical significance of these data showed that a high level of p27KIP1 expression in tumors of this type is a bad prognostic sign as in the univariate and multivariate analysis. These results show that in diffuse B-both lymphoma (DLBCL) have abnormal expression of p27KIP1, with adverse clinical significance, on the assumption that this abnormal protein p27KIP1 may operate in a non-functional state upon interaction with other proteins that regulate the cell cycle (Br. J. Cancer. 1999 Jul; 80(9): 1427-34. p27KIP1 abnormally expressed in diffuse B-both lymphoma, and it is associated with adverse clinical outcome. Saez A, Sanchez E, Sanchez-Beato M, Cruz MA, Chacon I, Munoz E, Camacho FI, Martinez-Montero JC, Mollejo M, Garcia JF, Piris MA. Department of Pathology, Virgen de la Salud Hospital, Toledo, Spain).

Chronic lymphocytic leukemia

B-cell chronic lymphocytic leukemia (CLL) is the most common type of leukemia in the Western hemisphere, and are diagnosed each year approximately 10,000 new cases (Parker SL, Tong T, Bolden S, Wingo PA: Cancer statistics, 1997. Ca. Cancer. J. Clin. 47:5, (1997)). In relation to other forms of leukemia prognosis CLL in General, the om is favorable, even patients with the most advanced stage have a life expectancy equal to 3 years.

The inclusion of fludarabine in the primary therapy of patients with symptomatic CLL led to higher complete response (27% vs. 3%) and life expectancy without causing disease (33 months versus 17) compared to earlier methods of treatment based on alkylating funds. Although achieving a complete clinical response after therapy is the first step toward improved survival in CLL, the majority of patients either do not achieve complete remission, or do not respond to fludarabine. In addition, all patients with CLL, which were treated with fludarabine, in the end there was a relapse, making the role of fludarabine as the sole means of treatment is purely palliative care (Rai KR, Peterson B, Elias L, Shepherd L, Hines J, Nelson D, Cheson B, Kolitz J, Schiffer CA: A randomized comparison of fludarabine and chlorambucil for patients with previously untreated chronic lymphocytic leukemia. A SWOG CALGB, CTG/NCI-C and ECOG Inter-Group Study. Blood 88:141a, 1996 (abstr 552, suppl 1). Therefore, if must realize further progress in the treatment of this disease, it will be necessary to identify new tools with new mechanisms of action that complement cytotoxicity of fludarabine and eliminate the resistance of CLL to the action of medicines caused by internal factors.

Most sh is widely studied and permanent predictive power factor of poor response to therapy and low survival of CLL patients is abnormal p53 function, characterized by point mutations or deletions of chromosome 17p13. Really the actual lack of response to therapy as alkylating drugs, so-and purine analogues, such CLL-patients with abnormal p53 function has been documented in several separate set of histories. Development of a therapeutic agent, which is able to overcome drug resistance associated with p53 mutation in CLL, could potentially become a significant progress in the treatment of this disease.

Flavopiridol and CYC 202, i.e. inhibitors of cyclin-dependent kinases, induce in vitro apoptosis of malignant cells of B-cell chronic lymphocytic leukemia (B-CLL).

The effect of flavopiridol leads to the stimulation of the activity of caspase 3 and dependent on caspase cleavage of p27(kip1), i.e., a negative regulator of the cell cycle, which is redundantly expressed in B-CLL (Blood. 1998 Oct 15;92(10):3804-16 Flavopiridol dosage apoptosis in chronic lymphocytic leukemia cells via activation of caspase-3 without evidence of bcl-2 modulation or dependence on functional p53. Byrd JC, Shinn C, Waselenko JK, Fuchs EJ, Lehman TA, Nguyen PL, Flinn IW, Diehl LF, Sausville E, Grever MR).

Aurora kinase

Relatively recently been discovered a new family of serine/threonine kinases, known as Aurora kinases, which are involved in phase G2 and M cell cycle, and which are important regulators of mitosis. That is the main role of Aurora kinases have yet to be explained, but they participate in the regulation of checkpoint mitosis, the dynamics of chromosomes and cytokinesis (Adams et al., Trends Cell Biol., 11: 49-54 (2001)). Aurora kinases are at the centrosomes of interphase cells at the poles of the bipolar spindle and the cell plate in the plane of the equator of the mitotic spindle apparatus.

To date in mammals was discovered three members of the Aurora family of kinases (E.A. Nigg, Nat. Rev. Mol. Cell Biol. 2: 21-32, (2001)). Namely:

Aurora A (also referred to in the literature Aurora 2);

Aurora B (also referred to in the literature Aurora 1); and

Aurora C (also referred to in the literature Aurora 3).

Aurora kinases include vysokomolochnye catalytic domains, but differ significantly in their N-terminal parts (Katayama H, Brinkley WR, S. Sen; The Aurora kinases: role in cell transformation and tumorigenesis; Cancer Metastasis Rev. 2003 Dec; 22(4):451-64).

It was found that the substrates of Aurora kinases A and B include similar to kinesin motor proteins, proteins of the spindle apparatus, protein histone H3, a protein of kinetochore and suppressing tumor protein p53.

I believe that Aurora A kinase involved in spindle formation and during the early phase G2 localized at the centrosome, where they phosphorylate proteins associated with the spindle (Prigent et al., Cell 114:531-535 (2003). Hirota and co-authors (Hirota et al, Cell, 114: 585-598, (2003)) found that cells depleted of Aurora A protein kinase, were unable to start mitosis. In addition, it was found (Adams, 2001)that the mutation is whether the destruction of Aurora A, A gene in different species leads to mitotic abnormalities, including the separation of centrosomes and defects of maturation, spindle abnormalities and defects in the segregation of chromosomes.

Aurora kinase, usually expressed in small amounts in most normal tissues, and the exceptions are fabrics with a high proportion of dividing cells, such as the thymus gland and the testes. However, elevated levels of Aurora kinases have been detected in most cancer man (Giet et al., J. Cell. Sci. 112: 3591-361, (1999) and Katayama (2003)). In addition, Aurora A kinase displays a region of chromosome 20ql3, which, as is often detected, amplified in many cancers person.

For example, a significant overexpression of Aurora A has been detected in cancers of the breast, ovary and pancreas (see Zhou et al, Nat. Genet. 20: 189-193, (1998), Tanaka et al., Cancer Res., 59: 2041-2044, (1999) and Han et al, cancer Res., 62: 2890-2896, (2002).

In addition, Isola, American Journal of Pathology 147, 905-911 (1995) reported that the amplification of the locus Aurora A (20ql3) correlates with poor prognosis for patients with breast cancer without lymph nodes.

Amplification and/or overexpression of Aurora A is observed in cancerous tumors of the human urinary bladder, and amplification of Aurora A is associated with aneuploidy and aggressive clinical behavior, see Sen et al., J. Natl. Cancer Inst, 94: 1320-1329 (2002).

Increased expression of Aurora A was detected in more than 50% of cases Colo is challnege cancer (see Bischoff et al., EMBO J. 17: 3052-3065, (1998) and Takahashi et al., Jpn. J. Cancer Res., 91: 1007-1014 (2000)), ovarian cancer (see Gritsko et al., Clin. Cancer Res., 9: 1420-1426 (2003)) and cancer of the stomach (Sakakura et al., British Journal of Cancer, 84: 824-831 (2001)).

Tanaka et al., Cancer Research, 59: 2041-2044 (1999) found evidence of excessive expression of Aurora A in 94% of cases of invasive adenocarcinoma of the ducts of the breast.

In addition, high levels of Aurora A kinase were detected in cell lines tumors of the kidney, cervix, pancreas, and prostate, as well as cell lines, neuroblastoma, melanoma, lymphoma Bischoff et al. (1998), EMBO J., 17: 3052-3065 (1998); Kimura et al. J. Biol. Chem., 274: 7334-7340 (1999); Zhou et al., Nature Genetics, 20: 189-193 (1998); Li et al., Clin Cancer Res. 9 (3): 991-7 (2003)].

Aurora B in large quantities is expressed in many cell lines of human cancers, including leukemia cells [Katayama et al., Gene 244:1-7]. In primary colorectal cancers levels of this enzyme increase as a function of the stage at the Duke [Katayama et al., J. Natl Cancer Inst., 91: 1160-1162 (1999)].

High levels of Aurora-3 (Aurora-C) were detected in several lines of tumor cells, even though in normal tissues, the presence of this kinase tends to be limited to the germ cells (see Kimura et al., Journal of Biological Chemistry, 274: 7334-7340 (1999)). In addition, Takahashi et al., Jpn. J. Cancer Res., 91: 1007-1014 (2001) reported excessive expression of Aurora-3 approximately 50% of cases to lorescale cancer.

Other messages about the role of Aurora kinases in proliferative disorders can be found in Bischoff et al., Trends in Cell Biology 9: 454-459 (1999); Giet et al. Journal of Cell Science, 112: 3591-3601 (1999) and Dutertre, et al. Oncogene, 21: 6175-6183 (2002).

Royce and staff reported that the expression of Aurora 2 gene (known as STK15 or BTAK) was reported in approximately one-fourth of cases of primary breast tumors (Royce ME, Xia W, Sahin AA, Katayama H, Johnston DA, Hortobagyi G, Sen S, Hung MC; STK15/Aurora-A expression in primary breast tumours is correlated with nuclear grade but not with prognosis; Cancer. 2004 Jan 1; 100(1): 12-9).

Endometrial carcinoma (EC) includes at least two types of cancer: endometrial carcinoma (EEC) is a tumor associated with estrogen, which are often uploadname and have a favorable prognosis. Endometrioid carcinoma (NEEC; serous and clear forms) are not associated with estrogen, often aneuploidy and are clinically aggressive. It was also discovered that Aurora was increased by 55.5% at NEEC, but not at all EEC (P≤0,001) (Moreno-Bueno G, Sanchez-Estevez C, Cassia R, Rodriguez-Perales S, Diaz-Uriarte R, Dominguez O, Hardisson D, Andujar M, Prat J, Matias-Guiu X, Cigudosa JC, Palacios J. Cancer Res. 2003 Sep 15; 63(18): 5697-702).

Reichardt and co-authors (Oncol Rep. 2003 Sep-Oct; 10 (5): 1275-9) reported that quantitative DNA analysis using PCR to search for amplification of Aurora in gliomas showed that five of the 16 tumors (31%) in different stages according to the who classification (1 in stage II, 1 stage III, 3 in stage IV) show the if amplification DNA gene Aurora 2. We formulated the hypothesis that the Aurora 2 gene amplification can be non-random genetic changes in human gliomas, which plays a role in the genetic pathways of tumor development.

The results Hamada and employees (Br. J. Haematol. 2003 May; 121 (3): 439-47) also suggest that Aurora 2 is an effective candidate to identify not only the activity of the disease, but also ways of formation of non-Hodgkin lymphoma. Slowing the growth of tumor cells, which is a limitation of the functions of this gene could be a therapeutic approach to the treatment of non-Hodgkin lymphoma.

In the study carried out Gritsko and co-authors (Clin Cancer Res. 2003 Apr; 9 (4): 1420-6) in 92 patients with primary ovarian tumors were investigated activity of kinases and protein levels of Aurora A. Analysis of kinases in vitro revealed the increased activity of Aurora A kinase in 44 cases (48%). Elevated protein levels of Aurora A were found in 52 samples (57%). High levels of Aurora A, A protein correlated well with increased kinase activity.

The results obtained by Li and co-authors (Clin. Cancer Res. 2003 Mar; 9(3): 991-7), showed that the gene Aurora A is subjected to excessive expression in tumors of the pancreas and the lines of cancer cells, and suggested that increased expression of Aurora A may play a role in the formation of cancerous tumors of the pancreas.

Analogic what was shown that gene amplification of Aurora A and the associated increase in expression of mitotic kinases, which encodes the gene, is associated with aneuploidy and aggressive clinical behavior of bladder cancer man (J. Natl. Cancer Inst. 2002 Sep 4; 94 (17): 1320-9).

The research of several groups (Dutertre S, Prigent C., Aurora-A overexpression leads to override of the microtubule-kinetochore attachment checkpoint; Mol. Interv. 2003 May; 3(3): 127-30 and Anand S, Penrhyn-Lowe S, Venkitaraman AR., Aurora-A amplification overrides the mitotic spindle assembly checkpoint, inducing resistance to Taxol, a Cancer Cell. 2003 Jan; 3 (1): 51-62) suggest that overexpression of the activity of Aurora kinase is associated with resistance to some of these modern methods of cancer treatment. For example, overexpression of Aurora A in fibroblasts of mouse embryos can lower the sensitivity of these cells to the cytotoxic effect of derivative taxane. Therefore, inhibitors of Aurora kinases may find particular use in patients with developed resistance to existing therapies.

On the basis of research carried out to date, it appears that inhibition of Aurora kinases, in particular Aurora kinase A and Aurora kinase B, will provide an effective means of slowing down the development of tumors.

Harrington and employees (Nat Med. 2004 Mar; 10 (3) 262-7) demonstrated that the inhibitor of the Aurora kinases, suppresses tumor growth and induces tumor regression in vivo. In this study, the inhibitor of Aurora kinase BC who was Kirova proliferation of cancer cells, and, in addition, has launched the process of cell death in a number of lines of cancer cells, including leukemia and colorectal cell lines and cell line of breast cancer. In addition to this, the possibility of treatment of leukemia by inducing apoptosis of leukemia cells. VX-680 effectively kills resistant to treatment of cells with primary acute myelogenous leukemia (AML) patients (Andrews, Oncogene, 2005, 24, 5005-5015).

Cancers that may be particularly exposed to inhibitors of Aurora include cancer of the breast, bladder, colorectal cancer, pancreatic cancer, ovarian cancer, nahodkinskuju lymphoma, glioma and endometrioid endometrial carcinoma. Types of leukemia are particularly susceptible to the action of Aurora inhibitors include acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), B-cell lymphoma (mantle cell tissue) and acute lymphoblastic leukemia (ALL).

Kinase glycogen synthase

Kinase-3 glycogen synthase (GSK3) is a serine-threonine kinase, which is found in humans in the form of two isoforms expressed in all tissues (GSK3α and GSK3β). GSK3 were involved in the review as the enzyme having a specific role in embryonic development, protein synthesis, cell proliferation, cell differentiation, dynamics microcannulas, to emochnoy motility and cell apoptosis. As such, GSK3 is involved in the development of such illnesses as diabetes, cancer, Alzheimer's, stroke, epilepsy, amyotrophic lateral sclerosis and/or head injury. Phylogenetically GSK3 most closely associated with cyclin-dependent kinases (CDK).

The consensus peptide sequence of the substrate recognized by GSK3 is a sequence (Ser/Thr)-X-X-X-(pSer/pThr), where X represents any amino acid in position (n+1), (n+2), (n+3)) and pSer and pThr are phosphoserine and posttraining, respectively, in position (n+4). GSK3 phosphorylates the first serine or threonine at position (n). Phosphoserine or posttraining in position (n+4)is likely to be required for stimulation of GSK3 to achieve the maximum rate of conversion of substrate. The GSK3α phosphorylation at Ser21 or GSK3β at Ser9 leads to inhibition of GSK3. Studies of mutagenesis and competitive reactions with peptides led to a model in which the phosphorylated N-end GSK3 able to compete with phosphopeptide substrate (S/TXXXpS/pT) due to the mechanism of autoinhibitory. In addition, data is available, giving the opportunity to assume that GSK3α and GSK3β can marginally be regulated by phosphorylation of tyrosine residues 279 and 216, respectively. Mutation of these residues Phe caused a recovery of the activity of the kinase in vivo. The establishment of a crystal is graficheskiy patterns GSK3β using x-rays helped to shed light on all aspects of activation and regulation of GSK3.

GSK3 forms part of a signaling pathway response to insulin in mammals, the enzyme is able to fosforilirovanii and thereby inactivate glycogen synthase. Thus, the increased activity of glycogen synthase and glycogen synthesis, via inhibition of GSK3, was seen as a potential means of combating diabetes type II or non-insulin-dependent diabetes mellitus (NIDDM): a condition in which the body's tissues become resistant to stimulation by insulin. The response of cells to insulin in liver, adipose and muscular tissues is triggered by binding of insulin to the extracellular receptor insulin. This causes the phosphorylation and subsequent admission to the plasmatic membrane protein substrates of the insulin receptor (IRS). Further phosphorylation of IRS proteins triggers the appearance of phosphoinositide-3-kinase (PI3K) to the plasma membrane, where it is capable of releasing second messenger - phosphatidylinositol 3,4,5-triphosphate (PIP3). It promotes localization of 3-phosphoinositide-dependent protein kinase 1 (PDK1) and protein kinase B (PKB or Akt) on the membrane, where PDK1 activates PKB. PKB can fosforilirovanii and thereby to inhibit GSK3α and/or GSK3β by phosphorylation of Ser9 residue or Ser21, respectively. Then, the inhibition of GSK3 initiates an increase in activity and glycogen synthase. Thus, a therapeutic agent capable of inhibiting GSK3, can cause reactions of the cells, similar to the reactions observed upon stimulation with insulin. Additional in vivo substrate of GSK3 is the eukaryotic initiation factor of protein synthesis 2B (eIF2B). eIF2B is inactivated upon phosphorylation and, thus, acquires the ability to inhibit protein biosynthesis. Inhibition of GSK3, for example, by inactivation of protein, representing a "molecule-target of rapamycin in mammals" (mTOR), may, therefore, increase the synthesis of protein. Finally, there is evidence in favor of regulation of activity through GSK3 signaling pathway mitogen-activated protein kinase (MAPK) phosphorylation of GSK3 kinases, such as mitogen-activated protein kinase activated protein kinase (MAPKAP-K1 or RSK). These data suggest that GSK3 activity may be modulated mitogenic, insulin and/or amino acid stimulating effects.

In addition, it was shown that GSK3β is a key component in the Wnt signaling pathway in vertebrates. It was shown that this biochemical pathway is critical for normal embryonic development and regulates cell proliferation in normal tissues. GSK3 is subject to inhibition in response to stimulatory who is step Wnt. This may lead to dephosphorylating substrates GSK3, as, for example, Aksana, i.e. gene product adenomatous polyposis of the colon (APC), and β-catenin. Abnormal regulation of the Wnt pathway has been associated with many cancers. Mutations in APC and/or β-catenin are common in colorectal cancer and other tumors. In addition, it was shown that β-catenin plays an important role in cell adhesion. Thus, GSK3 also, to some extent, can modulate the processes of cell adhesion. Besides the already described biochemical pathways, there are data that suggest the involvement of GSK3 in the regulation of mitosis by phosphorylation cyclina-D1, in the phosphorylation of transcription factors such as c-Jun, CCAAT/enhancer-binding protein α (C/EBPα), c-Myc and/or other substrates, such as nuclear factor of activated T-cells (NFATc), factor heat shock-1 (HSF-1) and protein binding to the c-AMP dependent element (CREB). In addition, GSK3, apparently played a certain, although tissue-specific, role in regulation of apoptosis. The role of GSK3 in the modulation of apoptosis via Pro-apoptotic mechanism may be of particular importance for medical conditions that may be observed apoptosis of neurons. Examples of such conditions include head injury, stroke, epilepsy, Alzheimer's disease and the side Amiot epicheskii sclerosis, progressive supranuclear paralysis, kortiko-basal degeneration and disease Peak. It has been shown in vitro that GSK3 is able to hyperphosphorylate micro canals associated with Tau protein. Hyperphosphorylated Tau-protein stops its normal binding microcanonical and can also lead to the formation of intracellular filaments of Tau protein. I believe that the consistent accumulation of these fibers leads to possible dysfunction and degeneration of neurons. Inhibition of phosphorylation of the Tau protein by inhibiting GSK3 may, therefore, provide means of limiting and/or preventing neurodegenerative effects.

The LEVEL of TECHNOLOGY

In WO 02/34721 filed Du Pont, disclosed class inden[1,2-c]pyrazole-4-ones as inhibitors of cyclin-dependent kinases.

In WO 01/81348 filed by Bristol Myers Squibb, disclosed the use of 5-thio-, sulfinil and sulfonylurea[3,4-b]pyridines as inhibitors of cyclin-dependent kinases.

In WO 00/62778, also filed Bristol Myers Squibb, disclosed class of inhibitors tyrosinemia protein kinase.

In WO 01/72745A1 filed Cyclacel described 2-substituted 4-heteroarylboronic and receive them, containing their pharmaceutical compositions and their use as inhibitors of cyclin-dependent kinases (CDK) and, therefore, their use in the treatment of proliferative disorders such as cancer, leukemia, psoriasis, etc.

p> In WO 99/21845 filed Agouron described derivatives of 4-aminothiazole intended for inhibition of cyclin-dependent kinases (cdks), such as CDK1, CDK2, CDK4 and CDK6. This invention is also directed to therapeutic and prophylactic use of pharmaceutical compositions containing such compounds, and methods of treatment of malignant tumors and other disorders by introducing effective amounts of such compounds.

In WO 01/53274 filed Agouron, as inhibitors of CDK kinase disclosed class of compounds, which may include amazonienne benzene ring is associated with N-containing heterocyclic group.

In WO 01/98290 (Pharmacia & decision Upjohn) disclosed class of derivatives of 3-aminocarbonyl-2-carboxamidine as inhibitors of protein kinases. It is established that these compounds possess activity against a large number of protein kinases.

In WO 01/53268 and WO 01/02369 filed Agouron, discloses compounds that mediate or inhibit cell proliferation by inhibiting protein kinases, such as cyclin-dependent kinases or tyrosine kinases.

In WO 00/39108 and WO 02/00651 (both filed Du Pont Pharmaceuticals) described extensive classes of heterocyclic compounds, which are inhibitors of trypsin-like enzymes proteolytic enzymes, especially factor Xa and thrombin. It is established that these compounds when enemy as anticoagulants or prevention of thromboembolic disorders.

In each of the applications US 2002/0091116 (Zhu et al.), WO 01/1978 and WO 01/64642 revealed different group of heterocyclic compounds that have activity against factor Xa.

In WO 03/035065 (Aventis) revealed extensive class of benzimidazole derivatives as inhibitors of protein kinases, but not disclosed activity against CDK kinases or GSK kinase.

In WO 97/36585 and US 5874452 (application and a patent owned by Merck) disclosed bilaterally compounds that are inhibitors farnesyl transferase.

In WO 03/066629 (Vertex) revealed benzimidazolecarbamic as inhibitors of GSK-3.

In WO 97/12615 (Warner Lambert) discloses the benzimidazole as inhibitors of 15-lipoxygenase.

In WO 2004/54515 (SmithKline Beecham Corporation) disclosed class of benzimidazole as mimetics of thrombopoetin.

In WO 2004/41277 (Merck) disclosed class of aminobenzimidazole as modulators of the androgen receptor.

In WO 2005/028624 (Plexxikon) revealed the skeletons of molecules of compounds with activity against protein kinases.

In the submitted earlier by the authors of the present invention to international patent application PCT/GB2004/002824 (WO 2005/002552) discovered a class of substituted [(1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]ureas as inhibitors of CDK, Aurora kinase and GSK kinase. One of the compounds specifically named and given as an example in WO 2005/002552 is 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea. the experimental section WO 2005/002552 describes the obtaining of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form.

The INVENTION

In the present invention developed compounds that have activity against inhibition or modulation of cyclin-dependent kinases, the activity against inhibition or modulation of kinase-3 glycogen synthase (GSK3), as well as activity against inhibition or modulation of Aurora kinase, and which seem to be relevant in the prevention or treatment of diseases or conditions mediated by these kinases.

For example, it is envisaged that the compounds of the present invention will be applicable to relieve symptoms or reduce the number of cancer cases.

In the first aspect of the invention relates to the compound of formula (I):

or its salts, MES, tautomer or N-oxide,

where M is selected from group D1 group D2:

and where(A) if M is a group D1:

X is selected from O, NH and NCH3;

A is selected from communication and group NR2where R2represents hydrogen or methyl;

E is selected from CH2CH(CN) and C(CH3)2;

R1chosen from:

(i) 3-5-membered cycloalkyl group, optionally substituted by hydroxy, fluorine, amino, methylamino, stands or ethyl;

(ii) 4-6-membered of NASA the military heterocyclic group, comprising 1 or 2 heteroatoms selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4the alkyl, amino or hydroxy; but excluding unsubstituted 4-morpholinyl, unsubstituted tetrahydropyran-4-yl, unsubstituted 2-pyrrolidinyl, and unsubstituted and 1-substituted piperidine-4-yl;

(iii) a 2,5-substituted phenyl group of the formula:

in which: (a) if X represents NH or N-CH3, R3selected from chlorine and cyano; and (b) if X represents O, R3represents CN;

(iv) group CR6R7R8in which each of R6and R7selected from hydrogen and methyl, and R8selected from hydrogen, methyl, C1-4alkylsulfonyl, hydroxymethyl and cyano;

(v) pyridazin-4-ilen group, optionally substituted by one or two substituents selected from methyl, ethyl, methoxy, ethoxy;

(vi) substituted imidazothiazole group in which the substituents are selected from methyl, ethyl, amino, fluorine, chlorine, amino and methylamino; and

(vii) optionally substituted 1,3-dihydroindol-2-ilen, or 2,3-dihydroindol-1-ilen groups, which are optionally present substituents in each case selected from halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, the de C 1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino;

(viii) 3-pyridyl, optionally substituted by one or two substituents selected from hydroxy, halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino; but excluding compounds such as [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amide of 2-oxo-1,2-dihydropyridines-3-carboxylic acid and 2,6-dimethoxy-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]nicotinamide;

(ix) thiomorpholine or its S-oxide or S,S-dioxide, optionally substituted by one or two substituents selected from halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino; and

if E-A represents NR2, R1additionally selected from:

(x) 2-ftoheia, 3-ftoheia, 4-ftoheia, 2,4-dipthera, 3,4-dipthera, 2,5-dipthera, 3,5-dipthera, 2,4,6-tryptophanyl, 2-methoxyphenyl, 5-chloro-2-methoxyphenyl, cyclohexyl, unsubstituted 4-tetrahydropyranyl and tert-butyl;

(xi) is the group NR 10R11where each of R10and R11represents a C1-4alkyl, or R10and R11connected so that NR10R11form a 4-6-membered saturated heterocyclic group optionally containing a second heteroatom in the cycle selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4the alkyl, amino or hydroxy;

(xii) pyridone, optionally substituted by one or two substituents selected from hydroxy, halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino;

when E-A is C(CH3)2NR2or CH2-NR2, R1additionally selected from:

(xiii) unsubstituted 2-furil and 2,6-diphthera; and

when E-A is C(CH3)2NR2, R1additionally selected from:

(xiv) unsubstituted phenyl; and

if E is a CH2, R1additionally selected from:

(xv) unsubstituted tetrahydropyran-4-yl; and

(B) if M is a group D2:

A is selected from communication and group NR2where R2represents hydrogen or methyl;

E is selected from communication, CH2CH(CN) and C(CH3 )2;

R1chosen from:

(xvi) a 2-substituted 3-shriley group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2and 5 - or 6-membered saturated cycle optionally substituted by hydroxy, fluorine, amino, methylamino, stands or ethyl;

(xvii) of 5-substituted 2-shriley group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2and 5 - or 6-membered saturated heterocyclic group optionally substituted by hydroxy, fluorine, amino, methylamino, stands or ethyl; provided that the compound is not [3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amidon 5-piperidine-1-ilmatieteen-2-carboxylic acid;

(xviii) a group of the formula:

/p>

in which R9represents hydrogen, methyl, ethyl or isopropyl; G represents CH, O, S, SO, SO2or NH, and the specified group optionally substituted by one, two or three substituents selected from C1-4hydrocarbide, hydroxy, C1-4hydrocarbonate, fluorine, amino, mono - and di-C1-4alkylamino, and where each of the C1-4hydrocarbonrich and C1-4hydrocarbonoxygen optionally substituted by hydroxy, fluorine, amino, mono - or di-C1-4alkylamino; and

(xix) of 3,5-disubstituted phenyl group of the formula:

in which X is selected from O, NH and NCH3;

(C) if M is a group D1:

and X represents O; A represents a group NR2where R2is hydrogen; E is a bond; and R1represents a 2,6-differenl; then the compound of formula (I) is an acid additive salt selected from salts formed by the acid that is selected from the group consisting of acetic, adipic, alginic, ascorbic (e.g. L-ascorbic), aspartic (e.g. L-aspartic), benzosulfimide, benzoic, camphoric (for example, (+)camphor), capric, Caprylic, carbonic, citric, reklamowa, dodecanol, modellserie, ethane-1,2-disulfonate, econsultancy, fumaric, galatarasay, hentaimovi, Glu is agatonovic, D-gluconic, glucuronic (for example, D-glucuronic), glutamic (for example, L-glutamic), α is oxoglutarate, glycolic acid, hippuric, hydrochloric, isetionate, isoalkanes, breast (e.g., (+)- L-lactic and (±)-DL-lactic), lactobionic, laurylsulphate, maleic, malic, (-)-L-malic, malonic, methansulfonate, mucus, naphthalenesulfonates (for example, naphthalene-2-sulphonic, naphthalene-1,5-disulfonate, nicotine, oleic, orotovoy, oxalic, palmitic, pambou, phosphoric, propionic, sebacinales, stearic, succinic, sulfuric, tartaric (for example, (+)-L-tartaric), titanoboa, toluensulfonate (for example, p-toluensulfonate), valerianic and xinfulai acids.

In one embodiment, the implementation of the group M is a group D1 or D2, defined in the description of the subgroups (A) and (B) of the above formula (I).

In another embodiment, the group M is a group D1 and the compound of formula (I) is an acid additive salt of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea as defined in the description of the subgroup (S) of the above formula (I).

In a separate embodiment, the invention relates to salts of the free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and the spiral is but the lactate specified connection.

In addition, the invention relates to new applications 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its salts (for example, an acid additive salt), solvate, tautomers or N-oxides.

Among other things, the invention also applies to:

The use of compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, for the manufacture of a medicine for the prevention or treatment of a disease or condition mediated by a cyclin dependent kinase or kinase-3 glycogen synthase.

The method of prevention or treatment of a disease or condition mediated by a cyclin dependent kinase or kinase-3, glycogen synthase, and this method includes the introduction to the subject, if necessary, compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application.

- To relieve symptoms or reduce the frequency of occurrence of a disease or condition mediated by a cyclin dependent kinase or kinase-3, glycogen synthase, and this method includes the introduction to the subject, if necessary, compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual p is x examples of compounds of the above formulas, defined in this application.

The method of treatment of a disease or condition comprising or arising from abnormal cell growth in a mammal, and the method includes the administration to a mammal the compound of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, in an amount effective for inhibiting abnormal cell growth.

- To relieve symptoms or reduce the frequency of occurrence of a disease or condition comprising or arising from abnormal cell growth in a mammal, and the method includes the administration to a mammal the compound of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, in an amount effective for inhibiting abnormal cell growth.

The method of treatment of a disease or condition comprising or arising from abnormal cell growth in a mammal, and the method includes the administration to a mammal the compound of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, in an amount effective for inhibiting the activity of CDK kinase (such as CDK1 or CDK2) or kinase-3 is glycogen-synthase.

- To relieve symptoms or reduce the frequency of occurrence of a disease or condition comprising or arising from abnormal cell growth in a mammal, and the method includes the administration to a mammal the compound of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, in an amount effective for inhibiting the activity of CDK kinase (such as CDK1 or CDK2) or kinase-3 glycogen synthase.

- Method of inhibiting cyclin-dependent kinases or kinase-3, glycogen synthase, and the method includes bringing in the interaction of the indicated kinase C inhibitory kinase compound of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application.

- Method of modulating a cellular process (for example cell division) by inhibiting the activity of cyclin-dependent kinases or kinase-3 glycogen synthase using compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application.

- The use of compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, for the manufacture of Les is artenova means for the prevention or treatment of diseases or conditions, characterized by increasing the regulation of Aurora kinase (e.g. Aurora A kinase and/or Aurora B kinase).

- The use of compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, for the manufacture of a medicine for the prevention or treatment of cancer, and cancer is cancer that is characterized by increasing regulation of Aurora kinase (e.g. Aurora A kinase and/or Aurora B kinase).

- The use of compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, for the manufacture of a medicinal product for preventing or treating cancer in a patient selected from the subgroups with the Ile31 variant of the gene Aurora A.

- The use of compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, for the manufacture of a medicinal product for preventing or treating cancer in a patient, which results, is a member of the subgroup with the Ile31 variant of the gene Aurora A.

The method of prevention or treatment of a disease or condition characterized by an increase of regulacje the Aurora kinase (e.g., Aurora A kinase and/or Aurora B kinase), and the method includes introducing the compound of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application.

- To relieve symptoms or reduce the frequency of occurrence of diseases or conditions characterized by increasing regulation of Aurora kinase (e.g. Aurora A kinase and/or Aurora B kinase), and the method includes introducing the compound of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application.

The method of prevention or treatment of (or alleviating symptoms or frequency of occurrence) of cancer patients suffering from cancer or who suggest the presence of cancer; and the method includes (i) conducting diagnostic testing of the patient to determine whether the patient Ile31 variant of the gene Aurora A; and (ii) if the patient really has the specified option, the introduction of the patient is then compounds of formulas (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, having inhibitory activity against Aurora kinase.

The method of prevention or treatment of (or alleviating symptoms or reduce the frequency of appearance), the TB is evani or conditions, characterized by increasing the regulation of Aurora kinase (e.g. Aurora A kinase and/or Aurora B kinase); and the method includes (i) conducting diagnostic testing of the patient to detect a marker characteristic of increasing regulation of Aurora kinase; and (ii) if the diagnostic test indicates increasing regulation of Aurora kinase, introduction to the patient after the connection formulas (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, having inhibitory activity against Aurora kinase.

The method of prevention or treatment of (or alleviating symptoms or frequency of occurrence) of diseases or conditions characterized by (a) excessive activation of CDK kinase; and/or (b) increase the sensitivity of the signaling pathway to the normal activity of CDK; and/or (c) increasing regulation cycline E; moreover, this method includes (i) conducting diagnostic testing of the patient to detect a marker characteristic (a) and/or (b) and/or (c); and (ii) if the diagnostic test indicates (a) and/or (b) and/or (c), introduction to the patient after the connection formulas (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, having inhibitory activity against CDK kinases.

The method of treatment, use in medicine or connection for applications where the compound of formula (I), (II), (III) or (XXX) or any sub-groups or examples of compounds of the above formula, are defined in this application, enter (for example, in therapeutically effective amounts) subgroup of patients with one or more of the diagnostic tests described in this application, identified as having a disease or condition that should be treated the specified connection.

- The use of compounds of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, for the manufacture of a medicinal product for preventing or treating painful conditions described in this application.

The compound of formula (I), (II), (III) or (XXX) or any sub-group or individual examples of compounds of the above formula, are defined in this application, for use in the prevention or the treatment of painful conditions described in this application.

The method of prevention or treatment of (or alleviating symptoms or frequency of occurrence) of diseases or conditions described in this application, and the method comprises the administration to a mammal a therapeutically effective amount of a compound of formula (I), (I), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application.

Pharmaceutical compositions comprising a compound of formula (I), (II), (III) or (XXX) or any sub-groups or examples of compounds of the above formula, are defined in this application, as well as pharmaceutically acceptable carrier.

Pharmaceutical compositions intended for administration in the form of an aqueous solution, and a pharmaceutical composition includes the compound of formula (I), (II), (III) or (XXX) or any sub-groups or examples of compounds of the above formula, are defined in this application, in the form of a salt having a solubility in water of more than 1 mg/ml, typically, more than 5 mg/ml, more typically, more than 15 mg/ml, more typically, more than 20 mg/ml and preferably greater than 25 mg/ml

The compound of formula (I), (II), (III) or (XXX) or any sub-group or individual examples of compounds of the above formula, are defined in this application, for use in medicine.

Connection defined in this application, designed for any application and method, from among the above, and as described elsewhere in this application.

The compound of formula (I), (II), (III) or (XXX) or any sub-group or individual examples of compounds of the above formula, are defined in this application, or their salts (e.g. the R, acid additive salts, solvate, tautomers or N-oxides for use in the treatment of B-cell lymphoma.

The compound of formula (I), (II), (III) or (XXX) or any sub-group or individual examples of compounds of the above formula, are defined in this application, or their salts (for example, an acid additive salt), solvate, tautomers or N-oxides for use in the treatment of chronic lymphocytic leukemia.

The compound of formula (I), (II), (III) or (XXX) or any sub-group or individual examples of compounds of the above formula, are defined in this application, or their salts (for example, an acid additive salt), solvate, tautomers or N-oxides for use in the treatment of diffuse B-both lymphoma.

- Method of treating B-cell lymphoma, diffuse B-both lymphoma or chronic lymphocytic leukemia by administering to a patient in case of need of such treatment the compound of formula (I), (II), (III) or (XXX) or any sub-groups or individual examples of compounds of the above formula, are defined in this application, or their salts (for example, an acid additive salt), solvate, tautomers or N-oxides.

The compound of formula (I), (II), (III) or (XXX) or any sub-group or individual examples of compounds of the above formula, are defined in this application, or their salts (for example, acid-additive with the NML), the solvate, tautomers or N-oxides for use in the treatment of leukemia, in particular re or refractory acute myelogenous leukemia, myelodysplastic syndrome, acute lymphocytic leukemia and chronic myelogenous leukemia.

Acid additive salts of the free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and, specifically, lactate, intended for all applications and methods set forth above and described elsewhere in this application.

Preferred options and definitions

To each of the fragments D1, D2, A, E, X, Xaand R1-R9and to their various subgroups, defined within sub-groups, examples and options for implementation will apply the following General preferences and definitions unless the context does not indicate otherwise.

All references to formula (I) in this application will also apply to the formulas (II)-(VIII) and any other sub-group of compounds among the compounds of formula (I), unless the context requires a different kind of understanding.

The term "increasing regulation of Aurora kinase" in this description is defined as including increased expression or over-expression of Aurora kinases, including gene amplification (i.e. the multiplication of copies of the gene) and the increased expression under the action of transcriptio the nogo effect, as well as hyperactivity and activation of Aurora kinases, including activation under the influence of mutations.

The term "saturated" in the present description refers to cycles in which there are no multiple bonds between cyclic atoms.

The term "hydrocarbon" in the present description, regardless of whether he used separately or as part of a complex term, such as "hydrocarbonate"is a General term covering the aliphatic and alicyclic groups, the skeleton of which is composed entirely of carbon. Examples hydrocarbonrich groups include alkyl, cycloalkyl, cycloalkenyl, alkenyl, quinil, cycloalkenyl, cycloalkenyl. Separate hydratability groups are saturated groups such as alkyl and cycloalkyl group.

Examples of hydrocarbonate groups include alkoxy, cycloalkane, cycloalkene, alkenylacyl, alkyloxy, cycloalkylation, cycloalkylation. Separate hydrocarbonate groups are saturated groups such as alkoxy.

The prefix “C1-n” (where n indicates an integer) in the present description refers to the number of carbon atoms in the group. So, C1-4gidrolabilna group contains from 1 to 4 carbon atoms, whereas C1-3gidrolabilna group contains from 1 to 3 carbon atoms, etc.

Examples of C1-4hydrocarbonrich groups in luchot C 1-3gidrolabilna group or a C1-2gidrolabilna group, and specific examples include any individual, group or combination of groups selected from hydrocarbonrich groups C1C2C3and C4.

The term "alkyl" includes both alkyl groups with a linear chain and branched alkyl groups. Examples of alkyl groups are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl and tert-butyl.

Examples cycloalkyl groups are groups derived from cyclopropane, CYCLOBUTANE and cyclopentane.

Examples alkenyl groups are ethynyl (vinyl), 1-propenyl, 2-propenyl (allyl), Isopropenyl, butenyl and buta-1,4-dienyl.

Examples cycloalkenyl groups are cyclopropyl and cyclobutyl.

Examples etkinlik groups are etinilnoy and 2-proponila (propargyl) group.

Examples cycloalkylation and cycloalkenyl groups include cyclopropylmethyl.

Examples of alkoxygroup are methoxy, ethoxy, n-propyloxy, isopropoxy, n-butoxy, isobutoxy and tert-butoxy.

If an alkyl group is an integral part of monoalkylamines or dialkylamino groups, the alkyl group may be any of the alkyl groups given above as examples. Specific alkylamino, dialkylamino groups awsomeeeee, dimethylamino, ethylamino, diethylamino, n-propylamino, isopropylamino, butylamino, isobutylamino and tert-butylamino. Particular alkyl -, dialkylamino groups are methylamino and dimethylamino.

The term "saturated heterocyclic group" in the present description refers to a heterocyclic group containing no multiple bonds between adjacent atoms in the cycle. Saturated heterocyclic group may include 1 or 2 cyclic heteroatom selected from O, S and N.

Depending on the context of the heterocyclic groups can contain, for example, cyclic fragments of simple ether (such as tetrahydrofuran or dioxane), cyclic fragments tiefer (e.g. as in tetrahydrothiophene and dithiane), cyclic amine fragments (such as pyrrolidine), cyclic amide fragments (such as pyrrolidone), cyclic thioamides, cyclic complex thioethers, cyclic urea (for example, as in imidazolidin-2-Ohe), cyclic fragments of ester (such as butyrolactone), cyclic sulfones (for example, as in sulfolane and sulfolane), cyclic sulfoxidov, cyclic sulfonamides and combinations of the above fragments (for example, thiomorpholine).

Saturated heterocyclic group, as a rule, are monocyclic and usually contain 4, 5 or 6 atoms of the cycle, if not specified otherwise.

Specific examples of saturated heterocyclic groups containing 4 atoms in the cycle, is azetidinone group.

Examples of saturated heterocyclic groups containing 5 atoms in the cycle, include pyrrolidine (for example, 1-pyrrolidinyl, 2-pyrrolidinyl and 3-pyrrolidinyl), pyrrolidone, tetrahydrofuran, and tetrahydrothiophene.

Examples of saturated heterocyclic groups containing 6 atoms in the cycle, include morpholine, thiomorpholine, S-oxide thiomorpholine, S,S-dioxide thiomorpholine, piperidine (e.g. 1-piperidinyl, 2-piperidinyl, 3-piperidinyl and 4-piperidinyl), piperidine, dioxane, tetrahydropyran (for example, 4-tetrahydropyranyl), piperazine, piperazine, and N-alkylpiperazine, as, for example, N-methylpiperazine.

Specific embodiments of and preferences for D1, D2, A, E, R1-R9and X in the subgroups (A) and (B) of the formula (I)

In one of the General embodiments M is a group D1.

In another General embodiment, M is a group D2.

X is selected from O, NH and NCH3. In one particular embodiment, X represents O.

A is selected from communication and group NR2where R2is hydrogen or stands.

In one of the embodiments A is a connection.

In another embodiment, A represents the Wallpaper group NR 2where R2is hydrogen or stands.

E is selected from communication, CH2CH(CN) and C(CH3)2.

In one subgroup of compounds E is a connection.

In another subgroup of compounds E is CH2.

In another subgroup of compounds E represents CH(CN).

In another subgroup of compounds E is C(CH3)2.

If M is a group D1, R1may be selected from groups (i), (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi) and (xii).

If M is a group D1, and E-A is C(CH3)2NR2or CH2-NR2, R1may additionally be selected from:

(xiii) unsubstituted 2-furil and 2,6-diphthera.

If M is a group D1, and E-A is C(CH3)2NR2, R1may additionally be selected from:

(xiv) unsubstituted phenyl.

If M is a group D1, and E represents CH2, R1may additionally be selected from:

(xv) unsubstituted tetrahydropyran-4-yl.

Each group in the list of groups (i)-(xv) is a specific variant of the implementation of the present invention.

In the embodiment, (i) R1is 3-5-membered cycloalkyl group, optionally substituted by hydroxy, fluorine, amino, methylamino, stands or E. the mud.

Separate cycloalkyl group optionally substituted cyclopropenes and cyclobutyl groups, more typically optionally substituted cyclopropylamine groups. In a preferred embodiment, R1is unsubstituted cyclopropane group.

In the embodiment, (ii) R1is a 4-6-membered saturated heterocyclic group containing 1 or 2 heteroatoms in the cycle selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4the alkyl, amino or hydroxy; but from among these groups are excluded unsubstituted 4-morpholinyl, unsubstituted tetrahydropyran-4-yl, unsubstituted 2-pyrrolidinyl, and unsubstituted and 1-substituted piperidine-4-yl.

Examples of saturated heterocyclic groups described above under "General preferences and definitions".

Some examples of saturated heterocyclic groups include:

- membered cycles containing only part of the cycle heteroatom selected from O, N and S (other than unsubstituted 2-pyrrolidinyl);

- six-membered ring containing two included in the cycle heteroatoms selected from O, N and S (other than unsubstituted 4-morpholinyl).

Saturated heterocyclic group may be substituted or unsubstituted. In one of the embodiments, they are unsubstituted. Drugom embodiment, they are replaced by one or two C 1-4alkyl groups, for example one or two methyl groups.

One of the specific saturated heterocyclic groups is optionally substituted group tetrahydrofuran (e.g. tetrahydrofuran-2-yl or tetrahydrofuran-3-yl), more preferably unsubstituted group tetrahydrofuran.

In the embodiment, (iii) R1represents a 2,5-substituted phenyl group of the formula:

in which (a) if X represents NH or N-CH3, R3selected from chlorine and cyano; and (b) if X represents O, R3represents CN.

In one of the subgroups of compounds included in an implementation option (iii), X represents N-CH3and R3selected from chlorine and cyano.

In another subgroup of compounds included in an implementation option (iii), X represents O and R3represents CN.

In the embodiment, (iv) R1is a group of CR6R7R8in which each of R6and R7selected from hydrogen and methyl, and R8selected from hydrogen, methyl, C1-4alkylsulfonyl, hydroxymethyl and cyano.

In the embodiment, (iv) specific examples R1are methyl, cyanomethyl, HOCH2C(CH3)2- and 2-methylsulfonylmethyl.

In the embodiment, (iv) other specific the examples R 1are methyl and isopropyl.

In the embodiment, (v) R1is pyridazin-4-ilen group, optionally substituted by one or two substituents selected from methyl, ethyl, methoxy, ethoxy. Pyridazinyl group can be pyridazin-3-ilen or pyridazin-4-strong group, but, as a rule, is pyridazin-4-yl. Specific substituents are metoxygroup, and, for example, pyridazinyl group can carry two methoxy substituent.

In the embodiment, (vi), R1represents a substituted imidazothiazole group in which the substituents are selected from methyl, ethyl, amino, fluorine, chlorine, amino and methylamino. The particular Deputy is methyl.

In the embodiment, (vii), R1represents an optionally substituted 1,3-dihydroindol-2-ilen, or 2,3-dihydroindol-1-ilen group in which optionally present substituents in each case selected from halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino.

Specific substituents selected from methyl, ethyl, fluorine, chlorine, preferably only on the aryl cycle dihydroindole or dihydroindole), CONH2 , amino, methylamino, dimethylamino and methoxy.

In one subset of compounds in the embodiment, (vii), each of the fragments of dihydroindole or dihydroindole is unsubstituted.

In the embodiment, (viii) R1represents 3-pyridyl, optionally substituted by one or two substituents selected from hydroxy, halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino; provided that not formed such a connection, as 2,6-dimethoxy-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]nicotinamide or such a connection, as [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amide of 2-oxo-1,2-dihydropyridines-3-carboxylic acid.

In one embodiment, the implementation of R1represents 3-pyridyl, optionally substituted by one or two substituents selected from hydroxy, halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino, but if R1represents 3-pyridyl, X is O, a is a bond and E is a bond, iridal has one or two substituent, selected from halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino.

Specific substituents selected from methyl, ethyl, fluorine, chlorine, CONH2, amino, methylamino, dimethylamino and methoxy. More specifically, the substituents selected from methyl, ethyl, fluorine, chlorine, CONH2, amino, methylamino and dimethylamino.

In one subgroup of compounds 3-Peregrina group is unsubstituted.

In the embodiment, (ix) R1represents thiomorpholine or its S-oxide or S,S-dioxide, optionally substituted by one or two substituents selected from halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2or CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino.

In one of the subgroups of compounds thiomorpholine or its S-oxide or S,S-dioxide are unsubstituted.

In the embodiment, (x) E-A represents NR2and R1selected from: 2-ftoheia, 3-ftoheia, 4-ftoheia, 2,4-dipthera, 3,4-dipthera, 2,5-dipthera, 3,5-dipthera, 2,4,6-tryptophanyl, 2-methoxyphenyl, 5-chloro-2-methoxyphenyl, cyclohexyl unsubstituted 4-tetrahydropyranyl and tert-butyl;

In the embodiment, (xi) E-A represents NR2and R1represents a group NR10R11where each of R10and R11represents a C1-4alkyl, or R10and R11connected so that NR10R11form a 4-6-membered saturated heterocyclic group optionally containing in the cycle of the second heteroatom selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4the alkyl, amino or hydroxy.

In this embodiment, one of the subgroups of compounds is that group of compounds in which each of R10and R11represents a C1-4alkyl, in particular methyl.

Another subgroup of compounds is that group of compounds in which R10and R11connected so that NR10R11form a 4-6-membered saturated heterocyclic group optionally containing in the cycle of the second heteroatom selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4the alkyl, amino or hydroxy. The saturated heterocyclic group may be any of the nitrogen-containing saturated heterocyclic groups listed above in the section "preferred options and definitions", but the specific saturated heterocyclic groups include the feast of original, morpholinyl, piperazinil and N-C1-4alkylpiperazine. These groups usually are unsubstituted or substituted by one or two methyl groups, and, in one particular embodiment, are unsubstituted.

In the embodiment, (xii) E-A represents NR2and R1represents Spiridonova group, optionally substituted by one or two substituents selected from hydroxy, halogen, cyano, amino, C1-4mono - and dialkylamino, CONH2, CONH-C1-4of alkyl, C1-4the alkyl and C1-4alkoxy, where C1-4alkyl and C1-4alkoxygroup optionally substituted by hydroxy, methoxy or amino.

Spiridonova group can be N-substituted, for example an alkyl group such as methyl, and in another case may be unsubstituted.

In the embodiment, (xiii) E-A is C(CH3)2NR2or CH2-NR2and R1selected from unsubstituted 2-furil and 2,6-diphthera.

In the embodiment, (xiv) E-A is C(CH3)2NR2and R1is unsubstituted phenyl.

In the embodiment, (xv) E represents CH2and R1is unsubstituted tetrahydropyran-4-yl.

If M is a group D2, R1may be selected from groups (xvi), (xvii), (xviii) and (xix).

Each individual GRU the PA in the list of groups (xvi)-(xix) represents a separate variant embodiment of the invention.

In the embodiment, (xvi) R1represents a 2-substituted 3-follow group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2and 5 - or 6-membered saturated cycle optionally substituted by hydroxy, fluorine, amino, methylamino, stands or ethyl. In one embodiment, the implementation of R1represents a 2-substituted 3-follow group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2and 5 - or 6-membered saturated cycle optionally substituted by hydroxy, fluorine, amino, methylamino, stands or ethyl, but where a is a relation and E is a bond, R4and R5linked such that NR4R5forms unsubstituted piperidine.

P is emery saturated heterocyclic groups correspond to the above in the section "preferred options and definitions", but the specific saturated heterocyclic group include pyrrolidinyl, morpholinyl, piperazinil and N-C1-4alkylpiperazine. Such groups usually are unsubstituted or substituted by one or two methyl groups, and, in one particular embodiment, are unsubstituted.

Specific examples of compounds in which R4and R5selected from hydrogen and C1-4of alkyl, are methylamino and dimethylamino group, more typically a dimethylamino group.

In the embodiment, (xvii) R1represents a 5-substituted 2-follow group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2and 5 - or 6-membered saturated heterocyclic group optionally substituted by hydroxy, fluorine, amino, methylamino, stands or ethyl; provided that the compound is not [3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amidon 5-piperidine-1-ilmatieteen-2-carboxylic acid.

Examples of saturated heterocyclic groups correspond izlojennaya in the preferred options and definitions", but the specific saturated heterocyclic group include pyrrolidinyl, morpholinyl, piperazinil and N-C1-4alkylpiperazine. Such groups usually are unsubstituted or substituted by one or two methyl groups, and, in one particular embodiment, are unsubstituted.

In the embodiment, (xviii) R1represents a group of the formula:

in which R9represents hydrogen, methyl, ethyl or isopropyl; G represents CH, O, S, SO, SO2or NH, and the group is optionally substituted one, two or three substituents selected from C1-4hydrocarbide, hydroxy, C1-4hydrocarbonate, fluorine, amino, mono - and di-C1-4alkylamino, and where each of the groups C1-4hydrocarbon and C1-4hydrocarbonate optionally substituted by hydroxy, fluorine, amino, mono - or di-C1-4alkylamino.

In one subgroup of compounds included in an implementation option (xviii), G is selected from O and CH.

In the embodiment, (xviii) a group R1as a rule, is unsubstituted or substituted by one or two methyl groups, and more typically is unsubstituted.

In the embodiment, (xix) R1represents a 3,5-disubstituted phenyl group of the formula:

in which Xaas X, select the n from O, NH and NCH3.

Preferably Xarepresents N-CH3.

Specific examples of fragment R1-A - is shown in table 1, and the asterisk indicates the attachment point to the carbonyl group C=O group, R1-E-A-C(=O)-NH-.

Table 1
Examples of fragment R1-E-A-

In table 1, the preferred groups R1-E-A - include A1, A4, A10, A11, A13, A20, A22, A23, A24, A29, A30, A31, A32, A38, A42, A43, A44, A46, A47, A49, A54 and A56.

In another embodiment, the group R1-E-A represents the A57, A58 or A59.

A preferred subset of the groups R1-E-A - includes A1, A4, A20, A24, A30, A44, A46 and A54. In this subset of one particular group, R1-E-A - is a group A24.

One subgroup of compounds according to the present invention represented by formula (II):

where R1E, A and X correspond to the one presented in the present description definitions.

In the formula (II) one of the subsets of compounds is the subset in which X represents O.

One subgroup of compounds of formula (II) can be represented by the formula (III):

and their salts, in particular lactate.

In the formula (III) one of the subsets of compounds is the subset in which E is the link.

Another subset of compounds among the compounds of the formula (III) is a subset in which E represents CH2or C(CH3)2.

In one particularly preferred embodiment, compounds of formula (III) E is a bond, R2represents H and R1is cycloalkyl group (i), as defined in this application. In one of the embodiments cycloalkyl group can be cyclopropyl or cyclobutyl. More preferably, R1represents cyclopropyl group.

To avoid with the developments it should be understood that each General and specific preferred option, option exercise and example of the groups R1can be combined with each General and specific preference, embodiment and example of the groups R2and/or R3and/or R4and/or R5and/or R6and/or R7and/or R8and/or R9and/or R10and/or R11and/or D1 and/or D2 and/or A and/or E and/or X and/or Xaand any of its subgroups, defined in this application, unless the context indicates otherwise, and that all such combinations are embraced by this application.

Various functional groups and substituents, which is made up of compounds of formula (I)is generally selected so that the molecular weight of the compounds of formula (I) does not exceed 1000. Often molecular weight compounds is less than 750, for example, less than 700, or less than 650 or 600, or less than 550. More preferably, the molecular weight has a value of less than 525, for example, is 500 or less.

Specific compounds of the present invention are compounds that are shown to illustrate in the examples below.

One preferred compound of the present invention is 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its salts, solvate, and tautomers.

Salt, solvate, the tautomers, the isomers, N-oxides, esters, prodrugs and forms containing isotopes, compounds of groups (A) and (B) compounds of formula (I)and their subgroups and implementation options for

Unless otherwise specified, references to a particular compound also includes ionic forms, salts, solvate and their protected form, for example, as discussed below.

Many of the compounds of formula (I) can exist in the form of salts, for example, acid additive salts or, in some cases, salts of organic and inorganic bases, for example in the form of carboxylate, sulphonate and phosphate salts. All such salts are included within the scope of the present invention, and references to compounds of formula (I) include the salt forms of these compounds. As in the previous section of this application, all references to formula (I) should be taken as referring to formula (II), (III) and their subgroups, as defined in this application, unless the context indicates otherwise.

Salt of the present invention can be synthesized from starting compounds which contain a basic or acid fragment using standard chemical methods, such as methods described in Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, hardcover, 388 pages, August 2002. Generally, such salts can be floor is obtained by the interaction of a free acid or base of the considered compounds with the appropriate base or acid in water or an organic solvent, or mixtures thereof; as a rule, apply nonaqueous media like ether, ethyl acetate, ethanol, isopropanol or acetonitrile.

Acid additive salts can be formed with a wide range of acids, both inorganic and organic. Examples of the acid additive salts include salts formed with acids selected from the group consisting of acetic, 2,2-dichloracetic, adipic, alginic, ascorbic (e.g. L-ascorbic), L-aspartic, benzosulfimide, benzoic, 4-acetamidobenzoyl, butane, (+)camphor, camphorsulfonate, (+)-(1S)-camphor-10-sulphonic, capric, Caproic, Caprylic, cinnamon, lemon, reklamowa, modellserie, ethane-1,2-disulfonate, econsultancy, 2-hydroxyethanesulfonic, formic, fumaric, galatarasay, hentaimovi, glucoheptonate, D-gluconic, glucuronic (for example, D-glucuronic), glutamic (for example, L-glutamic), α is oxoglutarate, glycolic acid, hippuric, Hydrobromic, hydrochloric, idiscovered, isetionate, (+)-L-lactic, (±)-DL-lactic, lactobionic, maleic, malic, (-)-L-malic acid, malonic acid, (±)-DL-almond, methansulfonate, naphthalene-2-sulphonic, naphthalene-1,5-disulfonate, 1-hydroxy-2-naphthoic, nicotine, nitrogen, oleic, orotovoy, oxalic, palmitic, pambou, phosphoric, propionic, L-pioglit the speech, salicylic, 4-aminosalicylic, sebacinales, stearic, succinic, sulfuric, tanning, (+)-L-tartaric, titanoboa, p-toluensulfonate, undecylenoyl and valerianic acids, as well as acylated amino acids and cation exchange resins.

In addition, the acid-salt additive can be selected from salts of aspartic (e.g., D-aspartic), carboxylic, dodecanol, isoalkanes, laurylsulphate, mucus, naphthalenesulfonates (for example, naphthalene-2-sulfonic), toluensulfonate (for example, p-toluensulfonate) and xinfulai acids.

A separate group of salts consisting of the salts formed with hydrochloric, idiscovered, phosphoric, nitric, sulfuric, citric, lactic, succinic, maleic, malic, isetionate, fumaric, benzosulfimide, toluensulfonate, methanesulfonate, econsultancy, naphthalenesulfonates, valerianic, acetic acid, propane, butane acid, malonic acid, glucuronic and lactobionic acids.

One group of salts consisting of the salts formed with hydrochloric, acetic, adipic, L-aspartic, and DL-lactic acids.

Another sub-group of salts consisting of acetate, nelfinavir, aconsultant, DL-lactate, adipate, D-glucuronate, D-gluconate or hydrochloride.

Salt, as, for example, an acid additive salt, have several advantages over suitable the mi-free basis. For example, the salt will be different by one or more of the following advantages over the free bases so that they will be:

- greater solubility and, therefore, will be better suited for intravenous injection (e.g., via infusion) and will have better pharmacokinetics;

- greater stability (for example, extended shelf life);

having the best thermal stability;

less basic and therefore will be better for intravenous injection;

to have advantages for obtaining;

to have a better metabolic properties; and

- to show a smaller change clinical properties from patient to patient.

Preferred salts for use in the production of the liquid (e.g. water) compositions of compounds of formula (I), (II), (III), (XXX)and sub-groups and specific examples of these compounds described in this application are salts having a solubility in a given liquid carrier (e.g. water)in excess of 25 mg/ml of a liquid medium (e.g. water), more typically greater than 50 mg/ml and preferably greater than 100 mg/ml

In other embodiments, implementation of the preferred salts for use in the production of the liquid (e.g. water) compositions of compounds of formula (I), (II), (III), (XXX)and sub-groups and individual examples pointed to by the x compounds described in this application are salts having a solubility in a given liquid carrier (e.g. water or buffer systems)exceeding 1 mg/ml, typically greater than 5 mg/ml of a liquid medium (e.g. water), more typically greater than 15 mg/ml, more typically greater than 20 mg/ml and preferably greater than 25 mg/ml

In another preferred embodiment, the acid additive salts are mesilate, aconsultant, D - or L-lactate or hydrochloride. In a specific embodiment, the acid additive salt is a lactate, in particular L-lactate or D-lactate, preferably L-lactate.

In one of the embodiments of the present invention the pharmaceutical composition comprising an aqueous solution, which contains compounds of formulas (I), (II), (III), (XXX)and sub-groups and examples of such compounds described in the present application, in the form of salts, in concentrations exceeding 25 mg/ml, typically greater than 50 mg/ml and preferably greater than 100 mg/ml

In another embodiment of the present invention the pharmaceutical composition comprising an aqueous solution, which contains compounds of formulas (I), (II), (III), (XXX)and sub-groups and examples of such compounds described in the present application, in the form of a salt, the concentration is in excess of 1 mg/ml, typically exceeding 5 mg/ml of a liquid medium (e.g. water), more typically greater than 15 mg/ml, more typically greater than 20 mg/ml and preferably greater than 25 mg/ml

If the compound is anionic, or comprises a functional group which may be anionic (e.g.,- COOH may be in the form-COO-), then can be formed salt with a suitable cation. Examples of suitable inorganic cations include, but are not limited to the above, ions of alkali metals such as, for example, Na+and K+the cations of alkaline earth metals such as, for example, Ca2+and Mg2+and other cations, such as Al3+. Examples of suitable organic cations include, but are not limited to the above, ammonium ion (i.e. NH4+) and substituted ammonium ions (e.g., NH3R+, NH2R2+, Other3+, NR4+). Examples of some suitable substituted ammonium ions are ammonium ions derived from: ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, Ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylendiamine, choline, meglumine and tromethamine, as well as amino acids such as lysine and arginine. An example of a conventional Quaternary ammonium ion is N(CH3)4+.

If the compounds of formula (I) contain the amino group, they may form a Quaternary ammonium salt, for example by reaction with an alkylating reagent, in accordance with methods which are well known in the art. Such Quaternary ammonium compounds covered by formula (I).

Salt forms of the compounds of the present invention, as a rule, are pharmaceutically acceptable salts, and examples of pharmaceutically acceptable salts are discussed in Berge et al., 1977, “Pharmaceutically Acceptable Salts”, J. Pharm. Sci., Vol. 66, pp.1-19. However, salts which are not pharmaceutically acceptable may also be obtained as an intermediate form, which can then be converted into pharmaceutically acceptable salts. Such forms of pharmaceutically unacceptable salts, which can be used, for example, when cleaning or separation of the compounds of the present invention, also included in the scope of the invention.

The compounds of formula (I)containing an amino group, may also form N-oxides. In this application references to the compounds of formula (I)which contain an amino group include N-oxides.

If the compound contains several amino groups, one or more nitrogen atoms can be oxidized with the formation of N-oxides. Specific examples of the N-oxides are the N-oxides of tertiary amines or nitrogen atoms asado is containing a series of heterocycles.

N-oxides can be obtained by treating the corresponding amines oxidizing reagent such as hydrogen peroxide or peroxynitrate (for example, peroxocarbonate acid), see, for example, Advanced Organic Chemistry, by Jerry March, 4thEdition, Wiley Interscience, pp. More specifically, the N-oxides can be obtained by the method L.W. Deadfly (Syn. Comm. 1977, 7, 509-514) in which the compound containing the amino group, enter into interaction with m-chloroperoxybenzoic acid (MCPBA), for example, in an inert solvent such as dichloromethane.

The compounds of formula (I) may exist in the form of a number of different geometrically isomeric and tautomeric forms, and the mention of the compounds of formula (I) include all such forms. For the avoidance of doubt, if a connection can exist in one of several geometrically isomeric or tautomeric forms, and specifically described or shown only one of them, all the other forms, however, covered by formula (I).

For example, in compounds of formula (I) benzimidazole group may take any one of the following two tautomeric forms A, A', B and B'. For simplicity, the General formula (I) shows the form A and A', but you should accept this formula as encompassing all four tautomeric forms.

Pyrazol cycle can also be tautomerism, and things he can in order to act in two tautomeric forms C or D, below:

Other examples of tautomeric forms include, for example, ketone form enol form and shape enolates, as, for example, the following tautomeric pairs: ketone/enol (illustrated below), Imin/enamine, amide/iminspect, amicin/amidin, nitroso/oxime, thioketone/ential and nitro/ACI-nitro:

If the compounds of formula (I) contain one or more chiral centers and may exist in the form of two or more optical isomers, references to compounds of the formula (I) include all optical isomeric forms (e.g., enantiomers, epimere and diastereoisomers), either as individual optical isomers, or mixtures (e.g. racemic mixtures) of two or more optical isomers, unless the context requires otherwise.

For example, group A may include one or more chiral centers. So, if E and R1both attached to the same carbon atom linking group A, the indicated carbon atom, typically, is chiral and therefore, the compound of formula (I) will exist as pairs of enantiomers (or more than one pair of enantiomers, if the connection is more than one chiral center).

The optical isomers can be characterized and identified by their optical activity (i.e. to the + and - isomers, or d and l isomers) or they can be characterized in terms of their absolute stereochemical configuration, using the item “R and S”, developed by kann, ingoldo and Prelogon, see Advanced Organic Chemistry, by Jerry March, 4thEdition, John Wiley & Sons, New York, 1992, pages 109-114, and, in addition, see Cahn, Ingold & Prelog, Angew. Chem. Int. Ed. Engl., 1966, 5, 385-415.

The optical isomers can be separated using a number of techniques, including chiral chromatography (chromatography on chiral stationary phase), and such techniques are well known to the person skilled in the technical field.

Alternatively, chiral chromatography, optical isomers can be separated by formation of diastereomeric salts with chiral acids, such as (+)-tartaric acid, (-)-pyroglutamic acid, (-)-ditolyl-L-tartaric acid, (+)-almond acid, (-)-malic acid and (-)-camphorsulfonic acid, separation of the diastereomeric salts by selective crystallization, and then the decomposition of the salts with obtaining individual enantiomers of the free base.

If the compounds of formula (I) exist as two or more optical isomeric forms, one of the enantiomers enantiomeric pairs can exhibit advantages over the other enantiomer, for example, from the point of view of biological activity. Thus, to determine the certain circumstances it may be desirable for use as a therapeutic means of only one enantiomer of a pair or only one of the many diastereomers. Accordingly, the invention relates to compositions containing a compound of the formula (I)which has one or more chiral centers and in which not less than 55% (for example, not less than 60, 65, 70, 75, 80, 85, 90 or 95%) of the compounds of formula (I) is present in the form of a single optical isomer (e.g., enantiomer or diastereoisomer). One of the main options for the implementation of 99% or more of the total amount of compounds of formula (I) (for example, virtually all connection) may be present in the form of a single optical isomer (e.g., enantiomer or diastereoisomer).

Compounds of the present invention include compounds in which one or more atoms replaced by their isotopes, and the mention of specific element includes all isotopes of this element. For example, the mention of hydrogen include1H,2H(D) and3H(T). Similarly, references to carbon and oxygen include, respectively12C,13C and14C and16O and18O.

These isotopes can be radioactive or non-radioactive. In one of the embodiments of the present invention compounds do not contain radioactive isotopes. Such compounds are preferred for therapeutic use. However, in another embodiment, the compounds can contain one or more radio is active isotopes. Compounds containing such radioactive isotopes may be useful in diagnostic applications.

Esters, such as esters of carboxylic acids and allactivity compounds of formula (I)comprising a carboxylic acid group or hydroxyl group, is also covered by formula (I). Examples of esters are compounds containing the group-C(=O)OR where R is a Deputy of the ester group, for example, C1-7alkyl group, a C3-20heterocyclic group or a C5-20aryl group, preferably C1-7alkyl group. Specific examples of ester groups include, but are not limited to the above, -C(=O)OCH3, -C(=O)OCH2CH3, -C(=O)OC(CH3)3and-C(=O)OPh. Examples of acyloxy (converted ether) groups represented by the formula-OC(=O)R, where R is a Deputy alloctype, for example, C1-7alkyl group, a C3-20heterocyclic group or a C5-20aryl group, preferably C1-7alkyl group. Specific examples of acyloxy include, but are not limited to the above, -OC(=O)CH3(acetoxy), -OC(=O)CH2CH3, -OC(=O)C(CH3)3, -OC(=O)Ph and-OC(=O)CH2Ph.

In addition, formula (I) covered by any of the polymorphic forms of the compounds, solvate (e.g., hydrates), complexes (e.g. inclusion complexes illutrate with such compounds as cyclodextrins, or complexes with metals) compounds or prodrugs of the compounds. By "prodrug" refers to, for example, any compound in vivo is converted to the biologically active compound of the formula (I).

For example, one of the types of prodrugs are esters of the active compounds (e.g., physiologically acceptable, capable of metabolism esters). In the process of metabolism ester group (-C(=O)OR) is cleaved resulting in the formation of active medicinal substances. Such esters can be obtained by etherification, for example, any of the groups, carboxylic acid (-C(=O)OH) in the original connection, provided preliminary protection, if possible, any other reactive groups present in the initial compound, with subsequent removal of the protection if necessary.

Examples of such are capable of metabolism of esters include esters of the formula-C(=O)OR in which R represents:

C1-7alkyl

(for example, -Me, -Et, -n-Pr-ISO-Pr, n-Bu, -sec-Bu, -ISO-Bu, t-Bu);

C1-7aminoalkyl

(for example, aminoethyl; 2-(N,N-diethylamino)ethyl; 2-(4-morpholino)ethyl); and

acyloxy-C1-7alkyl

(for example, acyloxymethyl;

acyloxyacyl;

pivaloyloxymethyl;

acetoxymethyl;

1-acetoxyethyl;

1-(1-methoxy-1-methyl)ethylcarbonate ITIL;

1-(benzoyloxy)ethyl; isopropoxycarbonyloxymethyl;

1-isopropoxycarbonyloxymethyl; cyclohexylcarbodiimide;

1-cyclohexylcarbodiimide;

cyclohexyloxycarbonyloxy;

1-cyclohexyloxycarbonyloxy;

(4 tetrahydropyranyloxy)carbonylmethyl;

1-(4-tetrahydropyranyloxy)carbonylmethyl;

(4-tetrahydropyranyl)carbonylmethyl; and

1-(4-tetrahydropyranyl)carbonyloxy).

In addition, some prodrugs are activated by the action of enzymes, forming the active compound or compounds, which result in further chemical reactions to form active compounds (for example, as in the case of ADEPT, GDEPT, LIDEPT etc). For example, the prodrug may be derived sugar or other conjugated glycoside or ester can be derived amino acids.

1-Cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its salts

One of the specific compounds of formula (I), which belongs to the subgroup (A)is 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

Accordingly, in one of the preferred embodiments the invention relates to a free base form or acid additive salt 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]m is chevigny.

The form of the free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, from which the formed salt has the formula (XXX):

The compound of formula (XXX) may be named in the present application is its chemical name, ie 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, or, for convenience, "connect XXX", "compound of formula (XXX)or compound of example 24. Each of these synonyms refers to the compound shown by the formula (XXX) above, called 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

References of the form of the free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and the acid additive salts include all its solvate, tautomers and isotopes, and where permitted by the context, N-oxides, other ionic forms, and prodrugs. Therefore, references to other tautomer the compounds of formula (XXX), i.e., 1-cyclopropyl-3-[3-(6-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, it should be understood, as mentioned compound (XXX).

Acid additive salts of the compounds of formula (XXX) can be selected from salts formed wide range of acids, both inorganic and organic. Examples of the acid additive salts is clucalc salt, educated acids selected from the group consisting of acetic, 2,2-dichloracetic, adipic, alginic, ascorbic (e.g. L-ascorbic), aspartic (e.g. L-aspartic), benzosulfimide, benzoic, 4-acetamidobenzoyl, butane, (+)camphor (for example, (+)-camphor), camphorsulfonate, (+)-(1S)-camphor-10-sulphonic, capric, Caproic, Caprylic, carbonic, cinnamic, citric, reklamowa, dodecanol, modellserie, ethane-1,2-disulfonates, econsultancy, 2-hydroxyethanesulfonic, formic, fumaric, galatarasay, hentaimovi, glucoheptonate, D-gluconic, glucuronic (for example, D-glucuronic), glutamic (for example, L-glutamic), α is oxoglutarate, glycolic acid, hippuric, Hydrobromic, hydrochloric, idiscovered, isetionate, breast (e.g., (+)-L-lactic and (-)-D-lactic), lactobionic, laurylsulphate, maleic, malic, (-)-L-malic acid, malonic acid, (±)-DL-almond, methansulfonate, mucus, naphthalenesulfonates (for example, naphthalene-2-sulphonic, naphthalene-1,5-disulfonate, 1-hydroxy-2-naphthoic, nicotine, nitrogen, oleic, orotovoy, oxalic, palmitic, pambou, phosphoric, propionic, L-Pyroglutamate, salicylic, 4-aminosalicylic, sebacinales, stearic, succinic, sulfuric, tanning, wine (for example, (+)-L-tartaric), t is cyanide, toluensulfonate (for example, p-toluensulfonate), undecylenate, valerianic and xinfulai acids, as well as acylated amino acids and cation exchange resins.

A separate group of salts consisting of the salts formed with hydrochloric, idiscovered, phosphoric, nitric, sulfuric, citric, lactic, succinic, maleic, malic, isetionate, fumaric, benzosulfimide, toluensulfonate, methanesulfonate, econsultancy, naphthalenesulfonates, valerianic, acetic acid, propane, butane acid, malonic acid, glucuronic and lactobionic acids.

One group of salts consisting of the salts formed with hydrochloric, acetic, adipic, L-aspartic and D - or L-lactic acid.

Another sub-group of salts consisting of acetate, nelfinavir, aconsultant, D - or L-lactate, adipate, D-glucuronate, D-gluconate or hydrochloride. In another preferred embodiment, the acid additive salts are mesilate, aconsultant, D - or L-lactate or hydrochloride.

In a specific embodiment, the acid additive salt is DL-lactate, in particular L-lactate or D-lactate, preferably L-lactate.

In another embodiment, the free base or the salt of the compounds of formula (XXX) is selected from L-lactate, dihydrate free base, esylate free the warping and hydrochloride.

In another and preferred embodiment, the salt of the compounds of formula (XXX) is selected from lactate and citrate, and mixtures thereof and more preferably selected from L-lactate and citrate and mixtures thereof, and L-lactate is particularly preferred. Particular and preferred embodiments of the present invention, related to L-lactate and citrate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, selected and described in more detail below.

In another embodiment, the compound of formula (XXX) is a freelance basis.

Salt of the present invention, such as lactates (e.g., L-lactate and citrate, can be obtained from the starting compound, i.e. a 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea by conventional chemical methods, such as methods described in Pharmaceutical Salts: Properties, Selection, and Use, P.Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, hardcover, 388 pages, August 2002. Generally, these salts can be obtained by the interaction of the parent compound, i.e. a 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea with the appropriate acid in water or an organic solvent, or a mixture of water and an organic solvent; mainly used non-aqueous environment, such as, for example, ether, etilize is at, ethanol, isopropanol or acetonitrile.

In another aspect, the present invention relates to a method for producing an acid additive salt 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, such as lactate (for example, L-lactate and citrate, and this method includes obtaining a solution of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form in a solvent (typically an organic solvent or solvent mixture, and processing the resulting solution of the acid sludge acid additive salt.

The acid may be added in the form of a solution in a solvent which is able to be mixed with the solvent in which is dissolved free base. The solvent in which initially dissolve the free base may be one of the solvents in which the insoluble acid additive salt of the free base. On the other hand, the solvent in which initially dissolve the free base may be a solvent in which the acid additive salt is soluble, at least partially, and later add another solvent in which the acid additive salt is less soluble, so that the salt falls out of solution.

In an alternative method according to the teachings of the acid additive salt, such as lactate (for example, L-lactate) or citrate, 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea are dissolved in a solvent containing a volatile acid, and optionally additional solvent, thereby obtaining a solution of an acid additive salt with a volatile acid, and then the resulting solution is concentrated or evaporated to highlight the salt. Another example of an acid additive salt, which can be obtained in this way is acetate.

In another aspect the invention relates to a method for producing an acid additive salt 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea as defined in the present description, such as lactate (for example, L-lactate and citrate, and this method includes the handling of compounds of formula (XXX):

organic or inorganic acid, specified in the present description, in an organic solvent to obtain an acid additive salt 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea with organic or inorganic acid, and, optionally, the allocation thus obtained acid additive salt.

Salt usually precipitates from the organic solvent after its formation and, therefore, may be adelena separation of solids from solution, for example by filtration.

One salt form of the compounds of the present invention can be converted into the free base and, optionally, to another salt form using methods well known to the specialist. For example, the free base can be obtained by passing a salt solution through the column with a stationary phase containing an amino group (for example, column Strata-NH2). On the other hand, the solution of salt in water can be treated with sodium bicarbonate to decompose the salt and precipitation of the free base. Then the free base can be introduced into the reaction with another acid by one of the methods described above or elsewhere in this application.

Preferred salts such as acid additive salts, such as lactate (for example, L-lactate and citrate, have several advantages. For example, the salt will be different by one or more of the following advantages, in that they:

- will have greater solubility, in particular, will have a better solubility in aqueous solution and, therefore, will be better for intravenous injection (e.g., infusion);

it will give the opportunity to adjust the pH of the solution and, therefore, will be better for intravenous injection;

- you can have the best anti-cancer activity; and

- may have a better therapeutic index.

The other item is akusherstvo salts are what they are:

- will have the best stability, such as thermal stability (for example, extended shelf life);

- will have advantages in the manufacture; and

- will have the best physico-chemical properties.

Lactate (for example, L-lactate) of the present invention has special advantages, because it has good solubility in water and has a better solubility in buffer systems.

Preferred salts for use in the preparation of liquid (e.g. water) pharmaceutical compositions are acid additive salts (such as lactate) having a solubility in a given liquid carrier (e.g. water) more than 1 mg/ml, typically more than 5 mg/ml of a liquid medium (e.g. water), more typically more than 15 mg/ml, more typically more than 20 mg/ml and preferably greater than 25 mg/ml

Aqueous solutions of salts (for example, in the form of pharmaceutical compositions) are another aspect of the present invention. Such solutions can be buffered or nezabyvaemymi. In the solution of these salts, as a rule, will be dissociate, forming protonated form of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea together with one or more counterions. Therefore, in another aspect, the present invention also relates to podnimetsia 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in protonated form together with one or more counterions and, optionally, one or more counterions (e.g., counterions formed from other salts, such as sodium chloride or buffering agents).

Salt 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea as a rule, are pharmaceutically acceptable salts, and examples of pharmaceutically acceptable salts are discussed in Berge et al., 1977, “Pharmaceutically Acceptable Salts”, J. Pharm. Sci., Vol. 66, pp. 1-19. However, salts which are not pharmaceutically acceptable may also be obtained as an intermediate form, which can then be converted into pharmaceutically acceptable salts. Therefore, such forms of pharmaceutically unacceptable salts are also included in the scope of the present invention.

In addition, 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea can form N-oxides. N-oxides can be obtained by the methods discussed above.

Like other compounds of the present invention, 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its acid additive salts can exist in various tautomeric forms, and the mention of these compounds in this application includes all these forms.

More specifically, 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]much the fault and its salts benzimidazole group can take any of the two tautomeric forms A and B”, shown below. For simplicity, the General formula (I) shows the shape of A”, but this formula should be understood as covering all tautomeric forms

Hence the mention of alternative tautomer, i.e. the 1-cyclopropyl-3-[3-(6-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea obviously refers to the same connection 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

Pyrazol cycle can also demonstrate the tautomerism and can exist in two tautomeric forms C and D, shown below

In addition, as shown below, the possible CIS - and TRANS-conformations of urea

References 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its salts also include variants of compounds with one or more isotopic substitutions, and mention one or another specific element includes all isotopes of this element. For example, the mention of hydrogen include1H,2H(D) and3H(T). Similarly, references to carbon and oxygen include, respectively12C,13C and14C and16O and18O.

These isotopes can be radioactive or non-radioactive. In one embodiment, assests the of the present invention compounds do not contain radioactive isotopes. Such compounds are preferred for therapeutic use. However, in another embodiment, the compounds can contain one or more radioactive isotopes. Compounds containing such radioactive isotopes may be useful in diagnostic applications.

In addition, the reference 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its salts covered by any polymorphic form, a solvate (e.g. hydrate) and complexes (e.g. inclusion complexes or clathrates with compounds, such as cyclodextrins, or complexes with metals) of this connection.

The lactates and citrates, and mixtures thereof crystals

As should be clear from previous sections of this application, the preferred salts of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea are acid additive salts formed lactic acid (more preferably L-lactic acid), citric acid or mixtures thereof.

For convenience salts formed lactic acid, L-lactic acid and citric acid can be named in this application lactate, L-lactate and citrate, respectively.

In one specific embodiment, the salt is an L-lactate or D-lactate, preferably L-lactate.

In other variants of the e implementation of the salt is a salt, formed citric acid.

More specifically, the salts are mixtures of L-lactate and citrate.

In the solid state the lactate (particularly the L-lactate) or citrate of the present invention may be crystalline or amorphous, or a mixture of these two forms.

In one embodiment, the implementation of the lactate (particularly the L-lactate) or citrate are amorphous.

In an amorphous solid substance no three-dimensional structure, which usually exist in crystalline form, and position of the molecules relative to each other in amorphous form are essentially random (see, for example, Hancock et al. J. Pharm. Sci. (1997), 86, 1).

In another embodiment, the lactate (particularly the L-lactate) or citrate are mostly crystalline, i.e. they can be 50-100% of the crystal and, more specifically, they can be crystalline at least 50%, or crystalline at least 60%, or crystalline at least 70%, or crystalline at least 80%, or a crystalline, at least 90%, or crystalline at least 95%, or crystalline at least 98%, or a crystalline at least 99%, or crystalline at least 99.5%, or crystalline at least 99.9%, for example crystalline 100%.

In another embodiment, lacto is (in particular, L-lactate) or citrate selected from the group consisting of lactate (particularly the L-lactate) or citrate, which are crystalline at 50-100%, for example crystalline at least 50%crystalline, at least 60%crystalline, at least 70%crystalline, at least 80%crystalline, at least 90%crystalline, at least 95%crystalline, at least 98%crystalline, at least 99%crystalline, at least 99.5% crystalline, at least 99.9%, for example crystalline 100%.

More preferably, the lactate (particularly the L-lactate) or citrate can be like salt (or to be selected from the group consisting of such salts), which are crystalline at 95-100%, for example crystalline, at least 98%, or crystalline at least 99%, or crystalline at least 99.5%, or crystalline at least 99.6 percent, or crystalline at least 99.7%, or crystalline at least 99.8%, or crystalline at least 99.9%for example crystalline 100%.

One example of mainly crystalline salt is a crystalline salt formed of L-lactic acid.

Another example of a mainly crystalline salt is kristallicheskaja, educated citric acid.

Salt of the present invention in the solid state can be solvated (e.g., hydrated) or resolutionunit (for example, anhydrous).

In one of the embodiments of salt are resolutionunit (for example, anhydrous).

Another example resolutional salt is a crystalline salt formed lactic acid (in particular, L-lactic acid), as defined in this application.

The term "anhydrous" in this application does not preclude the presence of a quantity of water on or within salt (for example, salt crystals). For example, water may be present in some quantity on the surface of a salt (for example, salt crystals), or a slight amount of water may be present in the amount of salt (e.g., crystals). Typically, the anhydrous form contains less than 0.4 molecules of water per molecule of the compound, more preferably contains less than 0.1 molecule of water per molecule of the compound, for example 0 water molecules.

In another embodiment, the lactate (particularly the L-lactate) or citrate are solvated. If salt gidratirovana, they may contain, for example, up to three molecules of water of crystallization, more usually up to two water molecules, for example, one water molecule and two water molecules. Also mo is ut to form non-stoichiometric hydrates, where available, the number of water molecules is less than one or, in another case, not an integer. For example, in the case of less than one molecule of water may be present, for example, 0.4, or 0.5, or 0.6, or 0.7, or 0.8, or 0.9 molecules of water per molecule of the compound.

Other solvate include alcoholate, such as ethanolate and isopropanolate.

In one embodiment, the implementation of the salt of lactic acid (in particular, a salt of L-lactic acid) solvated, for example, water and/or ethanol.

L-lactate or citrate can be obtained by methods described in the previous sections of this application and in any place of this application.

Benefits of L-lactates and citrates include General benefits outlined above in the previous sections of this application. However, crystalline lactate according to the present invention has special advantages in that it:

- non-hygroscopic;

- is anhydrous and does not form hydrates;

- exist in the same crystalline form and, as is, does not show polymorphism;

- is crystalline;

is stable during storage;

- has a distinct melting point and does not show changes forms, the analysis using DSC (differential scanning calorimetry);

- possesses good solubility in water; and

- demonstrates flexibility the solubility in buffer systems.

Thus, L-lactate exists in a stable crystalline form, which does not form hydrates, and not subject to change shape during standard operations, processing and storage conditions.

The term "stable" ("stable"or "stability" ("sustainability") when used in this description includes the chemical stability and physical stability of the solid state. The term "chemical stability" means that the compound can be stored in pure form or in the form of a composition in which it is included in the mixture, for example, pharmaceutically acceptable carriers, diluents or excipients described in the present application, in normal storage conditions, with little or no chemical decay or decomposition. "The stability of a solid state" means that the compound can be stored in a dedicated solid form or in the form of a solid composition in which it is included in the mixture, for example, pharmaceutically acceptable carriers, diluents or excipients described in the present application, in normal storage conditions, with little or no change in the solid state (e.g., hydration, dehydration, solvation, desolately, crystallization, recrystallization or phase transition in the solid is the state).

L-lactate and citrate, and mixtures thereof have good solubility in water and, therefore, can be used to prepare aqueous solutions containing relatively high concentrations of salts. Accordingly, in another embodiment, the developed aqueous solution (for example, in the form of pharmaceutical compositions)containing L-lactate or citrate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, or a mixture in concentrations greater than 1 mg/ml, typically greater than 5 mg/ml of a liquid medium (e.g. water or buffer systems), more typically greater than 15 mg/ml, more typically exceeding 20 mg/ml and preferably greater than 25 mg/ml In this embodiment, aqueous solutions (for example, in the form of a pharmaceutical composition containing (i) L-lactate or (ii) a mixture of L-lactate and citrate, are particularly preferred.

Aqueous solutions of L-lactate or citrate, or mixtures thereof can be obtained in the form of aqueous solutions having a pH in the range from 2 to 6, for example from 2 to 5, and more particularly from 4 to 6, such as from 4 to 5.

Aqueous solutions of L-lactate or citrate, or mixtures thereof can be buffered or nezabyvaemymi, but, in one of the embodiments, they are buffered, for example, to a pH in the above range.

Preferred the buffers are, capable of supererogate solution to a pH of approximately 4.5 and which are not volatile under the conditions used for lyophilization of a solution.

In the situation when the salt is formed L-lactic acid, the preferred buffer is a buffer obtained from citric acid and brought to the desired pH with NaOH or HCl, for example, until the pH value of the solution is approximately 4.5. At this pH and citrate buffer free base has a solubility of about 80 mg/ml

Aqueous solutions of L-lactate or citrate, or mixtures thereof will contain 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in protonated form together with L-lactate and/or citrate counterions. There can be other counterions, and they can be formed, for example, reagents that regulate toychest, such as saline solution (i.e. chlorine counterions) and/or buffering agents, such as citrate buffers. For example, if L-lactate is mixed in aqueous solution with citrate buffer, will be present as counterions L-lactic and citric acid, and the nature citrate counterion depends on the pH of the solution. In addition, aqueous solutions of L-lactate or citrate or mixtures thereof may contain one or more other fillers, as a rule, Eleusis the in compositions for intravenous injection, as, for example, reagents for regulating toychest, examples of which are described in detail in the United States Pharmacopeia and the national pharmaceutical formulary and include hexose, such as glucose, for example, dextrose (D-glucose).

Therefore, in another embodiment, the invention relates to a water solution of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in protonated form, containing one or more counterions selected from L-lactate, citrate, and mixtures thereof; and optionally (i) one or more other counterions, such as chloride ions and/or (ii) one or more fillers for intravenous formulations, such as, for example, means regulating toychest (e.g., hexoses, such as glucose, preferably D-glucose).

Aqueous solutions, among other methods, can be obtained by dissolution of lactate in the solution of citrate ions (for example, citrate buffer) or dissolution of citrate in the solution of lactate ions. Lactate and citrate ions may be present in solution in the ratio of lactate:citrate from 10:1 or less, e.g. from 10:1 to 1:10, more preferably less than 8:1, or less than 7:1, or less than 6:1, or less than 5:1, or less than 4:1, or less than 3:1, or less than 2:1, or less than 1:1, more specifically from 1:1 to 1:10. In one embodiment, the OS is enforced lactate and citrate ions are present in solution in the ratio of lactate:citrate from 1:1 to 1:10, for example, from 1:1 to 1:8 or 1:1 to 1:7 or 1:1 to 1:6 or 1:1 to 1:5, for example about 1:4,4.

Each of the aqueous solutions described in this section of the application, as well as anywhere in the application, may be subjected to lyophilization to obtain a solid composition, which optionally can be easily restored with the formation of an aqueous solution (preferably sterile solution) by adding water (preferably sterile water or aqueous medium containing the filler intravenous, such as saline and/or dextrose.

Accordingly, the present invention also relates to liofilizovannye composition (for example, in the form of a pharmaceutical composition)comprising L-lactate or citrate, or mixtures thereof, as defined in this application, for example, where the composition when dissolved in water has a pH of from 2 to 6, for example, from 2 to 5 and more particularly from 4 to 6, such as from 4 to 5.

In another embodiment, the invention relates to liofilizovannye composition (for example, in the form of a pharmaceutical composition comprising 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in protonated form together with one or more counterions selected from L-lactate, citrate and mixtures thereof; and optionally (i) one or more other counterions such as chloride-IO is s and/or (ii) one or more fillers for intravenous injection, such as the regulation of toychest (for example, hexose, such as glucose, preferably D-glucose).

The ratio of L-lactate ions to citrate ions in each of liofilizovannyh compositions may be in the limits established above for ionic solutions.

Crystal structure of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its salts

As mentioned above, the lactate (particularly the L-lactate) or citrate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea can be amorphous or substantially crystalline. In a specific embodiment, the lactate (particularly the L-lactate) or citrate are essentially crystalline, the term "essentially crystalline" has the meaning given above. In particular, lactate (including L-lactate) of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea is essentially crystalline.

1-Cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form may also be amorphous or crystalline creature. In a specific embodiment, the free base is essentially crystalline, the term "essentially crystalline" has the meaning given above. In one of the variations is tov the implementation of the free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea exists in the form of crystalline dihydrate.

The crystals described in this application, and the structure of crystals form another aspect of the present invention.

As noted above, it is believed that lactate according to the present invention exists in a single crystalline form, which has the characteristics set forth in this description. This crystalline form is a preferred embodiment of the invention. However, if other crystalline forms do exist, they are not excluded from the scope of the present invention.

Thus, if the lactate (particularly the L-lactate) of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea is essentially crystalline, can prevail only one form (e.g., crystalline form, as defined and described in the present description), although other crystalline forms may be present in minor and, preferably, is negligibly small amounts.

Crystalline forms (for example, crystalline forms that are defined and described in the present description) 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (or its salts) contain other crystalline forms of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (or its salts in to the number, less than or equal to about 5% by weight, in particular the content of other crystalline forms (or their salts) less than or equal to about 1% by weight.

In a preferred embodiment, the invention relates to a mainly crystalline salt (e.g., lactate, such as L-lactate, as described in this application) 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, containing a single crystalline form (e.g., crystalline form as defined and described in the present description) salt and not more than 5% by weight of other crystalline forms of salts.

Preferably along with a single crystalline form (e.g., crystalline form, as defined and described in the present description) is less than 4% or less 3%or less than 2% of other crystalline forms and, in particular, a number of other crystalline forms of less than or equal to about 1% by weight. More preferably a single crystalline form (e.g., crystalline form, as defined and described in the present description) is accompanied by less than 0.9%, or less than 0.8%, or 0.6%, or less than 0.5%, or less than 0.4%, or less than 0.3%, or less than 0.2%, or less than 0.1%, or less than 0.05%, or less than 0.01% by weight of other crystalline forms, for example, 0% by weight of other crystalline forms.

Chris is Alla and their crystal structure can be characterized using several techniques, including x-ray analysis of single crystals, x-ray diffraction on the powder (XRPD), differential scanning calorimetry (DSC) and infrared spectroscopy, such as infrared spectroscopy with Fourier transform (FTIR). The behavior of the crystals in the conditions of changing humidity can be analyzed by gravimetric studies on the absorption of water vapor and, in addition, XRPD.

The crystal structure of compounds can be carried out by x-ray diffraction analysis, which can be carried out according to standard methods, as described in this application and described in Fundamentals of Crystallography, C. Giacovazzo, H.L. Monaco, D. Viterbo, F. Scordari, G. Gilli, G. Zanotti and M. Catti, (International Union of Crystallography/Oxford University Press, 1992 ISBN 0-19-855578-4(p/b), 0-19-85579-2(h/b)). These techniques include the analysis and interpretation of x-ray diffraction on a single crystal.

The crystal structure of the dihydrate free base and L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea were determined by x-ray analysis (see below examples 69 and 71, respectively).

In tables 2 and 4 examples 69 and 71, respectively, the data on the coordinates of the atoms in the crystals of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and L-LACT is in the format of the Crystallographic Information File (CIF) (see Hall, Allen & Brown, Acta Cryst. (1991). A47, 655-685; http://www.iucr.ac.uk/iucr-top/cif/home.html). Other specialists in this field of technology can be used or preferred alternative file formats such as PDB format (for example, a format compatible with the format of the EBI Macromolecular Structure Database (Hinxton, UK). However, it should be clear that the application to real data in another file format, or convert the coordinates in tables are in the scope of the present invention. In the tables the numbers in parentheses represent the deviation (s.u., standard uncertainty). Crystal structure of lactate shown in Fig. 4 and 5.

In one of the embodiments the invention relates to a dihydrate free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea which is crystalline and (i) has a crystalline structure defined by the coordinates in table 2 of this application; and/or (ii) whose crystals belong to the monoclinic space group P2l/n (#14) with crystal lattice parameters of a=7,66(10), b=15,18(10), c=17,71(10)Å, β=98,53(2)°, α=γ=90°.

In another embodiment, the invention relates to L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea which is crystalline and has a crystal structure defined by the coordinates in table the goal 4 of the present description.

In another embodiment, the invention relates to L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea which is crystalline and has a crystal structure shown in figure 4 and 5.

In another embodiment, the invention relates to L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea which is crystalline and has a crystal structure that belongs to the orthorhombic space group P2l2l2l(#19) and has the lattice parameters at 97(2)K a=9,94(10), b=15,03(10), c=16,18(10)Å, α=β=γ=90°.

In another embodiment, the invention relates to L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea which is crystalline and has lattice parameters at room temperature a=10,08(10), b=15,22(10), c=16,22(10)Å, α=β=γ=90°.

According to another variant implementation of the present invention relates to L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea which is crystalline and:

(a) has a crystalline structure, shown in Fig. 4 and 5; and/or

(b) has a crystalline structure defined by the coordinates in table 4 of the present description; and/or

(c) has the lattice parameters at 97(2)K a=9,94(10), b=15,03(10), c=16,18(10)Å, α=β=γ=90°; and/or

(d) has the lattice parameters at room temperature a=10,08(10), b=15,22(10), c=16,22(10)Å, α=β=γ=90°; and/or

(e) has a crystal structure that belongs to the orthorhombic space group P2l2l2l(#19).

On the other hand, the crystal structure of compounds can be analyzed using applicable for solids methods of x-ray diffraction on the powder (XRPD). XRPD can be carried out according to conventional methods, as for example described in this application (see examples 70 and 72), and is described in Introduction to X-Ray Powder Diffraction, Ron Jenkins and Robert L. Snyder (John Wiley & Sons, New York, 1996). The presence of pronounced peaks (as opposed to random background noise) in the diffraction pattern indicates that the connection has a certain degree of crystallinity.

Powder x-ray connection is characterized by the parameters of the x-ray diffraction spectrum, the diffraction angle (2θ) and interplanar distance (d). These parameters are related by the Bragg equation nλ=2dsin θ (where n=1; λ=wavelength of the applied cathode, d=interplanar distance; and θ=diffraction angle). In this way the interplanar distance, the diffraction angles and the General appearance of the picture are important for the identification of the crystal powder x-ray, through the I character of these data. The relative intensity must not be an accurate interpretation, as it may change depending on the direction of crystal growth, particle size and conditions of registration of the spectrum. In addition, the diffraction angles usually means the angles that coincide in the range of 2θ±0,2°. The peaks indicate the main peaks and include peaks not exceeding the average at diffraction angles different from the angles specified above.

As L-lactate, and the form of the free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea were characterized by XRPD. In each case, the powder x-ray is expressed in the values of the diffraction angles (2θ), interplanar spacings (d) and/or relative intensities. In tables 3, 5 and 6 in examples 70 and 72 are shown values of interplanar distances (d) spectrum of x-ray diffraction, which correspond to the values of the diffraction angles in a free base, L-lactate and dihydrate free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

Therefore, 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its salts have a picture of the x-ray diffraction, the main features shown in figure 3, 6, 7, or 8 and/or in tables 3, 5, or 6 in examples 70 and 72.

In accordance with one of the three options for implementing the present invention relates to crystals of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form and L-lactate, showing powder x-ray that includes peaks at the same diffraction angles as peaks in the powder x-ray shown in Fig. 3, 6, 7, or 8 and/or table 3 and/or table 5 and/or table 6, and these peaks do not necessarily have the same relative intensity. More specifically, the crystals of these salts are such that they have a powder x-rays, basically similar to the one shown in Fig. 3, 6, 7, or 8.

In a preferred embodiment, the invention relates to crystals of salt 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and L-lactic acid, powder x-ray which is essentially shown in Fig.6.

In another embodiment, the invention relates to a mainly crystalline L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, which shows peaks corresponding to the same diffraction angles as in the powder x-ray shown in Fig.6. Preferably, the peaks have the same relative intensity as the peaks of figure 6.

The invention also relates to essentially crystalline salt of L-lactic acid and 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, powder x-ray which in the main corresponds shown in Fig.6.

X ray powder salt of L-lactic acid can be characterized by the presence of peaks corresponding to the diffraction angles (2θ) and interplanar distances (d) and, preferably, the intensities shown in table 5 of example 72.

Consequently, the invention relates to crystals of L-lactate cyclopropyl-3-[3-(6-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, which show the powder x-ray having characteristic peaks corresponding to the diffraction angles (2θ±1,0°, as, for example, ±0,2°, in particular of ±0.1°) of table 5 in example 72.

In addition, the invention relates to crystals of L-lactate cyclopropyl-3-[3-(6-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, x-ray diffraction on the powder which shows main peaks corresponding to the diffraction angles 2θ equal 17,50, 18,30, 19,30, 19,60 and 21.85±1,0°, as, for example, ±0,2°, in particular of ±0.1°. These crystals can be further characterized by peaks in the pattern of diffraction of x-rays at angles 2θ 12,40, 15,20, 15,60, 17,50, 18,30, 18,50, 19,30, 19,60, 21,85 and 27,30±1,0°.

Crystals of L-lactate cyclopropyl-3-[3-(6-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea also differ in that the characteristic powder x-ray presents the distances between the planes of the crystal lattice de), specified in table 5 in example 72.

In another embodiment, the invention relates to crystals of lactate cyclopropyl-3-[3-(6-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, x ray powder which comprises characteristic peaks appearing at distances in the crystal lattice (d) of 5.06, 4,85, 4,60, 4,53 and 4,07, and more specifically includes an additional characteristic peaks appearing at distances in the crystal lattice (d) 7,13, 5,83, 5,68, 5,06, 4,85, 4,79, 4,60, 4,53, 4,07 and 3.26 Angstrom.

In another embodiment, the invention relates to essentially crystalline L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, x ray powder which is characterized by the presence of major peaks corresponding to the diffraction angles (2θ) 17,50, 18,30, 19,30, 19,60 and 21,85°, more specifically 12,40, 15,20, 15,60, 17,50, 18,30, 18,50, 19,30, 19,60, 21,85 and 27,30° and the interplanar distances (d)equal to is 5.06, 4,85, 4,60, 4,53 4,07 and, more specifically 7,13, 5,83, 5,68, 5,06, 4,85, 4,79, 4,60, 4,53, 4,07 and 3.26 Angstrom.

In another embodiment, the invention relates to essentially crystalline L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, x ray powder which is characterized by the presence of peaks corresponding to the diffraction angles (2θ), milosc the local distances (d) and preferably, the intensities shown in table 5 in example 72.

In addition, the invention relates to crystals of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form, powder x-ray which includes the characteristic peaks corresponding to the diffraction angles (2θ±1,0°, as, for example, ±0,2°, in particular of ±0.1°) of table 2.

In another embodiment, the invention relates to crystals of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form, in powder x-ray which has characteristic peaks appearing at interplanar distances (d) from table 2.

Crystalline salts of the present invention can also be characterized by differential scanning calorimetry (DSC).

L-lactate was analyzed by DSC method and showed a peak (melting point and decomposition) at 190°C.

Accordingly, in another aspect the invention relates to L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, which is anhydrous and shows an endothermic peak at 190°C in the study by DSC method.

Another aspect of the present invention is L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-Ben is imidazol-2-yl)-1H-pyrazole-4-yl]urea which shows peaks corresponding to the same diffraction angles, which are present in the film x-ray diffraction shown in Fig. 6, 7, or 8, and, in addition, exhibits an endothermic peak accompanying the decomposition, near 190°C according to thermal analysis (DSC).

The form of the free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea shows peaks at the same diffraction angles as peaks in the powder x-ray shown in figure 3 and/or table 2 and, in addition, exhibits an exothermic peak accompanying the decomposition, near 193°C according to thermal analysis (DSC).

The behavior of the salts of the present invention in high humidity conditions can be investigated by standard methods gravimetric analysis absorption of vapors (GVS), for example, as described in section E of example 68.

L-lactate can exist in a stable anhydrous crystalline form under conditions of high relative humidity, and under such conditions, the crystalline structure is not modified.

Salt 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea can be also characterized by infrared spectroscopy, such as FTIR. The infrared spectrum of L-lactate (with KBr tablet) includes features and advantages of the systematic peaks at 3229, 2972 and 1660 cm-1.

Accordingly, in another embodiment, the invention relates to (preferably essentially crystalline) L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, which shows the infrared spectrum, using the method with a KBr tablet, including characteristic peaks at 3229, 2972 and 1660 cm-1.

As should be clear from the previous paragraphs, L-lactate according to the present invention can be characterized in a number of different physico-chemical parameters. Accordingly, in the preferred embodiment, the present invention relates to L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea which is crystalline and characterized by one or more (in any combination)or all parameters from among the following, namely that the salt:

(a) has a crystalline structure, shown in Fig. 4 and 5; and/or

(b) has a crystalline structure defined by the coordinates in table 4 of example 71 of the present description; and/or

(c) has the lattice parameters at 97(2)K a=9,94(10), b=15,03(10), c=16,18(10)Å, α=β=γ=90°; and/or

(d) has the lattice parameters at room temperature a=10,08(10), b=15,22(10), c=16,22(10)Å, α=β=γ=90°; and/or

(e) has a crystalline structure, to the which belongs to the orthorhombic space group P2 l2l2l(#19); and/or

(f) has a powder x-ray characterized by the presence of major peaks corresponding to the diffraction angles (2θ) 17,50, 18,30, 19,30, 19,60 and 21,85°, and more particularly advanced when 12,40, 15,20, 15,60, 17,50, 18,30, 18,50, 19,30, 19,60, 21,85 and 27,30° and/or the interplanar distances (d)equal to is 5.06, 4,85, 4,60, 4,53 and 4,07, and more specifically advanced 7,13, 5,83, 5,68, 5,06, 4,85, 4,79, 4,60, 4,53, 4,07 and 3.26 Angstrom; and/or

(g) shows peaks corresponding ibid diffraction angles that are in the film x-ray diffraction, is shown in Fig.6 or table 5 of example 72 and, optionally, where the peaks have the same relative intensity as the peaks of figure 6 or table 5; and/or

(h) has a powder x-ray are essentially corresponds shown in Fig.6; and/or

(i) is anhydrous and shows an endothermic peak at 190°C in the study by DSC method; and/or

(j) in the study applying the methodology with KBr tablet has an infrared spectrum that contains characteristic peaks at 3229, 2972 and 1660 cm-1.

Compounds of subgroup (C) of the formula (I)

In one of the subgroups of compounds of formula (I) (i.e. the subgroup of (C) of the formula (I), M is a group D1; X represents O; a is a group NR2where R2is hydrogen; E is a bond; R1JW is aetsa 2,6-differenial; and the connection is an acid additive salt, formed a select group of acids.

Accordingly, in one of the embodiments the invention relates to an acid additive salt of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, which is a salt formed by the acid selected from the group consisting of acetic, adipic, alginic, ascorbic (e.g. L-ascorbic), aspartic (e.g. L-aspartic), benzosulfimide, benzoic, camphoric (for example,

(+)camphor), capric, Caprylic, carbonic, citric, reklamowa, dodecanol, modellserie, ethane-1,2-disulfonate, econsultancy, fumaric, galatarasay, hentaimovi, glucoheptonate, D-gluconic, glucuronic (for example, D-glucuronic), glutamic (for example, L-glutamic), α is oxoglutarate, glycolic acid, hippuric, hydrochloric, isetionate, isoalkanes, breast (e.g., (+)- L-lactic and (±)-DL-lactic), lactobionic, laurylsulphate, maleic, malic, (-)-L-malic, malonic, methansulfonate, mucus, naphthalenesulfonates (for example, naphthalene-2-sulphonic, naphthalene-1,5-disulfonate, nicotine, oleic, orotovoy, oxalic, palmitic, pambou, phosphoric, propionic, sebacinales, stearic, succinic, sulfuric, tartaric (for example,(+)-L-tartaric), titanoboa, toluensulfonate (for example, p-toluensulfonate), valerianic and xinfulai acids.

In one embodiment, the implementation of the acid additive salt formed by the acid selected from the group consisting of adipic, alginic, ascorbic (e.g. L-ascorbic), aspartic (e.g. L-aspartic), benzoic acid, camphor (for example, (+)camphor), capric, Caprylic, carbonic, reklamowa, dodecanol, modellserie, ethane-1,2-disulfonate, galatarasay, hentaimovi, glucoheptonate, D-gluconic, glutamic (for example, L-glutamic), α is oxoglutarate, glycolic acid, hippuric, isoalkanes, laurylsulphate, mucus, naphthalene-1,5-disulfonate, nicotine, oleic, orotovoy, oxalic, palmitic, pambou, sebacinales, stearic, wine (for example, (+)-L-tartaric), titanoboa and xinfulai acids.

In another embodiment, the acid additive salt formed by the acid selected from the group consisting of acetic, adipic, ascorbic, aspartic, citric, DL-lactic, fumaric, gluconic, glucuronic, the hippuric, hydrochloric, glutamic, DL-malic, p-toluensulfonate, methanesulfonate (Sol-mesilate), econsultancy (Sol-Eilat), sebacinales, stearic, succinic, and tartaric acids.

In another embodiment, the acid-Additi the Naya salt formed by the acid, selected from the group consisting of adipic, ascorbic, aspartic, gluconic, the hippuric, glutamic, sebacinales, stearic and tartaric acids.

In another separate embodiment, the compound is an acid additive salt formed with hydrochloric acid.

Preferred salts are salts having a solubility in a given liquid carrier (e.g. water)in excess of 25 mg/ml of a liquid medium (e.g. water), more typically greater than 50 mg/ml and preferably greater than 100 mg/ml. Such salts are especially preferred for administration in liquid form, for example, by injection or infusion.

In another aspect of the present invention is designed composition (e.g., pharmaceutical composition)comprising an aqueous solution containing the above-described Sol, in concentrations greater than 25 mg/ml, typically greater than 50 mg/ml and preferably greater than 100 mg/ml

Salt of the present invention, which have a solubility in excess of 25 mg/ml, include D-glucuronate, mesilate, Eilat and DL-lactate, and the last three have a solubility of more than 100 mg/ml

Accordingly, in one specific embodiment, designed mesilate 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

In another specific options the ante implementation developed Eilat (econsultant) 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

In another specific embodiment, designed DL-lactate 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea. In one embodiment, the implementation of lactate is an L-lactate.

Free base or source connection of these compounds (i.e. acid-salt additive) subgroup (C) of the formula (I)has the formula (IA):

Salts of the compounds (IA) can be amorphous or crystalline.

In one embodiment, the implementation of the salt is an amorphous form.

In another embodiment, the compound has a crystalline form.

The connection can be nonsolvated (for example, anhydrous or solvated.

In one of the embodiments of salt resolutionary.

In another embodiment, the salt solvated, for example gidratirovana.

If compounds are hydrated, they may contain, for example, up to three molecules of water of crystallization, often up to two water molecules, for example, one water molecule and two water molecules.

Salt of the present invention have advantages over the form of the free base, i.e. the 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

For example, the salts have a high solubility in water and therefore, better suited for use in the preparation of parenteral compositions for injection or infusion (e.g., intravenous infusion). Salt of the present invention, in addition, have one or more benefits selected from:

best pharmacokinetics;

- higher stability, for example, increased retention;

- lower basicity, which makes them more suitable for intravenous administration;

- advantages for production;

the best metabolic properties; and

- smaller differences in clinical effectiveness from patient to patient.

Salts can be obtained by any of the methods outlined in the previous sections of this proposal describing the salts of the compounds of groups (A) and (B) of the formula (I).

Compounds of subgroup (C) of the formula (I)are pharmaceutically acceptable salts. However, salts which are not pharmaceutically acceptable may also be obtained as an intermediate form, which can then be converted into pharmaceutically acceptable salts. Such forms of pharmaceutically unacceptable salts, which can be useful, for example, when cleaning or separation of the compounds of the present invention, also included in the scope of the invention.

Compounds of subgroup (C) of the formula (I) can exist in the form of a number of different taut the situations forms, and references to compounds of the present invention include all such forms. For the avoidance of doubt, if a connection can exist in one of several geometrically isomeric or tautomeric forms, and specifically described or shown only one of them, all of them, however, are also covered by the present application.

For example, the compounds of the present invention benzimidazole group can take one of two tautomeric forms A and B shown above.

Pyrazol cycle can also be tautomerism and can exist in two tautomeric forms, C' and D'shown below.

Compounds of the present invention include compounds with one or more isotopic substitutions, and mentions of a particular item include all isotopes of this element. For example, the mention of hydrogen include1H,2H(D) and3H(T). Similarly, references to carbon and oxygen include, respectively12C,13C and14C and16O and18O.

These isotopes can be radioactive or non-radioactive. In one of the embodiments of the present invention compounds do not contain radioactive isotopes. Such compounds are preferred for therapeutic use. However, in the other embodiment, the compounds can contain one or more radioactive isotopes. Compounds containing such radioactive isotopes may be useful in diagnostic applications.

In addition, the present invention covered by any of the polymorphic forms of the compounds and complexes (e.g. inclusion complexes or clathrates with substances such as cyclodextrins, or complexes with metals) compounds and prodrugs of the compounds. By "prodrugs" refers to, for example, any compounds that are transformed in vivo to the biologically active compounds of the present invention.

Biological activity

Compounds of the present invention have inhibitory or modulating activity against cyclin-dependent kinases and inhibiting or modulating activity against kinase-3 glycogen synthase (GSK3), and/or inhibiting or modulating activity against Aurora kinase, and this activity seems to be useful in the prevention or treatment of conditions or diseases mediated by these kinases.

For example, it appears that the compounds of the present invention will be applicable to relieve symptoms or reducing the frequency of cancer cases.

More specifically, the compounds of formula (I) and its subgroups, are inhibitors of cyclin-dependent kinases. For example, the compounds of the present invention have the activity against kinases CDK1, CDK2, CDK3, CDK4, CDK5, CDK6 and CDK7, and, in particular, cyclin-dependent kinases selected from CDK1, CDK2, CDK3, CDK4, CDK5 and CDK6.

Preferred compounds are compounds that inhibit one or more CDK kinases selected from CDK1, CDK2, CDK3, CDK4 and CDK5, for example, CDK1 and/or CDK2.

In addition, interest can imagine CDK4, CDK8 and/or CDK9.

The lactates or citrates 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea possess activity against CDK2, CDK4, CDK5, CDK6 and CDK9 kinase, and in particular CDK2.

Compounds of the present invention also have activity against kinase-3 glycogen synthase (GSK-3).

In addition, the compounds of the present invention have activity against Aurora kinases. Preferred compounds of the present invention are compounds having the value of the IC50smaller than 0.1 ám.

In particular, lactates or citrates 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea are inhibitors of Aurora kinases and, for example, inhibit Aurora A and/or Aurora B.

Many of the compounds of the present invention show selectivity for Aurora A kinase in comparison with CDK1 and CDK2, and such compounds represent one preferred implementation of the present invention. For example, many compounds of the present invention have conducted the ins IC 50against Aurora A, which lie in the range from one-tenth to one-hundredth on the value of the IC50related to CDK1 and CDK2.

Due to the activity of the compounds of the present invention in modulating or inhibiting CDK, Aurora kinase and kinase glycogen synthase they expected to apply when creating a means to slow down the cell cycle or restore control over it in case of abnormally dividing cells. Therefore, it is expected that these compounds will indeed be applicable in the treatment or prevention of proliferative disorders such as cancer. In addition, it appears that the compounds of the present invention will be applicable in the treatment of conditions such as viral infections, diabetes type II or non-insulin-dependent diabetes mellitus, autoimmune diseases, head trauma, stroke, epilepsy, neurodegenerative disorders, such as Alzheimer's disease, motor neuron disease, progressive supranuclear paralysis, corticobasal degeneration and disease Peak, for example, in the treatment of autoimmune diseases and neurodegenerative diseases.

One group of diseases and medical conditions for which, it seems, will be applicable compounds of the present invention, consists of viral infecti is, autoimmune diseases and neurodegenerative diseases.

Cdks play a role in cell cycle regulation, apoptosis, transcription, differentiation and activity of the Central nervous system. Therefore, CDK inhibitors could be useful in the treatment of diseases which are disorders of cell proliferation, apoptosis or differentiation as, for example, cancer. In particular RB+ve tumors may be particularly sensitive to CDK inhibitors. RB-ve tumors can also be sensitive to CDK inhibitors.

Examples of cancers for which manifests inhibitory action, include, but are not limited to the above, a cancer, for example cancer of the bladder, breast, colon (e.g. colorectal cancers, such as adenocarcinoma of the colon and adenoma of the colon, kidney, epidermis, liver, lung, for example adenocarcinoma, small cell lung cancer and non-small cell cancers of the lung, esophagus, gall bladder, ovary, pancreas e.g. exocrine cancer of the pancreas, stomach, cervix, thyroid, prostate, or skin, for example squamous cancer cells; hematopoietic tumors of lymphoid origin, such as leukemia, acute lymphocytic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, nehodzhkin the th lymphoma, lymphoma hairy cell or lymphoma Burkett; hematopoietic tumors of myeloid origin, for example acute and chronic myelogenous leukemia, myelodysplastic syndrome, or promyelocytic leukemia; follicularly thyroid cancer; tumors of mesenchymal origin, for example fibrosarcoma or rabdomyosarcoma; tumors of the Central or peripheral nervous system, for example astrocytoma, neuroblastoma, glioma or sandamu; melanoma; seminoma; teratocarcinoma; osteosarcoma, pigmentary xeroderma; keratoacanthoma; follicularly thyroid cancer; or Kaposi's sarcoma.

Cancer may be the varieties of cancer that are sensitive to inhibition of any or all of several cyclin-dependent kinases selected from CDK1, CDK2, CDK3, CDK4, CDK5 and CDK6, for example one or more CDK kinases selected from CDK1, CDK2, CDK4 and CDK5, such as CDK1 and/or CDK2.

Whether or not a particular case of cancer, a disease sensitive to inhibition of cyclin-dependent kinase or Aurora kinase, can be determined by analysis of cell growth, as shown below in examples 79 and 80, or the way that is described in the section titled "methods of diagnosis".

In addition, it is known that cdks play a role in apoptosis, proliferation, differentiation and transcription and, consequently, inhibitors DK could also be applicable in the treatment of the following diseases, in addition to cancer: viral infections, for example herpes virus, pox virus, Epstein-Barr virus Sindbis, adenovirus, HIV, HPV (human papilloma virus), HCV (hepatitis C virus) and HCMV (cytomegalovirus); preventing the development of AIDS in HIV-infected patients; chronic inflammatory diseases, such as systemic lupus erythematosus, autoimmune-mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease and autoimmune diabetes mellitus; cardiovascular diseases such as hypertrophy of the heart, restenosis, atherosclerosis; neurodegenerative disorders, for example Alzheimer's disease, related AIDS dementia, Parkinson's disease, amyotrophic lateral sclerosis, pigmentary retinitis, spinal muscular atrophy and cerebellar degeneration; glomerulonephritis; myelodysplastic syndromes, myocardial infarction, associated with ischemic lesions, bumps and lesions associated with reperfusion injury, arrhythmia, atherosclerosis, liver disease caused by toxins or alcohol-related hematological diseases, such as chronic anemia and aplastic anemia; degenerative diseases of the musculoskeletal system, such as osteoporosis and arthritis, aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple sclerosis, ill the deposits in the kidney and cancer pain.

In addition, it was found that some of the inhibitors of the cyclin-dependent kinase may be used in combination with other anticancer agents. For example, an inhibitor of cyclin-dependent kinases, as flavopiridol used in combination therapy with other anticancer agents.

Thus, for pharmaceutical compositions, uses or methods according to the present invention for the treatment of a disease or condition comprising abnormal cell growth, the disease or condition comprising abnormal cell growth in one of the embodiments is a cancer.

One group of cancer includes cancers of the breast person (for example, primary breast tumors, breast cancer without lymph nodes, invasive adenocarcinoma of the ducts of the breast, endometrioid form of breast cancer); and lymphoma cells of the mantle tissue. In addition, other cancers are colorectal and endometrial cancers.

Another subset of cancer include breast cancer, ovarian cancer, cancer of the colon, prostate cancer, esophageal cancer, squamous cancer cells and non-small cell carcinoma of the lung.

For compounds with activity against Aurora kinase, specific examples of cancers in which case, as expected the use of the ima compounds of the present invention, inhibiting Aurora kinase include:

different types of breast cancer person (for example, primary breast tumors, breast cancer without lymph nodes, invasive adenocarcinoma of the ducts of the breast, endometrioid form of breast cancer);

different types of ovarian cancer (e.g., primary tumors of the ovaries);

different types of cancer of the pancreas;

different types of bladder cancer man;

different types of colorectal cancer (e.g., primary colorectal cancer);

tumors of the stomach;

different types of kidney cancer;

different types of cancer of the cervix;

neuroblastoma;

melanoma;

lymphoma;

different types of prostate cancer;

leukemia;

endometrioid cancer of the uterus;

gliomas; and

non-Hodgkin's lymphoma.

Cancers that can successfully resist the action of inhibitors of Aurora kinases, include cancers of the breast, bladder, colorectal cancer, pancreatic cancer, ovarian cancer, nahodkinskuju lymphoma, glioma and endometrioid cancer of the uterus.

A separate subset of cancers that can particularly well to resist the action of inhibitors of Aurora kinases consists of cancer of the breast, ovaries, colon, liver, stomach and prostate.

Another subset of cancer Zabol the requirements, for the treatment of which can be particularly well suitable inhibitors of Aurora kinases consists of hematological cancers, particularly leukemia. Therefore, in another embodiment, compounds of formula (I) is used to treat hematological cancer, specifically leukemia. Specific types of leukemia is selected from acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), B-cell lymphoma (mantle cell tissue) and acute lymphoblastic leukemia (ALL) (on the other hand known as acute lymphocytic leukemia). In one embodiment, the implementation of the varieties of leukemia selected from re or refractory acute myelogenous leukemia, myelodysplastic syndrome, and chronic myelogenous leukemia.

One group of cancers includes various types of cancer of the breast person (for example, primary breast tumors, breast cancer without lymph nodes, invasive adenocarcinoma of the ducts of the breast, endometrioid form of breast cancer); and lymphoma cells of the mantle tissue. In addition, other types of cancer is colorectal cancer and various types of cancer of the uterus.

Another subset of cancer includes hematopoietic tumors of lymphoid origin, such as leukemia, chronic lymphocytic leukemia, lymphoma is one of mantle tissue and B-cell lymphoma ( for example, diffuse B-both lymphoma).

One specific type of cancer is chronic lymphocytic leukemia.

Another specific type of cancer is lymphoma cells of the mantle tissue.

Another specific type of cancer is diffuse B-both lymphoma.

Further provides that the compounds of the present invention and, in particular, those compounds that have inhibitory activity against Aurora kinase, will be particularly applicable in the treatment or prevention of those types of cancer that are associated with, or characterized by the presence of elevated levels of Aurora kinases, such as cancer, referred to in this context in the introductory part of this application.

The activity of the compounds of the present invention as inhibitors of cyclin-dependent kinases, Aurora kinases and kinase-3 glycogen synthase can be measured using analytical methods described below in the examples, and the level of activity exhibited by a given compound can be expressed in terms IC50. Preferred compounds of the present invention are compounds having the values of the IC50less than 1 micron, more preferably less than 0.1 microns.

The advantages of the compounds according to the present invention

Compounds of the present invention (for example, the connection is in examples 24, 62, 63 and 64) have several advantages over the compounds of the prior art. For example, the compounds of the present invention (see table A) demonstrate improved selectivity and efficiency, in particular in respect of Aurora A and B kinases.

Table A
Inhibition of Aurora kinases in vitro 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea
Aurora kinaseIC50(nm)
Aurora-A52% at 3 nm
Aurora-B58% at 3 nm
The kinase activity in vitro was determined in accordance with the protocols described in examples 75 and 76.

Compounds of the present invention also have advantages over the compounds of the prior art in that they have a different sensitivity to P450 enzymes (see table B below and example 81).

Table B
Inhibition of expressed isoforms of cytochrome P450 1-cyclopropyl-3-[3-(5-morpho is in 4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in vitro
Isoform of P450IC50(µm)
CYP1A2>10
CYP2D6>10
CYP3A4>10
CYP2C9>10
CYP2C19>10

In addition, the compounds of the present invention offer advantages over the compounds of the prior art in that they demonstrate the improved properties related to drug metabolism and pharmacokinetic properties. In particular, the compounds of the present invention have reduced binding to plasma protein. The binding of the compounds of examples 24, 62, 63 and 64 with plasma proteins was relatively moderate for all investigated species, ranging from 61% in the plasma of rats to 82% in the plasma of mice. This could provide a benefit available in the General circulation more are available free medicines to achieve the proper venue for the purposes of manifestation of its therapeutic effect. Increased fraction of free medicines for the manifestation of pharmacological action is pwhash potentially leads to increased efficiency, therefore there is a possibility of reduction of the injected dose.

Compounds of the present invention (for example, the compounds of examples 24, 62, 63 and 64) also exhibit increased activity compared to cells in proliferation and count of clonogenic assays (e.g., assays described in examples 79 and 80) against a wider range of cell lines and solid tumors, showing, thereby, improved anticancer activity (table C). The data show that treatment with compounds of the present invention otherwise affects tumor cells compared with normal cells. Treatment of compounds of the present invention of tumor cells with abnormal checkpoint leads to the formation of many nuclei, due to interruption of mitosis, inhibition of cytokinesis and blocking checkpoints education spindle, due to the inhibition of Aurora kinase. It is the emergence of multiagency leads, apparently, to cell death. In contrast, normal cells with the correct control point, treated with compounds of the present invention, a smaller portion of the cells becomes multi-core or dies within 24 hours after treatment, whereas most of undergoing a reversible delay in G2/M and then re-enters the cell cycle, as soon as the connection will cease the t act. These differences in results probably reflect the fact that normal cells have a reference point in such places, to stop the cell cycle if there is accurate segregation of chromosomes, such as post-mitotic p-53 dependent checkpoint. In tumor cells, these control points are missing, which makes possible the continuation of mitosis and the onset of multidirect.

Table C
Inhibitory effects of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea to form colonies of tumor cells
OriginOriginIC50(nm)p53 status*
The colonHCT 11613+
HCT 116 N714-
HT-2911-
SW62014+
ionic A27807,7+
EasyA54912+
Mammary glandMCF720+
PancreasMIA-Pa-Ca-27,8-
* + indicates expression of p53 wild-type; - indicates no expression of p53 or that p53 is non-functional.

In addition, the salt forms of the compounds of the present invention exhibit enhanced solubility in an aqueous environment and improved physico-chemical properties, for example, lower logD.

Methods for obtaining compounds of formula (I)

In this section, as in all other sections of this application unless the context indicates otherwise, references to formula (I) include formula (II), (III), (XXX) and all other sub-groups and examples of compounds defined in the present application.

The compounds of formula (I) can be obtained in accordance with synthetic methods well known to the specialist.

For example, the compounds of formula (I), in the which A is a bond (i.e. where A and carbonyl group to form an amide bond), can be obtained by the coupling of compounds of formula (X):

with a carboxylic acid R1-E-CO2H or its reactive derivative in the standard conditions of formation of amides.

The condensation reaction between carboxylic acid and amine (X) may be conducted in the presence of some types of reagents commonly used in the formation of peptide bonds. Examples of such compounds include 1,3-dicyclohexylcarbodiimide (DCC) (Sheehan et al., J. Amer. Chem. Soc. 1955,77, 1067), 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide (EDC) (Sheehan et al, J. Org. Chem., 1961, 26, 2525), condensing reagents based bronevich compounds, as, for example, hexaflurophosphate O-(7-asobancaria-1-yl)-N,N,N',N'-tetramethylurea (HATU) (L.A. Caprino, J, Amer. Chem. Soc., 1993,115, 4397) and condensing reagents based fofanah compounds, as, for example, hexaflurophosphate 1-benzothiazolylthio(pyrrolidino)phosphonium (PyBOP) (Castro et al, Tetrahedron Letters, 1990,31, 205). Condensing reagents based on carbodiimide mainly used in combination with 1-hydroxydibenzofuran (HOAt) or 1-hydroxybenzotriazole (HOBt) (Konig et al., Chem. Ber., 103, 708, 2024-2034). The preferred condensing reagents include EDC and DCC in conjunction with HOAt or HOBt.

The condensation reaction typically is carried out in non-aqueous, aproton the second solvent, such as acetonitrile, dioxane, dimethyl sulfoxide, dichloromethane, dimethylformamide or N -, an organic or aqueous solvent, optionally in combination with one or more additional mixing solvents. The reaction can be conducted at room temperature, or if the reagents have lower reactivity (for example, in the case of depleted electron density anilines bearing dilatory electrons of the group, as, for example, sulfonamidnuyu group), at the proper elevated temperature. The reaction can be performed in the presence of a base, which does not prevent its occurrence, for example, tertiary amine, such as triethylamine or N,N-diisopropylethylamine.

Alternatively, you can apply reactive derivatives of carboxylic acids, for example anhydrides or acid chlorides. Interaction with reactive derivatives, such as anhydrides, mostly carried out under stirring amine and anhydride at room temperature in the presence of a base, such as pyridine.

Amines of formula (X) can be obtained by reduction of the corresponding nitro compounds of formula (XI) under standard conditions. Recovery can be carried out, for example, by catalytic hydrogenation in the presence of a catalyst such as palladium on charcoal, in a polar the solvent, as, for example, ethanol or dimethylformamide at room temperature.

Nitro compounds of formula (XI) can be obtained by the interaction nitropyrazole acid of formula (XII):

or with 4-morpholine-4-ylmethylene-1,2-diamine (to obtain compounds in which M represents D1)or 4,5-dimethoxybenzene-1,2-diamine (to obtain compounds in which M represents D2).

The interaction between the diamine and carboxylic acid (XII) can be carried out in the presence of such reagents as DCC or EDC in the presence of HOBt as described above, in the conditions of the condensation reaction to obtain the amides described previously, with education as the intermediate ortho-aminophenylamino (not shown), which is then subjected to cyclization with the formation of the benzimidazole cycle. The final stage cyclization, as a rule, is carried out by boiling under reflux in the presence of acetic acid.

Illustrative reaction scheme showing the formation of compounds of formula (X), where M stands for a group D1, shown in figure 1.

Typical conditions for each stage in figure 1 can be found in the "examples" section below.

Compounds in which M stands for a group D2 can be obtained than the ranks way but using 4,5-dimethoxybenzene-1,2-diamine instead of the diamine (XVI) in scheme 1.

In an alternative method of synthesis of compounds of formula (I)in which A represents a bond, diamines, i.e. the 4-morpholine-4-ylmethylene-1,2-diamine and 4,5-dimethoxybenzene-1,2-diamine, may also be introduced in the interaction with the carboxylic acids of the formula (XVII), where A denotes a bond, with formation of compounds of formula (I).

The interaction of a diamine with a carboxylic acid (XVII) can be carried out in conditions similar to those described above to obtain the nitro compound (XI). Carboxylic acids of the formula (XVII) can be obtained from the sequence of reactions shown in scheme 2.

As is evident from figure 2, substituted or unsubstituted 4-nitro-3-pyrazolylborate acid (XVIII) can be etherification interaction with thionyl chloride with the formation of the carboxylic acid as an intermediate and the subsequent interaction with ethanol, leading to obtaining the ethyl ester (XIX). On the other hand, the esterification can be carried out by the interaction of alcohol and carboxylic acid in the presence of an acid catalyst, one example of which is thionyl chloride. This reaction is usually carried out at room temperature using tarifitsiruemih alcohol (e.g. ethanol) to the operation of the solvent. Then the nitro-group can be restored with the use of palladium on coal according to the standard methods of obtaining amine (XX). The amine (XX) enter into a condensation reaction with the corresponding carboxylic acid R1-E-CO2H in the conditions of obtaining the amides, which coincide with the conditions described above, or similar, to obtain the amide (XXI). Then ester group, amide (XXI) can be hydrolyzed using alkali metal hydroxide such as sodium hydroxide, in a polar miscible with water, a solvent such as methanol, typically at room temperature.

The compounds of formula (I)in which A denotes the NR2can be obtained using standard methods of synthesis of ureas.

For example, such compounds can be obtained by the interaction derived aminopyrazole formula (X) with a suitably substituted isocyanate of formula R1-E-N=C=O in a polar solvent, such as DMF. The reaction is usually carried out at room temperature.

Another way to urea of formula (I) can be obtained by the interaction of the amine of formula (X) with an amine of formula R1-E-NH2in the presence of carbonyldiimidazole (CDI). The reaction is usually carried out in a polar solvent, such as THF, heating (for example, using microwave heating condition is the device) at a temperature of approximately 150°C.

Instead of using the CDI reaction of condensation of two amines with the formation of urea can be performed using triphosgene (bis(trichloromethyl)carbonate) in the presence of not participating in the reaction, the base, such as triethylamine, in a solvent such as dichloromethane at room or lower temperature.

As another alternative CDI instead of triphosgene you can apply phosgene.

In many of the above reactions may be necessary to protect one or more groups to prevent the passage of undesirable reactions fragments of molecules. Examples of protective groups and methods of protection of functional groups and remove protection can be found in Protective Groups in Organic Synthesis (T.Green and P.Wuts; 3rdEdition; John Wiley and Sons, 1999). The hydroxy-group may be protected, for example, the formation of ether (-OR) or an ester (OC(=O)R group, for example: the formation of tert-butyl simple ester; benzyl, benzhydryl (diphenylmethylene) or tretilova (triphenylmethyl) simple ether; trimethylsilyloxy or tert-butyldimethylsilyloxy simple ether; or acetylator of ester (-OC(=O)CH3, -OAc). Aldehyde or ketone group may be protected, for example, education, respectively, acetal (R-CH(OR)2)) or Catala (R2C(OR)2), in which the carbonyl group (>C=O) pravr who participate in fluids (> C(OR)2), the interaction of, for example, with a primary alcohol. Aldehyde or ketone group is easily regenerated by hydrolysis with the use of a large excess of water in the presence of acid. The amino group can be protected, for example, by the formation of amide (-NRCO-R) or a urethane (-NRCO-OR), for example, methylamine (-NHCO-CH3); benzylcyanide (-NHCO-OCH2C6H5, -NH-Cbz); tert-butoxide (-NHCO-OC(CH3)3, -NH-Boc); a 2-biphenyl-2-propoxyimino (-NHCO-OC(CH3)2C6H4C6H5, -NH-Bpoc), 9-fluorenylacetamide (-NH-Fmoc), 6-nitroferricyanide (-NH-Nvoc), 2-trimethylsilylacetamide (-NH-Teoc), 2,2,2-trichloroacetamide (-NH-Troc), allelochemical (-NH-Alloc) or 2-(phenylsulfonyl)ethylacetamide (-NH-Psec). Other protective groups for amines, such as cyclic amines and heterocyclic N-H groups include toluensulfonyl (tonilou) and methanesulfonyl (mesyl) groups, and benzyl groups, such as para-methoxybenzyl (PMB)group. The carboxyl group can be protected by formation of ester, for example, C1-7Olkiluoto ester (e.g. methyl ester, tert-butyl methyl ether); C1-7halogenosilanes ether (for example, C1-7trialgenericuh ether); Tris1-7alkylsilane-C1-7Olkiluoto ether; or C5-20aryl-C1-7Olkiluoto ether (for example, benzyl ether; bat benzilovogo ether); or amide formation, for example methylamide. Tolna group may be protected, for example, the formation of simple tiefer (-SR), for example benzyl tiefer; acetamidomethyl ether (-S-CH2NHC(=O)CH3).

Acid additive salts that form a subgroup of (C) compounds of formula (I)can be obtained during the synthesis of the compounds of formula (I), i.e., 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or a transformation of the parent compound in free base form into the desired salt, or converting one salt of the parent compound to another desired salt of the parent compound. The original connection, i.e. the 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (compound of formula (IA)) can be obtained by the method shown below in scheme 3.

As shown in scheme 3, 3,4-dinitrocresol acid (XIII), which is commercially available compound, and turn them into morpholide (XXI). Getting amide may be carried out by conversion of the acid (XIII) in active derivative, such as, for example, the acid chloride using standard methods. For example, the acid chloride can be obtained by heating with an excess of thionyl chloride at the temperature of its boiling point with the subsequent removal of excess thionyl chloride by azeotropic distillation with toluene.

Morpholide (XXI) which may be recovered in dinitrobenzaldehyde (XXIII) by treatment with a suitable regenerating reagent, as, for example, sodium borohydride in combination with boron TRIFLUORIDE. The reduction is usually carried out in an anhydrous solvent such as tetrahydrofuran at reduced temperature, for example at a temperature of 0-5°C. After this dinitrobenzaldehyde (XXIII) can be restored in diaminobenzidine (XXIV) under standard conditions, for example by catalytic hydrogenation in the presence of a catalyst such as palladium on coal in a polar solvent, such as ethanol at room temperature.

Then diaminobenzidine (XXIV) is injected into the reaction with commercially available 4-nitropyrazole-3-carboxylic acid to obtain nitroferricyanide (XXV). Getting nitroferricyanide (XXV) can be achieved, firstly, by the formation of amide bond between the carboxylic acid and diaminobenzanilide (XXIV) using the reagent for peptide condensation, as, for example, tetrafluoroborate O-(benzotriazol-1-yl)-N,N,N',N'-tetramethylurea (TBTU), can promote the formation of amide bond with an aromatic amino group. The intermediate amide (not shown) then cyclist in nitropropanediol (XXV) by heating in excess of glacial acetic acid, for example, at a temperature of approximately 65°C.

Nitropropanediol (XXV) can be restored in the appropriate AMI the (XXVI) under standard conditions. Recovery may be performed, for example, by catalytic hydrogenation in the presence of a catalyst, such as palladium on charcoal, in a polar solvent such as ethanol or dimethylformamide at room temperature.

Amine (XXVI) can be, in turn, converted into urea (IA) using standard methods of synthesis of ureas, for example, by the interaction of the amine (XXVI) with 2,6-diftorhinolonom in a polar solvent such as THF, at room or lower temperatures, for example at a temperature of 0-5°C.

Urea (IA) in free base form may be used to obtain the acid additive salts of the present invention.

Salt of the present invention can be obtained from the free bases by standard means, such as by the methods described in: Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, hardcover, 388 pages, August 2002. For example, salt can be obtained by the interaction of the free base with the appropriate acid in water or an organic solvent, or mixtures thereof; mainly used non-aqueous environment, such as ether, ethyl acetate, methanol, ethanol, isopropanol or acetonitrile.

In another aspect the invention relates to a method for producing an acid additive salts of 1-(2,6-deltorphin the l)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea moreover, the method includes obtaining a solution of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form in a solvent (typically an organic solvent or solvent mixture, and processing the resulting solution of the acid sludge acid additive salt.

The acid is usually added in the form of a solution in a solvent which is miscible with the solvent in which is dissolved free base.

The solvent in which the first dissolve the free base may be a solvent in which the insoluble acid additive salt. On the other hand, the solvent in which the first dissolve the free base may be a solvent in which the acid additive salt is soluble, at least partially, and add another solvent in which the acid additive salt is soluble to a lesser extent, so that the salt falls out of solution as a precipitate.

For example, in one of the ways to obtain salts of the present invention, the free base is dissolved in the first solvent (which may be an acetate or a mixture of ethyl acetate and alcohol, such as methanol) and then add a solution (e.g., concentrated or saturated solution) acid, such as hydrochloric acid, in the second dissolve the e (which may be ether, such as diethyl ether or dioxin), so that a precipitate of the acid additive salt, after which the precipitate is collected, for example, by filtering.

Methods of obtaining 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

In the examples earlier applications of the authors WO 2005/002552 and in schemes 1 and 3, above, shows that [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amide can be obtained by the sequence of stages, including:

(i) the interaction of 4-morpholine-4-ylmethylene-1,2-diamine with 4-nitro-1H-pyrazole-3-carboxylic acid in the presence of 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide (EDC) and 1-hydroxybenzotriazole (HOBt) in N,N-dimethylformamide (DMF) to obtain 5-morpholine-4-ylmethyl-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole; and

(ii) the restoration of the nitro-group by treatment with palladium on coal in hydrogen atmosphere;

or

(i) the interaction of 4-amino-1H-pyrazole-3-carboxylic ester with the appropriate carboxylic acid in the presence of 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide (EDC) and 1-hydroxybenzotriazole (HOBt) in N,N-dimethylformamide (DMF) or with an acid chloride of the appropriate acid in the presence of triethylamine, with the formation of 4-amide-1H-pyrazolylborate acid; and

(ii) the interaction of 4-morpholine-4-ylmethylene-1,2-diamine with the corresponding 4-amide-1H-Pires karbonovoi acid in the presence of 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide (EDC) and 1-hydroxybenzotriazole (HOBt) in N,N-dimethylformamide (DMF) to obtain [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amide.

In the present application discovered that instead of the interaction nitropyrazole connection with the diamine and subsequent recovery of the nitro group to the amine, or the reaction of aminopyrazole with diamine, aminopyrazole can be entered into interaction with the diamine, provided that the amino aminopyrazole adequately protected. The product of this reaction can then be cycletour with the formation of the benzimidazole. In addition, it was found that removing aminosidine group and cyclization with the formation of the benzimidazole can be carried out in one stage.

Accordingly, in another aspect, the present invention is a method for obtaining compounds of formula (XXVII)or (XXVIII), or their salts:

moreover, this method includes:

(i) the interaction of the compounds of formula (XXIX):

where PG denotes aminosidine group:

(ii) with the compound of the formula (XXXI):

in an organic solvent in the presence of a condensing reagent, such as EDC and HOBt:

The compound of formula (XXVIII) is regioisomers compound (XXVII).

Aminosidine group PG may be any known protecting group used to protect the amino groups under the conditions used in the above-described method, see, e.g., Green et al., mentioned above. T is to, for example, nitrogen may be protected by formation of the amide (NCO-R) or a urethane (NCO-OR), for example methylamide (NCO-CH3); benzylcyanide (NCO-OCH2C6H5, -NH-Cbz); tert-butoxide (-NCO-OC(CH3)3N-Boc); a 2-biphenyl-2-propoxyimino (NCO-OC(CH3)2C6H4C6H5N-Bpoc), 9-fluorenylacetamide (N-Fmoc), 6-nitroferricyanide (N-Nvoc), 2-trimethylsilylacetamide (N-Teoc), 2,2,2-trichloroacetamide (N-Troc), allelochemical (N-Alloc) or 2-(phenylsulfonyl)ethylacetamide (-N-Psec). Other protective groups for amines include benzyl groups as, for example, para-methoxybenzyl (PMB) group. Preferred aminosidine groups are urethanes (NCO-OR), for example benzylamine (NCO-OCH2C6H5, -NH-Cbz) or t-butoxide (-NCO-OC(CH3)3N-Boc); or Alliaceae (N-Alloc). In one embodiment, the implementation of the protective group PG is a protective group, APG, which is aminosidine group that can be removed under conditions of acidic environment. Such groups include urethanes. Particularly preferred urethane protecting group is tert-butyloxycarbonyl, which can be removed in an acidic environment.

In one embodiment, the implementation of the protective group PG is then removed from the compounds of formula (XXVII) or (XXVIII) and replace the protective group APG, obtaining the compounds of formula (XXVIIa) or (XXVIIa).

One particularly preferred compound of formula (XXIX) is a compound of the following formula (XXXII):

In addition, the invention has developed a new chemical intermediate compound of formula (XXXII) as such.

The invention also relates to new chemical intermediate compounds of formula (XXVII) or (XXVIII) as such, for example, new chemical intermediate compounds of formula (XXVIIa) or (XXVIIIa), below. Therefore, in the present invention developed (2-amino-4-morpholine-4-ylmethylene)amide of 4-amino-1H-pyrazole-3-carboxylic acid or (2-amino-5-morpholine-4-ylmethylene)amide of 4-amino-1H-pyrazole-3-carboxylic acid, and their protected form as a new chemical intermediates. One particularly preferred new chemical intermediate compound of formula (XXVII) is tert-butyl ester of [3-(2-amino-4-morpholine-4-iletilenlerin)-1H-pyrazole-4-yl]carbamino acid. One particularly preferred new chemical intermediate compound of formula (XXVIII) is tert-butyl ester of [3-(2-amino-5-morpholine-4-iletilenlerin)-1H-pyrazole-4-yl]carbamino acid.

If the protective group PG is tert-butoxycarbonyl group, the total yield of this process exceeds 85%. Furthermore, the method has the advantage that it PR is changing relatively simple and inexpensive reagents and solvents, and, furthermore, the method is advantageous in terms of ease of cleaning products.

In another aspect the invention relates to a method for producing [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amine or its salts, and the method includes:

(i) treatment of compounds of formula (XXVIIa) or (XXVIIIa):

acid in a solvent, optionally with heating; and

(ii) neutralizing the reaction mixture.

Aminosidine group APG can be any known protective group used to protect the amino group, as defined above in relation to compounds of formula (XXVII) or (XXVIII), and which can be removed under the conditions used in the previous method.

At stage (i) reaction with acid may be carried out by heating, for example, to a temperature in the range from 80 to 100°C. the Solvent in which the reaction is carried out in stage (i)is an alcohol-based solvent, and may be, for example, ethanol.

The protective group on stage (i) is preferably such protecting group, as the group is Boc, which can be removed by treatment with an acid, and the acid is chosen so that it is suited for protonation of the intermediate connection to the activation of the carbonyl group for the cyclization reaction. Suitable acids include strong acids such as sulfuric acid, methanesulfonic the th acid or hydrochloric acid, and one particular acid is hydrochloric acid.

After completion of the reaction in stage (i), which is judged, for example, the extinction of the original substance (XIIIa), it is possible to neutralize the reaction mixture.

At stage (ii) is used not interfering basis. The term "non-interfering basis" in this context refers to the base, such as sodium carbonate, which will not react with the received connection. Stage (ii) is usually carried out at room temperature.

At stage (ii), the reaction mixture is neutralized, for example, up to saturation of the reaction mixture by neutralizing reagent and at a pH of 8.5.

After stage (ii) compound may be introduced into interaction with carbonyliron reagent, such as, for example, 1,1'-carbonyl diimidazol (CDI) or the equivalent of phosgene, and then processed by cyclopropylamines. Equivalents of phosgene include triphosgene or phosgene. Preferred carbonyliron reagent is 1,1'-carbonyldiimidazole (CDI).

Alternatively, the urea can be obtained by the interaction of aminopyrazole, ie 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine with phenylcarbamates in the presence of a base, for example pyridine, in a solvent such as, for example, THF, to obtain the cyclic urea and subsequent processing by cyclopropylamino, or interaction is the influence of aminopyrazole with cyclopropylethanol, which can be obtained by rearrangement of azide cyclopropanecarbonyl acid in Kurzius (as described in US 4313755 and US 4299778).

Thus, another aspect of the present invention is a method of obtaining a 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or its salts, and the method includes

(i) treatment of compounds of formula (XXVIIa) acid in a solvent, optionally with heating;

(ii) neutralizing the reaction mixture;

(iii) the interaction of the product of stage (ii) with carbonyliron reagent;

(iv) the interaction of the product of stage (iii) with cyclopropylamine.

Stage (iii) is usually carried out by boiling under reflux, for example, at a temperature up to about 100°C, more typically up to 70-75°C. At stage (iii) the reaction can be carried out in a polar aprotic solvent such as tetrahydrofuran. Carbonyloxy reagent can be such a connection as 1,1'-carbonyldiimidazole (CDI) or the equivalent of phosgene, such as triphosgene or phosgene. Preferred carbonyliron reagent is 1,1'-carbonyldiimidazole (CDI).

Stage (iv), usually carried out by heating, for example, at temperatures up to about 100°C.

After stage (iv) the product may be subjected to transformation in Sol or recrystallization (for example, using 2-propanol or ethanol to the operation of the solvent) to increase the purity and the purpose of obtaining crystalline form.

Stage (iii) leads to the production of intermediate compounds of formula (XXXIII) and/or its regioisomer (XXXIIIa):

Intermediate compounds of formula (XXXIII) and (XXXIIIa), which if necessary can be allocated, then injected into the reaction cyclopropylamino, receiving the compound of formula (XXX).

Accordingly, in another aspect, the invention relates to a method for obtaining compounds of formula (XXX), as defined in this application, the method involves reacting the compounds of formula (XXXIII) or (XXXIIIa) cyclopropylamino and then an optional receipt acid additive salts of the compounds of formula (XXX). The reaction is usually carried out in a polar aprotic solvent such as N-an organic, preferably at elevated temperatures, such as temperatures greater than 80°C, more typically in excess of 90°C, for example from 95 to 105°C.

The previous method can also be used to obtain other compounds of formula (I) and their subgroups described in this application, in which A in the formula (I) represents a group NH.

Accordingly in another aspect the invention relates to a method for obtaining compounds of formula (I), as defined in this application, in which A in the formula (I) represents a group NH; moreover, this method involves the interaction of (i) the compounds of formula (XXXIII) and the and its regioisomer (XXXIIIa), or (ii) the compounds of formula (XXXIV) and/or its regioisomer (XXXIVa):

with the compound of the formula R1-E-NH2, preferably in a polar aprotic solvent such as N-an organic, preferably at elevated temperatures, such as temperatures greater than 80°C, more typically in excess of 90°C, for example, from 95 to 105°C, and then the optional formation of acid additive salts of the compounds of formula (I).

In addition, the invention has developed new chemical intermediate compounds of formula (XXXIII), (XXXIIIa), (XXXIV) and (XXXIVa).

In another embodiment, can be obtained compound of formula (XXVIIa) in the above-described method of obtaining 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine or its salt, or a method of producing 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or its salts, a method, which includes:

(i) the interaction of the compounds of formula (XXIX), in which PG represents aminosidine group which can be removed by acid, i.e. APG;

(ii) with the compound of the formula (XXXI) in an organic solvent in the presence of a condensing reagent such as EDC and HOBt.

The method described above, optionally includes the additional step of recrystallization of salt to get to istoricheskoi form, for example, crystalline forms described in this application.

How to clean

Compounds of the present invention can be isolated and purified Radom ways, well known to experts in the art, and examples of such methods include chromatographic techniques, such as, for example, column chromatography (e.g., flash chromatography) and HPLC. Preparative LC-MS is a standard and effective method used for the purification of small organic molecules, such as molecules of compounds described in this application. You can modify the technique of liquid chromatography (LC) and mass spectrometry (MS) to provide a better separation of the crude substances and better identification of the composition of the samples using MC. Optimization of preparative gradient LC methods will include a selection of speakers, volatile solvents and modifiers, as well as gradients. Optimization of preparative LC-MS techniques and their application for purification of compounds well known in the art. Such methods are described in Rosentreter U, Huber U.; Optimal fraction collecting in preparative LC/MS; J Comb Chem.; 2004; 6(2), 159-64 and Leister W, Strauss K, Wisnoski D, Zhao Z, Lindsley C., Development of a custom high-throughput preparative liquid chromatography/mass spectrometer platform for the preparative purification and analytical analysis of compound libraries; J Comb Chem.; 2003; 5(3); 322-9.

One such system for purification of the compounds of preparative LC-MS description the on below in the experimental section, although a specialist in the art will take into account that could be used for alternative systems and methods described for this purpose. In particular could have used methods based on preparative LC on normal phase, instead of the methods described here on reversed phase. Most preparative LC-MS systems used on LC reversed-phase and volatile acid modifiers, because this approach is very effective for the purification of small molecules and solvents compatible with the technique of mass spectrometry with elektrorazpredelenie positive ions. On the other hand, cleaning compounds could be applied to other solutions developed in chromatography, such as LC on the normal phase, alternative buffering of the mobile phase, key modifiers, etc. that are briefly described in the analytical methods described above.

Recrystallization

Recrystallization of compounds of formula (I) and their salts, in particular 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and its salts, can be performed by methods well known to the specialist - see, for example (P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Chapter 8, Publisher Wiley-VCH). The products obtained by organic reactions, are rarely pure privateline directly from the reaction mixture. If the connection (or its salt) is a solid, it can be cleaned and/or obtained in crystalline form by recrystallization from a suitable solvent. A good solvent for recrystallization should in moderate quantity to dissolve the subject recrystallization substance at elevated temperature and only a small amount of this substance at a lower temperature. It should be easy to dissolve impurities at low temperatures or not to dissolve them. Finally, the solvent should be easily removed from the purified product. This usually means that it has a relatively low boiling point, and a specialist in the art should know the solvents for recrystallization of a particular substance, or, if this information is unavailable, explores several solvents. To achieve a good yield of purified substance use minimum amount of hot solvent, capable of dissolving all of the contaminated substance. In practice, apply 3-5% more solvent than is necessary, so that the solution is unsaturated. If the contaminated substance contains a mixture, which is insoluble in the solvent, then it can be removed by filtration, and then leave the solution to crystallize. In addition, if the contaminated substance contains traces of Krashennikov, which are not your own coloring connections, they can be removed by adding to a hot solution of a small amount of decolorizing activated carbon followed by filtration and crystallization of the solution. Typically, the crystallization occurs spontaneously upon cooling of the solution. If this does not happen, the crystallization can cause cooling of the solution to a temperature below room temperature or by adding a single crystal of a pure substance (crystal seed). It is possible to perform recrystallization and/or to optimize yield with application of antibacterial. In this case, the compound is dissolved in a suitable solvent at elevated temperature, filtered and then to promote crystallization add additional solvent, which is subjected to cleaning compound has poor solubility. After that, typically using vacuum filtration, isolated crystals, washed and then dried, for example in an oven, or by drying.

Other examples of methods of crystallization include crystallization from a vapor state, which includes a step of evaporation, for example, in a sealed tube or in the air stream, and the crystallization from the melt (Crystallization Technology Handbook 2ndEdition, edited by A. Mersmann, 2001).

In particular, the compounds of formula (I) can is subjected to recrystallization (e.g., using 2-propanol or ethanol as solvent) to increase the purity and obtain in crystalline form.

Generally, the obtained crystals are exploring ways diffraction of x-rays, such as x-ray diffraction on the powder (XRPD) or x-ray analysis of crystals.

Pharmaceutical compositions

Although you can enter active compounds and their salts in pure form, it is preferable to apply them in the form of pharmaceutical compositions (e.g., compositions), comprising at least one active compound of the present invention in combination with one or more pharmaceutically acceptable carriers, excipients, fillers, diluents, buffer compounds, stabilizers, preservatives, lubricating means or other substances well known to the person skilled in the art and optionally other therapeutic or prophylactic methods, for example by means that reduce or alleviate some of the side effects associated with chemotherapy. Specific examples of such means include an antiemetic and tools that prevent or reduce the duration associated with chemotherapy-related neutropenia and prevent complications, the cat is who appear due to low levels of red blood cells, or leukocytes, for example, erythropoietin (EPO), granulocyte macrophage-colony stimulating factor (GM-CSF) and granulocyte-colony stimulating factor (G-CSF).

Thus, the present invention inter alia relates to pharmaceutical compositions, as defined above, and methods of producing pharmaceutical compositions, comprising mixing at least one active compound, as defined above, with one or more pharmaceutically acceptable carriers, fillers, buffers, excipients, stabilizers or other substances described in this application.

The term "pharmaceutically acceptable" in the present description refers to compounds, substances, compositions and/or dosage forms which are, within the reasonable opinion of medicine, suitable for use in contact with tissues of a subject (e.g. human)without causing excessive toxicity, irritation, allergic reactions, or other problems or complications, and meet the reasonable value of benefit/risk. Each of the carriers, fillers, etc. must also be "acceptable" in the sense of having compatibility with other ingredients of the composition.

Accordingly in another aspect the present invention relates to compounds of formula (I) and their subgroups, as, for example, compounds of formulas (II) and (III, and their subgroups, as defined in this application, in the form of pharmaceutical compositions.

The pharmaceutical composition may be of any form suitable for oral, parenteral, local, intranasal, ocular, aural, rectal, intravaginal or transdermal routes of administration. If the compositions are intended for parenteral administration, they can be intended for intravenous, intramuscular, intraperitoneal, subcutaneous injection, or for direct delivery to the target organ or tissue by injection, infusion or other means of delivery. Delivery can occur by bolus injection, intermittent infusion or longer-term infusion and can take the form of passive infusion or using a suitable pump for infusion.

Pharmaceutical compositions intended for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffer compounds, bacteriostatic and dissolved substances, which give the composition isotonicity with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions that can include suspendresume tools and thickeners. Their examples are described in R.G. Strickly, Solubilizing Excipients in oral and injectable formulations, Pharmacutical Research, Vol 21(2) 2004, p.201-230. In addition, they can contain additional solvents, mixtures of organic solvents, cyclodextrine complexing agents, emulsifying means (for the formation and stabilization of emulsion formulations), liposomal components for the formation of liposomes, gelling polymers for the formation of polymer gels, protective equipment for lyophilization and combination funds for, among other things, stabilize the active ingredient in a soluble form and giving the structure isotonicity with the blood of the intended recipient. These formulations can be placed into containers containing one or more doses, for example, sealed ampoules or vials and may be stored in dried by freezing (liofilizirovannom) condition requiring only the addition of sterile liquid carrier, for example water for injections, immediately prior to use.

The solubility of drug molecules which are susceptible of ionization, can be increased to the desired value by regulating the pH, if the pKa of the medicinal product is significantly different from the pH of the composition. The acceptable pH range for intravenous and intramuscular injection is 2-12, but for subcutaneous injection is in the range of 2.7-9.0 in. the pH of the solution is regulated or salts drug compounds with a strong acid is Tami/bases as, for example, hydrochloric acid or sodium hydroxide, or with buffer solutions of compounds which include, but are not limited to the above, buffer solutions, derived from glycine, citrates, acetates, malatov, succinate, histidine, phosphate, Tris(hydroxymethyl)aminomethane (TRIS) or carbonates.

In the compositions suitable for injection, often used a combination of aqueous and water-soluble organic solvent/surfactant(surfactant) (i.e. additional solvent). Water-soluble organic solvents and surfactants used in suitable for injection compositions, include, but are not limited to, propylene glycol, ethanol, polyethylene glycol 300, polyethylene glycol 400, glycerin, dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP; Pharmasolve), dimethylsulfoxide (DMSO), Solutol HS 15, Cremophor EL, Cremophor RH 60, and polysorbat 80. Such compositions usually, but not always, can be diluted before injection.

Propylene glycol, PEG 300, ethanol, Cremophor EL, Cremophor RH 60, and polysorbat 80 are completely organic miscible with water, solvents and surfactants used in commercially available compositions for injection, and they can be used in combinations with each other. The obtained organic compounds usually diluted at least twice before intravenous bolus injection or infusion.

On the other side is s, increased solubility can be achieved by the formation of molecular complexes with cyclodextrins.

Liposomes are nearly spherical vesicles consisting of an external lipid bilayer membrane and an inner aqueous core, with a total diameter of <100 μm. Depending on the level of hydrophobicity of moderately hydrophobic drugs can be solubilisation liposomes, if the drug is encapsulated or incorporated in a liposome. Hydrophobic drugs can also be solubilisation liposomes, if the molecule drugs is becoming an integral part of the lipid bilayer membrane, and in this case, the hydrophobic drug is dissolved in the lipid portion of the lipid double layer. Typical liposomal composition contains water with phospholipid at a concentration of 5-20 mg/ml, isotonic agent, a buffer with a pH of 5-8 and, optionally, cholesterol.

The compositions can be placed in containers designed for one or several doses, for example sealed ampoules or vials and may be stored in dried by freezing (liofilizirovannom) condition requiring only the addition of sterile liquid carrier, for example water for injections, immediately prior to use.

The pharmaceutical composition can be obtained by lyophilization connect the Oia formula (I) or its acid salt additive. Lyophilisation is called the method of drying the composition during freezing. Therefore, the terms "drying by freezing and lyophilization are used in this application interchangeably. The typical way is to solubilisate connection, after which the resulting composition, fined, filtered under sterile conditions and aseptically transferred to a tank suitable for lyophilization (e.g., bottles). As for the bottles, they are partially closed lyophilization covers. The composition can be cooled to freezing, subjected to freeze-drying under standard conditions and then sealed, resulting in a stable freeze dry composition. The composition typically will have a low residual water content, for example less than 5%, for example less than 1 mass%, by weight of liofilizirovannogo substances.

Liofilizovannye compounds may contain other fillers, such as thickeners, dispersing funds, buffer compounds, antioxidants, preservatives and regulators toychest. Typical buffer compounds include phosphates, acetates, citrate, and glycine. Examples of antioxidants include ascorbic acid, sodium bisulfite, metabisulfite sodium, monothioglycerol, thiourea, bottled hydroxytoluene, bottled hydroxyanisol and salt Ethylenediamine is strukturnoi acid. Preservatives may include benzoic acid and its salts, sorbic acid and its salts, alkalemia esters of para-hydroxybenzoic acid, phenol, chlorobutanol, benzyl alcohol, thimerosal, benzalkonium chloride and a chloride of cetylpyridinium. The previously mentioned buffer compounds, as well as dextrose and sodium chloride, can be used to control toychest if necessary.

The technology of freeze-drying, as a rule, are used to add volume to facilitate the process and/or volume and/or mechanical integrity of liofilizovannyh mass. Under the tool to add volume means freely soluble in water filler consisting of solid particles, which is co-lyophilized with the compound or its salt yields a physically stable liofilizovannyh mass, provides the most optimal process flow lyophilization drying, and quick and full recovery. Tool to add volume can also be used to make the solution isotonicity.

Water-soluble means to add volume can be any of the pharmaceutically acceptable inert solids, usually used for lyophilization. Such means for giving volume to include, for example, sugars such as glucose, maltose, sucrose and lactose; Isparta, such as sorbitol or mannitol; amino acids such as glycine; polymers, such as vinyl pyrrolidone; and polysaccharides, such as dextran.

The ratio of mass to add volume to the weight of active compound usually is in the range from about 1 to about 5, for example, from about 1 to about 3, for example, in the range of from about 1 to about 2.

On the other hand, it is possible to obtain compositions in the form of a solution, which can be concentrated and tightly Packed in suitable vessels. Sterilization dosage forms may be carried out by filtration or heating in an autoclave vessels together with their contents at a suitable stage of the process of obtaining compositions. When using compounds may require additional dilution or preparation of the compositions before the introduction of, for example dilution in a suitable sterile containers for infusion.

Solutions and suspensions for injection immediate preparation can be obtained from sterile powders, granules or tablets.

In one of the preferred embodiments of the present invention the pharmaceutical composition has a form suitable for intravenous administration, for example injection or infusion.

The pharmaceutical compositions of the present invention for parenteral injection may, in addition to the, include pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and sterile powders for recovery in sterile solutions or dispersions for injection immediately before use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or bases include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), carboxymethylcellulose and suitable mixtures, vegetable oils (such as olive oil) and suitable for injectable organic esters, such as etiloleat. Mobility can be supported, for example, the use of such materials to cover as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants.

The compositions of the present invention may also contain auxiliary substances such as preservatives, means for wetting, emulsifying agents and dispersing funds. Prevent the activity of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, etc. in Addition, it may be desirable inclusion isotonic means, such as sugars, sodium chloride, etc. longer absorb the Oia pharmaceutical forms, intended for injection can be achieved by the inclusion of means, which delay absorption, such as, for example, aluminum monostearate and gelatin.

If the connection is unstable or has a low solubility in water, its basis can be obtained concentrate in organic solvents. This concentrate can then be diluted to a lower concentration in the aqueous system, and it can be mostly stable over a short period of time after the injection. Therefore, in another aspect, the pharmaceutical composition comprising non-aqueous solution entirely based on one or more organic solvents, which can be entered by itself or, more often diluted before the introduction of appropriate intravenous filler (saline, dextrose; buffered or nezaloginennym) (Solubilizing excipients in oral and injectable formulations, Pharmaceutical Research, 21(2), 2004, p.201-230). Examples of solvents and surfactants are propylene glycol, PEG 300, PEG 400, ethanol, dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP; Pharmasolve), glycerin, Cremophor EL, Cremophor RH 60, and polysorbate. Specific non-aqueous solutions consist of 70-80% of propylene glycol and 20-30% ethanol. One of the specific non-aqueous solutions consists of 70% propylene glycol and 30% ethanol. Other includes 80% propylene glycol and 20% ethanol. Typically, these plants shall oriali used in combination and usually diluted, at least twice before intravenous bolus injection or intravenous infusion. Typical quantities for bolus intravenous formulations are ~50% glycerol, propylene glycol, PEG, PEG and ~20% for ethanol. Typical quantities for intravenous formulations for infusion are ~15% glycerol, 3% for DMA and ~10% for propylene glycol, PEG, PEG and ethanol.

In one of the preferred embodiments of the present invention the pharmaceutical composition has a form suitable for intravenous administration, for example, by injection or infusion. In the case of intravenous solution can be introduced in its original form or original, you can enter the solution in the bag for liquids (containing pharmaceutically acceptable filler, such as 0.9% saline solution or 5% dextrose), before the introduction of the patient.

In another preferred embodiment, the pharmaceutical composition has a form suitable for subcutaneous (s.c.) introduction.

Pharmaceutical dosage form appropriate for oral administration include tablets, capsules, pills in the form of capsules, pills, diamond-shaped tablets, syrups, solutions, powders, granules, elixirs and suspensions, sublingual tablets, pills or patches, as well as buccal patches.

Pharmaceutical compositions that contain Asia the compounds of formula (I), can be created in accordance with known techniques, see, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA.

For example, the composition of the tablets may contain a unit dose of the active compound together with an inert diluent or carrier, such as sugar or sugar alcohol, for example lactose, saccharose, sorbitol or mannitol; and/or diluent, which is not a sugar, such as sodium carbonate, calcium phosphate, calcium carbonate or cellulose or its derivatives, such as methylcellulose, ethylcellulose, hypromellose, as well as starches, for example corn starch. In addition, tablets may contain such standard ingredients such as binders or granulating means, as, for example, polyvinylpyrrolidone, dezintegriruetsja means (for example, capable of swelling the cross crosslinked polymers, such as cross crosslinked carboxymethylcellulose), means for sliding (for example, stearates), preservatives (e.g. parabens), antioxidants (such as BHT), buffer means (for example, phosphate or citrate buffers), as well as means for giving sipacate, such as a mixture of citrate/bicarbonate. Such fillers are well known, and there is no need to discuss them here.

Capsules can have varieties of hard gelatin and soft what about gelatin and may contain the active component in the solid, semi-solid or liquid form. Gelatin capsules can be obtained from animal gelatin or its equivalent synthetic or of plant origin.

Solid dosage forms (e.g. tablets, capsules etc) may or may not be covered, but, as a rule, are coated, for example, covered with a protective film (for example, wax or glaze), or coating, which regulates the release of the drug. This coating (for example, type polymer EudragitTMcan be designed to release the active component in the desired location in the gastrointestinal tract. For example, the coating can be chosen in such a way that it was destroyed at specific values of pH in the gastrointestinal tract, thereby selectively releasing the connection in the stomach or ileum, or in the duodenum.

Instead of a coating, or in addition thereto, the drug can be incorporated into a solid matrix containing the means of regulating the release of, for example, means that slows the release, which can be adapted for the selective release of connections in a changing acidity or alkalinity in the gastrointestinal tract. On the other hand, the matrix material or slow release coating may take the form of Atroshenko polymer (for example, polymer of maleic anhydride), which is subjected to continuous destruction during the passage of the dosage form through the gastrointestinal tract. As another alternative, the active compound may be incorporated into the delivery system, which provides the osmotic regulation of release of connection. Compositions with osmotic release, as well as other compositions with delayed or prolonged release can be obtained by methods well known to specialists in this field of technology.

Pharmaceutical compositions include from about 1% to about 95%, preferably from about 20% to about 90% of the active ingredient. The pharmaceutical compositions of the present invention can be obtained, for example, in the form of dosage forms, such as ampoules, vials, suppositories, pills, tablets or capsules.

Pharmaceutical compositions for oral administration can be obtained by mixing the active ingredient with solid carriers, optionally granulating the resulting mixture, and further processing the mixture, if desired or necessary after the addition of appropriate excipients, into tablets core tablets or capsules. In addition, there is the possibility of inclusion of the active ingredients in the polymeric carriers that which allows the active ingredients to diffuse or be released in the calculated quantities.

Compositions for topical application include ointments, creams, sprays, patches, gels, liquid drops and inserts (for example, intraocular insert). Such compositions can be created according to known methods.

Compositions for parenteral administration typically are sterile aqueous or oily solution or fine suspension, or they may be in the form of fine sterile powder intended for solution for injection in sterile water immediately before use.

Examples of formulations for rectal or intravaginal injection include pessaries and suppositories, which can be manufactured, for example, of molded plastic or waxy substance that contains the current connection.

Compositions for administration by inhalation can take the form suitable for inhalation powder compositions, or liquid, or powder sprays, and can be entered in standard form using powder inhalers or devices for dispensing aerosols. Such devices are well known. Powder formulations for administration by inhalation, typically include the active compound together with an inert solid powder diluent, such as lactose.

The pharmaceutical compositions can PE advetise patient in "packing of the patient, containing a complete course of treatment in one package, usually a blister pack. Packaging the patient has the advantage compared to traditional methods, where the pharmacist separates the consumption of patient drug from its total number, in that the patient always has access to the insert contained within the packaging of the patient, which is usually absent in the recipes of patients. It was shown that the inclusion of the liner in the package improves the readiness of the patient to follow the instructions of the doctor.

Compounds of the present invention will be mainly provided in dosage forms that are, as a rule, will contain sufficient compound to provide the desired level of biological activity. For example, a composition intended for oral administration may contain from 0.1 milligrams to 2 grams of active ingredient, for example, from 1 nanogram to 2 milligrams of the active ingredient. In this range the individual sub-bands of the content of the compound is from 0.1 milligrams to 2 grams of active ingredient (more usually from 10 milligrams to 1 gram, for example, from 50 milligrams to 500 milligrams, or from about 1 micrograms to 20 milligrams (for example, from 1 microgram to 10 milligrams, for example, from 0.1 milligrams to 2 milligrams of active ingredient)).

the La oral compositions, single unit dosage forms may contain from 1 milligram to 2 grams, more typically from 10 milligrams to 1 gram, for example, from 50 milligrams to 1 gram, for example from 100 milligrams to 1 gram of the current connection.

The current connection will be introduced to the patient if necessary (for example, the patient who is an animal or person) in a quantity sufficient to achieve the desired therapeutic effect.

Treatment

It is expected that the compounds of formula (I), (II), (III), (XXX) and sub-groups defined in this application will be applicable in the prevention or treatment of diseases or medical conditions mediated by cyclin-dependent kinases, kinase-3 glycogen synthase and Aurora kinases. Examples of such diseases and conditions above.

These compounds are typically administered to a subject such as a person or an animal, preferably a human, in case of need such an introduction.

Connection, as a rule, must be administered in amounts that are therapeutically or prophylactically useful and which are substantially non-toxic. However, in some situations (for example, in the case of life-threatening diseases), the benefits from the introduction of the compounds of formula (I) can be greater than the harm of any toxic or side effects, and in this case, it may be desirable to implement compounds in quantities, which is associated with a certain degree of toxicity.

The compounds can be administered over an extended period to maintain a favorable therapeutic results, or can be entered only during a short period. On the other hand, you can enter them in pulsed or continuous mode.

A typical daily dose of a compound can be in the range from 100 picograms to 100 milligrams per kilogram body weight, more typically 5 nanograms to 25 milligrams per kilogram of body weight and more usually 10 nanograms to 15 milligrams per kilogram (e.g. 10 nanograms to 10 milligrams, and more typically 1 microgram per kilogram to 20 milligrams per kilogram, as, for example, from 1 microgram to 10 milligrams) of body weight, although, if necessary, can be higher or lower doses.

The described compounds (for example, such a connection as 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or a salt thereof, such as the lactate or citrate) may be administered daily or the administration can be repeated, for example, every 2 or 3 or 4 or 5, or 6, or 7, or 10, or 14, or 21, or 28 days. However, ultimately, the number of input connections and the type of composition should reflect the nature of the disease or physiological condition being treated, and should be selected on the trigger the friction of the doctor.

An example of a daily dosage of such compounds as 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or its salts, such as the lactate (particularly the L-lactate) or citrate, includes the introduction of the specified connection (for example, in the form of L-lactate) at an initial dosage of 1 mg/m2/day to 100 mg/m2/day, in particular 1 mg/m2/day to 10 mg/m2/day, more specifically 3-6 mg/m2/day (equivalent to 2.5-5 mg free base/m2/day) or effective dosage of lactate, equal to 2.5 mg/m2/day 1.5 g/m2/day, in particular 25 mg/m2/day - 600 mg/m2/day, more specifically 200-500 mg/m2/day, as, for example, 250 mg/m2/day or 45-200 mg/m2/day, for example, 45-150 mg/m2/day or 56-185 mg/m2/day (equivalent 45-150 mg free base /m2/day), but when necessary can be higher or lower doses.

In one particular scheme introduction the patient will receive continuous intravenous infusion of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or a salt thereof, such as the lactate (particularly the L-lactate) or citrate for periods from 2 to 120 hours, for example from 2 to 96 hours, in particular from 24 to 72 hours, and the introduction is repeated over a desired period of time and, as, for example, once in one to three weeks.

More specifically, the patient may receive a continuous intravenous infusion of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or a salt thereof, such as the lactate (particularly the L-lactate) or citrate for periods of 24 hours daily for 5 days, and the introduction is repeated every week, or during periods of 48 hours, and the introduction is repeated every two weeks or during periods of 72 hours and the introduction is repeated every three weeks.

In another specific schema introduction the patient receives an infusion of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or a salt thereof, such as the lactate (particularly the L-lactate) or citrate, as an intravenous bolus over 2 hours one day a week every 1, 2 or 3 weeks, or once for 2 hours each 1, 2 or 3 weeks.

Higher doses, such as, for example, 1.5 g/m2/day, could be implemented with the use of dosage regimes with frequent interruptions, as, for example, 24 to 48 hour continuous intravenous infusion every one to two weeks. Lower doses could also be entered using dosage regimes with longer dosing (but still cyclic on/off), as, for example, 48-72 hour continuous intravenous infusion every who's two or three weeks.

In particular, the compounds of formula (I') or 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or all salts, such as the lactate or citrate, in particular, lactate, could be administered to the patient at a dose of 250 mg/m2/day for 72 hours by continuous intravenous infusion every three weeks.

In another embodiment, compounds of formula (I') or 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or all salts, such as the lactate or citrate, in particular lactate, could be administered to the patient during the five-day cycle of introduction.

Ultimately, the number of input connections and the type of composition should reflect the nature of the disease or physiological condition being treated, and to get at the discretion of the physician.

Compounds of formula (I), (II), (III), (XXX) and sub-groups of these compounds defined in the present application can be administered as the sole therapeutic agent or they can be administered in combination therapy with one or more other compounds for treatment of specific painful conditions, such as tumors, such as cancer, as defined above in the text of this application. Examples of other therapeutic agents or treatments that may enter the I or applied together (or at the same time, or in other time intervals) with the compounds of the present invention, include, but are not limited to, topoisomerase inhibitors, alkylating tools, antimetabolites, DNA-binding means, inhibitors microtubules (on target tubulin funds), and specific examples include cisplatin, cyclophosphamide, doxorubicin, irinotecan, fludarabine, 5FU, taxanes, mitomycin C and radiation therapy.

Other examples of therapeutic agents that can be administered, together (simultaneously or at other intervals) with the compounds of formula (I), (II), (III), (XXX) and their subgroups, defined in this application, include monoclonal antibodies and inhibitors of signal transduction.

For the case of CDK inhibitors or Aurora kinases, in combination with other therapies, the patient can be given two or more types of treatment for individual changing patterns of doses and different routes of administration.

If the compounds of formula (I) is administered in combination therapy with one, two, three, four or more other therapeutic agents (preferably one or two, more preferably one), these compounds can be administered simultaneously (either in the same or in different pharmaceutical compositions) or sequentially. In the case of sequentially what about the introduction they can be entered for short intervals (for example, over a period of 5-10 minutes) or at longer intervals (for example, 1, 2, 3, 4 or more hours, or, if necessary, even over longer periods of time), and the exact dosage regimes are consistent with the properties of a therapeutic agent (funds).

In addition, the compounds of the present invention can be used in conjunction with negitiations treatments such as radiation therapy, photodynamic therapy, gene therapy; surgery and regulated diet.

For use in combination therapy with other chemotherapeutics compound of formula (I) and one, two, three, four or more other therapeutic agents can be, for example, jointly included in a dosage form containing two, three, four or more therapeutic agents. Alternatively, individual therapeutic agent may be included in separate compositions, and may be provided together in the form of a kit, optionally with instructions for use.

Specialist in the art from its usual General education knows what dosage regimes and combined therapy should be used.

The diagnostic methods

Before the introduction of the compounds of formula (I), the patient may be subjected is issledovaniy to determine, whether a disease or condition that affects or is likely to suffer a patient, a disease or condition which would be susceptible to treatment with a compound having activity against Aurora and/or cyclin-dependent kinases.

For example, you can analyze the patient biological sample to determine whether a condition or disease such as cancer, which suffers or may suffer a patient's condition or disease that is characterized by a genetic abnormality or abnormal protein expression which leads to excessive activation of the CDK or to increase the sensitivity of the signaling pathway to the normal activity of the CDK. Examples of such anomalies, which lead to activation or increased sensitivity to the signal CDK2 include increasing regulation cycline E (Harwell RM, Mull BB, Porter DC, Keyomarsi, K.; J. Biol. Chem. 2004 Mar 26; 279(13): 12695-705) or loss of p21 or p27 or having options CDC4 (Ms. Rajagopalan H, Jallepalli PV, Rago C, Velculescu VE, Kinzler KW, Vogelstein B, Lengauer C.; Nature. 2004 Mar 4; 428(6978): 77-81). Tumors with CDC4 mutants or increasing regulation, in particular the excessive expression, cycline E, or loss of p21 or p27 may be particularly sensitive to CDK inhibitors. As an alternative or Supplement the biological sample obtained from the patient, can be analyzed to determine whether the status is e or disease, such as cancer, which suffers or may suffer a patient's condition or disease that is characterized by increasing regulation of Aurora kinase and, thus, may be particularly sensitive to inhibitors of Aurora kinases. The term "improving regulation" includes an increased or excessive expression, including gene amplification (i.e. multiple copies of the gene) and increased expression under the influence of transcriptional effect, as well as hyperactivity and activation, including activation as a result of mutations.

For example, the patient may be subjected to a diagnostic test to detect a marker characteristic of a redundant expression, increasing regulation or activation of Aurora kinase, or the patient may be subjected to a diagnostic test to detect a marker characteristic of increasing regulation cycline E, or loss of p21 or p27, or the presence of variants CDC4. The term "diagnosis" includes testing. Among the markers, the authors include genetic markers, including, for example, the study of the structure of DNA to identify mutations Aurora or CDC4. In addition, the term "marker" includes markers that are characteristic of increasing regulation of Aurora kinase or cycline E, including enzyme activity, enzymes, state of enzymes (e.g., phosphorylated or not)and mRNA levels is mentioned above proteins. Tumors with increasing regulation cycline E, or loss of p21 or p27 may be particularly sensitive to CDK inhibitors. Before treatment of the tumor can be mainly investigated to improve the regulation cycline E or loss of p21 or p27. Thus, the patient may be subjected to a diagnostic test for the detection of a marker characteristic of increasing regulation cycline E, or loss of p21 or p27.

Diagnostic tests typically performed on biological samples, selected samples from the tumor biopsy, blood samples (isolation and enrichment of the separated tumor cells), biopsy stool, sputum, chromosomal analysis, pleural fluid, peritoneal fluid, or urine.

It was found, see Ewart-Toland et al., (Nat Genet. 2003 Aug; 34(4): 403-12)that the individuals that form part of the groups with the Ile31 variant STK gene (gene Aurora kinase A), may have an increased susceptibility to some forms of cancer. Therefore, it seems that such individuals suffering from cancer, will be useful for the introduction of compounds with inhibitory activity against Aurora kinase. Therefore, a patient suffering from cancer or suspected cancer, can be tested to determine whether he or she is part of a group of people with Ile31 variant. In addition, Ms. Rajagopalan & co Nature. 2004 Mar ; 428 (6978): 77-81) found that when colorectal cancer and cancer of the uterus person, CDC4 (also known as Fbw7 or archipelago) are mutations (Spruck et al, Cancer Res. 2002 Aug 15; 62 (16): 4535-9). Identification of the individual with a mutation in the CDC4, may mean that the patient could well be suitable for treatment with CDK inhibitors. Before treatment the tumor mainly could be tested for the presence of variant CDC4. The method of study normally will include direct sequencing, oligonucleotide micrometrical analysis or mutant-specific antibodies.

Tumors with activating mutants Aurora or improving the regulation of Aurora, including any of the isoforms of the enzyme, can be particularly sensitive to inhibitors of Aurora kinases. Before treatment of the tumor can be mainly investigated to improve the regulation of Aurora or gene Aurora with the Ile31 variant (Ewart-Toland et al., Nat Genet. 2003 Aug; 34(4): 403-12). Ewart-Toland and collaborators have identified a common genetic variant in STK15 (leading to amino acid substitution F31I), which is mainly amplified and linked to the degree of aneuploidy in tumors of the colon of a person. These results are consistent with an important role option STK15 Ile31 in the susceptibility of people to cancer. In particular, it was assumed that this polymorphism in Aurora A is genetically the Kim modifier for developing breast carcinoma (Sun et al., Carcinogenesis, 2004, 25 (11), 2225-2230).

Gene Aurora displays A region of chromosome 20q13, which is frequently amplified in many cancers, such as breast, bladder, colon, ovarian, pancreas. Patients with tumors that have amplification of this gene could be particularly sensitive to treatment directed to the inhibition of Aurora kinases.

Methods of identification and analysis of mutations and increasing regulation of the protein, such as isoforms of Aurora, as well as amplification of chromosome 20q13 known to the person skilled in the art. Methods of testing could include, but is not limited to, standard methods, such as polymerase chain reaction with reverse transcriptase (RT-PCR) or in situ hybridization.

When tested by the method of RT-PCR the mRNA level in the tumor is determined by creating a cDNA copy of the mRNA, followed by amplification of cDNA by PCR. Methods PCR amplification, primers and amplification conditions known to the person skilled in the art. Operations with nucleic acids and PCR performed using standard techniques as described, for example, Ausubel, F.M. et al., eds. Current Protocols in Molecular Biology, 2004, John Wiley & Sons Inc. or Innis, M.A. et al., eds. PCR Protocols: a guide to methods and applications, 1990, Academic Press, San Diego. The reaction and operation, including methods that pertain to the nucleic acids described also is in Sambrook et al., 2001, 3rdEd, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press. On the other hand, can be used commercially available kit for RT-PCR (e.g., Roche Molecular Biochemicals), or methods described in U.S. patents 4666828, 4683202, 4801531, 5192659, 5272057, 5882864 and 6218529 and incorporated into the present application by reference.

Examples of methods of hybridization in situ to assess the mRNA expression could serve as fluorescent in situ hybridization (FISH) (see Angerer, 1987 Meth. Enzymol., 152: 649).

Generally, in situ hybridization comprises the following main stages: (1) fixation of tissue to be analyzed; (2) prehybridization processing of the sample to increase the availability of the target nucleic acid and reduce nonspecific binding; (3) hybridization of the mixture of nucleic acids with the nucleic acid in the biological structure of the tissue; (4) polarisation washing to remove nucleic acid fragments not bound in time, hybridization, and (5) identification of hybridized nucleic acid fragments. The probes used in such applications, as a rule, mark, for example, radioactive isotopes or fluorescent reporters. Preferred probes have considerable length, for example, from about 50, 100, or 200 nucleotides to about 1000, or more nucleotides, to allow specific hybridization with the target nucleotide sequence that is new acid (acids) in harsh environments. Standard methods for FISH described in Ausubel, F.M. et al., eds. Current Protocols in Molecular Biology, 2004, John Wiley & Sons Inc and Fluorescence In Situ Hybridization: Technical Overview by John M.S. Bartlett in Molecular Diagnosis of Cancer, Methods and Protocols, 2nded.; ISBN: 1-59259-760-2; March 2004, pps. 077-088; the Series Methods in Molecular Medicine.

On the other hand, the protein products expressed from mRNA, can be investigated by immunohistochemistry samples of tumors, a solid-phase immunoassay in microtiter tablets, Western blotting, 2-d electrophoresis on SDS-polyacrylamide gel, ELISA, flow cytometry and other methods known in the art to detect specific proteins. Detection methods could include the use of site-specific antibodies. The practitioner should be aware that all such well-known methods for the detection of increasing regulation cycline E, or loss of p21 or p27, or detection options CDC4, increasing regulation of Aurora and mutants Aurora could be applicable in this case.

Therefore, these techniques could also be used to identify tumors, especially suitable for treatment with compounds of the present invention.

Tumors with CDC4 mutants or increasing regulation, in particular the excessive expression of cycline E, or loss of p21 or p27, may be particularly sensitive to CDK inhibitors. Before treatment of the tumor can the be mainly investigated on increasing regulation, in particular, excessive expression cycline E (Harwell RM, Mull BB, Porter DC, Keyomarsi, K.; J. Biol. Chem. 2004 Mar 26; 279 (13): 12695-705) or loss of p21 or p27, or having options CDC4 (Ms. Rajagopalan H, Jallepalli PV, Rago C, Velculescu VE, Kinzler KW, Vogelstein B, Lengauer C.; Nature. 2004 Mar 4; 428 (6978): 77-81).

Patients with lymphoma mantle tissue (MCL) can be selected for treatment with compounds of the present invention with the use of diagnostic tests described in this application. MCL is a distinct clinico-pathological variant of non-Hodgkin lymphoma characterized by the proliferation of lymphocytes from small to medium in size with coexpressed CD5 and CD20, an aggressive and incurable clinical course and frequent translocation t(11; 14)(q13; q32). Overexpression of mRNA cycline D1 found in the lymphoma mantle cell (MCL)is the most important diagnostic marker. Yatabe and co-authors (Blood. 2000 Apr 1; 95 (7): 2253-61) suggested that the presence of cycline D1 should be considered as one of the standard criteria for the diagnosis of MCL, and the need to explore innovative ways of treatment of this incurable disease, based on the specified new criteria. Jones and co-authors (J. Mol. Diagn. 2004 May; 6 (2): 84-9) developed a quantitative PCR analysis reverse transcription real-time expression cycline D1 (CCND1) to aid in the diagnosis of lymphoma mantle cell (MCL). Howe and co-authors (Clin Chem. 2004 Jan; 50 (1): 0-7) used quantitative RT-PCR analysis real-time to assess the mRNA expression cycline D1 and found what quantitative RT-PCR for mRNA cycline D1, normalized by CD19 mRNA, can be used for the diagnosis of MCL in the blood, bone marrow and tissues. On the other hand, patients with breast cancer for treatment with CDK inhibitors could be selected using the above diagnostic tests. Tumor cells usually Express cyclin E in excessive amounts, and it has been shown that cyclin E redundantly expressed in breast cancer tumors (Harwell et al, Cancer Res, 2000, 60, 481-489). Therefore, breast cancer can be treated, in particular, inhibitors of CDK, developed in the present invention.

Antifungal use

In another aspect the present invention relates to the use of compounds of formula (I), (II), (III), (XXX) and their subgroups, as defined in this application as antifungal agents.

Compounds of formula (I), (II), (III), (XXX) and sub-groups defined in this application can be used in veterinary medicine (e.g., in the treatment of mammals, such as humans), or for processing plants (for example, in agriculture and horticulture), or as antifungal agents common actions, such as preservatives and disinfectants.

In one of the embodiments the present invention relates to compounds of formula (I), (II), (III), (XXX) and their subgroups, defined in nastoyasheva, for use in the prevention or treatment of fungal infections in mammals, such as humans.

Besides, the use of compounds of formula (I), (II), (III) and their subgroups, as defined in this application, for manufacturing a medicinal product intended for use in the prevention or treatment of fungal infections in mammals, such as humans.

For example, the compounds of the present invention can be administered to patients from the number of people who suffer or have the risk of local fungal infections caused by, among other organisms, species of Candida, Trichophyton, Microsporum or Epidermophyton, or infections of the mucous membranes caused by Candida albicans (e.g. thrush and vaginal candidiasis). In addition, the compounds of the present invention can be administered for the treatment or prophylaxis of systemic fungal infections caused by, for example, Candida albicans, Cryptococcus neoformans, Aspergillus flavus, Aspergillus fumigatus, Coccidiodies, Paracoccidiodies, Histoplasma or Blastomyces.

In another aspect the invention relates to fungicidal compositions for agricultural (including horticulture) applications containing compounds of formula (I), (II), (III), (XXX) and their subgroups described in this application, together with acceptable for agricultural use diluent or carrier.

In addition, the invention relates to what the manual treatment of animals (including mammals, as, for example, human), processing plant or seed having a fungal infection, which includes the processing of these mammals, plants or seeds or source of infection of these plants or seeds with an effective amount of compounds of formula (I), (II), (III), (XXX) and their subgroups described in this application.

In addition, the invention relates to a method of combating fungal infection of plants or seeds, which includes the treatment of the plants or seeds are effective against fungi number of fungicidal compositions containing the compound of formula (I), (II), (III), (XXX) and their subgroups described in this application.

To select compounds of the present invention, having specificity against inhuman CDK enzymes, can be used in various screening studies. Connection-specific effect on CDK enzymes of eukaryotic pathogens, can be used as anti-fungal and anti-parasitic substances. Inhibitors of CDK kinase Candida, i.e. CKSI, can be used in the treatment of candidiasis. Antifungal agents can be used against infections of the above types, or opportunistic infections, which are commonly found in debilitated and immunocompromised patients, such as patients with various types of leukemia and lymphoma, people receiving immunodepressive therapy, and patients with predisposing conditions, such as diabetes or AIDS, as well as neomonetarist patients.

Described in the technique methods of analysis can be applied for the selection of tools that can be useful for inhibition of at least one fungus involved in fungal infection, such as candidiasis, aspergillosis, mucormycosis, blastomycosis, geotechs, cryptococcosis, hromoblastomikoza, coccidioidomycosis, conidiospores, histoplasmosis, maduromycosis, rhinosporidiosis, Nocardia, para-actinomycosis, penicillin, magnolias or sporotrichosis. To identify antifungal agents, which may have therapeutic value in the treatment of aspergillosis, can be used various screening studies using CDK genes cloned from yeast fungi, such as Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus nidulans, or Aspergillus terreus, or, if a fungal infection is a mucon-nycosis, analysis of CDK can be carried out on the basis of such yeasts as Rhizopus arrhizus, Rhizopus oryzae, Absidia corymbifera, Absidia ramose or Mucorpusillus. Sources of other CDK enzymes include pathogen Pneumocystis carinii.

As an example, in vitro evaluation of antifungal activity of compounds can be performed by determining the minimum inhibitory concentration (M.I.C.), which isone of the concentration of the test compound in a suitable medium, when there is a weakening of the growth of a particular microorganism. In practice, seeded with a standard culture of, for example Candida albicans, a number of cups with agar, each of which contains the test compound at a certain concentration and then incubated each Cup over the relevant time period at 37°C. Then the Cup is examined for the presence or absence of growth of fungi and mark the appropriate M.I.C. value on the other hand, can be analyzed turbidity of liquid cultures, and the Protocol that describes an example of this analysis can be found in example 64.

Evaluation of the activity of compounds in vivo can be carried out by injection of the compounds at various dosage levels using intraperitoneal or intravenous injection, or oral administration, mice that were infected with fungi, for example, a strain of Candida albicans or Aspergillus flavus. The activity of compounds can be assessed by observing the development of fungal infection in groups of mice that were injected and not injected connections (using histology or by selection of fungi in infected animals). Activity can be measured by the dose level at which the compound provides 50% protection against lethal infection (PD50).

For antifungal use in humans, the compounds of formula (I), (II), (III), (XXX) and their functions uppy, defined in this application, can be administered by themselves or in a mixture with a pharmaceutical carrier selected in accordance with the intended route of administration and standard pharmaceutical practice. For example, they may be administered orally, parenterally, intravenously, intramuscularly or subcutaneously, in the form of compounds described above in the section entitled "pharmaceutical compositions".

For oral or parenteral administration to people the level of daily dosing of antifungal compounds of the present invention may range from 0.01 to 10 mg/kg (single dose) based, including the effectiveness of the compounds when introduced either orally or parenterally. Tablets or capsules of the compounds can contain, for example, from 5 mg to 0.5 g of active compound for administration of one, two or more tablets or capsules at the appropriate time. In any case, the physician should determine the actual dosage (effective amount), which is most likely to be appropriate for the patient, and the dosage will vary depending on the age, weight and response of the individual patient.

On the other hand, an antifungal compound can be administered in the form of a suppository or pessary, or they can be applied tapicerki in the form of a lotion, solution, cream, ointment or powder. For example, they can be incorporated into a cream consisting of an aqueous emulsion of polyethylene glycols or liquid paraffin; or they can be included in a concentration of from 1 to 10% in an ointment consisting of a base, for example bleached wax or white soft paraffin, as well as those stabilizers and preservatives that may be necessary.

In addition to the above-described therapeutic use of antifungal agents, selected using various screening assays can be used, for example, as preservatives in foods, food supplements to accelerate weight gain in cattle, in disinfectant compositions for treatment of inanimate matter, for example, to remove dirt hospital equipment and premises. Similarly a side by side comparison of the inhibition of mammalian CDK and CDK insects, such as CDK5 gene in Drosophila (Hellmich et al. (1994) FEBS Lett 356:317-21), will allow additional connections for this application the selection of inhibitors, which have different effects on enzymes humans/mammals and insects. Accordingly, the present invention specifically addresses the use and the inclusion of the compounds of the present invention in the composition of insecticides, such as, for example, for use in regulating the number of insects such as fruit fly.

In another VA who ianthe implement some of these CDK inhibitors can be selected on the basis of inhibitory specificity for cdks plants, relative to the corresponding enzymes of animals. For example, plant CDK can be submitted to various tests with one or more human enzymes, to select those compounds that have the greatest selectivity for the inhibition of the plant enzyme. Thus, in the present invention specifically addresses the structures on the basis of the described CDK inhibitors for use in agriculture, for example, in the form of defoliants, etc.

For the purposes of agriculture and horticulture compounds of the present invention can be applied in the form of compositions, the composition of which corresponds to a specific application and intended purpose. Thus, the compounds can be applied in the form of Farrukh Dustov or granules, Bates seed, aqueous solutions, dispersions or emulsions, impregnations, sprays, aerosols or means of fumigation. In addition, the composition can be supplied in the form of dispersible powders, granules or grains or concentrates for dilution before use. Such compositions may contain conventional carriers, diluents or excipients which are known and acceptable in agriculture and gardening, and they can be produced according to conventional methods. In addition, these compositions may include other active ingredients, for example, soybean is inane, with herbicide or insecticide activity, or other fungicide. Compounds and compositions can be used in a number of ways, for example they can be applied directly to the foliage, stems, branches, seeds or roots of plants, or to soil or other growth medium, and they can be used not only to eliminate the disease, but also for the prevention, to protect plants or seeds from defeats. As an example, the composition may contain from 0.01 to 1% of the mass. the active ingredient. For use in field conditions probable expenditure rate of the active ingredient can be from 50 to 5000 g/ha.

The present invention also provides the use of compounds of formula (I), (II), (III), (XXX) and their subgroups, as defined in this application to control the fungus that destroys wood, and in the treatment of the soil on which the plants grow, fields with seedlings of rice or water for irrigation. In addition, the present invention provides the use of compounds of formula (I), (II), (III), (XXX) and their subgroups, as defined in this application, to protect stored grain and other substances nereshitelno origin from fungal infection.

BRIEF DESCRIPTION of DRAWINGS

Figure 1 represents the image of the molecule dihydrate free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)1H-pyrazole-4-yl]urea with atoms in the form of thermal ellipsoids, as described in example 69 below.

Figure 2 shows a diagram of the crystal packing dihydrate free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea as described in example 69 below.

Figure 3 shows the XRPD pattern 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form, as described below in example 70.

Figure 4 shows the image of a molecule of L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea with atoms in the form of thermal ellipsoids, as described in example 71 below.

Figure 5 shows a diagram of the crystal packing of L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea as described in example 71 below.

Figure 6 shows the XRPD pattern of the original sample of L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and samples tested for stability as described below in example 72.

Figure 7 shows the XRPD pattern of the original sample 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form (FB1) and samples tested for stability as described below in example 72.

On Fig shows the XRPD pattern of the original sample 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)1H-pyrazole-4-yl]urea in the form of dihydrate free base (FB2) and sample tested for stability as described below in example 72.

EXAMPLES

In this section, the present invention will be illustrated but not limited by reference to specific embodiments of described in the following examples.

In the examples the following abbreviations are used:

AcOHacetic acid
BOCtert-butoxycarbonyl
CDI1,1-carbonyldiimidazole
DMAW90a mixture of solvents: DCM:MeOH, AcOH, H2O(90:18:3:2)
DMAW120a mixture of solvents: DCM:MeOH, AcOH, H2O(120:18:3:2)
DMAW240a mixture of solvents: DCM:MeOH, AcOH, H2O(240:20:3:2)
DCMdichloromethane
DMFdimethylformamide
DMSOthe sulfoxide
EDC1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide
Et3N the triethylamine
EtOActhe ethyl acetate
Et2Odiethyl ether
HOAt1-gidroksibenzotriazola
HOBt1-hydroxybenzotriazole
MeCNacetonitrile
MeOHmethanol
SiO2the silicon oxide
TBTUtetrafluoroborate N,N,N',N'-tetramethyl-O-(benzotriazol-1-yl)Urania
THFtetrahydrofuran

System analysis LC-MS and description techniques

In the examples, the compounds obtained were characterized by liquid chromatography and mass spectroscopy, using the following system and working conditions. If the connection was part of the atoms of elements with different isotopes, and are the only mass value provided for the connection weight corresponds to the weight of a connection with one isotope (i.e.35Cl;79Br and so on). Used more of the following systems, and they were equipped and configured to function in a very similar operating modes. Used modes of operation are also described below.

System LC-MS platform Waters
The HPLC system:Waters 2795
Mass spectroscopic detector:Micromass Platform LC
The PDA detector:Waters 2996 PDA

Acidic conditions analysis
Eluent A:H2O with 0.1% formic acid)
Eluent B:CH3CN (0,1% formic acid)
Gradient:5-95% eluent B over 3.5 minutes
Feed speed:0.8 ml/min
Column:Phenomenex Synergi 4µ MAX-RP 80A, a 2.0×50 mm

Basic conditions for analysis:
Eluent A:H2O (10 mm NH4HCO3/sub> buffer, the pH increased to 9.2 by addition of NH4OH)
Eluent B:CH3CN
Gradient:5-95% eluent B over 3.5 minutes
Feed speed:0.8 ml/min
Column:Phenomenex Luna C18(2) 5 μm to 2.0×50 mm

Polar conditions analysis
Eluent A:H2O with 0.1% formic acid)

Eluent B:CH3CN (0,1% formic acid)
Gradient:0-50% of eluent B over 3 minutes
Feed speed:0.8 ml/min
Column:Phenomenex Synergi 4µ MAX-RP 80A, a 2.0×50 mm

Lipophilic conditions analysis
Eluent A:H2O with 0.1% formic acid)
Eluent B:CH3CN (0,1% formic acid)
Gradient:55-95% of eluent B over 3.5 min
Feed speed:0.8 ml/min
Column:Phenomenex Synergi 4µ MAX-RP 80A, a 2.0×50 mm

Acidic conditions for sustained analysis
Eluent A:H2O with 0.1% formic acid)
Eluent B:CH3CN (0,1% formic acid)
Gradient:5-95% eluent B over 15 minutes
Feed speed:0.4 ml/min
Column:Phenomenex Synergi 4µ MAX-RP 80A, a 2.0×150 mm

Basic conditions for long-term analysis
Eluent A:H2O (10 mm NH4HCO3buffer, the pH increased to 9.2 by addition of NH4OH)
Eluent B: CH3CN
Gradient:5-95% eluent B over 15 minutes
Feed speed:0.8 ml/min
Column:Phenomenex Luna C18(2) 5 μm to 2.0×50 mm

Conditions of work platforms MS
The voltage on the capillary:3.6 kV (3,40 kV negative elektrorazpredelenie)
The voltage on the cone:25
The source temperature:120°C
Scanning range:100-800 atomic unit of mass
The method of ionization:Positive elektrorazpredelenie or negative elektrorazpredelenie, or positive and negative elektrorazpredelenie

System LC-MS platform Waters Fractionlynx
The HPLC system:2767 autosampler - 2525 binary gradient pump
Mass spectroscopic de the sector: Waters ZQ
The PDA detector:Waters 2996 PDA

Acidic conditions analysis
Eluent A:H2O with 0.1% formic acid)
Eluent B:CH3CN (0,1% formic acid)
Gradient:5-95% eluent B over 4 minutes
Feed speed:2.0 ml/min
Column:Phenomenex Synergi 4µ MAX-RP 80A, a 4.6×50 mm

Polar conditions analysis
Eluent A:H2O with 0.1% formic acid)
Eluent B:CH3CN (0,1% formic acid)
Gradient:0-50% solvent B over 4 minutes
Feed speed:2.0 ml/min
Column:Phenomenex Synergi 4µ MAX-RP 80A, a 4.6×50 mm

Lipophilic conditions analysis
Eluent A:H2O with 0.1% formic acid)
Eluent B:CH3CN (0,1% formic acid)
Gradient:55-95% of eluent B over 4 min
Feed speed:2.0 ml/min
Column:Phenomenex Synergi 4µ MAX-RP 80A, a 4.6×50 mm

Conditions of work platform Fractionlynx MS
The voltage on the capillary:3.5 kV (3,20 kV negative elektrorazpredelenie)
The voltage on the cone:25 (30 In negative elektrorazpredelenie)
The source temperature:120°C
Scanning range:100-800 atomic emassy
The method of ionization:Positive elektrorazpredelenie or negative electric is sputtering, or positive and negative elektrorazpredelenie

LC-MS purification system, controlled molecular weight

Preparative LC-MS is a standard and effective method used for the purification of organic compounds with small molecules such as compounds described in this application. Methods of liquid chromatography (LC) and mass spectrometry (MS) can be changed to provide better separation of the crude substances and the best detection samples using MC. Optimization of preparative gradient LC methods will include a selection of speakers, volatile eluents and modifiers, as well as gradients. In the technique of well-known methods of optimization of preparative LC-MS techniques and their subsequent use for cleansing compounds. Such methods are described in Rosentreter U, Huber U.; Optimal fraction collecting in preparative LC/MS; J Comb Chem.; 2004; 6(2), 159-64 and Leister W, Strauss K, Wisnoski D, Zhao Z, Lindsley C, Development of a custom high-throughput preparative liquid chromatography/mass spectrometer platform for the preparative purification and analytical analysis of compound libraries; J Comb Chem.; 2003; 5(3); 322-9.

One such system for purification of the compounds of preparative LC-MS as described below, although the specialist in the art will understand that could be applied alternative described systems and methods. In particular, could be applied methodology, based on preparative LC on a normal phase in the seat described herein methods for reversed phase. Most preparative LC-MS systems used on LC reversed-phase and volatile acid modifiers, because this approach is very effective for the purification of molecules of small size and because the solvents are compatible with mass-spectroscopy with elektrorazpredelenie positive ions. Alternatively, cleaning compounds could be applied to other chromatographic techniques, such as LC on the normal phase, other buffering of the mobile phase, key modifiers, etc. as shown in the above-described methods of analysis.

Preparative system LC-MS:

System Waters Fractionlynx:

-Equipment:

2767 Dual Loop Autosampler/Fraction Collector

2525 preparative pump

CFO (column fluidic organiser) to select columns

RMA (Waters reagent manager) as feed pump

Waters ZQ Mass Spectrometer

Waters 2996 Photo Diode Array detector

Waters ZQ Mass Spectrometer

-Software:

Masslynx 4,0

Conditions of work platforms MS Waters
The voltage on the capillary:3.5 KV (3,20 KV negative elektrorazpredelenie)
The voltage on the cone:25
The source temperature:120 the C
Multiplier:500
Scanning range:100-800 atomic emassy
The method of ionization:Positive elektrorazpredelenie or negative elektrorazpredelenie

LC-MS preparative system Agilent 1100

-Equipment:

The device autodate samples: 1100 series “prepALS”

Pump: 1100 series “PrepPump for preparative feed gradient solvent and 1100 series “QuatPump” for filing a modifier in a stream of solvent

UV detector: 1100 series “MWD” Multi Wavelength Detector

MS detector: 1100 series LC-MSD VL”

Collector fractions: 2 × “Prep-FC”

Feed pump: Waters RMA”

Agilent Active Splitter

-Software:

Chemstation: Chem 32

Conditions of work platforms MS Agilent
The voltage on the capillary:4000 (3500 In negative elektrorazpredelenie)
Fragmentation/gain:150/1
The supply of the drying gas:13,0 l/min
Gas temperature:350°C
Pressure sprayer:50 psi
Scanning range:125-800 atomic unit of mass
The method of ionization:Positive elektrorazpredelenie or negative elektrorazpredelenie

Conditions for chromatography:

-Column:

1. Chromatography at low pH values:

Phenomenex Synergy MAX-RP, 10µ, 100×21,2 mm (alternatively applied Thermo Hypersil-Keystone HyPurity Aquastar, 5µ, 100 × 21,2 mm for more polar compounds)

2. Chromatography at high pH values:

Phenomenex Luna C18(2), 10µ, 100×21,2 mm (alternatively used Phenomenex Gemini, 5µ, 100×21,2 mm)

-Eluent:

1. Chromatography at low pH values:

Solvent A: H2O + 0.1% of formic acid, pH~1,5

Solvent B: CH3CN + 0.1% of formic acid

2. Chromatography at high pH values:

Solvent A: H2O + 10 mm NH4HCO3+ NH4OH, the pH of 9.2

Solvent B: CH3CN

3. Additional solvent

MeOH + 0.2% of Formic acid (for chromatography of both types)

-Methods:

In accordance with the analytical features were selected the most suitable type of preparative chromatography. The typical process is ur consisted of the implementation of the analytical LC-MS, using the most appropriate for the connection structure type chromatography (at low or high pH values). If analytical signs pointed to a good result chromatography, chose a suitable preparative methods of the same type. Typical conditions for the implementation of chromatographic methods at low and high pH values were as follows:

The flow rate: 24 ml/min

Gradient: Basically, all the gradients included the original 0,4-min stage when fed a mixture of 95% A + 5% B. Then, in accordance with the analytical characteristics were chosen 3,6-min gradient to achieve good separation (for example, from 5 to 50% B for poorly retained compounds, from 35 to 80% for sredneagressivnyh connections etc)

FlushingAt the end of the gradient was performed 1,2-min stage leaching

The return to equilibrium: to prepare the system for the next run was performed 2,1-min stage of return to equilibrium

The additional speed flow: 1 ml/min

-The solvent is:

All compounds are usually dissolved in 100% MeOH or 100% DMSO

On the basis of the information provided by any of the experts in the art could clean up the connections described in this application, using preparative LC-MS.

The initial substance in each of the examples are commercially available, unless otherwise specified.

Example 1

Synthesis of 5-cyano-2-methoxy-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazo the l-2-yl)-1H-pyrazole-4-yl]benzamide

1A.Synthesis of (3,4-dinitrophenyl)morpholine-4-ylmethanone

A mixture of 3,4-dinitrobenzoic acid (10.0 g) and thionyl chloride (30 ml) was boiled under reflux for 2 hours, cooled to room temperature and remove excess thionyl chloride by azeotropic distillation with toluene. The residue was dissolved in THF (100 ml) and at 0°C were simultaneously added to a mixture of morpholine (4,1 ml) and Et3N (7.2 ml). The mixture was stirred for 3 hours, adding water (100 ml) and then was extracted with EtOAc. The organic fraction was washed with a saturated solution of salt, dried (MgSO4) and was evaporated in vacuum. Recrystallization of the residue from MeOH gave (3,4-dinitrophenyl)morpholine-4-ylmethanol (8,23 g) as a solid yellow color. (1H NMR (300 MHz, DMSO-d6) δ 8.3 (l, 1H), and 8.3 (s, 1H), 8.0 a (d, 1H), 3,7-3,5 (m, 8H)).

1B.Synthesis of (3,4-diaminophenyl)morpholine-4-ylmethanone

A mixture of (3,4-dinitrophenyl)morpholine-4-ylmethanone (1.0 g) and 10% Pd/C (150 mg) in MeOH (30 ml) was shaken in a hydrogen atmosphere at room temperature for 10 hours, then filtered through a layer of celite and was evaporated in vacuum, obtaining (3,4-diaminophenyl)morpholine-4-ylmethanol (900 mg) (1H NMR (300 MHz, DMSO-d6) δ 6,6 (s, 1H), 6,5 (s, 2H), 4,8 (s, 1,5H), 4,6 (s, 1,5H), 4,1 (s, 1H), 3,6 (m, 4H), 3,4 (m, 4H)).

1C.Synthesis of 4-morpholine-4-ylmethylene-1,2-diamine

To a mixture of (3,4-dinitrophenyl)morpholine-4-ylmethanone (2,84 g) and dry THF (50 ml) was added NaBH4(954 mg) and then dropwise BF3.Et2O (3.2 ml). The mixture was stirred at room temperature for 3 hours and then was suppressed by the addition of MeOH. The mixture was evaporated in vacuum, was distributed between EtOAc and water, the organic fraction was washed with a saturated solution of salt, dried (MgSO4) and was evaporated in vacuum. The residue was purified column flash chromatography, elwira EtOAc, and has obtained 4-(3,4-dinitrobenzyl)morpholine (1.08 g).

A mixture of 4-(3,4-dinitrobenzyl)of the research (550 mg) and 10% Pd/C (75 mg) in MeOH (10 ml) was shaken in a hydrogen atmosphere at room temperature for 4 hours, then filtered through a layer of celite and was evaporated in vacuum, obtaining 4-morpholine-4-ylmethylene-1,2-diamine (483 mg) as a main component of the mixture.

1D.Synthesis of 5-morpholine-4-ylmethyl-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole

A mixture of 4-morpholine-4-ylmethylene-1,2-diamine (2.3 g, 11.1 mmol), 4-nitro-1H-pyrazole-3-carboxylic acid (of 1.57 g, 10.0 mmol), EDC (2,13 g, 11.1 mmol) and HOBt (1.50 g, 11.1 mmol) in dry DMF (25 ml) was stirred at room temperature for 24 hours. The mixture was evaporated in vacuo, the crude residue was dissolved in AcOH (40 ml) and boiled under reflux for 3 hours the Solvent was removed in vacuum and the residue cleaned the Lee column flash chromatography, elwira 0-20% MeOH in EtOAc, receiving 5-morpholine-4-ylmethyl-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole in the form of a yellow solid substance (1.0 g, 61%). (LC/MS: Rt1,83, [M+H]+329).

1E.Synthesis of 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamineand

Palladium on coal (10%, 0.08 g) was added to a solution of 5-morpholine-4-ylmethyl-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole (0,82 g, 2.5 mmol) in DMF (30 ml) under nitrogen atmosphere. The mixture was shaken in a hydrogen atmosphere for 4 hours, then filtered through celite, rinsing with methanol. The filtrate was concentrated in vacuum, obtaining 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine in the form of a brown solid substance (530 mg, 71%). (LC/MS: Rt1,94, [M+H]+299).

1F.Synthesis of 5-cyano-2-methoxybenzoic acid

To a mixture of methyl 2-hydroxy-5-cyanobenzoate (2 g, 5.6 mmol) and K2CO3(4.68 g, a 16.8 mmol) in acetone (50 ml) was added methyliodide (0.7 ml, 5.6 mmol). After that the reaction mixture was heated at 65°C during the night, getting a solid, which was filtered while it was hot, and washed with acetone, obtaining the methyl ester of 5-cyano-2-methoxybenzoic acid (0.45 g). The crude product was dissolved in THF (5 ml), then treated with a solution of LiOH (to 0.108 g, 0.26 mmol) in water (5 ml) and stirred at room temperature is f during the night. The reaction mixture was acidified using 2M HCl and was extracted with EtOAc (×2). The organic fraction was dried (MgSO4) and was evaporated in vacuum, obtaining 5-cyano-2-methoxybenzoic acid (0,277 g). (LC/MS kislotno.: Rt2,92, [M+H]+178).

1G.Synthesis of 5-cyano-2-methoxy-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]benzamide (method using carboxylic acid)

5-Cyano-2-methoxybenzoic acid (example 1F) (40 mg, 0.22 mmol) was dissolved in dichloromethane (5 ml), then was added dropwise oxalicacid (to 34.4 mg, 0,264 mmol) and then DMF (1 drop). The reaction mixture was stirred at room temperature for 1 hour, evaporated in vacuo, then was evaporated using toluene (×2). A mixture of 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (100 mg, 0.33 mmol), 5-cyano-2-methoxybenzylamine and diisopropylethylamine (1,83 μl, 0.9 mmol) in THF (5 ml) was stirred at 0°C and then allowed to warm to room temperature for 2 hours. Then the reaction mixture was concentrated in vacuum. The residue was purified column flash chromatography (SiO2, 5-7% MeOH-DCM)to give 5-cyano-2-methoxy-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]benzamide (12 mg). (LC/MS kislotno.: Rt2,02 min, [M-H]+458).

Example 2

Synthesis of [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-piraso the-4-yl]amide of 6-Mei[2.1-b]thiazole-5-carboxylic acid

A mixture of 6-Mei[2.1-b]thiazole-5-carboxylic acid (Bionet) (61 mg, 0.33 mmol), 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (100 mg, 0.33 mmol), EDC (77 mg, 0,39 mmol) and HOAt (54 mg, 0,39 mmol) was stirred in DMF (3 ml) at 80°C for 1 h, then at room temperature for 20 hours the Mixture was evaporated in vacuum and the residue was distributed between EtOAc and saturated NaHCO3. The organic portion was washed with a saturated solution of salt, dried (MgSO4) and was evaporated in vacuum. The residue was purified preparative LC/MS, receiving [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amide of 6-methylimidazo[2.1-b]thiazole-5-carboxylic acid (29 mg). (LC/MS VI.: Rt2,56, [M+H]+463).

Example 3

Synthesis of 2-cyano-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]ndimethylacetamide

The mixture tsianuksusnogo acid (23 mg, 0.28 mmol), 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (70%, 100 mg, 0.23 mmol), TBTU (89 mg, 0.28 mmol) and DMF (2 ml) was stirred at 25°C during the night. Then the mixture was evaporated in vacuum. After purification with flash chromatography, the elution of DCM to 6% MeOH/DCM, received 2-cyano-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]ndimethylacetamide in a solid yellow color (65 mg, 77%). (LC/MS (acidic method/end connection): Rtbr4.61, [MH] +366).

Example 4

2-Cyano-2-cyclopropyl-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]ndimethylacetamide

4A.Synthesis cyanocyclohexane acid

1H. NaOH (3,26 ml, 3,26 mmol) was added to a solution of ethyl ester cyanocyclohexane acid (0.5 g, 3,26 mmol) in THF (15 ml). After 4 hours stirring at 25°C. the reaction mixture was evaporated in vacuo, re-dissolved in water (20 ml) and neutralized by adding 1N. HCl solution (3,26 ml). Then the mixture was extracted with EtOAc (3×20 ml) and the combined, dried (Na2SO4) organic fraction was evaporated in vacuum, obtaining contaminated cyanocyclohexane acid in the form of a clear oil. The resulting substance without any further purification used in the synthesis of example 4B.

4B.2-Cyano-2-cyclopropyl-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]ndimethylacetamide

The product of example 4A was introduced in the interaction with 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-aluminum according to the method described in example 3, except that the crude product was distributed between dichloromethane and saturated aqueous NaHCO3and then cleaned by rubbing with Et2O. LC/MS (kislotno. way): Rt1,79, [M+H]+406.

Examples 5-14

Received preprogram the Oia examples 5-14, following the methods of examples 1-3, modified in those cases, which are shown in the table below.

td align="center"> 14
ExampleStructureA common way to obtainDifferences from the common wayLC/MS
5Example 1G[M+H]+425
Rt1,77 kislotno.
6Example 1GCleaning prep.
LC/MS
[M+H]+365
Rt2,45 VI.
7Example 1GCleaning prep.
LC/MS
[M+H]+339
Rt2,21 VI.
8Example 2Cleaning prep.
LC/MS
[M+H]+399
Rt1,74 kislotno.
9Example 2Cleaning prep.
LC/MS
[M+H]+397
Rt1,64 kislotno.
10Example 3Purification of column chromatography
[SiO2the elution DMAW 240-120]
[M+H]+381
Rt1,85
kislotno.
11Example 3Purification of column chromatography
[SiO2the elution DMAW 240-120]
[M+H]+397
Rt1,76
kislotno.
12Example 3Purification of column chromatography
[SiO2the elution DMAW 240-120]
[M+H]+397
Rt1,76
kislotno.
13Example 3Purification of column chromatography
[SiO2the elution DMAW 240-120]
[M+H]+397,24
Rt1,79
kislotno.
Example 3Purification of column chromatography
[SiO2the elution DMAW 240-120]
[M+H]+465,3
Rt1,99
kislotno.

Example 15

Synthesis of triptoreline N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-2-morpholinylcarbonyl

15A.Synthesis of 5,6-dimethoxy-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole

To a solution of EDC (to 4.81 g, 25 mmol), HOBt (3,40 g, 25 mmol) and triethylamine (4,67 g, 46 mmol) in DMF (100 ml) was added 4-nitro-1H-pyrazole-3-carboxylic acid (3,63 g, 23,09 mmol) and the dihydrochloride 4,5-dimethoxybenzene-1,2-diamine (of 5.06 g, 20,99 mmol) and the mixture was stirred at room temperature overnight. The solvent was removed in vacuum and the remaining solid substance was distributed between EtOAc (50 ml) and saturated aqueous NaHCO3(50 ml). The precipitate was removed by filtration. The filtrate was washed with water, diethyl ether and then subjected to the azeotropic distillation with MeOH and toluene, receiving (2-amino-4,5-acid)amide of 4-nitro-1H-pyrazole-3-carboxylic acid (2.35 g, 36%). (2-Amino-4,5-acid)amide of 4-nitro-1H-pyrazole-3-carboxylic acid (2.35 g, the 7.65 mmol) was dissolved in acetic acid (150 ml) and boiled under reflux at 140°C for 5 hours. R is the target left to cool and the solvent was removed in vacuum. The obtained solid substance was distributed between EtOAc (25 ml) and saturated salt solution (25 ml). The organic layer was separated, dried (MgSO4), filtered and solvent was removed in vacuum, obtaining 5,6-dimethoxy-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole (2,08 g, 94%).

15B.Synthesis of 3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine

A mixture of 5,6-dimethoxy-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole (2,08 g, 7.2 mmol) and 10% palladium on coal (200 mg) in ethanol (150 ml) and DMF (50 ml) was first made at room temperature and pressure overnight. The reaction mixture was filtered through celite and solvent was removed in vacuum. The obtained solid substance was subjected to the azeotropic distillation with methanol and toluene and the solvent was removed in vacuum. The crude substance was purified flash chromatography, elwira a mixture of DCM:MeOH:acetic acid:water(120:18:3:2) [DMAW120] and then with a mixture of DCM:MeOH:acetic acid:water(90:18:3:2) [DMAW90]. Then combined fractions containing the product, and the solvent was removed in vacuum, obtaining 3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (~1 g, 53%).

15C. Synthesis of N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-4-BOC-2-morpholine carboxamide

To a solution of EDC (125 mg, 0.54 mmol) and HOAt (74 mg, 0.54 mmol) in DMF (2 ml) was added 3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-the Lamin (117 mg, 0.45 mmol) (example 15B) and (rac)-BOC-2-carboxymaltose (125 mg, 0.54 mmol) and the mixture was stirred over night at room temperature. Then the mixture was distributed between EtOAc and water. After that, the organic layer was sequentially washed with saturated aqueous sodium bicarbonate solution, saturated salt solution and then dried (MgSO4). The solution is evaporated in vacuum to dryness and the residue was purified column flash chromatography [SiO2, gradient elution: EtOAc-hexane (1:1)to EtOAc-MeOH (80:20)], obtaining N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-4-BOC-2-morpholine carboxamide (65 mg) as colorless solids. (LC/MS (kislotno. way): Rtto 2.65 min, [M+H]+473).

15D.Synthesis of triptoreline N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-2-morpholinylcarbonyl

N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-4-BOC-2-morpholine carboxamide (65 mg, 0.14 mmol) and anisole (60 μl, of 0.56 mmol) was dissolved in a mixture triperoxonane acid and dichloromethane (1:1; 2 ml). After three hours at room temperature the mixture was evaporated to dryness, getting triptorelin N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-2-morpholinylcarbonyl (73 mg) as a colourless solid (LC/MS (kislotno. way): Rtof 1.42 min, [M-H]+371).

Example 16

Synthesis of N-[3-(5,6-dime the hydroxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-4-isopropyl-2-morpholine carboxamide

To triptoreline N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-2-morpholinylcarbonyl (example 15D) (34 mg, 0.07 mmol) and K2CO3(20 mg, 0.14 mmol) in MeCN (1 ml) was added 2-iodopropane (17 μl, 0.15 mmol). The mixture was stirred at 80°C for approximately 48 hours, after which the mixture was concentrated and the residue was purified flash chromatography [SiO2the elution gradient: DCM:MeOH (98:2) to DCM:MeOH:conc. aq. NH3(90:10:1)], obtaining N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-4-isopropyl-2-morpholinylcarbonyl (12 mg) as a colourless resin (LC/MS (VI. way): Rt2,52 min, [M+H]+415).

Example 17

Synthesis of N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-1-methylpiperidin-3-carboxamide

To a solution of 3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (65 mg, 0.25 mmol) (example 15B), the hydrochloride (rac)-1-methylpiperidin-3-carboxylic acid (50 mg, 0.27 mmol) and diisopropylethylamine (50 μl, 0.27 mmol) in DMF (1 ml) was added TBTU (97 mg, 0.30 mmol). The mixture was stirred at room temperature for about 16 hours, after which was added 1N. aqueous NaOH (1 ml) and the mixture was stirred for another 1 hour. Then the mixture was evaporated to dryness in vacuo, the residue was purified column flash chromatography (SiO2the elution gradient: DCM:MeOH (98:2) to DCM:MeOH:conc. aq. NH3(70:30:3)), receiving N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-rac-1-methylpiperidin-3-carboxamide (20 mg) as a colourless resin (LC/MS (VI. way): Rt2,35 min, [M+H]+385).

Example 18

Synthesis of 3-chloro-N-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl-5-(4-methylpiperazin-1-yl)benzamide

18A.Synthesis of 3-chloro-5-(4-methylpiperazin-1-yl)benzonitrile

5-fluoro-3-chlorobenzonitrile (1 g, 6.4 mmol) was dissolved in DMSO (20 ml) followed by addition of K2CO3(1.3 g, 9.6 mmol) and 1-methylpiperazine (1,4 ml, 12.8 mmol). The reaction mixture was heated at 80°C for 20 hours. To the crude substance was added diethyl ether (10 ml), then acidified using 1N. HCl. From the crude reaction mixture was filtered precipitate, getting 3-chloro-5-(4-methylpiperazin-1-yl)benzonitrile (1.4 g, yield 93%) as a white solid (LC/MS: Rt1,83 min, [M+H]+236, kislon. way).

18B.Synthesis of 3-chloro-5-(4-methylpiperazin-1-yl)benzoic acid

To 3-chloro-5-(4-methylpiperazin-1-yl)benzonitrile (1.4 g, 5.9 mmol), dissolved in ethanol (10 ml), was added 2M NaOH (20 ml) and the reaction mixture is boiled under reflux for 20 hours. The mixture was evaporated in vacuo, acidified crude product is 1H. HCl to pH 6 and was distributed between EtOAc and H2O. the Organic layer was evaporated in vacuo the e dry, receiving 0.7 g specified in the title compound as a white solid (LC/MS: Rt1,67 min, [M+H]+256, kislon. way).

18C.Synthesis of [3-chloro-N-[3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-5-(4-methylpiperazin-1-yl)benzamide

The connection is produced by way similar to example 15C, but using 3-chloro-5-(4-methylpiperazin-1-yl)benzoic acid (200 mg, 0.78 mmol) as a reagent in the method of example 15C instead of (rac)-BOC-2-carboxymaltose. The crude product was purified column flash chromatography [SiO2, elwira DMAW 240-90]getting 92 mg (yield 25%) specified in the title compound as light brown solid (LC/MS: Rt2,07, [M+H]+496).

Examples 19-21

Following the method described in example 15, modified in those cases, where indicated, were obtained compounds of examples 19-21.

ExampleStructureA common way to obtainDifferences from the General methodsLC/MS
19Example 15CUsed 5-pyrrolidin-1-ilmatieteen-2-carboxylic acid [M+H]+437
Rt2,62
VI.
20Example 15CUsed 2-dimethylamino-methylfuran-3-carboxylic acid
Purification preparative LC/MS
[M+H]+411
Rt1,6
kislotno.
21Example 15CWithout treatment water environment
Purification column
chromatography
[SiO2the elution DMAW 240] further purification preparative LC/MS
[M+H]+453,17
Rt1,75
kislotno.

Example 22

Synthesis of 5-chloro-2-methoxy-N-{3-[5-(4-methylpiperazin-1-ylmethyl)-1H-benzimidazole-2-yl]-1H-pyrazole-4-yl}benzamide

22A.Synthesis of (3,4-dinitrophenyl)-(4-methylpiperazin-1-yl)methanone

3,4-Dinitrobenzoic acid (50 g, 0.24 mol) was boiled under reflux in SOCl2(160 ml) for 6 hours. Then the mixture was evaporated to dryness in vacuum. The product was dissolved in THF and was cooled to 5°C. To this solution was added dropwise N-methylpiperazine (26,2 ml, 0.24 mol) and Et3N (42 ml) in solution in THF (50 ml). After stirring over night at whom atoi temperature the solution was poured into water (1.5 l) and stirred at approximately 5°C for 0.5 hours. The resulting solid precipitate was collected and dried, obtaining (3,4-dinitrophenyl)-(4-methylpiperazin-1-yl)methanon (40 g) in a solid yellow color.

22B.Synthesis of 1-(3,4-diaminophenyl)-4-methylpiperazine

To a cooled solution (5°C) (3,4-dinitrophenyl)-(4-methylpiperazin-1-yl)methanone (12.2 g, 0,041 mol) in THF was added powdered NaBH4then adding dropwise BF3·Et2O supporting all this time the temperature below 5°C. Gave rise the temperature of the mixture to room for 2 hours, then stirred another 2 hours at room temperature. Then to the reaction mixture was carefully added MeOH (causing a heated gas), continued the stirring for 10 minutes and then concentrated mixture. The residue was distributed between EtOAc and saturated aqueous NaHCO3. The organic layer was washed with water, saturated salt solution and then dried (MgSO4). The solution was evaporated in vacuo, then the residue was purified flash chromatography on SiO2, elwira gradient of DCM:MeOH (98:2) to DCM:MeOH:conc. aq. NH3(90:10:1)to give an orange crystalline solid (3.7 g). Recrystallization from MeOH allowed to obtain 1-(3,4-dinitrobenzyl)-4-methylpiperazine (1 g) in the form of a solid crystalline substances orange color.

22C.Synthesis of 1-(3,4-diami benzyl)-4-methylpiperazine

1-(3,4-dinitrobenzyl)-4-methylpiperazine (1 g) was dissolved in a mixture of DMF:MeOH (1:1, 20 ml) and stirred with 10% Pd/C (50 mg) in an atmosphere of H2within 6 hours. Then the mixture was filtered and evaporated, receiving a dark solid, which was immediately used in the next stage without any further purification.

22D.Synthesis of 5-chloro-2-methoxy-N-{3-[5-(4-methylpiperazin-1-ylmethyl)-1H-benzimidazole-2-yl]-1H-pyrazole-4-yl}benzamide

4-(5-Chloro-2-methoxybenzylamine)-1H-pyrazole-3-carboxylic acid (1,17 g), the crude diamine, i.e 1-(3,4-diaminophenyl)-4-methylpiperazine (0.87 g) and TBTU (1.52 g) was dissolved in DMF (15 ml) and stirred for approximately 16 hours. Then the mixture was evaporated to dryness obtaining a dark solid. This is a dark solid (100 mg) was dissolved in AcOH (4 ml) and the mixture was heated at 80°C for 3 hours. The reaction mixture was evaporated in vacuum and the residue was purified flash chromatography (SiO2, elwira DMAW 120)to give 5-chloro-2-methoxy-N-{3-[5-(4-methylpiperazin-1-ylmethyl)-1H-benzimidazole-2-yl]-1H-pyrazole-4-yl}benzamide as luxushotels salt (35 mg). (LC/MS(acidic method/end connection): Rt6,63 [M+H]+480).

Example 23

Synthesis of 1-(2,6-diferensial)-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

A mixture of 3(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (example 1E), (100 mg, 0.33 mmol) and CDI (217 mg, of 1.34 mmol) in THF (2 ml) was irradiated with microwave irradiation (150°C, 150 W) for 15 minutes. After this was added 2,6-differentiatin (384 mg, 2.68 mmol)and the reaction mixture was again irradiated in the same conditions for a further 15 minutes. After cooling the heterogeneous mixture was filtered, the filtrate was concentrated, the residue was purified column chromatography (SiO2, elwira gradient of DCM:MeOH:AcOH:H2O(240:20:3:2) (DMAW240) - (120:18:3:2) (DMAW120)) and got 1-(2,6-diferensial)-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (30 mg, 19%). (LC/MS kislotno.: Rt1,84 [M+H]+468).

Examples 24-34

Following the General method outlined in example 23, but with the modifications shown in the table below were obtained compounds of examples 24-34.

ExampleStructureDifferences from the General methodsLC/MS
24As the amine used cyclopropylamine[M+H]+382,24
Rt1,59
kislotno.
25[M+H] +440,31
Rt1,84
kislotno.
26Purification preparative LC/MS[M+H]+440,34
Rt2,20
polarn.
27Purification preparative LC/MS[M+H]+426,27
Rt1,57
kislotno.
28The crude product was distributed between EtOAc and .NaHCO3.
Was cleaned by column flash chromatography on silica, elwira 100% EtOAc To 10% MeOH
[M+H]+444
Rt6,67
kislotno.
29The crude product was distributed between EtOAc and .NaHCO3.
Was cleaned by column flash chromatography on silica, elwira 100% EtOAc To 10% MeOH
[M+H]+444
Rt6,98
kislotno.
30Added 2.68 mmol DIPEA. The mixture was stirred with 2M NaOH and was extracted with dichloromethane.
PTS is looking in column flash chromatography 2-5% MeOH-dichloromethane, then LC/MS
[M+H]+411
Rt2,45
VI.
31Added 2.68 mmol DIPEA.
Was purified preparative LC/MS
[M+H]+385
Rt1,60
kislotno.
32Added 2.68 mmol DIPEA.
Was purified preparative LC/MS
[M+H]+427
Rt1,69
kislotno.
33Added 2.68 mmol DIPEA.
Handled distribution between EtOAc and feast upon. NaHCO3.
Was purified column chromatography on silica (DMAW240-120), then preparative LC/MS
[M+H]+448
Rt2,07
VI.
34Added 2.68 mmol DIPEA.
Handled distribution between EtOAc and feast upon. NaHCO3.
Was purified preparative LC/MS
[M+H]+396
Rt1,74
VI.

Example 35

Synthesis of 1-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-3-pyridin-3-rocephine

A mixture of 3-aminopyridine (31.5 mg, 0.33 mmol), Et3N (of € 0.195 ml, of 1.32 mmol) in dichloromethane (3 ml) was cooled to 0°C and then was treated with triphosgene (85 mg, 0.28 mmol). The reaction mixture was stirred at room temperature for 1 hour, then added 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (100 mg, 0.33 mmol) and stirred at room temperature until completion of the reaction. The mixture was treated with 2M NaOH in MeOH for 30 minutes and then was evaporated in vacuum. The residue was purified column flash chromatography [SiO2, 2-20% MeOH/DCM], then rubbing with dichloromethane and then with diethyl ether there was obtained 1-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]-3-pyridin-3-Ilocano (20 mg). (LC/MS VI.: Rtto 2.29, [M+H]+419).

Example 36

Synthesis of [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amide thiomorpholine-4-carboxylic acid

Phosgene (20% in toluene) (0.3 ml) at 0°C was added to a solution of 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (100 mg, 0.33 mmol) in a mixture of toluene/dichloromethane (1:1). The reaction mixture was stirred at room temperature for 1 hour, then the excess phosgene was blown out by a stream of nitrogen. Added thiomorpholine (35 mg, 0.33 mmol) and the reaction mixture was stirred at room temperature in the course is 1 hour then at 60°C for 1 hour. Then the reaction mixture was concentrated in vacuo and the residue was purified preparative LC/MS, receiving [3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amide thiomorpholine-4-carboxylic acid. (LC/MS polarn.: Rt2,58, [M+H]+428).

Example 37

Synthesis of 1-(4-forfinal)-1-methyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

The method used for the synthesis specified in the connection header, similar to the method described in example 35, but using 4-fluoro-N-methylamine instead of 3-aminopyridine, in addition, the reaction was conducted at 50°C for 2 hours. The crude product was isolated as a precipitate from the cooled reaction mixture and then purified column flash chromatography [SiO2, elution with a mixture of DCM:MeOH:AcOH:water(240:20:3:2)]. The obtained product was washed with diethyl ether, obtaining 1-(4-forfinal)-1-methyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (3 mg) as a colorless solid substance. (LC/MS kislotno.: Rt2,12, [M+H]+450).

Examples 38-43

Following the procedures described in examples 35 and 37, modified in the cases indicated in the table below were obtained compounds of examples 38-43.

Example StructureA common way to obtainDifferences from the General methodsLC/MS
38Example 35Handled distribution between EtOAc and feast upon. NaHCO3.
Was purified preparative LC/MS
[M+H]+460
Rt2,03
kislotno.
39Example 35Handled distribution between EtOAc and feast upon. NaHCO3.
Was purified preparative LC/MS
[M+H]+449
Rt2,32
polarn.
40Example 35The reaction mixture was heated at 60°C. was Treated distribution between EtOAc and feast upon. NaHCO3.
Was purified preparative LC/MS
[M+H]+460
Rt2,22
VI.
41Example 37[M+H]+450,24
Rt2,09
kislotno./td>
42Example 37[M+H]+468,38
Rt1,99
kislotno.
43Example 37The crude product was isolated from the filtrate, and not from the precipitate. Further purification preparative LC/MS[M+H]+450,41
Rt2,68
VI.

Example 44

Synthesis of 1-(4-forfinal)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

To 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (example 1E) (100 mg, 0.33 mmol) in THF (2 ml) was added 4-forgenerations and the mixture was stirred for about 16 hours at room temperature. Added Tris-amine on the polymer resin (800 mg, 4 mmol/g) and which mixture for another 4 hours. The resin was removed by filtration, the filtrate was treated with 1H. KOH (2 ml, MeOH:THF 1:3) and the solution was stirred for approximately 16 hours. Then the mixture was distributed between EtOAc and H2O. Then was extracted aqueous layer with EtOAc and then the combined organic fractions were washed with a saturated solution of salt, dried (MgSO4 ) and was evaporated to dryness. The crude solid was dissolved in dichloromethane and was washed with hexane, obtaining a solid substance was collected by filtration. This solid was purified column flash chromatography [SiO2, EtOAc-MeOH (90:10)], obtaining 1-(4-forfinal)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (30 mg, 20%) as a solid yellow (LC/MS (kislotno. way): Rt2,01 min, [M-H]+434).

Examples 45-56

Following the procedure described in example 44, but modifying the conditions in the cases indicated in the table below were obtained compounds of examples 45-56.

ExampleStructureDifferences from the General methodsLC/MS
45The reaction required heating (80°C, 4 h)
Chromatography was not required
[M+H]+398
Rt1,79
kislotno.
46[M+H]+454
Rt1,95
kislotno.
47
48After chromatography on silica, impurities were removed by precipitation from solution in MeOH[M+H]+452
Rt2,09
kislotno.
49Was purified preparative LC/MS[M+H]+436
Rt2,68
VI.
50Was purified preparative LC/MS[M+H]+436
Rt2,77
VI.
51[M+H]+422
Rt1,89
kislotno.
52[M+H]+422
Rt1,65
kislotno.
53After chromatography on silica, impurities were removed by precipitation from solution in MOH [M+H]+452
Rt2,21
kislotno.
54Tris-Amin is not required. Was purified preparative LC/MS[M+H]+472
Rt1,89
kislotno.
55The reaction was continued for 1.5 hours at 0°C. the Mixture was concentrated and purified directly by preparative LC/MS.[M+H]+448
Rt6,28
kislotno.
56The reaction was continued for 1.5 hours at 0°C. the Mixture was concentrated and purified directly by preparative LC/MS.[M+H]+482
Rt7,28
kislotno.

Examples 57-59

Following the General method described in example 23, but modified in the cases indicated in the table below were obtained compounds of examples 57-59.

ExampleStructureA common way to obtainDifferences from the General methodsLC/MS
57Example 23, using dimethylamine[M+H]+368
Rt2,39
(VI. method)
58Example 23, using cyclobutylamine[M+H]+396
Rt2,48
(VI. method)
59Example 23, using Isopropylamine[M+H]+384
Rt2,40
(VI. method)

Example 60

Synthesis of hydrochloride of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

60A.Synthesis of (3,4-dinitrophenyl)morpholine-4-ylmethanone

A mixture of 3,4-dinitrobenzoic acid (1 mol. EQ.) and thionyl chloride (9,2 mol. EQ.) boiled under reflux for 6 hours, cooled to room temperature and excess thionyl chloride was removed by azeotropic distillation with toluene. The residue was dissolved in THF (8 volumes), then simultaneously to the mixture was added morpholine (1,0 mol. EQ.) and Et3N (1,1 pray EQ.) p and 0-5°C. The resulting mixture was stirred for 1 hour at room temperature, then poured into water (25 volumes). The mixture was cooled to 3-7°C and left to stand for 0.5 hours, and during this time the product was separated as a precipitate. The precipitate was collected by filtration, washed with water and dried, obtaining (3,4-dinitrophenyl)morpholine-4-ylmethanol (75%) in a solid yellow color (1H NMR(300 MHz, DMSO-d6) δ 8.3 (l, 1H), and 8.3 (s, 1H), 8.0 a (d, 1H), 3,7-3,5 (m, 8H).

60B.Synthesis of 4-(3,4-dinitrobenzyl)research

To a mixture of (3,4-dinitrophenyl)morpholine-4-ylmethanone (1 mol. EQ.) and dry tetrahydrofuran (THF) (25 volumes) at 0-5°C was added NaBH4(2 mol. EQ.) and then dropwise BF3·Et2O (1,01 mol. EQ.) so that the temperature remained in the range 0-5°C. Then the mixture was stirred at room temperature for 3 hours and then was suppressed by the addition of methanol. Then the mixture was evaporated in vacuum, was distributed between ethyl acetate and saturated aqueous NaHCO3. The resulting mixture before separation of the layers was rapidly stirred for 30 minutes. The organic layer was sequentially washed with water and saturated salt solution, and then evaporated in vacuum. The product was led from methanol, receiving 4-(3,4-dinitrobenzyl)morpholine (85%). (LC/MS (VI. way): Rt2,80, [M+H]+ 268).

60C.Synthesis of 4-morpholine-4-ylmethylene-1,2-diamine

A mixture of 4-(3,4-dinitrobenzyl)research (1 mol. EQ.) and 5% Pd/C (0.05 mass. EQ.) in IMS (33 volume) was stirred at 0-5°C and simultaneously filed in the vessel hydrogen. The temperature of the mixture was carefully raised to 15-20°C with stirring until the reaction was completed (<24 hours). The mixture was filtered and the filtrate was evaporated to dryness, obtaining 4-morpholine-4-ylmethylene-1,2-diamine (90%). The resulting material was immediately used in the next stage. (LC/MS (VI. way): Rt1,64, [M-N(CH2CH2)2O-]+121).

60D.Synthesis of 5-morpholine-4-ylmethyl-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole

4-Morpholine-4-ylmethylene-1,2-diamine (1 mol. EQ.) 4-nitro-1H-pyrazole-3-carboxylic acid (1 mol. EQ.) was dissolved in dimethylformamide (DMF) (10 volumes). Added tetrafluoroborate O-(benzotriazol-1-yl)-N,N,N',N'-tetramethylurea (TBTU) (1,2 mol. EQ.) and the mixture was stirred at room temperature for 24 hours. The mixture was concentrated in vacuum until no further distillation of the solvent ceased to be visible. Then the precipitate was dissolved in glacial acetic acid (10 volumes) and was heated at 65°C for ~12 hours. The mixture was concentrated in vacuo and then dissolved in water (6 volumes) at 75°C. the resulting solution of Chernogoria was cooled to 0-5°C for 2 hours, and at this time was formed solid. This solid substance was removed by filtration and the aqueous filtrate was diluted with ethyl acetate (4 volume) and tetrahydrofuran (2 volume). To the stirred mixture was slowly added solid NaHCO3until then, until he ceased to be a gas and not achieved the pH to 6.8. The mixture is then stirred up sediment. The mixture was left to stand for 2 hours at 0-5°C, then were collecting the solid by filtration and washed with water (2 volume), ethyl acetate (2 volume) and dried, obtaining 5-morpholine-4-ylmethyl-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole in the form of a solid brown color (40%). (LC/MS (VI. way): Rt1,93, [M-H+]-327).

60E. Synthesis of 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine

To 5-morpholine-4-ylmethyl-2-(4-nitro-1H-pyrazole-3-yl)-1H-benzimidazole (1 mol. EQ.) in DMF (36 volumes) under nitrogen atmosphere was added 5% Pd/C (0.1 masses. EQ.). The reaction vessel was filled with hydrogen and stirred at room temperature for 24 hours. Then the mixture was filtered through celite, rinsing with methanol. The filtrate was concentrated in vacuum, obtaining 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine as a brown solid (90%). (LC/MS (VI. way): Rt1,94, [M-H+] -297). The product was used without further purification.

60F. Synthesis of hydrochloride of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

To a mixture of 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (1 mol. EQ.) and THF (10 volumes) at 0-5°C and stirring was added 2,6-differentiational (1,3 mol. EQ.). Then the mixture was stirred for 16 hours at room temperature and after this time was treated with 1M aq. KOH (4 volume). After stirring for 2 hours the mixture was concentrated in vacuum and distributed between ethyl acetate and saturated aqueous NaHCO3. The organic layer was washed with a saturated solution of salt, dried (MgSO4), was evaporated to dryness and then the residue was purified column flash chromatography [SiO2, elwira gradient of CH2Cl2-MeOH(98:2)-(90:10)], getting 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

60G.Recrystallization and characteristics of the free base

After chromatography on silica described in example 60E, the product was dissolved in minimum amount of hot ethyl acetate, filtered and left to cool. In this way received a free base in the form of pure crystalline solid.

the received connection i.e. 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, had the following physico-chemical parameters.

the values of pKa - 3,42, 6,92 and 10.97
logP - 3,24
logPion- 0,36
logD(pH 6) of 2.27
(pH 6.5) 2,68
(pH 7,4) 3,11

60H.Getting hydrochloric salt

The obtained product was dissolved in ethyl acetate and treated with excess saturated solution of HCl in diethyl ether. The precipitate was collected by filtration, washed with diethyl ether, and dried, obtaining the hydrochloride of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (59%) as a colourless solid. (LC/MS (kislotno. way): Rt1,80, [M+H]+454).

Replacing hydrogen chloride other acids (e.g., DL lactic acid, econsultancy acid and methanesulfonic acid) and changing if necessary the nature of the solvent, it is possible to get other salts of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pee the azole-4-yl]urea.

60J.Comparison of the solubilities of the free base and hydrochloride

We measured and compared the solubility of the free base and hydrochloride. It was found that the solubility of the free base with a pH of 7.4 (buffer aqueous solution) was <0.001 mg/ml, whereas the solubility of the hydrochloride at a pH of 7.1 (in buffered aqueous solution), as it was discovered, was 0,093 mg/ml Thus, from the viewpoint of solubility, hydrochloric salt had a significant advantage relative to the free base.

Example 61

Determination of the solubilities of the acid additive salts of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

1-(2,6-Differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form was introduced in interaction with various acid by the method described below to assess the solubilities of the resulting acid additive salts.

Method

In a test tube with a volume of 8 ml was added to the free base (59 mg, 0.13 mmol) and water (0,59 ml). In a test tube was added the appropriate acid (1 EQ, 0.13 mmol) and was dissolved in a test tube at room temperature for 16 hours. At the end of this time was subjected to a test tube visual inspection. If we observed formation of a homogeneous solution,the experiment was stopped and made a conclusion, what obtained as described Sol had a solubility in excess of 100 mg/ml

If the remained solid was added 0,59 ml of water and the tubes were shaken for 4 hours. If at this stage formed a homogeneous solution, came to the conclusion that the salt has a solubility exceeding 50 mg/ml

If a solid substance remained in this situation, adding more of 1.18 ml of water and shake the test tube at room temperature. If this led to the formation of a homogeneous solution, came to the conclusion that the solubility of greater than 25 mg/ml If the solid still remained, it was concluded that the solubility of salt is less than 25 mg/ml Free base was regenerated by passing a salt solution through a column of Strata-NH2.

The results of the described experiments are given in the table below.

Solubility of salts of 1-(2,6-differenl)-N-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea
>100 mg/ml>50 mg/ml>25 mg/ml<25 mg/ml
Mesilate
Econsultant
DL-lactate
D-glucuronate Acetate
Adipat
L-(+)-aspartate
D-gluconate
L-glutamate
hydrochloride
toilet
free base

Based on the results shown in the table, one can conclude that mesilate, aconsultant and DL-lactate should be particularly suitable for the production of liquid compositions, water-based, for example, for parenteral administration.

Example 62

Free base and salts of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

The compound of example 24, i.e. the 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, can be isolated in free base form or acid additive salts of the following or similar methods.

Free base

After chromatography on silica (see example 24) the product of example 24 was dissolved in minimum amount of hot MeOH, filtered and left to cool. After about ~16 h were collected product as a colourless crystalline solid.

Hydrochloric salt (General method)

After chromatography on silica product (2,05 g) was dissolved in a mixture of MeOH:EtOAc (1:10, 100 ml) and was treated with 4n. HCl in dioxane (1,1 mol. EQ.). The precipitate was collected by filtration and dried, obtaining the hydrochloride of 1-cyclopropyl-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (1.5 g). The product was dissolved in minimum amount of MeOH and then triturated with Et2O to turbidity, which in a few seconds. After standing over night in cold conditions collected product as a colourless crystalline solid.

Mesilate

Received the product as a colorless crystalline solid using the above General method, but using methansulfonate acid instead of hydrochloric.

Other salts

It is expected that other interest salts could be obtained by the above General method.

Example 63

Determination of solubility of the free base and salts of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

The compound of example 24, i.e. the 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, was introduced in interaction with various acid by the method described below to assess the solubilities of the resulting acid additive salts.

Method

In a test tube with a volume of 8 ml) was added the compound of example 24 in the form of a free base (50 mg, 0,131 mmol) and water (0,50 ml). In a test tube was added the appropriate acid (1 EQ., 0,131 mmol) and was dissolved in a test tube at room temperature for 14-16 hours. In the end this is about the time were subjected to a test tube visual inspection. If we observed formation of a homogeneous solution, the experiment was stopped and made a conclusion that obtained in the described manner Sol had a solubility in excess of 100 mg/ml

If remained solid, was added 0.5 ml of water and the tubes were shaken for 6 hours. If at this stage formed a homogeneous solution, came to the conclusion that the salt has a solubility exceeding 50 mg/ml

If a solid substance remained in this situation, was added 1 ml of water and shake the test tube at room temperature. If this led to the formation of a homogeneous solution, came to the conclusion that the solubility of greater than 25 mg/ml If the solid still remained, it was concluded that the solubility of salt is less than 25 mg/ml

The free base was regenerated by passing a salt solution through a column of Strata-NH2.

The results of the described experiments are given in the table below.

Solubility of salts of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea
>100 mg/ml>50 mg/ml>25 mg/ml<25 mg/ml
acetate
mesilate
econsult is at
DL-lactate
adipat
D-glucuronate
D-gluconate
hydrochloride
toiletL-(+)-aspartate
L-glutamate
free base

Based on the results shown in the table, one can conclude that acetate, mesilate, aconsultant, DL-lactate, adipate, D-glucuronate, D-gluconate and hydrochloride should be particularly suitable for the production of liquid compositions, water-based, for example, for parenteral administration.

From the data collected to date, it appears that the compounds of the present invention and, in particular, 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in the form of a free base and salts (particularly the L-lactate), will have several advantages over the compounds of the prior art. In particular, such benefits include one or more of the following:

- large solubility in aqueous solution;

the best physico-chemical properties, in particular lower logD;

- different sensitivity to P450 enzymes;

- improved drug metabolism and pharmacokinetic properties;

- better stability, such as longer shelf life and/or better thermal stability;

- you want to lower Osinovka;

the best effectiveness against targets of therapy and, in particular Aurora A and B;

- best activity against cells, for example, in proliferation and count of clonogenic assays;

- higher anticancer activity; and

the best therapeutic index.

Example 64

Obtaining lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

To a solution of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (0.7 g, to 1.83 mmol) in EtOAc-MeOH was added L-lactic acid (166 mg, of 1.85 mmol). The mixture was stirred at room temperature, then was evaporated in vacuum. The obtained solid was purified by recrystallization from boiling EtOH (20 ml)to give after drying L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (0,48 g).

Example 65

Synthesis of L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea can be obtained by the method shown in the diagram below.

Stage 1: Synthesis of (3,4-dinitrophenyl)morpholine-4-ylmethanone

A solution of 3,4-dinitrobenzoic acid (10 g, 47 mmol, 1 EQ.) and DMF (0.1 ml) in THF (100 ml) education is atively by thionyl chloride (4.5 ml, 62 mmol, 1.3 EQ.) and then boiled under reflux for 2.5 hours the Mixture was cooled in ice, then for 20 min was added triethylamine (10 ml, 71 mmol, 1.1 equiv.) maintaining an internal temperature <5°C. To the resulting thick yellow suspension for 15 minutes was added morpholine (6.2 ml, 71 mmol, 1.5 equiv.) maintaining an internal temperature <10°C. bath was Removed with ice and gave the mixture to warm to room temperature. After 15 minutes, was added an additional portion of the research (1 ml, 11 mmol, 0.24 equiv.) and the mixture was stirred over night. Diluted mixture of water (250 ml) and cooled on ice. The beige solid was filtered in vacuo, washed with an additional portion of cold water (25 ml) and dried in vacuum, obtaining the connection specified in the header of the example (12.7 g, 96%).

Stage 2: Synthesis of 4-(3,4-dinitrobenzyl)research

Sodium borohydride (3,36 g, 89 mmol, 2.1 EQ.) crushed, placed in a nitrogen-filled flask and suspended in THF (120 ml). After cooling to ~0°C through a syringe was added to the complex of boron TRIFLUORIDE in diethyl ether (11.3 ml, 89 mmol, 2.1 EQ.). This reaction is moderately exothermic, and there was a slight evolution of hydrogen. Was added 4-(3,4-dinitrobenzene)morpholine (11,91 g, 42 mmol, 1.0 EQ.) one portion in the form of a solid substance, and the vessel was washed the additional portion of THF (20 ml). Deleted the bath with ice and stirred suspension at room temperature for 3 hours, then cooled on ice. Caution was added methanol (100 ml) (hydrogen gas), then the mixture is boiled under reflux for 1 h the Mixture was concentrated in vacuum, and then distributed the residue between ethyl acetate (100 ml) and the mixture of 1:1 saturated sodium bicarbonate solution/water (100 ml). Separated organic phase was washed with water (50 ml), then saturated salt solution (100 ml) and dried (MgSO4). Solution of bicarbonate, which was carried out by first washing, the second time was extracted with ethyl acetate (50 ml)and this extract was subsequently washed with the same water structures, which was used for the first extract, and then dried (MgSO4), combine the extracts and concentration resulted in 10,97 g of the crude substance. Recrystallization from methanol (45 ml, 10 ml rinse) resulted in obtaining specified in the connection header (9,34 g, 83%).

Stage 3: Synthesis of 4-morpholine-4-ylmethylene-1,2-diamine

4-(3,4-Dinitrobenzyl)morpholine (21 g, 101 mmol) suspended in ethanol (0.9 liters) and blew the vessel with nitrogen. 10% palladium on charcoal (1,05 g) suspended in ethanol (25 ml) and was added to the substrate. The mixture was cooled on ice, then the gas in the vessel was replaced by the hydrogen. The mixture was allowed to warm up to 15-20°C and continued hydrogenation at atmospheric pressure for 2 days. The vessel was purged with nitrogen, and then the mixture was filtered through celite, washing with ethanol (0.3 l), divided into portions. Concentration gave the opportunity to receive the specified header connection (15,8 g, 97%).

Stage 4: Synthesis of methyl ester 4-nitro-1H-pyrazole-3-carboxylic acid

In a reaction vessel with a volume of 20 liters, equipped with a digital thermometer and a stirrer, was loaded with 4-nitro-1H-pyrazole-3-carboxylic acid (1,117 kg, 7,11 mol, 1 parts by weight) and methanol (8,950 l, 8 volumes). The reaction mixture was stirred in nitrogen atmosphere, cooled to a temperature of 0-5°C for 180 minutes was added thionyl chloride (0,581 l, 8.0 mol, 0.52 volume), the mixture was allowed to warm up to 18-22°C and was stirred overnight, after which analysis by spectroscopy1H NMR (d6-DMSO) showed completion of the reaction. The reaction mixture was concentrated under reduced pressure at a temperature of 40-45°C, the residue was treated with toluene and re-concentrated (3×2,250 l, 3×2 volumes) under reduced pressure at a temperature of 40-45°C, receiving methyl ester 4-nitro-1H-pyrazole-3-carboxylic acid in the form of not-quite-white solid (1,210 kg, 99.5%pure).

Stage 5: Synthesis of methyl ester of 4-amino-1H-pyrazole-3-carboxylic acid

In a reaction vessel with a volume of 20 liters, equipped with a digital thermometer and a stirrer, under nitrogen atmosphere was loaded palladium on coal (10% wet paste, 0,170 kg to 0.14 parts by weight). In a separate vessel was heated suspension of methyl ester 4-nitro-1H-pyrazole-3-carboxylic acid (1,210 kg, 7,07 mol, 1 parts by weight) in ethanol (12,10 l, 10 volumes) to 30-35°C, seeking the dissolution, and the solution was added to the catalyst in an atmosphere of nitrogen. After purging the vessel with gases in the sequence of the nitrogen-hydrogen in a reaction vessel were introduced hydrogen and the reaction mixture is kept at 28-30°C until completion of the reaction (5-10 hours), which was determined by spectroscopy1H NMR (d6-DMSO). After the purge cycle, the reaction mixture was filtered under nitrogen atmosphere and then concentrated liquid residue under reduced pressure, obtaining the methyl ester of 4-amino-1H-pyrazole-3-carboxylic acid (0,987 kg, 98.9 per cent).

Step 6: Synthesis of 4-tert-butoxycarbonylamino-1H-pyrazole-3-carboxylic acid

To a mixture of methyl ester of 4-amino-1H-pyrazole-3-carboxylic acid (50.0 g, 355 mmol) in dioxane (500 ml) was added 2M aqueous NaOH solution (213 ml, 426 mmol), the mixture was heated to 50°C and was stirred for 5 hours Then to this mixture was added (BOC)2O (81,4 g, 373 mmol), by washing with dioxane (100 ml)and the mixture was heated at 50°C for another 5 h and then stirred at room themes is the temperature value within 14 PM The dioxane was removed in vacuo and added water (1 l). the pH of the mixture was brought to ~2 using concentrated aqueous HCl solution, the resulting solid substance was collected by filtration and dried on the filter. The obtained solid was further dried by azeotropic distillation with toluene (×3) and in a vacuum furnace, receiving 4-tert-butoxycarbonylamino-1H-pyrazole-3-carboxylic acid (70,0 g, 87%) as a solid purple color.

Step 7: Synthesis of tert-butyl methyl ether [3-(2-amino-4-morpholine-4-iletilenlerin)-1H-pyrazole-4-yl]carbamino acid

A mixture of 4-tert-butoxycarbonylamino-1H-pyrazole-3-carboxylic acid (10.0 g, 44.1kHz mmol), 4-morpholine-4-ylmethylene-1,2-diamine (10.0 g, to 48.5 mmol), EDC (10,14 g of 52.9 mmol) and HOBt (7,15 g of 52.9 mmol) in DMF (150 ml) was stirred at room temperature for 20 h and then removed most of the solvent in vacuo. The residue was distributed between EtOAc (150 ml) and saturated aqueous NaHCO3(150 ml), the layers were separated and the organic portion was washed with saturated salt solution, dried over MgSO4and was evaporated in vacuum, obtaining tert-butyl ester of [3-(2-amino-4-morpholine-4-iletilenlerin)-1H-pyrazole-4-yl]carbamino acid (17.6 g, 96%) as a solid brown color. As shown by the analysis of LC/MS, the product contained ~15% diamide. This paragraph shall oduct showed approximately 5% level in the spectrum of 1H NMR. Diamid was cut in subsequent stages.

Step 8: Synthesis of 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine

A mixture of tert-butyl methyl ether [3-(2-amino-4-morpholine-4-iletilenlerin)-1H-pyrazole-4-yl]carbamino acid (12.0 g, 28.8 mmol) and 2M aqueous HCl (50 ml) was heated at 85°C for 14 hours, then allowed to cool to room temperature. Caution was added solid Na2CO3to achieve the pH of the mixture ~8.5 and the solution became saturated. Formed viscous liquid dark color. The mixture was allowed to settle and the solvent decantation. To the residue was added EtOH (60 ml), the mixture was boiled under reflux for 1 h and then filtered in hot condition, washing EtOH (C ml) to remove residual inorganic compounds. The filtrate was evaporated in vacuum, obtaining a glassy solid, which was then stirred Et2O (60 ml) for 1 hour, the obtained powder purple color, was collected by filtration and dried in vacuum, obtaining 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (6.8 g, 80%, ~90% purity).

Step 9: Synthesis of 7-morpholine-4-ylmethyl-2,4-dihydro-1,2,4,5a,10-pentaethylene[a]fluoren-5-it

To a mixture of 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-Il is in (3.2 g, of 10.7 mmol) in anhydrous THF (50 ml) at room temperature and stirring was added 1,1'-carbonyldiimidazole (1.78 g, 11 mmol). The mixture was boiled under reflux for 14 h and then cooled to room temperature. The formed solid substance was collected by filtration, washed with THF (20 ml) and dried in vacuum, obtaining 7-morpholine-4-ylmethyl-2,4-dihydro-1,2,4,5a,10-pentaethylene[a]fluoren-5-he (2,34 g, 67%) as a solid pink substance.

Step 10: Synthesis of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

To a mixture of 7-morpholine-4-ylmethyl-2,4-dihydro-1,2,4,5a,10-pentaethylene[a]fluoren-5-she (10.7 g, from 32.9 mmol) and N-methylpyrrolidone (65 ml) was added cyclopropylamine (6.9 ml, 99 mmol). The mixture was heated at 100°C for 5 hours. LC/MS analysis showed the degree of transformation of the original substance in the product ~75%, so I added an additional portion of cyclopropylamine (2.3 ml, 33 mmol), the mixture was heated at 100°C for 4 h and then cooled to room temperature. The mixture was diluted with water (100 ml) and was extracted with EtOAc (100 ml). The organic fraction was washed with a saturated aqueous solution of NH4Cl (2×50 ml), saturated salt solution (50 ml) and then the aqueous fraction was re extracted with EtOAc (3×100 ml). The combined organic fractions were dried over MgSO4and upar is Wali in vacuum, getting 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea as an orange glassy solid (9,10 g).

Stage 11: Synthesis of L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

To a solution of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (9,10 g, 24 mmol) in a mixture of EtOAc-iPrOH (1:1, 90 ml) was added L-lactic acid (2.25 g, 25 mmol). The mixture was stirred at room temperature for 24 h and then evaporated in vacuum. The residue successively suspended in toluene (100 ml) and Et2O (100 ml) and the resulting solid was collected and dried (8,04 g).

This solid was purified by recrystallization from boiling iPrOH (200 ml)to give after drying L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (5.7 g) as a solid beige color.

Example 66

Stage 1: Receive (3,4-dinitrophenyl)morpholine-4-ylmethanone

In a flask under nitrogen atmosphere was loaded 3,4-dinitrobenzoic acid (1,000 kg, 4,71 mol, 1 parts by weight), tetrahydrofuran (10,00 l, 10.0 volumes) and dimethylformamide (0,010 l, 0.01 volume). Added thionyl chloride (0,450 l, 6,16 mol, 0.45 volume) at a temperature of from 20 to 30°C and the reaction mixture was heated is about 65-70°C. The completion of the reaction, which usually occurred after 3 hours was determined by spectroscopy1H NMR (d6-DMSO). The reaction mixture was cooled to 0-5°C and at 0-10°C was added triethylamine (1.25 l, 8,97 mol, 1.25 times the volume). At 0-10°C in the reaction mixture was added morpholine (0,62 l, 7,07 mol, of 0.62 volume) and stirred the resulting suspension for 30 minutes at 0-10°C. Completion of the reaction was determined by spectroscopy1H NMR (d6-DMSO). The reaction mixture was heated up to 15-20°C and was added water (4,00 l, 4.0 volume). This mixture was then loaded into a 40 liter flask with a flange containing water (21,00 l, 21,0 volume) at a temperature of 15-25°C to precipitate the product. The contents of the flask was cooled to 0-5°C, kept at this temperature for 1 hour and collected solids by filtration. Pellet the precipitate on the filter was washed with water (4×5,00 l, 4×5.0 volumes), and it has been found that the pH of the last portion of the wash water was 7. The wet cake of the precipitate on the filter was analyzed1H NMR in the presence of triethylamine hydrochloride. Then the filtered substance was dried at 40-45°C under vacuum until the water content by KF (Karl-Fischer) <0.2% wt./mass., receiving (3,4-dinitrophenyl)morpholine-4-ylmethanol (1,286 kg, 97%, KF 0,069% wt./mass.) in a solid yellow color.

Stage 2: Obtain 4-(3,4-dinitrobenzyl)research

<> In a flask under nitrogen atmosphere was downloaded (3,4-dinitrophenyl)morpholine-4-ylmethanol (0,750 kg, to 2.67 mol, and 1.0 parts by weight) and tetrahydrofuran (7,50 l, 10.0 volume) and was cooled to 0-5°C. At a temperature of 0-5ºC added apirat boron TRIFLUORIDE (0,713 l, 5,63 mol, 0.95 volume), and the suspension was stirred at the same temperature for 15-30 minutes. For 90-120 minutes was added sodium borohydride into six equal portions (0,212 kg, the ceiling of 5.60 mol, 0,282 parts by weight). (Delayed start an exothermic reaction was observed in 10-15 minutes after adding the first portion. If an exothermic reaction started and the reaction mixture was again cooled, the following portions of the reagent was added with 10-15-min intervals, allowing the reaction mixture to cool down between additions). The reaction mixture was stirred at 0-5°C for 30 minutes. Completion of reaction was determined by analyzing the reaction mixture spectroscopy1H NMR (d6-DMSO). At 0-10°C was added dropwise methanol (6,30 l, 8.4 volume) for damping the reaction mixture (rapid evolution of gas, the formation of a certain amount of foam). After quenching the reaction mixture was stirred at 0-10°C for 25-35 minutes, then was heated to 20-30°C and stirred at this temperature (exothermic reaction, the gas/vapour of ether, when dissolved solids) up until the evolution of gas has not slowed down. The resulting mixture was naked evali to 65-70°C and stirred at this temperature for 1 hour. The mixture was cooled to 30-40°C and then concentrated in vacuum at 40-45°C, obtaining the crude 4-(3,4-dinitrobenzyl)morpholine (0,702 kg, 98,4%) as a yellow/orange solid.

4-(3,4-Dinitrobenzyl)morpholine (2,815 kg, 10,53 mol, and 1.0 parts by weight) and methanol (12,00 l, 4.3 volume) were loaded into a flask under nitrogen atmosphere and was heated to 65-70°C. This temperature was maintained until complete dissolution. Then the mixture was cooled to 0-5°C and kept at this temperature for 1 hour. The solids were isolated by filtration. Pellet the precipitate on the filter was washed with methanol (2×1,50 l, 2×0.5 volume) and dried in vacuum at 35-45°C, receiving 4-(3,4-dinitrobenzyl)morpholine (2,353 kg, 83.5 per cent relative to the educt stage 2, 82,5% of the total output, relative to the educt stage 1) in a solid yellow color.

Stage 3: Obtain 4-morpholine-4-ylmethylene-1,2-diamine

4-(3,4-Dinitrobenzyl)morpholine (0.800 to kg, 2,99 mol, and 1.0 parts by weight) and ethanol (11,20 l, 14.0 volume) was loaded into a suitable flask, was stirred at 15-25°C and three times was carried out by the cycle vacuum/filling with nitrogen. 10% palladium on coal (10% Pd/C, pasta 50% humidity, 0,040 kg, dry weight of 0.05 parts by weight) suspended in ethanol (0,80 l, 1.0 vol) and added to the reaction mixture. The mixture was cooled to 10-20°C and three times was carried out by the cycle vacuum/filling with nitrogen. Three times done by ulali cycle vacuum/filling with hydrogen and stirred the reaction mixture in an atmosphere of hydrogen at 10-20°C. Completion of reaction was determined by analyzing the reaction mixture spectroscopy1H NMR (d6-DMSO)as a rule, it took 14-20 hours. Three times was carried out by the cycle vacuum/filling with nitrogen, and the reaction mixture under nitrogen atmosphere was filtered through a paper from microtechnology. The filter cake was washed with ethanol (3×0,80 l, 3×volume of 1.0), the combined filtrate and washing liquid were concentrated in vacuum at 35-45°C to dryness, obtaining 4-morpholine-4-ylmethylene-1,2-diamine (0,611 kg, 98,6%) as a solid brown color.

Stage 4: obtain the methyl ester 4-nitro-1H-pyrazole-3-carboxylic acid

Into the flask with a flange, provided with a mechanical stirrer, reflux condenser and thermometer, was loaded with 4-nitro-1H-pyrazole-3-carboxylic acid (1.00 kg, 6,37 mol, and 1.0 parts by weight) and methanol (8,00 l, 8.0 volumes). The suspension was cooled under nitrogen atmosphere to 0-5°C and at this temperature was added thionyl chloride (0.52 in l, 7,12 mol, 0.52 volume). The temperature of the mixture was raised to 15-25°C for 16-24 hours. Completion of reaction was determined by analyzing the reaction mixture spectroscopy1H NMR (d6-DMSO). The mixture was concentrated in vacuum at 35-45°C. To the residue was added toluene (2.00 l, 2.0 volume) and was removed in vacuum at 35-45°C. the Azeotropic distillation was repeated twice with toluene (2.00 l, 2,0 about what Yama), and obtained the methyl ester 4-nitro-1H-pyrazole-3-carboxylic acid (1,071 kg, 98,3%) as not quite white solid.

Stage 5: obtain the methyl ester of 4-amino-1H-pyrazole-3-carboxylic acid

A suspension of methyl ester 4-nitro-1H-pyrazole-3-carboxylic acid (1,084 kg, 6,33 mol, and 1.0 parts by weight) and ethanol (10,84 l, 10.0 volumes) was heated to 30-35°C and kept at this temperature until complete dissolution. 10% palladium on coal (10% Pd/C, wet paste, 0,152 kg to 0.14 parts by weight) were introduced in a separate flask under nitrogen atmosphere and three times was carried out by the cycle vacuum/filling with nitrogen. The catalyst solution was added methyl ester 4-nitro-1H-pyrazole-3-carboxylic acid in ethanol and three times was carried out by the cycle vacuum/filling with nitrogen. Three times was carried out by the cycle vacuum/filling with hydrogen and the reaction mixture was placed in an atmosphere of hydrogen. The reaction mixture was stirred at 28-30°C to estimated according to spectroscopy1H NMR (d6-DMSO) completion of the reaction. The resulting mixture was filtered under nitrogen atmosphere and then concentrated in vacuum at 35-45°C, obtaining the methyl ester of 4-amino-1H-pyrazole-3-carboxylic acid (0,883 kg, 98,9%) as a solid purple color.

Step 6: Obtain 4-tert-butoxycarbonylamino-1H-pyrazole-3-carboxylic acid

Methyl ester of 4-amino-1H-pyrazole-3-carboxylic acid (1,024 kg, 7,16 mol, and 1.0 parts by weight) and dioxane (10,24 l, 10.0 volume) were loaded into a flask with a flange, provided with a mechanical stirrer, reflux condenser and thermometer. At a temperature of 15-25°C was added a solution of sodium hydroxide (4,36 l, 8,72 mol, 4.26 deaths volume), and the mixture was heated to 45-55°C. the Temperature was maintained at 45-55°C until completion of the reaction, which was determined by spectroscopy1H NMR (d6-DMSO). At 45-55°C was added di-tert-BUTYLCARBAMATE (Boc anhydride, 1,667 kg of 7.64 mol, 1,628 parts by weight), and the mixture was stirred 55-65 minutes. Analysis1H NMR IPC (d6-DMSO) indicated the presence of 9% unreacted intermediate compounds. At 55°C was added an additional portion of di-tert-BUTYLCARBAMATE (Boc anhydride, 0,141 kg, 0.64 mol, of 0.14 parts by weight), and the mixture was stirred for 55-65 minutes. Completion of reaction was determined by spectroscopy1H NMR (d6-DMSO). Remove the dioxane under vacuum at 35-45°C and to the residue was added water (17,60 l, 20.0 volumes). Using 2M aqueous hydrochloric acid solution (4,30 l, 4,20 volumes), brought the pH to 2 and the mixture was filtered. The filtered substance suspended in water (10,00 l, 9.7 volumes) for 20-30 minutes and the mixture was filtered. The filter cake was washed with heptane (4,10 l, 4.0 volume) and dried on the strip for 16-20 hours. The obtained t is ardoe substance was subjected to the azeotropic distillation with toluene (5×4,00 l, 5×4.6 volume), then dried under vacuum at 35-45°C, receiving 4-tert-butoxycarbonylamino-1H-pyrazole-3-carboxylic acid (1,389 kg, with 85.4%) as a solid purple color.

Step 7: Obtain tert-butyl ester [3-(2-amino-4-morpholine-4-iletilenlerin)-1H-pyrazole-4-yl]carbamino acid

4-tert-Butoxycarbonylamino-1H-pyrazole-3-carboxylic acid (0,750 kg 3,30 mol, and 1.0 parts by weight), 4-morpholine-4-ylmethylene-1,2-diamine (0,752 kg, 3.63 mol, and 1.0 parts by weight) and N,N'-dimethylformamide (11,25 l, 15,0 volume) in the atmosphere of hydrogen was loaded into a flask with a flange, provided with a mechanical stirrer and thermometer. At 15-25°C was added 1-hydroxybenzotriazole (HOBt, 0,540 kg of 3.96 mol, 0,72 parts by weight). At 15-25°C was added N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC, 0,759 kg of 3.96 mol, 1,01 parts by weight), and the mixture was stirred at this temperature for 16-24 hours. Completion of reaction was determined by spectroscopy1H NMR. The reaction mixture was concentrated in vacuum at 35-45°C. the Residue was distributed between ethyl acetate (7,50 l, 10.0 volumes) and saturated aqueous sodium hydrogen carbonate (8,03 l, 10.7 volumes) and the layers were separated. The organic phase was washed with a saturated aqueous solution of salt (3,75 l, 5.0 volumes), dried over magnesium sulfate (1.00 kg, of 1.33 parts by weight) and filtered. The residue of the substance on the filter was washed with ethyl acetate (1,50 l, 2,0 amount is). The combined filtrate and washing liquid were concentrated in vacuum at 35-45°C, receiving tert-butyl ester of [3-(2-amino-4-morpholine-4-iletilenlerin)-1H-pyrazole-4-yl]carbamino acid (1,217 kg, 88,6%) as a dark brown solid.

Step 8: Obtain 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine

tert-Butyl ether [3-(2-amino-4-morpholine-4-iletilenlerin)-1H-pyrazole-4-yl]carbamino acid (1,350 kg, 3,24 mol, and 1.0 parts by weight) and ethanol (6,75 l, 5.0 volume) were loaded into a flask with a flange, provided with a mechanical stirrer, reflux condenser and thermometer. At 15-30°C in nitrogen atmosphere was added concentrated aqueous hydrochloric acid (1,10 l, 13,2 mol, 0.80 volume), then the contents of the flask were heated to 70-80°C and held at this temperature for 16-24 hours. At 70-80°C was added to the second portion of hydrochloric acid (0.11 litres, 1,32 mol, 0,080 volume) and the reaction mixture was heated for another 4 hours. Completion of reaction was determined by HPLC analysis. The reaction mixture was cooled to 10-20°C and at this temperature portions was added potassium carbonate (1,355 kg, the remaining 9.08 mol, and 1.0 parts by weight). The resulting suspension was stirred until the gas evolution stops and then filtered. Pellet substances on the filter was washed with ethanol (1,35 l, 1.0 vol) and the filtrates were preserved. Otherthrow the TES substance suspended in ethanol (4,00 l, 3.0 volume) at 15-25°C for 20-40 minutes and the mixture was filtered. Pellet substances on the filter was washed with ethanol (1,35 l, 1.0 volume), all the filtrates were combined and concentrated in vacuum at 35-45°C. To the residue was added ethanol (4,00 l, 3.0 volume) and was removed in vacuum at 35-45°C. To the residue was added tetrahydrofuran (5,90 l, 4,4 volume) and was stirred for 10-20 minutes at 15-25°C. the resulting solution was filtered, the filter cake was washed with tetrahydrofuran (1,35 l, 1.0 volume), the filtrates were combined and concentrated in vacuum at 35-45°C. To the concentrate was added tetrahydrofuran (5,40 l, 4.0 volume) and was removed in vacuum at 35-45°C. To the concentrate was added tetrahydrofuran (5,40 l, 4.0 volume) and was removed in vacuum at 35-45°C, obtaining the desired product, i.e. the 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (0,924 kg, or 95.5%, 82,84% by area on the chromatogram HPLC) as a purple foam.

Step 9: Obtain 7-morpholine-4-ylmethyl-2,4-dihydro-1,2,4,5a,10-pentaethylene[a]fluoren-5-it

3-(5-Morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine (0,993 kg of 3.33 mol, and 1.0 parts by weight) and tetrahydrofuran (14,0 l, 15,0 volume) were loaded into a flask with a flange, provided with a mechanical stirrer, reflux condenser and thermometer. The content was stirred in nitrogen atmosphere at 15-25°C and was added 1,1'-carbonyldiimidazole (0,596 kg, to 3.67 mol, of 0.60 parts by weight). P the following contents were heated to 60-70°C and stirred at this temperature for 16-24 hours. Completion of reaction was determined by TLC analysis. The mixture was cooled to 15-20°C and filtered. Pellet substances on the filter was washed with tetrahydrofuran (4,00 l, 4.0 volume) and dried for 15-30 minutes. The obtained solid was dried in vacuum at 35-45ºC, receiving 7-morpholine-4-ylmethyl-2,4-dihydro-1,2,4,5a,10-pentaethylene[a]fluoren-5-he (0,810 kg, 75,05%, 92,19% by area on the chromatogram HPLC) in a solid purple color.

Step 10: Obtain 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

7-Morpholine-4-ylmethyl-2,4-dihydro-1,2,4,5a,10-pentaethylene[a]fluoren-5-he (0,797 kg, 2,46 mol, and 1.0 parts by weight) and 1-methyl-2-pyrrolidinone (2,40 l, 3.0 volume) were loaded into a flask with a flange, provided with a mechanical stirrer, reflux condenser and thermometer. At 15-30°C in nitrogen atmosphere was added cyclopropylamine (0,279 kg, 4,88 mol, 0,351 parts by weight). The reaction mixture was heated to 95-105°C and stirred at this temperature for 16-24 hours. Completion of reaction was determined by spectroscopy1H NMR. The reaction mixture was cooled to 10-20°C, was added ethyl acetate (8,00 l, 10.0 volumes) and a saturated aqueous solution of sodium chloride (2,50 l, 3.0 volume), the mixture was stirred for 2-5 minutes and the layers were separated. The organic phase was stirred with saturated aqueous sodium chloride (5,00 l, 6.0 volumes) t is the within 25-35 minutes, the mixture was filtered and the filter cake was washed with ethyl acetate (0,40 l, 0.5 volume). The filtered precipitate was retained, the filtrate was transferred into a separating funnel and the layers were separated. This procedure was repeated three more times, the saved portion of the solids was combined with the organic phase and the mixture was concentrated to dryness in vacuum at 35-45°C. the Concentrate was dissolved in propan-2-OLE (8,00 l, 10.0 volumes) at 45-55°C and was added activated carbon (0.080 kg, of 0.1 parts by weight). The mixture was stirred at 45-55°C for 30-40 minutes and then filtered in hot condition at 45-55°C. the filter cake was washed with propan-2-I (0,40 l, 0.5 volume). To the combined filtrate and wash liquid were added activated carbon (0.080 kg, of 0.1 parts by weight), and the mixture was stirred at 45-55°C for 30-40 minutes. The mixture was filtered in hot condition at 45-55°C and the precipitate on the filter was washed with propan-2-I (0,40 l, 0.5 volume). The filtrate and the wash liquid was concentrated in vacuum at 35-45°C. To the concentrate at 25-35°C was added ethyl acetate (8,00 l, 10.0 volumes) and water (2,20 l, 3.0 volume) and the mixture was stirred for 1-2 minutes. The layers were separated and the organic phase was concentrated in vacuum at 35-45°C. To the residue was added ethyl acetate (4,00 l, 5.0 volumes) and concentrated in vacuum at 35-45°C. To the residue was added ethyl acetate (4,00 l, 5.0 volumes) and the mixture was stirred for 2-20 hours at 15-25°C. the Mixture ohla is given to 0-5°C, held at this temperature for 90-120 minutes and then filtered. Pellet substances on the filter was washed with ethyl acetate (0,80 l, 1.0 vol) and dried for 15-30 minutes. The obtained solid substance was dried under vacuum at 35-45°C, receiving 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (of 0.533 kg, 56,8%, 93,20% of the peak area on the chromatogram HPLC) as a brown solid substance.

Several portions of the product of stage 9 was treated in the described manner, and the details regarding the amounts of the original substance and the product for each load are shown in table 1A.

Table 1A
The outputs of stage 10 - stage receiving urea
Room
download
Number 7-morpholine-4-ylmethyl-2,4-dihydro-1,2,4,5a,10-pentaethylene[a]fluoren-5-it (g)Output 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (g)Chemical purity by area HPLC
1680442
55,2%, 64,9%wt./mass.
91,80
282 487
47,0%, 56.6%of mass./mass.
91,21
3879445
43,0%, and 50.6%of the mass./mass.
91,66
4797533
56,8%, 66.8%of mass./mass.
93,20

Stage 11: Obtain L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

1-Cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (1,859 kg, 4,872 mol, and 1.0 parts by weight), propane-2-ol (9,00 l, 5.0 volume) and ethyl acetate (8,00 l, 4.5 volume) were loaded into a flask with a flange, provided with a mechanical stirrer and thermometer. The contents of the flask was stirred in nitrogen atmosphere and at 15-25°C was added L-lactic acid (0,504 kg, 5,59 mol, 0,269 parts by weight), after which the installation was washed with ethyl acetate (0,90 l, 0.5 volume). The mixture was stirred at 15-25°C for 120-140 minutes. A solid substance was separated by filtration, the cake of the precipitate on the filter was washed with ethyl acetate (2×2,00 l, 2×1,0 volume) and dried 20-40 minutes. The filtered substance was dissolved in ethanol (33,00 l, 17.7 per volume) at 75-85°C, cooled to 65-70°C and was purified solution by filtration through paper from microtechnology. The filtrate was cooled to 15-25°C is kept at this temperature for 2-3 hours. Crystallized solid was isolated by filtration, the material on the filter was washed with ethanol (2×1,00 l, 1×0.5 volume) and dried for at least 30 minutes. The obtained solid substance was dried under vacuum at 35-45°C, receiving L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea (1,386 kg, 58,7%, 99,47% by area on the chromatogram HPLC) as a dark pink homogeneous solids.

The infrared spectrum of lactate (with KBr tablet) contains characteristic peaks at 3229, 2972 and 1660 cm-1.

Not wanting to be limited by any theory, I believe that peaks in the infrared spectrum can be attributed to the structural fragments of the salt obtained in the following way:

Peak:Source:
3229 cm-1N-H
2972 cm-1aliphatic C-H
1660 cm-1group C=O, urea

Example 67

Obtaining crystalline free base and crystalline forms of salts of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

A.Getting 1-cyclopropyl-3[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

Was given a sample of crude 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form, as described in example 60, and the first step was to clear the connection column chromatography on silica gel, elwira gradient EtOAc-MeOH (98:2-80:20). Received a sample of the free base is then recrystallized from hot methanol, obtaining the free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in the form of a crystalline substance.

B.Getting dihydrate free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

A sample of crude 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in the form of a free base was dissolved in THF and then concentrated in vacuo to a minimum volume (~4 volumes). To the solution was added dropwise water (2-4 volume), until it became turbid. Added a small amount of THF to restore transparency and the mixture was left to stand over night, getting a crystalline substance, which was dried in the air and got dihydrate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form.

C.Getting hydrochloride 1-cyclopropyl-3-[3-(5-morpholine-4-and the methyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

A sample of crude 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in the form of a free base was dissolved in minimum amount of MeOH and was then diluted with EtOAc. To the solution at 0°C was slowly added 1.1 equivalent of HCl (4M solution in dioxane). After addition of the solution was precipitated solid substance was collected by filtration. This solid matter was added methanol and the mixture was evaporated in vacuum. To remove traces of residual methanol, the residue was dissolved in water, evaporated and then dried at 60°C/0.1 mbar, getting hydrochloride.

D.Getting econsultant 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

To a solution of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in the form of the free base in a mixture of MeOH-EtOAc was added 1 equivalent econsultancy acid. The mixture was stirred at room temperature and then was evaporated in vacuum. The residue was dissolved in methanol and to the solution was added Et2O. the Mixture was left to stand for 72 hours, the resulting solid substance was collected by filtration and dried, obtaining econsultant 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

E.Getting methansulfonate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzoni the azole-2-yl)-1H-pyrazole-4-yl]urea

To a solution of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea free base form (394 mg) in a mixture of MeOH-EtOAc was added 1 equivalent methanesulfonic acid (67 ml). Formed solid substance was collected by filtration, washing EtOAc. This solid was dissolved in minimum amount of hot MeOH, giving to cool, and then triturated with Et2O. the Solid was left to stand for 72 hours and then collected by filtration, washing MeOH, and obtained the methanesulfonate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea.

Example 68

Characterization of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in the form of a free base and salts

Defined characteristics of different forms of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea. The form selected for characterization, have been identified through research that were originally dedicated to the study of the degree of polymorphism and stability of salts. Salts selected for further characterization, was L-lactate, dihydrate free base, Eilat, free base and hydrochloride.

A.Differential scanning calorimetry (DSC)

is aerogramme was detected on device TA instrument Q1000, equipped with automatic document feeder 50 samples. Calibration standard for energy and temperature served as the Indies. Samples were heated at a rate of 10°C/min from 10 to 250°C. Above the sample passed a stream of nitrogen 30 ml/min was Used from 2 to 10 mg samples (unless otherwise specified), and all samples were placed in aluminum pans with point hole in the lid.

SubstanceMelting point (°C)
L-lactate190°C
The dihydrate free baseWas dehydration (peak at 110°C)
EilatWas observed (up to 350°C)
Free base193°C
Hydrochloride190°C

B.Thermogravimetric analysis (TGA)

Thermograms were recorded on a device TA instruments Q500. Samples were heated at a rate of 10°C/min. Over the sample passed a stream of nitrogen at a rate of 100 ml/min is Usually suspended in an open aluminum sample cell was placed 5-20 mg of sample.

Link is on The observed phenomena
L-lactateLoss of 1.7% unbound solvent, melting with decomposition at 190°C
The dihydrate free baseWeight loss (before decomposition) of 4.1% of the mass. (corresponded to 1 equivalent of water)
EilatThe loss of 4% unbound solvent, others are clearly visible features were absent
Free baseLoss of 1.7% unbound solvent, melting with decomposition at 193°C
HydrochlorideLoss of 5.4% unbound solvent, melting with decomposition at 190°C

C.Microscopy in polarized light

The specimens were examined on a Leica microscope LM/DM with a digital camera for image capture. A small amount of sample in immersion oil was placed on a glass slide and covered the cover glass. As can be better separated individual particles and observed at magnifications 50-500× and partially rotated relative to each other polarizing filters, jointly mounted on λ-wave plate.

Substance
The result of observations
L-lactateCrystalline particles of irregular shape
The dihydrate free baseCrystalline particles of irregular shape
EilatCrystalline particles of irregular shape
Free baseNeedle-like crystalline particles
HydrochlorideCrystalline particles of irregular shape

D. XRPD (x-ray Diffraction on the powder)

D5000

Research XRPD was performed on a Siemens diffractometer D5000, using CuKα radiation (40 kV, 40 mA), θ-θ goniometer, automatic deflecting and receiving slits, a graphite secondary monochromator and a scintillation counter. Data were recorded in the angular range 2θ from 2 to 30° in continuous scanning mode using a step size or 0.02° 2θ, or of 0.005° 2θ and a step time of 1 second.

The samples that were investigated in ambient conditions, were prepared in the form of flat plates, using powders, in the form in which they were received, without grinding. Approximately 25-50 mg of sample was carefully placed the depression with a diameter of 12 mm and a depth of 0.5 mm polished silicon wafer with zero substrate (510) (The Gem Dugout, 1652 Princeton Drive, Pennsylvania State College, PA 16803, USA). All studies according to the method XRPD was carried out with application of the Diffrac Plus XRD Commander v.2.3.1.

Diffractometer Bruker AXS C2 GADDS (used for samples returned from GVS)

Picture of x-ray diffraction on powder samples received on the diffractometer Bruker AXS C2 GADDS, using CuKα radiation (40 kV, 40 mA), automated XYZ XY table, laser video microscope for auto-positioning of samples and 2-dimensional detector surface HiStar. Optics for x-rays consisted of one multilayer mirror Goebel, coupled with the point of the collimator with a diameter of 0.3 mm

The divergence of the beam, i.e. the effective size of the x-ray beam on the sample was approximately 4 mm was Used for continuous scanning θ-θ, when the distance from the sample to the detector, is equal to 20 cm, giving an effective range of 2θ equal to 3,2-29,8°. Typical check-diffraction pattern of a sample was 120 C.

Samples were prepared in the form of flat plates, using powders in the form as they were received, without grinding. Approximately 1-2 mg of sample was subjected to a light pressure on a glass slide to obtain a flat surface.

Picture XRPD was recorded for L-lactate and free base. On the diffraction pattern was observed good is the rate of signal/noise, and she pointed to the crystalline nature of the substances.

E.Gravimetric adsorption pair (GVS)

All the specimens were examined on the analyzer adsorption of moisture Hiden IGASorp using the program CFRSorp. The sample mass was approximately 10-25 mg Isotherm adsorption/desorption of water vapor was detected as described below. The samples were loaded and unloaded at room temperature and humidity (approximately 40% humidity, 25°C) and later analyzed by the method XRPD (using the system Bruker AXS C2 GADDS).

The standard method of registering isotherms were single cycle, commencing at a relative humidity of 40%.

Humidity stepwise changed as follows:

40, 50, 60, 70, 80, 90

85, 75, 65, 55, 45, 35, 25, 15, 5, 0

10, 20, 30, 40

(i) L-lactate

Isotherm GVS L-lactate showed that the sample did not show hygroscopic properties and does not form a hydrate. X ray powder diffraction XRPD for samples registered after experiments on GVS, are consistent with the x-ray of the original matter, by showing the absence of phase changes during the experiment.

(ii) Free base

During the experiment, the mass of the sample at a humidity of 0% and a humidity of 95% differed by about 9%. These data showed that the sample in nature was hygroscopic.

Example 69

Defined the E. the crystal structure of the dihydrate free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea x-ray analysis

The crystal used for x-ray analysis, was colorless and had irregular shape with dimensions of 0.2×0.2 to×0.2 mm3. It was obtained by precipitation of an aqueous solution of Eilat using THF in the experiment on the diffusion of liquid-liquid. The equivalence of this sample and the sample of the same crystalline form obtained from the free base (when using water as antibacterial in conjunction with a number of solvents, such as alcohols, e.g. ethanol, ketones such as methyl ethyl ketone and ethers such as THF and dioxane) was established by comparison of the x ray powder patterns of both samples. Crystallographic data were recorded at 101(2)K using CuKα radiation (λ=1,5418 Å) rotating anode Rigaku RU3HR, blue confocal optics Osmic, the goniometer AFC9 1/4χ and detector Rigaku Jupiter CCD. The image was recorded in the four scans in ω, one at 2θ=30° and three at 2θ=90°, when the distance from the crystal to the detector 67 mm data recording took place under program control CrystalClear, and images were processed and scaled textures using Dtrek. Although the absorption coefficient was moderate (µ=0,82 mm-1), the data is corrected using the correction absorption Fourier 4-th order to compensate for the absorption of glue and cristallografia (microterraces). It was found that cu is the growth belong to the monoclinic space group P2 l/n (#14) with crystal lattice parameters of a=7,66(10), b=15,18(10), c=17,71(10) Å, β=98,53(2)°, α=γ=90°. The numbers in brackets mean deviation (s.u., standard uncertainty).

The crystal structure was solved using direct methods included in SHELXS-97. Data on intensity, in total, 2822 unique reflections in the range resolution of 11.5 to 0.89 Å (3,85<θ<60,01) used to identify 274 crystallographic parameters using SHELXL-97. The resulting statistical parameters were as follows: wR2=0,2416 (all data), RF=0,0866 (data with I > 2σ(I)) and the degree of agreement S=1,145.

It was found that one molecule of free base and two water molecules are located in the asymmetric cell. The elemental composition of asymmetric cell C19H26N7O4and the calculated density of the crystals was 1.36 mg/m3. The hydrogen atoms were generated on geometric grounds, while the position of the hydrogen atoms associated with heteroatoms, was confirmed by the study of maps of the difference Fo-Fc. Positional and thermal parameters of the hydrogen atoms were restricted depending on the corresponding non-hydrogen atoms. thermal motion of the non-hydrogen atoms were simulated anisotropic thermal factors (see figure 1).

Crystal structure included one intramolecular (N22-H...N14 2,898 Å)and seven intermolecular hydrogen bonds (see 2). Molecule 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea are connected to each other in chains along the crystallographic b axis by two hydrogen bonds: N7-H...O24 2,761 Å and N25-H...N2 3,310 Å. Benzimidazole fragments of the two chains are stacked in the crystal structure at a distance of 3.5 and 3.6 Å from each other. The crystalline lattice of molecules 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea forms pockets occupied by four water molecules, which are cross-connected with the centre of symmetry. Three hydrogen bonds connect the molecules 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea with water molecules, one of them with the first water molecule (O1W1-H...N16 2,845 Å), and the remaining two with the second water molecule (N1-H...O1W2 OF 2.875 Å and O1W2-H...O19 2,746 Å). Water molecules interact with each other at the expense of the other two hydrogen bonds: O1W1-H...O1W2 2,884 Å and O1W2-H...O1W1 2,771 Å.

The image patterns obtained from the results of x-ray analysis, in the form of thermal ellipsoids shown in figure 1, and the scheme of the crystal packing is shown in figure 2.

The coordinates of the atoms that make up the structure of the dihydrate free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea are shown in table 2. Numbers in parentheses represent the QCD is onanie (s.u., standard uncertainty).

Table 2
the size of the cell a7,662(10)
the size of the cell b15,184(10)
the size of the cell c17,711(10)
the angle alpha cells90,00
the angle beta cell98,53(2)
the angle gamma cell90,00
temperature measurement cell101(2)

Position atomata atom

Position atomise symbol

Position atomaton part x

Position atomaton part y

Position atomaton part z

Position _U ISO or equivalent

Position atomated adp

Position atomatically

Position ATOMKRAFT symmetry

Position atomate calculation

Example 70

XRPD diffraction pattern 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea is in the form of a free base

Samples for recording data on an x-ray diffraction on the powder (XRPD) gently crushed in a marble mortar and placed in crystallographic capillary (Hampton Research, Quartz or Glass Type 10, a diameter of 0.4 or 0.7 mm). Diffraction patterns were recorded at room temperature using CuKα radiation (λ=1,5418 Å) rotating anode Rigaku RU3HR, blue confocal optics Osmic, 1/4χ the goniometer and detector signal in the anode circuit Rigaku HTC. Two-dimensional images recorded during the rotation around the axis φ, when the distance of the detector crystal 250 mm Registration data managed program CrystalClear, and two-dimensional images were converted into one-dimensional curve (in the coordinates of the 2θ - intensity) using Datasqueeze (intensity were averaged over the azimuth angle of 0<x<360° for 2θ range of 3 to 30° with a step of 0.01 or 0.02°). For processing and visualization of the one-dimensional diffraction patterns XRPD used independently created program AstexXRPD.

Picture XRPD and the relative signal intensity was not changed for various parties crystalline samples, which is consistent with the presence of only one crystalline form.

XRPD diffraction pattern for form FB1 free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea is shown in figure 3, and details regarding the main peaks are summarized in table the CE 3.

Table 3
2θ, the interplanar distance d and the relative intensity of the main peaks
2θ/°d/EI
7,9711,09100
or 10.608,3526
11.87 per7,4623
12,137,3015
13,306,6616
15,04of 5.896
15,975,559
16,855,2624
18,684,758
19,404,5814
20,10 4,4242
21,404,1572
21,924,0513
22,813,9013
23,923,728
24,623,629
24,98of 3.568
26,783,339
27,523,2415

Example 71

The crystal structure of lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

Identified single-crystal form of L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea. The crystal used for the diffraction study was colorless prism with dimensions of 0.1×to 0.1×0.1 mm3obtained by precipitation from ethanol during its evaporation. Crystallographic data were recorded at 97 K, ISOE is isua CuKα radiation (λ=1,5418 Å) rotating anode Rigaku RU3HR, blue confocal optics Osmic, the goniometer AFC9 1/4χ and detector Rigaku Jupiter CCD. Images were recorded in the five scans in ω, one at 2θ=15° and four at 2θ=90°, when the distance from the crystal to the detector 67 mm data recording took place under program control and CrystalClear images were processed and scaled textures using Dtrek. Although the absorption coefficient was moderate (µ=0,78 mm-1), the data is corrected using the correction absorption Fourier 4-th order to compensate for absorption in the adhesive and crystallochemical (microkeratome). It was found that the crystals belong to the orthorhombic space group P2l2l2l(#19) with lattice parameters a=9,94(10), b=15,03(10), c=16,18(10) Å, α=β=γ=90°. The numbers in brackets mean deviation (s.u., standard uncertainty). Had one short scan at room temperature to verify the parameters and symmetry of the crystal lattice. Found that symmetry was the same as at 97(2)K, and the lattice parameters were similar (a=10,08, b=15,22, c=16,22 Å).

The crystal structure was solved using direct methods included in SHELXS-97. The absolute configuration was chosen so that it matches the configuration of L-lactic acid used in the experiment for obtaining the foi is of metal. Data on intensity, in total, 3417 unique reflections in the range resolution 11-0,9 Å (4,01<θ<58,92) used to identify 308 crystallographic parameters using SHELXL-97. The resulting statistical parameters were as follows: wR2=0,2275 (all data), RF=0,0817 (data with I > 2σ(I)) and the degree of agreement S=1,076.

It was found that in asymmetric cell are one molecule of free base and one anion of L-lactic acid. The elemental composition of asymmetric cell C22H29N7O5and the calculated density of the crystals was of 1.30 mg/m3. The hydrogen atoms were generated on geometric grounds, while the position of the hydrogen atoms associated with heteroatoms, was confirmed by the study of maps of the difference Fo-Fc. Positional and thermal parameters of the hydrogen atoms was limited, so they depended on the corresponding non-hydrogen atoms. thermal motion of the non-hydrogen atoms were simulated anisotropic thermal factors (see figure 4).

Crystal structure included one intramolecular (N22-H...N14 2,852 Å) and seven intermolecular hydrogen bonds, forming a complex three-dimensional lattice (see figure 5). Two intermolecular hydrogen bonds, namely N7-H...O24 2,800 Å and N25-H...N2 3,004 Å binding molecules 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-p is razol-4-yl]urea in chains along the crystallographic c axis. Anions L-lactic acid bound in chains along the crystallographic axis a by hydrogen bonds O3L-H...O1L 2,626 Å. Two bifurcated hydrogen bonds link the cations 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea and anions L-lactic acid. Protonated nitrogen atom morpholino fragment interacts with the two carboxyl oxygen atoms (N16-H...O1L 3,125 Å and N16H...O2L 2,625 Å), whereas pyrazol the nitrogen atom N1 is a donor H atoms O2L and O3L (N1-H...O2L 2,882 Å, N1-H...O3L 2,740 Å).

The image patterns obtained from the results of x-ray analysis, in the form of thermal ellipsoids shown in figure 4, and the scheme of the crystal packing is depicted in figure 5.

The coordinates of the atoms that make up the structure of lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea are shown in table 4. Numbers in parentheses represent the deviation (s.u., standard uncertainty).

Table 4
the size of the cell a9,941(10)
the size of the cell b15,034(10)
the size of the cell c16,175(10)
at the ol cell alpha 90,00
the angle beta cell90,00
the angle gamma cell90,00
temperature measurement cell97(2)

Position atomata atom

Position atomise symbol

Position atomaton part x

Position atomaton part y

Position atomaton part z

Position _U ISO or equivalent

Position atomated adp

Position atomatically

Position ATOMKRAFT symmetry

Position atomate calculation

Example 72

Stability of salt 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea at 40°C and relative humidity 75%

Approximately 15 mg of the samples to study the stability gently crushed in a marble mortar and transferred into a Petri dish in a thin layer. Then the samples were placed in tightly closed containers containing a saturated solution of NaCl with an excess of undissolved NaCl. This capacity, in turn, were placed in the incubator, where he supported the 40°C to obtain the environmental temperature is Oh 40°C and relative humidity (RH)≈75%. Samples were regularly analyzed by the method of x-ray diffraction on the powder (XRPD).

To obtain data XRPD samples were placed in a crystallographic capillary (Hampton Research, made from quartz, diameter=0.4 mm). Diffraction patterns were recorded at room temperature using CuKα radiation (λ=1,5418 Å) rotating anode Rigaku RU3HR, blue confocal optics Osmic, 1/4χ the goniometer and detector signal in the anode circuit Rigaku HTC. Two-dimensional images recorded during the rotation around the axis φ, when the distance of the detector crystal 250 mm Registration data managed program CrystalClear, and two-dimensional images were converted into one-dimensional curve (in the coordinates of the 2θ - intensity) using Datasqueeze (the intensity was averaged over the azimuth angle of 0<x<360° for 2θ range of 3 to 30° with a step of 0.01°). For processing and visualization of the one-dimensional diffraction patterns XRPD used independently created program AstexXRPD.

XRPD diffraction pattern of lactate, free base (FB1) and dihydrate free base (FB2) did not change over the period of 1-2 months, when the environment temperature of 40°C and relative humidity 75%. Diffraction pattern of the original samples as well as samples of lactate, free base (FB1) and dihydrate free base (FB2), the resulting stability, is shown in Fig. 6-8.

XRPD diffraction pattern of L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea is shown in Fig.6 and detailed information about the main peaks are summarized in table 5.

Table 5
2θ/ºd/ÅI
8,0011,053
10,308,587
10,508,4215
11,557,668
11,857,4623
12,407,1335
12,906,8611
14,006,3215
14,60the 6.066
15,20of 5.83 27
15,60of 5.6830
16,005,549
17,50of 5.0681
18,304,8554
18,504,7936
19,304,6041
19,604,5340
20,404,3516
20,754,2814
to 21.154,2020
21,604,1122
21,854,07100
22,503,9523
22,753,91 15
23,703,7512
24,153,6814
24,40the 3.6515
of 24.903,5713
25,603,4816
26,503,3610
27,303,2629
28,303,156
29,00is 3.089
29,503,0315

Diffraction XRPD pattern of form FB2, i.e. dihydrate free base 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, shown in Fig, and detailed information about the main peaks are summarized in table 6.

Table 6
2θ/ºd/ÅI
7,5011,788
10,008,845
11,50of 7.699
11,90the 7.4313
12,806,9148
13,206,7044
14,206,239
15,40of 5.7517
16,205,4724
16,605,3413
17,00to 5.2111
17,405,0952
18,00is 4.9320
19,004,6748
20,00of 4.4431
20,804,2776
to 21.154,2030
a 21.754,0822
22,603,9320
23,103,85100
23,553,7822
23,953,7122
of 24.903,5726
25,303,5235
26,653,3434
27,003,3024
27,803,2122
28,853,0918
29,353.04 from14

BIOLOGICAL ACTIVITY

Example 73

Measurement of activated complex CDK2/cyclin A in the study of inhibitory activity against kinase (IC50)

Compounds of the present invention were tested for inhibitory activity against the kinase activity using the following Protocol.

Activated complex CDK2/cyclin A (Brown et al, Nat.Cell Biol., 1, pp.438-443,1999; Lowe, E.D., et al, Biochemistry, 41, pp. 15625-15634, 2002) was diluted to 125 PM in analytical buffer of 2.5× concentration (50 mm MOPS pH of 7.2, 62.5 mm β-glycerol, 12.5 mm EDTA, 37.5 mm MgCl2, 112,5 mm ATP, 2.5 mm DTT, 2.5 mm of orthovanadate sodium, 0.25 mg/ml bovine serum albumin) and 10 μl mixed with 10 μl of the substrate mixture containing histone (60 μl bovine histone H1 (Upstate Biotechnology, 5 mg/ml), 940 μl of H2O, 35 mccoury γ33P-ATP) and added to 96-well tablets together with 5 μl of a solution of the analyte in DMSO at various dilutions (up to 2.5%). Given the reaction to go for 2-4 hours, then stopped her excess phosphoric acid (5 μl conc. 2%). γ33P-ATP, which are not connected to histone H1, was separated from phosphorylated histone H1 on the filter plate Millipore MAPH. ACAC is on the plate MAPH moistened with 0.5% phosphoric acid and then through these cells were filtered, reaction products with apparatus for vacuum filtration Millipore. After filtration the residue was twice washed with 200 µl of 0.5% phosphoric acid. When the filters were dried, was added 20 ml of scintillator Microscint 20 and then made calculations on a Packard Topcount for 30 seconds.

Expected inhibition of CDK2 activity in % and put it on a graph to determine the concentration of test compound required to inhibit 50% activity of CDK2 (IC50).

Connection examples 1, 10, 11, 18, 20, 22, 30, 31, 32, 46, 47 and 54 in the analysis of inhibition of CDK2 mattered IC50less than 1 µm, whereas the compounds of examples 44, 45, 48, 51 and 53 mattered IC50less than 10 microns.

Example 74

Measurement of activated complex of CDK1/cyclin B in the study of inhibitory activity against kinase (IC50)

Analysis of complex CDK1/cyclin B is identical to that described above for the analysis of complex CDK2/cyclin A, except that used CDK1/cyclin B (Upstate Discovery) and the enzyme was diluted to 6.25 nm.

Connection examples 1, 4, 6, 10, 11, 13, 22, 42, 47 and 54 in the analysis of inhibition of CDK1 mattered IC50less than 1 µm, whereas the compounds of examples 3, 8, 9, 16, 17, 20, 24, 28, 29, 31, 32, 34, 39, 41, 45, 46, 48, 49, 50, 51, 52, 53 and 56 mattered IC50less than 10 microns and connection examples 2, 23, 26, 27, 33, 37 and 43 mattered IC50less than 50 microns.

Example 75

Research Aurora A kinase

The activity of Aurora A kinase can the be determined using dissociatively-enhanced lanthanide immunoassay (kits are used) received from GSK-3 biotinylated peptide. The number of the resulting phosphorylated peptide was measured using phospho-specific primary antibodies and labeled with europium anti-rabbit IgG antibody using fluorescence with a time resolution at λex=337 nm, λcm=620 nm.

The kinase reaction

Analytical reactions were performed in 96-well tablets for a total reaction mixture of 25 µl with 0.5 nm Aurora A (Upstate Discovery), 3 μm Biotin-CGPKGPGRRGRRRTSSFAEG, mm ATP and various dilutions of test compounds in a mixture of 10 mm MOPS, pH 7.0, 0.1 mg/ml BSA, about 0.001% Brij-35, 0.5% glycerol, 0.2 mm EDTA, 10 mm MgCl20.01% of β-mercaptoethanol and 2.5% DMSO. The reaction was let go for 60 minutes at room temperature, then stopped her by adding 100 ál of buffer STOP, containing 100 mm EDTA, 0.05% of Surfact-Amps20 (Pierce) and 1× BlockerTMBSA in TBS (Pierce).

Phase detection

After the reaction mixture was transferred into a 96-well plate coated with Neutravidin (Pierce) and incubated for 30 minutes to capture the biotinylated peptide. After 5 times washing with TBST buffer with 200 μl per well in each well was added a mixture of antibodies anti-phospho(Ser/Thr)-AKT substrate (Cell signaling Technology) and Eu-N1anti-rabbit IgG (Perkin Elmer) and left for 1 hour. After additional stage wash all wells were added kits are used amplifying solution (Perkin Elmer). After incubation for 5 min is t count the holes on the reader tablet Fusion.

In the above analysis of the compounds of examples 1-56 mattered IC50less than 1 μm. Hydrochloric salt of example 60H mattered IC50equal 0,0025 mm.

Example 76

Research Aurora B kinase

The kinase reaction

Analytical reactions were performed in 96-well tablets, for a total reaction mixture of 25 µl with 0.5 nm Aurora B (ProQinase), 3 μm Biotin-CGPKGPGRRGRRRTSSFAEG, 15 μm ATP and various dilutions of test compounds in a mixture of 25 mm TRIS pH 8.5, 0.1 mg/ml BSA, 0.025% of Surfact-Amps20, 5 mm MgCl2, 1 mm DTT and 2.5% DMSO. The reaction was let go for 90 minutes at room temperature, then stopped her by adding 100 ál of buffer STOP, containing 100 mm EDTA, 0.05% of Surfact-Amps20 (Pierce) and 1× BlockerTMBSA in TBS (Pierce).

Phase detection led, as described for Aurora A.

In the analysis of inhibition of Aurora B hydrochloric salt of example 60H demonstrated a 57% inhibition at a concentration of 0.003 microns.

Example 77

Analysis of the inhibitory activity against the kinase GSK3-B

GSK3-β (Upstate Discovery) was diluted to a concentration of 7.5 nm in a mixture of 25 mm MOPS, pH 7.0, 25 mg/ml BSA, 0,0025% Brij-35, 1,25% glycerol, 0.5 mm EDTA, 25 mm MgCl2are 0.025% β-mercaptoethanol, 37.5 mm ATP and 10 μl mixed with 10 μl of the substrate mixture. Substrate mixture for GSK3-β was a 12.5 mm peptide-2 phospho-glycogen synthase (Upstate Discovery) in 1 ml of water with 35 mccoury γ33P-ATP. Enzyme and substrate were placed in a 96-well tablets together with 5 μl of a solution of test compound in DMSO at various dilutions (up to 2.5%). The reaction was let go for 3 hours (GSK3-β), then stopped her excess phosphoric acid (5 μl, conc. 2%). Method of filtering consistent with the above method of analysis of the activated complex of CDK2/cyclin A.

Example 78

Research selectivity against CDK

78A. Protocol A

Compounds of the present invention can be tested for inhibitory activity against kinases for example, a number of different kinases using the General Protocol described above in example 3, but modified as described below.

Kinase was diluted to 10× working backup solution in a mixture of 20 mm MOPS, pH 7.0, 1 mm EDTA, 0,1% β-mercaptoethanol, 0.01% of Brij-35, 5% glycerol, 1 mg/ml BSA. One unit corresponded to the incorporation of 1 nmol of phosphate per minute at 0.1 mg/ml histone H1 or a peptide which is a substrate for CDK7, at 30°C with a final ATP concentration of 100 μm.

The substrate for all research CDK (except CDK7) was histone H1, before using diluted to 10× spare working solution in 20 mm MOPS buffer pH of 7.4. A substrate for CDK7 was a specific peptide, diluted to 10× spare working solution in deionized water.

Methods of analysis for CDK1/C the wedge B, CDK2/cyclin A, CDK2/cyclin E, CDK3/cyclin E, CDK5/p35, CDK6/cyclin D3:

At final volume of 25 µl reaction mixture, the enzyme (5-10 min.) incubated with 8 mm MOPS, pH 7.0, 0.2 mm EDTA, 0.1 mg/ml histone H1, 10 mm Mg acetate and [γ33P-ATP] (specific activity approximately 500 pulses/min/pmol, concentration as required by the conditions of the experiment). The reaction was initiated by addition of Mg2+33P-ATP]. After incubation for 40 minutes at room temperature the reaction was stopped by adding 5 μl of a 3% solution of phosphoric acid. 10 μl of reaction mixture was applied on a flat filter P30 and three times for 5 minutes, washed with 75 mm phosphoric acid and once with methanol, and then dried and made the count.

Methods of analysis for CDK7/cyclin H/MAT1

When the final volume of the reaction mixture of 25 μl of enzyme (5-10 min.) incubated with 8 mm MOPS, pH 7.0, 0.2 mm EDTA, 500 μm peptide, 10 mm Mg acetate and [γ33P-ATP] (specific activity approximately 500 pulses/min/pmol, concentration as required by the conditions of the experiment). The reaction was initiated by addition of Mg2+33P-ATP]. After incubation for 40 minutes at room temperature the reaction was stopped by adding 5 μl of a 3% solution of phosphoric acid. 10 μl of reaction mixture was applied on a flat filter P30 and three times for 5 minutes, washed with 75 mm phosphoric acid and one R is h with methanol, then dried and made the count.

W. Protocol B

Inhibitory activity against these enzymes investigated in Upstate Discovery Ltd. Received solutions of the enzyme at a final concentration of 10× enzyme buffer (composition listed in the table below). Then the enzymes were incubated in the analytical buffer with different substrates and33P-ATP (~500 pulse/min/pmol), as shown in the table.

The reaction was initiated by addition of Mg/ATP. Then the reaction was let go for 40 minutes at room temperature, then was stopped by adding 5 μl of a 3% solution of phosphoric acid. Ten μl of the reaction mixture was applied on a flat filter or A P30, three times washed with 75 mm phosphoric acid and once with methanol, and then dried and produced scintillation counting.

The test compounds were tested at the concentrations described in detail below in duplicate, with all kinases, and hoped existing activity compared with control. If the compounds showed significant inhibition was determined IC50.

EnzymeEnzyme bufferAnalytical bufferSubstrateThe concentration of ATP (μm)
CDK3AA0.1 mg/ml histone H1200
CDK6AA0.1 mg/ml histone H1200
CDK7AA500 mm peptide90
CDK9AA100 μm KTFCGTPEYLAPEVRREPRI
LSEEEQEMFRDFDYIADWC
45

Used enzyme buffer had the following composition:

A: 20 mm MOPS, pH 7.0, 1 mm EDTA, 0.1% of β-mercaptoethanol, of 0.01% Brij-35, 5% glycerol, 1 mg/ml BSA.

Used analytical buffer had the following composition:

A: 8 mm MOPS, pH 7.0, 0.2 mm EDTA, 10 mm Mg acetate.

Example 79

Antiproliferative activity

Antiproliferative activity of the compounds of the present invention can be determined by measuring the ability of compounds to inhibit the growth of cells of different cell lines. Measured the inhibition of cell growth using the analysis with the test system Alamar Blue (Nociari, M.M., Shalev, A., Benias, P., Russo, C. Journal of Immunological Methods 1998, 213, 157-167). The method is based on the ability of viable cells in ustanavlivat resazurin to its fluorescent product resorufin. For each study cell proliferation were applied to 96-well plates and gave them to recover for 16 hours, after which the following 72 hours was added inhibiting compounds. At the end of the incubation period was added 10% (volume/volume) Alamar Blue and incubated for another 6 hours, after which determined the content of the fluorescent product when the wavelength 535 nm excitation/590 nm emission. In the case studies nonproliferative cells accumulation of cells kept for 96 hours, then added inhibiting compounds for the next 72 hours. The number of viable cells was determined by assay with Alamar Blue as described above. In addition, recorded all of the morphological changes. The cell line can be obtained from ECACC (European collection of cell cultures).

In the study, which used cell line HCT-116, hydrochloric salt of example 60H had IC50equal 0,070 mm.

In particular, it was tested the activity of the compounds of the present invention against cell line HCT-116 (ECACC, reg. number: 91091005)obtained from cancer of the colon of a person.

It was found that many of the compounds of the present invention in this analysis are the values of the constants IC50less than 25 microns, and the preferred compounds are the values of the IC50less than 1 μm. On the other hand, was found who, for many compounds, the minimum concentration at which there is polyploidy or multicore cells is less than 10 μm, and the preferred compounds are the values of the IC50less than 100 nm.

It has been found that such a connection as 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea, in the above analysis is the value of the IC50less than 1 μm. In addition, for this connection, it was found that the minimum concentration at which there is polyploidy or multiple cores is less than 100 nm.

Example 80

A.General Protocol for the study of colonies

The effects of various compounds on line adhesion of tumor cells was evaluated in a count of clonogenic analysis.

Cells were sown at a concentration of 75-100 cells/ml of the appropriate culture medium in 6 - or 24-hole plates to tissue cultures and allowed to recover for 16 hours.

The connection or the media in the case of the control experiment (DMSO) was added in paired wells to achieve a final concentration of DMSO of 0.1%. After adding the connection to the colonies allowed to grow for 10-14 days for optimal discrete counting of colonies. Colonies were fixed with 2 ml of fixative composition Carnoys (25% acetic acid, 75% methanol and stained in 2 ml of 0.4% weight/volume of the m crystal violet. Counting the number of colonies in each well. The values of the IC50was calculated using a sigmoidal IC50curves dose-response (variable slope)using the program Graphpad Prism.

B.The Protocol for the study of colonies for 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea

The effects of 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea on cell lines A, A549, HCT 116, HCT 116 N7, HT-29, MCF7, MIA-Pa-Ca-2, SW620 were evaluated in the count of clonogenic analysis.

Cells were sown at a concentration of 75-100 cells/ml of the appropriate culture medium in 6 - or 24-hole plates to tissue cultures and allowed to recover for 16 hours.

Cell lineWednesdayComments
HCT 116DMEM + 10% FBS + GLUTAMAX I
HCT 116 N7DMEM + 10% FBS + GLUTAMAX I + 0.4 mg/ml G418
HT-29McCoy'5a + 10% FBS +2mm L-glutamine
SW620L-15 + 10%FBS + GLUTAMAX IAtmos the EPA CO 2
A2780RPMI 1640 + 2 mm glutamine + 10% FBS
A549DMEM + 10% FBS + GLUTAMAX I
MCF7EMEM + 10% FBS + 2 mm L-glutamine + 1% NEAA
MIA-Pa-Ca-2DMEM + 10% FBS + GLUTAMAX I

1-Cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or, in the case of the control experiment, the carrier (DMSO) was added in paired wells to achieve a final concentration of DMSO of 0.1%. After adding the connection to the colonies allowed to grow for 10-14 days for optimal discrete counting of colonies. Colonies were fixed with 2 ml of fixative composition Carnoys (25% acetic acid, 75% methanol and stained in 2 ml of 0.4% weight/volume of crystal violet. Counting the number of colonies in each well. Take into account only multicellular colonies, which included approximately 50 or more cells, which showed proliferation from single cells to colonies of many cells (i.e. complete cell cycle, including the successful cytokinesis). Separate multi-core (polyploid) cells did not take into account. The values of the IC50was calculated using a sigmoidal IC 50curves dose-response (variable slope)using the program Graphpad Prism.

The results of this research are shown in table C in the section entitled "advantages of the compounds of the present invention".

Example 81

The definition of efficiency in relation to cytochrome P450

The effectiveness of the compounds of example 24 in respect of CYP450 1A2, 2C9, 2C19, 3A4 and 2D6 were determined with the use of research set Pan Vera Vivid Cyp450, which can be purchased from Invitrogen (Paisley, UK). CYP was introduced into the reaction mixture in the form of maculosum containing CYP450 and NADPH reductase. Substrates were fluorescent substrates Vivid.

The final reaction mixture contained the following components:

1A2

100 mm potassium phosphate, pH 8, 1% methanol, the substrate is 2 μm 1A2 vivid blue, 100 μm NADP+, 4 nm CYP450 1A2, to 2.66 mm glucose-6-phosphate, of 0.32 U/ml glucose-6-phosphate dehydrogenase.

2C9

50 mm potassium phosphate, pH 8, 1% methanol, the substrate is 2 μm vivid green, 100 μm NADP+, 8 nm CYP450 2C9, to 2.66 mm glucose-6-phosphate, of 0.32 U/ml glucose-6-phosphate dehydrogenase.

2C19

50 mm potassium phosphate, pH 8, 1% methanol, the substrate 8 μm vivid blue, 100 µm

NADP+, 4 nm CYP450 2C19, to 2.66 mm glucose-6-phosphate, of 0.32 U/ml glucose-6-phosphate dehydrogenase.

3A4

100 mm potassium phosphate, pH 8, 1% methanol, the substrate 10 μm 3A4 vivid blue, 100 μm NADP+, 2.5 nm CYP450 3A4, to 2.66 mm glucose-6-phosphate, of 0.32 U/ml glucose-6-phosphate Degi the rogenaza deficit.

2D6

100 mm potassium phosphate, pH 8, 1% methanol, the substrate is 5 μm 2D6 vivid blue, 100 μm NADP+5 nm CYP450 2D6, of 2.66 mm glucose-6-phosphate, of 0.32 U/ml glucose-6-phosphate dehydrogenase.

Fluorescence was recorded for 20 min after 30 s intervals on a Molecular Devices Spectramax Gemini reader. The wavelengths of excitation and emission was equal to 390 nm and 460 nm for 1A2, 2C19 and 3A4, 390 nm and 485 nm for 2D6 and 485 nm and 530 nm for 2C9. Initial velocities were determined from the curves of the progress of the reaction.

Prepared solutions of test compounds in methanol and determined their effectiveness against CYP450 at a concentration of 10 μm. The results are presented in Table B.

PHARMACEUTICAL COMPOSITIONS

Example 82

(i) Compounding tablets

Composition of tablets containing a compound of the formula (I)obtained by mixing 50 mg of the compounds with 197 mg of lactose (BP) as a diluent and 3 mg of magnesium stearate as a means to slip, and compressing the mixture in a known manner to form tablets.

(ii)Recipe capsules

The composition in the form of capsules is obtained by mixing 100 mg of the compounds of formula (I) with 100 mg of lactose and filling the resulting mixture standard opaque hard gelatin capsules.

(iii)Suitable for injection part I

Parenteral composition for administration by injection can be obtained by dissolving the compounds of formula (I) (e.g. in a salt form) in water is, containing 10% propylene glycol, to obtain the concentration of the active compounds of 1.5% by weight. Then the resulting solution is sterilized by filtration, filled them ampoules and sealed.

(iv)Suitable for injection composition II

Parenteral composition for injection is obtained by dissolution in water of the compounds of formula (I) (e.g. in a salt form) (2 mg/ml) and mannitol (50 mg/ml), sterile filtering the solution and filling this solution tightly closed 1 ml vials or ampoules.

(v)Suitable for injection part III

Composition for intravenous injection or infusion may be obtained by dissolving the compounds of formula (I) (e.g. in a salt form) in water (20 mg/ml Then the bottle tightly closed and sterilized by autoclave.

(vi)Suitable for injection composition IV

Composition for intravenous injection or infusion may be obtained by dissolving the compounds of formula (I) (e.g. in a salt form) in water containing a buffer (e.g., 0.2m acetate buffer pH 4.6), in the amount of 20 mg/ml Then the bottle tightly closed and sterilized by autoclave.

(vii)Liofilizovannye part I

Aliquots of the compounds of formula (I) or salts thereof, as defined in this application, mixed with other necessary components, placed in 50 ml vials and lyophilized. While l is utilizacii composition is frozen, using the one-step method of freezing when (-45°C). The temperature was raised to -10°C for annealing, then reduce to freezing at -45°C, followed by primary drying at +25ºC for approximately 3400 min, followed by secondary drying with step raising the temperature to 50°C. During primary and secondary drying install a pressure of 80 millitorr.

(viii)Liofilizovannye part II

Aliquots of the compounds of formula (I) or salts thereof, as defined in this application, mixed with other necessary components, placed in 50 ml vials and lyophilized. During lyophilization of the frozen composition using single-stage technique of freezing when (-45°C). The temperature was raised to -10°C for annealing, then reduce to freezing at -45°C, followed by primary drying at +25°C for approximately 3400 min followed by secondary drying with step raising the temperature to 50°C. During primary and secondary drying install a pressure of 80 millitorr.

(ix)Liofilizovannye composition used for intravenous III

Buffered aqueous solution obtained by dissolving L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea concentration 12,86 mg/ml in 0,02M citrate buffer, pH of which was adjusted to 4.5 by adding sodium hydroxide or x is olistostromes acid.

Received buffered solution filled in, with intermediate filtration to remove particles, capacity (as, for example, glass vial of class 1), which is then partially closed (for example, through a tube Florotec). If the connection and the composition is generally stable, the composition is sterilized by autoclave at 121°C for the required time period. If the composition is not sufficiently stable to be processed in the autoclave, it can be sterilized by use of a suitable filter and sterile conditions to fill them sterile vials. The solution is subjected to lyophilization using a suitable cycle, for example:

Freeze - cooled to -40°C within 2 hours and maintained at -40°C for 3 hours.

Primary drying - raise the temperature from -40°C to -30°C for 8 hours and maintained at -30°C for 7 hours.

Secondary drying - raise the temperature to +30°C for 4 hours and incubated at 30°C for 8-10 hours.

Upon completion of the lyophilization cycle the tank again filled with nitrogen to atmospheric pressure, cover and protect the cover (for example, aluminum crimp cap). For intravenous administration liofilizovannye solid composition can be restored using a pharmaceutically acceptable excipient, such as, for example, 0.9% saline or 5% dextrose. R is the target, you can enter in this form, or before entering the patient to inject it into the bag for storage of liquids (containing pharmaceutically acceptable filler, as, for example, 0.9% saline or 5% dextrose).

(x)Composition for subcutaneous injection

Composition for subcutaneous injection is obtained by mixing the compounds of formula (I) with corn oil, pharmaceutical grade, reaching a concentration of 5 mg/ml of the Composition is subjected to sterilization and fill her suitable container.

Example 83

Determination of antifungal activity

Antifungal activity of the compounds of formula (I) can be determined using the following Protocol.

Tested the activity of compounds against a set of fungi, including Candida parpsilosis, Candida tropicalis, Candida albicans-ATCC 36082 and Cryptococcus neoformans. The studied microorganisms included on the beveled nutrient media Sabourahd Dextrose Agar at 4°C. Singlet suspension of each microorganism was obtained by growing the fungus overnight at 27°C on a rotating drum in nutrient broth for nitrogen-yeast base (YNB) with amino acids (Difco, Detroit, Mich.), pH 7.0 with 0,05M morpholinepropanesulfonic acid (MOPS). After that, the suspension was centrifuged and washed twice 0,85% NaCl, after which the washed cell suspension was treated with ultrasound for 4 seconds (Branson Sonifier, model 350, Danbury, Conn.). Singlet of blastospore counted in hemocytometer, and brought up to the desired concentration of 0.85% NaCl.

The activity of test compounds was determined by the, using a modification of the method of microdesmidae in the broth. Test compounds were diluted in DMSO to 1 mg/ml, then diluted to 64 µg/ml in YNB broth pH 7.0 with MOPS (as a control) was used fluconazole), getting a working solution of each compound. When using a 96-hole tablet wells 1 and 3-12 filled YNB broth, holes 2-11 did ten dilutions of a solution of test compounds (concentration range from 64 to 0.125 mg/ml). Hole 1 was the control of sterility and no experience for spectrophotometric analysis. Hole 12 served as a growth control. Microtiter tablets were seeded in 10 μl into each of the holes 2-11 (final seeded number was 104microorganisms/ml). Sowed the plates were incubated for 48 hours at 35°C. the values of the IC50was determined spectrophotometrically by measuring the absorption at 420 nm (Automatic Microplate Reader, DuPont Instruments, Wilmington, Del.) after stirring the contents of the tablets for 2 minutes using a vortex mixer (Vorte-Genie 2 Mixer, Scientific Industries, Inc., Bolemia, N.Y.). Endpoint IC50was defined as the lowest concentration of drug that causes about 50% (or more strong) slower growth compared with the control hole. Using turbidimetric analysis it was determined as the lowest drug concentration at which turbidity in lunk who was < 50% of the control (IC50). Minimal cytolytic concentrations (MCC) was determined by reseeding all wells of the 96-well plate onto the tablet with Sabourahd Dextrose Agar (SDA), incubation for 1-2 days at 35°C and then checking viability.

Example 84

The Protocol of the biological assessment regulation in vivo fungal infection of the whole plant

The compounds of formula (I) was dissolved in acetone and then performed serial dilutions in acetone to obtain the desired concentration range. Finite volume formulations for the treatment was obtained by adding nine volumes of 0.05% aqueous Tween-20TMor 0.01% Triton X-100TMdepending on the pathogen.

Then the obtained compositions were used for studying the activity of the compounds of the present invention against the disease of tomato (Phytophthora infectans), using the following Protocol. Tomatoes (cultivar Rutgers) were grown from seeds on soilless potting mix based on peat until the seedlings reached a height of 10-20 see Then the plants were sprayed to runoff solution from the leaves of the test compounds at a concentration of 100 parts per million 24 hours of the investigated plants were infected by spraying with an aqueous suspension of the sporangium of Phytophthora infectans and held in a cell with irrigation during the night. Then the plants transferred to the greenhouse until disease progression on the raw counter is selected plants.

Similar protocols were used to test activity of compounds of the present invention against brown rust of wheat (caused by Puccinia), powdery mildew of wheat (Ervisiphe vraminis), wheat (cultivar Monon), causing spot blotch of wheat (Septoria tritici and Septoria Koloskova scales of wheat (Leptosphaeria nodorum).

Equivalents

The above examples are given to illustrate the invention and should not be construed as imposing any limitation on the scope of the invention. Should be easily understood that the specific embodiments of the invention described above and illustrated in the examples, can be made of numerous modifications and changes without derogating from the principles underlying the invention. It is assumed that all such modifications and changes are covered by the present application.

1. The compound of formula (I):

or its pharmaceutically acceptable salt, MES or tautomer,
where M is selected from group D1 group D2:


and where (a) if M is a group D1:
X is selected from O, NH and NCH3;
And selected from communication and group NR2where R2represents hydrogen or methyl;
E is selected from communication, CH2CH(CN) and(CH3)2;
R1selected from
(i) 3--membered cycloalkyl group;
(ii) 4-6-membered saturated heterocyclic groups containing 1 or 2 heteroatoms selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4by alkyl; but excluding unsubstituted 4-morpholinyl, unsubstituted tetrahydropyran-4-yl, unsubstituted 2-pyrrolidinyl, and unsubstituted and 1-substituted piperidine-4-yl;
(iii) a 2,5-substituted phenyl group of the formula:

in which (a) if X represents NH or N-CH3, R3represents chlorine; and (b) if X represents Oh, R3represents CN;
(iv) group CR6R7R8in which each of R6and R7selected from hydrogen and methyl, and R8selected from hydrogen, methyl,1-4alkylsulfonyl, hydroxymethyl and cyano;
(v) pyridazin-4-ilen group, optionally substituted by one or two substituents selected from methoxy, ethoxy;
(vi) substituted imidazothiazole group in which the substituents are selected from methyl and ethyl; and
(vii) 1,3-dihydroindol-2-ilen, or 2,3-dihydroindol-1-ilen groups;
(viii) 3-pyridyl;
(ix) thiomorpholine or its S-oxide or S,S-dioxide; and
if E-h is a NR2, R1additionally, selected from
(x) 2-ftoheia, 3-ftoheia, 4-ftoheia, 2,4-dipthera, 3,4-dipthera, 2,5-dipthera, 3,5-dipthera, 2,,6-tryptophanyl, 2-methoxyphenyl, 5-chloro-2-methoxyphenyl, cyclohexyl, unsubstituted 4-tetrahydropyranyl and tert-butyl;
(xi) the group NR10R11where each of R10and R11represents a C1-4alkyl, or R10and R11connected so that NR10R11form a 4-6-membered saturated heterocyclic group optionally containing a second heteroatom in the cycle selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4by alkyl;
(xii) pyridone, optionally substituted C1-4by alkyl;
if E-a represents C(CH3)2NR2or CH2-NR2, R1additionally, selected from
(xiii) unsubstituted 2-furil and 2,6-diphthera; and
if E-is(CH3)2NR2, R1additionally, selected from
(xiv) unsubstituted phenyl; and
if E is a CH2, R1additionally, selected from
(xv) unsubstituted tetrahydropyran-4-yl; and
(C) if M is a group D2:
And that is the link;
E represents a bond;
R1selected from
(xvi) a 2-substituted 3-shriley group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5connected this about what atom, that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2;
(xvii) of 5-substituted 2-shriley group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2; provided that the compound is not [3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amidon 5-piperidine-1-ilmatieteen-2-carboxylic acid;
(xviii) a group of the formula:

in which R9represents hydrogen, methyl, ethyl or isopropyl; and G represents CH, O, S, SO, SO2or NH; and
(xix) of 3,5-disubstituted phenyl group of the formula:

in which X is selected from NH and NCH3;
(C) if M is a group D1:
and X represents O; a represents a group NR2where R2is hydrogen; E is a bond; and R1is 2,6-differenial; then the compound of formula (I) is a pharmaceutically acceptable acid-addition the Yu salt, selected from salts formed with acids, which are selected from the group consisting of acetic, adipic, aspartic (e.g. L-aspartic), benzosulfimide, econsultancy, D-gluconic, glucuronic (such as D-glucuronic), glutamic (e.g. L-glutamic), hydrochloric, lactic (for example, (+)- L-lactic and (±)-DL-lactic), methansulfonate and toluensulfonate (for example p-toluensulfonate) acids.

2. The compound according to claim 1 or its pharmaceutically acceptable salt, MES or tautomer, in which M is selected from group D1 group D2:


and where
(I) if M is a group D1:
X is selected from O, NH and NCH3;
And selected from communication and group NR2where R2represents hydrogen or methyl;
E is selected from communication, CH2CH(CN) and(CH3)2;
R1selected from
(i) 3-5-membered cycloalkyl group;
(ii) 4-6-membered saturated heterocyclic groups containing 1 or 2 heteroatoms selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4by alkyl; but excluding unsubstituted 4-morpholinyl, unsubstituted tetrahydropyran-4-yl, unsubstituted 2-pyrrolidinyl, and unsubstituted and 1-substituted piperidine-4-yl;
(iii) a 2,5-substituted phenyl group of the formula:

in which (a) if X represents NH or N-CH3, R3represents chlorine; and (b) if X represents Oh, R3represents CN;
(iv) group CR6R7R8in which each of R6and R7selected from hydrogen and methyl, and R8selected from hydrogen, methyl,1-4alkylsulfonyl, hydroxymethyl and cyano;
(v) pyridazin-4-ilen group, optionally substituted by one or two substituents selected from methoxy, ethoxy;
(vi) substituted imidazothiazole group in which the substituents are selected from methyl and ethyl; and
(vii) 1,3-dihydroindol-2-ilen, or 2,3-dihydroindol-1-ilen groups;
(ix) thiomorpholine or its S-oxide or S,S-dioxide; and
if E-h is a NR2, R1additionally, selected from
(x) 2-ftoheia, 3-ftoheia, 4-ftoheia, 2,4-dipthera, 3,4-dipthera, 2,5-dipthera, 3,5-dipthera, 2,4,6-tryptophanyl, 2-methoxyphenyl, 5-chloro-2-methoxyphenyl, cyclohexyl, unsubstituted 4-tetrahydropyranyl and tert-butyl;
(xi) the group NR10R11where each of R10and R11represents a C1-4alkyl, or R10and R11connected so that NR10R11forms a saturated 4-6 membered heterocyclic group optionally containing a second cyclic heteroatom selected from O, N, S and SO2/sub> and heterocyclic group optionally substituted C1-4by alkyl;
(xii) pyridone, optionally substituted C1-4by alkyl;
if E-a represents C(CH3)2NR2or CH2-NR2, R1additionally, selected from
(xiii) unsubstituted 2-furil and 2,6-diphthera; and
if E-a represents C(CH3)2NR2, R1additionally, selected from
(xiv) unsubstituted phenyl; and
if E is a CH2, R1additionally, selected from
(xv) unsubstituted tetrahydropyran-4-yl; and
(II) if M is a group D2:
And that is the link;
E represents a bond;
R1selected from
(xvi) a 2-substituted 3-shriley group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2;
(xvii) of 5-substituted 2-shriley group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR 4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2;
(xviii) a group of the formula:

in which R9represents hydrogen, methyl, ethyl or isopropyl; and G represents CH, O, S, SO, SO2or NH; and
(xix) of 3,5-disubstituted phenyl group of the formula:

in which X is selected from NH and NCH3.

3. The compound according to claim 1 or its pharmaceutically acceptable salt, MES or tautomer, where M is selected from group D1 group D2:


and where (a) if M is a group D1:
X is selected from O, NH and NCH3;
And selected from communication and group NR2where R2represents hydrogen or methyl;
E is selected from communication, CH2CH(CN) and(CH3)2;
R1selected from
(i) 3-5-membered cycloalkyl group;
(ii) 4-6-membered saturated heterocyclic groups containing 1 or 2 heteroatoms selected from O, N, S and SO2and heterocyclic group optionally substituted C1-4by alkyl; but excluding unsubstituted 4-morpholinyl, unsubstituted tetrahydropyran-4-yl, unsubstituted 2-pyrrolidinyl, and unsubstituted and 1-substituted piperidine-4-yl;
(iii) a 2,5-substituted phenyl group forms the crystals:

in which (a) if X represents NH or N-CH3, R3represents chlorine; and (b) if X represents Oh, R3represents CN;
(iv) group CR6R7R8in which each of R6and R7selected from hydrogen and methyl, and R8selected from hydrogen, methyl,1-4alkylsulfonyl, hydroxymethyl and cyano;
(v) pyridazin-4-ilen group, optionally substituted by one or two methoxy substituents;
(vi) substituted imidazothiazole group, in which the Deputy represents methyl; and
(vii) 1,3-dihydroindol-2-ilen, or 2,3-dihydroindol-1-ilen groups;
(viii) 3-pyridyl;
(ix) thiomorpholine or its S-oxide or S,S-dioxide; and
if E-h is a NR2, R1additionally, selected from
(x) 2-ftoheia, 3-ftoheia, 4-ftoheia, 2,4-dipthera, 3,4-dipthera, 2,5-dipthera, 3,5-dipthera, 2,4,6-tryptophanyl, 2-methoxyphenyl, 5-chloro-2-methoxyphenyl, cyclohexyl, unsubstituted 4-tetrahydropyranyl and tert-butyl;
(xi) the group NR10R11where each of R10and R11represents a C1-4alkyl, or R10and R11connected so that NR10R11form a 4-6-membered saturated heterocyclic group optionally containing a second heteroatom in the cycle, wybran the th of O N, S and SO2and heterocyclic group optionally substituted C1-4by alkyl;
(xii) pyridone, optionally substituted C1-4by alkyl;
if E-a represents C(CH3)2NR2or CH2-NR2, R1additionally, selected from
(xiii) unsubstituted 2-furil and 2,6-diphthera; and
if E-is(CH3)2NR2, R1additionally, selected from
(xiv) unsubstituted phenyl; and
if E is a CH2, R1additionally, selected from
(xv) unsubstituted tetrahydropyran-4-yl; and
(C) if M is a group D2:
And that is the link;
E represents a bond;
R1selected from
(xvi) a 2-substituted 3-shriley group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5linked such that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2;
(xvii) of 5-substituted 2-shriley group of the formula:

in which R4and R5are the same or different and selected from hydrogen and C1-4the alkyl, or R4and R5are connected in the way, that NR4R5forms a 5 - or 6-membered saturated heterocyclic group optionally containing a second heteroatom or group selected from O, NH, NMe, S or SO2; provided that the compound is not [3-(5,6-dimethoxy-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]amidon 5-piperidine-1-ilmatieteen-2-carboxylic acid;
(xviii) a group of the formula:

in which R9represents hydrogen, methyl, ethyl or isopropyl; and G represents CH, O, S, SO, SO2or NH; and
(xix) of 3,5-disubstituted phenyl group of the formula:

in which X represents NCH3.

4. The compound according to claim 1 of formula (II):

where R1E , a and X are such as defined in claim 1.

5. The compound according to claim 3 of formula (III):

or its pharmaceutically acceptable salt, MES or tautomer, where R1, R2and E such as defined in item 3.

6. The compound according to claim 5 of the formula (III):

or its pharmaceutically acceptable salt, MES or tautomer, in which E is a bond, R2represents hydrogen, and R1represents cycloalkyl group.

7. The connection according to claim 6, in which R1is cyclopropylidene group, with the specified connection is a 1-cyclopropyl-3-[3-(5-what orfelin-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or its pharmaceutically acceptable salt, MES or tautomer.

8. The connection according to claim 7 where the compound is a pharmaceutically acceptable salt, selected from acetate, nelfinavir, aconsultant, DL-lactate, adipate, D-glucuronate, D-gluconate or hydrochloride.

9. The connection according to claim 7 in free base form.

10. The connection according to claim 7 or its pharmaceutically acceptable salt, which is essentially crystalline.

11. The connection according to claim 9, which is a crystalline dihydrate crystals belong to the monoclinic space group P21/n (#14) with the following lattice parameters at 101(2)K: the cell size a=7,66(10), the cell size b=15,18(10), the cell size C=17,71(10)Å, angle of cell α=90°, the angle β cell=98,53(2)°, the angle of the cell γ=90°.

12. The connection according to claim 7 in the form of a pharmaceutically acceptable salt, selected from lactate and citrate, and mixtures thereof.

13. The connection section 12, which is an L-lactate.

14. The connection section 12, which is citrate.

15. The connection section 12, which is a mixture of L-lactate and citrate.

16. The connection 13, which represents an L-lactate 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea which is crystalline and characterized by one or more (in any combination) or all of the parameters of the following, namely, that this salt
a) has a crystalline structure, shown in figure 4 and 5; and/or
(b) has a crystalline structure, which belongs to the orthorhombic space group P212121(#19) with the following lattice parameters at 97(2)K: the cell size a=9,94(10), the cell size b=15,03(10), the cell size C=16,18(10)Å, angles cells α=β=γ=90°; and/or
(c) has a powder x-ray characterized by the presence of major peaks corresponding to the diffraction angles (2θ) 17,50, 18,30, 19,30, 19,60 and 21.85 degrees, and more specifically later on in 12,40, 15,20, 15,60, 17,50, 18,30, 18,50, 19,30, 19,60, 21,85 and 27,30 degrees, and/or the interplanar distances (d)equal to is 5.06, 4,85, 4,60, 4,53 and 4,07, and more specifically advanced 7,13, 5,83, 5,68, 5,06, 4,85, 4,79, 4,60, 4,53, 4,07 and 3.26 Angstrom; and shows peaks at the same diffraction angles as peaks in the powder x-ray shown in Fig.6, and not necessarily where the peaks have the same relative intensity as the peaks of figure 6;
moreover, salt is anhydrous and shows an endothermic peak at 190°C. when examined by DSC method; and/or in the study applying the methodology with KBr tablet has an infrared spectrum that contains characteristic peaks at 3229, 2972 and 1660 cm-1.

17. Optionally buffered aqueous solution for intravenous injection containing L-lactate or citrate, or a mixture of these salts 1-cyclopropyl-3-[3-(5-Mohali the-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea concentration, in excess of 1 mg/ml, with a pH in the range from 2 to 6.

18. Aqueous solution for intravenous injection containing 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in protonated form together with one or more counterions selected from L-lactate and citrate, and mixtures thereof; and optionally (i) one or more other counterions such as chloride ions and/or (ii) one or more fillers for intravenous drugs.

19. Antiproliferative pharmaceutical composition comprising 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or its pharmaceutically acceptable salt, MES or tautomer and a pharmaceutically acceptable carrier.

20. The pharmaceutical composition according to claim 19 in dried form, intended for dissolution in water.

21. The pharmaceutical composition according to claim 19 in liofilizovannyh form, including 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea in protonated form together with one or more counterions selected from L-lactate and citrate, and mixtures thereof; and optionally (i) one or more other counterions such as chloride ions and/or (ii) one or more fillers for intravenous drugs.

22. The compound according to any one of claims 1 to 16 or Pharma is efticiency acceptable salt, MES or tautomer for use in the treatment of mammalian diseases or conditions, and (i) the disease or condition involves abnormal growth of cells or (ii) a specified disease or condition occurs due to abnormal growth of cells.

23. The use of compounds according to any one of claims 1 to 16 or its pharmaceutically acceptable salt, MES or tautomer for the manufacture of a medicinal product for the treatment of a disease or condition comprising abnormal cell growth or arising from abnormal cell growth, in a mammal.

24. The application of item 23, where the disease or condition is a cancer.

25. The use of the compounds according to paragraph 24, where the cancer is characterized by increasing regulation of Aurora kinase.

26. The application of item 23, where the disease or condition is a cancer selected from bladder cancer, breast, colon, kidney, epidermis, liver, lung, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, or skin; hematopoietic tumors of lymphoid origin, selected from leukemia, acute lymphocytic leukemia, b-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, lymphoma, hairy cells or lymphoma of Burkett; hematopoietic op what Holy myeloid origin, selected from acute and chronic myelogenous leukemias, myelodysplastic syndrome, or promyelocytic leukemia; follicularis thyroid cancer; a tumour of mesenchymal origin, selected from fibrosarcoma or rabdomyosarcoma; a tumour of the Central or peripheral nervous system selected from astrocytoma, neuroblastoma, glioma or sanomi; melanoma; seminoma; teratocarcinoma; osteosarcoma; pigment xeroderma; keratoacanthoma; follicularis thyroid cancer; or Kaposi's sarcoma.

27. Use p, where the disease or condition is a leukemia.

28. The application of item 27, where the leukemia is selected from re or refractory acute myelogenous leukemia, myelodysplastic syndrome, acute lymphocytic and chronic myelogenous leukemia.

29. The use of compounds according to any one of claims 1 to 16 for the manufacture of a medicinal product for preventing or treating cancer in a patient, which results, is included in the subgroup of people with the option I1e31 gene Aurora A.

30. The use of compounds according to any one of claims 1 to 16 for the manufacture of a medicine for the prevention or treatment of cancer, characterized by increasing regulation of Aurora kinase.

31. The method of obtaining the compounds of formula (I)defined in claim 2, g is e a is a bond, including the interaction of the compounds of formula (X)

with a carboxylic acid R1-E-CO2H or its reactive derivative in the conditions of formation of amides.

32. The method of obtaining the compounds of formula (I), as defined in any of claim 2 to 16, where a represents NH, including interaction of the compounds of formula (X)

with an isocyanate of formula R1-E-N=C=O in the conditions of formation of urea derivatives.

33. The method of obtaining the compounds of formula (I), as defined in any of claim 2 to 16, where a represents NR2that includes the interaction of the compounds of formula (X)

with an amine of formula R1-E-NR2H in the presence of the reagent, forming a carbonyl-containing urea, such as 1,1'-carbonyldiimidazole (CDI), phosgene or triphosgene.

34. The method of obtaining the compounds of formula (XXVII) or (XXVIII) or its salt:
or
including the interaction of the compounds of formula (XXIX)

where PG is aminosidine group, with a compound of formula (XXXI):

in an organic solvent in the presence of a condensing reagent such as 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide (EDC) and 1-hydroxy-benzotriazole (HOBt).

35. Spasibo 34, which compound of formula (XXIX) is a compound of formula (XXXII) below:

36. Method for the preparation of 3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-ylamine or its salts, including
(i) treatment of compounds of formula (XXVIIa) or (XXVIIIa):
or
where APG is aminosidine group which can be removed in acidic conditions,
acid in a solvent, optionally with heating; and
(ii) neutralizing the reaction mixture.

37. A method of obtaining a 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or a salt thereof, MES or tautomer, including:
(i) treatment of compounds of formula (XXVIIa) p acid in a solvent, optionally with heating;
(ii) neutralizing the reaction mixture;
(iii) the interaction of the product of stage (ii) with carbonyliron reagent; and
(iv) the interaction of the product of stage (iii) with cyclopropylamine.

38. The method according to clause 37, where carbonyloxy reagent is a 1,1'-carbonyldiimidazole (CDI), triphosgene or phosgene.

39. A method of obtaining a 1-cyclopropyl-3-[3-(5-morpholine-4-ylmethyl-1H-benzimidazole-2-yl)-1H-pyrazole-4-yl]urea or a salt thereof, MES or tautomer, including interaction of the compounds of formula (XXXIII) or (XXXIIIa):

with cyclopropylamine and then the optional formation of an acid additive salt.

40. The intermediate compound of formula (XXXII), (XXVIIa), (XXVIIIa), (XXXIII) or (XXXIIIa):

or

where APG is a tert-butoxycarbonyl group.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: described are new compounds of general formula (I): het-X-AB (I) where het is pentamerous N-heteroaryl, additionally containing one O, S or N atom as a heteroatom with heteroaryl with additional O atom being condensed with a benzene ring, or hexamerous N-heteroaryl; X means S; and where N-atom of N- heteroaryl residue and an X group are separated by one carbon atom; AB means 1,2,3-triazolo[4,5-d] pyrimidine-7-yl radical of general formula (II): where R3 is C1-8alkyl, phenyl, benzyl optionally substituted; R5-H, C1-8alkyl or phenyl.

EFFECT: production of new compounds for preparing a pharmaceutical composition either effective for treating cardiovascular, cancer, autoimmune diseases, stroke, neurodegenerative diseases, cystic fibrosis, or used in antithrombotic therapy.

9 cl, 11 ex, 3 tbl

FIELD: chemistry.

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.

EFFECT: more effective use of the compounds.

12 cl, 87 tbl, 1057 ex

FIELD: chemistry.

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9 cl, 6 tbl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention describes novel polymorphic modification N-{5-[3-(thiophene-2-carbonyl)-pyrazolo[1,5-a]pyrimidin-7-yl]-2-fluoro-phenyl}-N-methyl-acetamide, methods of its obtaining, its application as medication, its application for preparation of medication and pharmaceutical compositions, including novel polymorphic modification.

EFFECT: obtaining novel polymorphic modification for preparation of medication and pharmaceutical compositions.

30 cl, 2 tbl, 24 ex, 7 dwg

FIELD: chemistry.

SUBSTANCE: present invention relates to novel pyrazole derivatives of formula (I) or pharmaceutically acceptable salts thereof, having tyrosine kinase Trk inhibiting properties and used for treating or preventing malignant growths accompanied by high level of Trk, to a method of producing said derivatives, use thereof to prepare a medicinal agent, pharmaceutical compositions based on said derivatives, a method of inhibiting Trk activity and a method of obtaining antiproliferative action. where A denotes a single bond or C1-2alkylene; where the said C1-2alkylene can be optionally substituted with one R22; ring C is a phenyl or a 5-6-member heterocyclic ring with 1-2 heteroatoms selected from N or S. Values of R1-R7, R22 and n are given in the formula of invention.

EFFECT: obtaining pharmaceutically acceptable salts having tyrosine kinase Trk inhibiting properties and used for treating or preventing malignant growths.

20 cl, 5 dwg, 193 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel acetylenyl-pyrazole-pyrimidine derivatives of general formula (I), having mGluR2 (metabotropic glutamate receptor) antogonist properties). In compounds of general formula (I): either E and J denote N, G denotes C and L denotes N, M denotes CH, or M denotes N, L denotes CH; or L and G denote N, E denotes C, and J and M denote CH; or J, G and L denote N, E denotes C, and M denotes CH; or E and L denote N, J and M denote CH, and G denotes C; R1 denotes H, halogen, CF3, CHF2 or C1-6alkyl; R2 denotes H, halogen, C1-6-alkyl, C1-6-alkoxy, CF3 or CHF2, wherein R1=R2≠H; R3 denotes H; -C(CH3)2OH; linear C1-4-alkyl or C3-4-cycloalkyl, which are possibly substituted with one or more substitutes selected from a group comprising 1-3 F and 1-2 OH; A is selected from a group comprising phenyl or a 5- or 6-member heteroaryl having in the ring 1-2 heteroatoms selected from nitrogen, sulphur or nitrogen and sulphur in the 5-member ring, and 1-2 nitrogen atoms i the 6-member ring, and possibly substituted with 1-3 Ra; Ra denotes halogen; hydroxy; cyano; CF3; NReRf; C1-C6-alkyl, possibly substituted amino or hydroxy; ; C1-6-alkoxy; C3-4-cycloalkyl; CO-NRbRc, SO2-NRbRc; or SO2-Rd-; Rb and RC can be identical or different and are selected from a group comprising H; normal or branched C1-6-alkyl, possibly substituted with one or more substitutes selected from a group comprising F, cyano, hydroxy, C1-6-alkoxy, -NH-C(O)-O-C1-6-alkyl, amino, (C1-6-alkyl)amino, di(C1-6-alkyl)amino, heterocycloalkyl having 6 ring atoms, from which 1-2 heteroatoms are selected from nitrogen or nitrogen and oxygen, or a 6-member heteroaryl with one nitrogen heteroatom in the ring; or a 6-membeer heteroaryl with one nitrogen heteroatom in the ring; or Rb and Rc, together with the nitrogen atom with which they are bonded, can form a heterocyclic ring having 6 members in the ring, from which 1-2 atoms are selected from nitrogen and/or oxygen, and which can be substituted with C1-6-alkyl; Rd denotes OH or C1-6-alkyl; Re and Rf denote H, C1-6-alkyl, possibly substituted hydroxy, -C(O)- C1-6-alkyl; S(O)2-C1-6-alkyl.

EFFECT: compounds can be used in preparing medicinal agents for treating central nervous system (CNS) disorders, such as Huntington's chorea, amyotrophic lateral sclerosis, dementia caused by AIDS, parkinsonism etc.

55 cl, 6 dwg, 321 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds of the 2,9-disubstituted imidazo[1,2-a]benzimidazole family, specifically to water-soluble salts of 9-aminoethyl-substituted 2-(4-fluorophenyl)imidazo[1,2-a]benzimidazole of general formula I:

,

where NR2 = pyrrolidine-, piperidine-, morpholine-; Y=HBr, H2SO4, (CH2COOH)2 and [CH(OH)COOH]2; n=1, 2.

EFFECT: novel compounds have analgesic action.

2 cl, 2 tbl, 15 ex

Pyrazolepyrimidines // 2412186

FIELD: chemistry.

SUBSTANCE: compounds can be used to treat tumourous diseases, such as solid tumours, breast, lung, large intestine or prostate gland tumours. In compounds of formula

: R1 is selected from a group comprising: (a) saturated cyclic radical containing 3-8 ring atoms, from which 1-3 atoms are N atoms, which can contain up to four substitutes independently selected from a group comprising: (i) lower alkyl; and (ii) CO2R3, OR7 or S(O)nR8; (b) C6-C10aryl, which can contain up to four substitutes independently selected from a group comprising: (i) S(O)nR8, lower alkyl; OR7 and halogen; (c) C3-C8cycloalkyl, which can be substituted with NR5R6; and (d) lower alkyl, which can be substituted: (i) OR7, NR5R6; R2 is selected from a group comprising: (i) H; (ii) lower alkyl; (iii) C6-C10aryl which can be substituted with a halogen, lower alkyl, lower alkoxy group; R3 is selected from a group comprising: (i) H; (ii) lower alkyl; (iv) C3-C8cycloalkyl; R5 and R6 are independently selected from a group comprising: (i) H; (ii) lower alkyl; (iii) C3-C8cycloalkyl; (v) SO2R3; and (vi) CO2R3; R7 is selected from a group comprising H and lower alkyl; R8 is selected from a group comprising: (iii) NR5R6; (iv) lower alkyl; and n equals 1 or 2.

EFFECT: capacity to inhibit activity of cyclin-dependant kinase.

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula [I-D1] or pharmaceutically acceptable salt thereof,

,

where each symbol is defined in the claim. The invention also relates to pharmaceutical compositions containing said compound and having HCV polymerase inhibiting activity.

EFFECT: disclosed compound exhibits anti-HCV activity, based on HCV polymerase inhibiting activity and is useful as an agent for preventing and treating hepatitis C.

32 cl, 497 tbl, 1129 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new compounds of formula I and to their pharmaceutically acceptable salts exhibiting inhibitory activity in relation to kinases chosen from Abl, Bcr-Abl, Bmx, BTK, b-RAF, c-RAF, CSK, cSRC, Fes, FGER3, Elt3, 1KKα, 1KKβ, JNK1α1, JNK2α2, Lck, Met, MKK4, MKK6, p70S6K, PAK2, PDGFRα, PKA, PKCα, PKD2, ROCK-II, Ros, Rsk1, SAPK2α, SAPK2β, SAPK3, SAPK4, SGK, Syk, Tie2 and TrkB. In compounds of formula I , n is equal to 1, m is equal to 0, Y1 is chosen from N and CR5, and R5 represents hydrogen, Y2 represents O, R1 represents hydrogen, R2 is chosen from hydrogen and C1-C6alkyl, R3 is chosen from a group including hydrogen, C1-C6alkyl, C1-C6alkoxy, R4 is chosen from NR5C(O)R6 and -C(O)NR5R6 where R5 is chosen from hydrogen and C1-C6 alkyl, and R6 represents phenyl optionally substituted with 1-3 radicals chosen of a group including NR3R3, halogen-substituted C1-C6alkyl, C5-C6heteroaryl(C0-C4)alkyl where heteroaryl contains 1-2 heteroatoms chosen from N and O, C5-C6heterocyclo(C0-C4)alkyl, where heterocyclyl contains 1-2 heteroatoms of N, and C5-C6heterocyclo(C0-C4)alkoxy where heterocyclyl contains 1-2 heteroatoms of N, and any heteroaryl or heterocyclyl contained in R6 is optionally substituted by 1-3 radicals independently chosen from a group including C1-C6alkyl and hydroxy(C1-C6)alkyl.

EFFECT: producing the compounds which can find application for treatment or prevention of diseases or disorders associated with abnormal or unregulated kinase activity, such as proliferative diseases, diseases of immune and nervous system.

8 cl, 26 ex

Aromatic compound // 2416608

FIELD: chemistry.

SUBSTANCE: invention describes a novel compound of general formula (1), where radicals R1, R2, X1, Y and A are as described in claim 1 of the invention. The invention also describes a method of obtaining compounds of formula (1), as well as a pharmaceutical composition based on said compounds, for treating fibrosis.

EFFECT: novel compounds with excellent collagen formation suppression, cause fewer side-effects and which are safer are obtained.

62 cl, 2717 ex, 432 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: compound of formula pharmaceutically acceptable salt or solvate of a compound or salt (I), ring Q represents optionally substituted monocyclic or condensed (C6-C12)aryl or optionally substituted monocyclic or condensed heteroaryl where said substitutes are chosen from: halogen; (C1-C6)alkyl optionally substituted by 1-3 halogen atoms; (C1-C6)alkylsulphonyl; phenyl optionally substituted by 1 or 2 substitutes chosen from halogen, (C1-C6)alkyl which can be substituted by 1-3 halogen atoms, groups (C1-C6)alkylamino, di(C1-C6)alkylamino, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl and (C1-C6)alkylthio; monocyclic or condensed heteroaryl optionally substituted by halogen; or oxo; Y1 represents a bond or -NR6-CO-, where R6 represents hydrogen, ring A represents optionally substituted a nonaromatic heterocyclyldiyl where said substitutes are chosen from (C1-C6)alkyl optionally substituted by groups hydroxy, (C1-C6)alkylamino, di(C1-C6)alkylamino, morpholino, (C1-C6)alkylaminocarbonyl, di(C1-C6)alkylaminocarbonyl; cyano; (C3-C6)cycloalkyl; (C1-C6)alkoxy; (C1-C6)alkoxy(C1-C6)alkyl; phenyl; benzyl; benzyloxymethyl; thienyl; 4-8-members monocyclic nonaromatic heterocycle having 1 or 2 heteroatoms chosen from N or O, and optionally substituted by 1 or 2 substitutes chosen from (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl and oxo; (C1-C6)alkylamino; di(C1-C6)alkylamino; a group of formula: -Y2Z'- represents a group of formula: [Formula 2] each R7 independently represents hydrogen, (C1-C6)alkyl or (C3-C6)cycloalkyl, each of R8 and R9 independently represents hydrogen or (C1-C6)alkyl, n is equal to an integer 0 to 3, Z1 represents a bond, -O-, -S- or-NR9 - where R9 represents hydrogen, (C1-C6)alkyl, acyl or (C1-C6)alkylsulphonyl, ring B represents optionally substituted aromatic carbocyclediyl or optionally substituted aromatic heterocyclediyl where said substitutes are chosen from (C1-C6)alkyl, halogen, (C1-C6)alkoxy and oxo; Y3 represents a bond optionally substituted (C1-C6)alkylene or (C3-C6)cycloalylene, optionally interrupted -O- or optionally substituted (C2-C6)alkenylene where said substitutes are chosen from (C1-C6)alkyl, (C3-C6)cycloalkyl, halogen and (C1-C6)alkoxycarbonyl; Z2 represents COOR3; R3 represents hydrogen or (C1-C6)alkyl.

EFFECT: preparation of new compounds.

30 cl, 9 tbl, 944 ex

FIELD: medicine.

SUBSTANCE: invention refers to the compound 3-{[5-(azetidine-1-ylcarbonyl)pyrazine-2-yl] oxy}-5-{[(1S)-1-methyl-2-(methyloxy)ethyl]oxy}-N-(5-methylpyrazine-5-yl)benzamide or to its pharmaceutically acceptable salt. Also, it refers to a pharmaceutical composition for treating insulin-independent diabetes or obesity containing said compound.

EFFECT: there is produced and described a new compound which can be effective in treating insulin-independent diabetes and obesity.

5 cl, 64 ex

FIELD: chemistry.

SUBSTANCE: invention relates to oxazolidinone derivatives of formula (I) or pharmaceutically acceptable salts thereof, synthesis method thereof and pharmaceutical compositions containing said derivatives which are used as an antibiotic. Oxazolidinone derivatives, where R1 and R1' independently denote hydrogen or fluorine; R2 denotes -OR7, fluorine, monophosphate or metal phosphate; and R7 denotes hydrogen, C1-3alkyl or an acylated amino acid group, where the amino acid is alanine, glycine, proline, proline, isoleucine, leucine, phenylalanine, β-alanine or valine; R3 denotes hydrogen, a C1-4alkyl group which is unsubstituted or substituted cyano, , -(CH2)m-OR7 (m equals 0, 1, 2, 3, 4) or a ketone group. Oxazolidinone derivatives of formula (I) have antibacterial activity against different human and animal pathogens.

EFFECT: oxazolidinone derivatives, having inhibiting activity towards a wide range of bacteria and having low toxicity.

27 cl, 4 tbl, 73 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new pyrrolidine derivatives of general formula (1) or its pharmaceutically acceptable salts where R101 and R102 values are described by the patent claim. The compounds inhibit serotonin and/or norepinephrine and/or dopamine reabsorption thereby allowing to be used for treating depression and anxiety disorder. A method for preparing thereof is described.

EFFECT: preparation of new pyrrolidine derivatives.

10 cl, 162 tbl, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to compounds of formula I and to their pharmaceutically acceptable salts. In formula I p is integer, equal to 0-1; L2 is selected from group including -XOX-, -XSX- and -XSXO-; where X is independently selected from group, including bond and C1-C4alkylene; R13 is selected from group, including halogen, C1-C6alkyl, C1-C6alkoxygroup, -C(O) C1-C6alkyl; R14 is selected from group, including -XOXC(O)OR17 and -C1-C4alkylene-C(O)OR17; where X represents bond or C1-C4alkylene; and R17 is selected from group, including hydrogen and C1-C6alkyl; R15 and R16 are independently selected from group, including -R18 and -YR18; where Y represents C2-C6alkenylene, and R18 is selected from group, including C6-C10aryl, benzo[1,3]dioxolyl, pyridinyl, pyrimidinyl, quinolyl, phenoxatiinyl, benzofuranyl, dibenzofuranyl, benzoxasolyl, 2,3-dihydrobenzofuranyl, 2-oxo-2,3-dihydrobenzooxasolyl, indolyl, 3-oxo-3,4-dihydro-2H-benzo[1,4]oxazinyl, 2,3-dihydrobenzo[1,4]dioxinyl, 3,4-dihydro-2H-benzo[b][1,4]dioxepinyl, where any C6-C10aryl, pyridinyl, benzoxasolyl, indolyl in R18 is optionally substituted by 1-2 radicals, independently selected from group, including halogen, nitrogroup, cyanogroup, C1-C6alkyl, C1-C6alkoxygroup, C1-C6alkylthiogroup, hydroxy-C1-C6alkyl, halogen-substituted C1-C6alkyl, halogen-substituted C1-C6alkoxygroup, piperidinyl, morpholinyl, pyrrolidinyl, phenyl, XS(O)0-2R17, -XNR17R17, -XNR17S(O)2R17, -XNR17C(O)R17, -XC(O)NR17R17, -XC(O)NR17R19, -XC(O)R17, -XC(O)R19 and -XOXR19, where X represents bond; R17 is selected from group, including hydrogen, C1-C6alkyl, halogen-substituted C1-C6alkyl, and R19 is selected from group, including C3-C12cycloalkyl, phenyl, piperidinyl, morpholinyl.

EFFECT: ensuring application of invention compounds for production of medication, modulating activity of activated receptors of peroxisome proliferators δ (ARPPδ), to pharmaceutical composition, possessing properties of ARPPδ activity modulator, including therapeutically efficient quantity of invention compound and to application of pharmaceutical composition for medication manufacturing.

8 cl, 1 tbl, 301 ex

FIELD: chemistry.

SUBSTANCE: invention describes compounds of formula I

, where R1 is selected from a group comprising hydrogen, lower alkyl, lower hydroxyl, lower alkoxyalkyl, lower halogenalkyl, lower cyanoalkyl; unsubstituted or substituted phenyl; lower phenylalkyl, where the phenyl ring can be unsubstituted or substituted; and heteroaryl, selected from pyridyl and pyrimidinyl; R2 denotes hydrogen or halogen; G denotes a group selected from

, where m equals, 0, 1; R3 is selected from lower alkyl, cycloalkyl and lower cycloalkylalkyl; n equals 0, 1; R4 denotes lower alkyl, as well as pharmaceutical compositions.

EFFECT: said compounds are used to treat or prevent diseases associated with histaminase receptor modulation.

19 cl, 1 tbl, 24 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new compounds of formula 1, its pharmaceutically acceptable salts and stereoisomers: $ (1), where: R1 means H, amidino, C1-C4-alkyl amidino, C1-C4alkanoylamidino, C1-C10-alkyl, C3-C7-cycloalkyl, C6-C10-aryl, 6-members heterocycle with O atom, 5-members heterocycle with two N atoms, 6-members heteroaryl with one or two N atoms, 5-members heteroaryl with two heteroatoms, one of which is N, and the other is S, C1-C6-alkylcarbonyl, C3-C7-cycloalkylcarbonyl, C1-C4-alkoxycarbonyl, C6-C10-aryl-C1-C4-alkoxycarbonyl, -SO2-C1-C4-alkyl, -C(O)-N(R6)(R7) or -C(S)-N(R6)(R7); and, R6, R7 means H, C1-C6-alkyl, C3-C7-cycloalkyl; alkyl, cycloalkyl, heterocycle, aryl or heteroaryl are unsubstituted or substituted; R2 means C6-C10-aryl which is unsubstituted or mono- or disubstituted; R3 means H, CN, C1-C6-alkyl, C3-C7-cycloalkyl, C2-C6-alkenyl, monocyclic 5-members heterocycle with N and O, monocyclic 5-members heteroaryl with two heteroatoms, one of which is N, and the other is O or S, C(O)-R8 or -C(S)-R8; and R8 means OH, C1-C4-alkyl, C1-C4-alkyloxy or N(R9)(R10); R9, R10 mean N, C1-C6-alkyl, C3-C7-cycloalkyl, C1-C4-alkyloxy, phenyl or 5-members heteroaryl with two heteroatoms, one of which is N, and the other is S, 6-members heteroaryl with N; R9, R10 together with N whereto attached can form a single 4-6-members ring which can include in addition O or S; and alkyl, cycloalkyl, heterocycle, phenyl or heteroaryl are unsubstituted or substituted. R4 means C3-C8-cycloalkyl, C6-C10-aryl, 5-members heteroaryl with two heteroatoms, one of which is N, and the other is S, 6-members heteroaryl with N, 6-members heterocycle with O, and C6-C10-aryl or heteroaryl are unsubstituted or mono- or polysubstituted. R5 means N, C1-C6-alkyl, -C(O)-R11, C1-C6-alkylsulphonyl, C6-C10-arylsulphonyl, -(CH2)p-C6-C10-aryl, -(CH2)p-heteroaryl or -(CH2)p-C3-C8-cycloalkyl where heteroaryl means 5-members heteroaryl with O or with N or with S which can contain in addition N. p is equal to 1 or 2; R11 means C1-C10-alkyl, C1-C6-alkenyl, C3-C8-cycloalkyl, C3-C8-cycloalkenyl, NH2, C1-C4alkylamino, (C1-C4-alkyl)(C1-C4-alkyl)amino, C6-C10-aryl, 5-members heteroaryl with N or with O or with 8 which can contain in addition N, 6-members heterocycle with N and O, 5- or 6-members heterocycle with O, and alkyl is unsubstituted or substituted with one substitute. Aryl, heteroaryl, cycloalkyl, cycloalkenyl or heterocycle are unsubstituted or mono- or disubstituted.

EFFECT: compounds are melanocortin receptor agonists so presented to be used in a pharmaceutical composition for treatment and prevention of obesity, diabetes, inflammation, erectile dysfunction.

19 cl, 18 tbl

FIELD: chemistry.

SUBSTANCE: present invention relates to novel derivatives of cis-2,4,5-triarylimidazoline of general formula I and pharmaceutically acceptable salts thereof, where X1 is selected from a group comprising lower alkoxy; X2 and X3 are independently selected from a group comprising hydrogen, halogen, cyano, lower alkyl, lower alkoxy, piperidinyl, -NX4X5, -SO2NX4X5, -C(O)NX4X5, -C(O)X6, -SOX6, -SO2X6, -NC(O)-lower alkoxy, -C≡C-X7, provided that both X2 and X3 do not denote hydrogen, lower alkyl or lower alkoxy, provided that when X2 or X3 denote hydrogen, the other does not denote lower alkyl, lower alkoxy or halogen, provided that when X2 denotes -HX4X5, X3 does not denote hydrogen, X2 and X3 together can form a ring selected from 5-7-member unsaturated rings which can contain three heteroatoms selected from S, N and O, X4 and X5 are independently selected from a group comprising hydrogen, lower alkyl, lower alkoxy, lower alkyl, substituted by a lower alkoxy, -SO2-lower alkyl, -C(O)piperazinyl-3-one; X6 is selected from a group comprising lower alkyl, morpholine, piperidine, pyrrolidine; X7 is selected from a group comprising hydrogen, lower alkyl, trifluoromethyl; Y1 and Y2 are independently selected from a group comprising halogen; R is selected from a group comprising lower alkoxy, piperidinyl substituted with a five-member heterocyclic ring which contains one nitrogen heteroatom, piperidinyl substituted with a hydroxy, -CH2OH or -C(O)NH2, piperazinyl substituted with one or two R1 [1,4]diazepanyl, substituted R1, R1 can denote one or two substitutes selected from a group comprising oxo, lower alkyl substituted with one R2, -C(O)R3, -SO2-lower alkyl, -SO2-five-memer heterocyclyl, which is selected from isoxazolyl, dimethylisoxazolyl, pyrrolidinyl, pyrrolyl, thiophenyl, imidazolyl, thiazolyl, thiazolidinyl, imidazolidinyl; R2 is selected from a group comprising -SO2-lower alkyl, hydroxy, lower alkoxy, -NH-SO2-lower alkyl, -cyano, -C(O)R4; R3 is selected from a group comprising a five-member heterocyclyl which is selected from isoxazolyl, dimethylisoxazolyl, pyrrolidinyl, pyrrolyl, thiophenyl, imidazolyl, thiazolyl, thiazolidinyl, imidazolidinyl, lower alkyl, lower alkenyl, lower alkyl substituted with a six-member heterocyclyl selected from piperidinyl, piperazinyl, 3-oxopiperazinyl, morpholinyl, C3-cycloalkyl; R4 is selected from a group comprising hydroxy, morpholine, piperidine, 4-acetylpiperazinyl, -NR5R6; R5 and R6 are independently selected from a group comprising hydrogen, lower alkyl, lower alkyl substituted with lower alkoxy or cyano, lower alkoxy and C3-cycloalkyl. The invention also relates to a pharmaceutical composition based on the formula I compound, use of the formula I compound in preparing a medicinal agent and a method for synthesis of the formula I compound.

EFFECT: novel derivatives of cis-2,4,5-triarylimidazoline of general formula I are obtained, which can be used to treat diseases, based on reaction of the MDM2 protein with p53-like protein, particularly as anticancer agent.

54 cl, 412 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to compounds of formula (I), where R1 denotes a 5- or 6-member ring of formulae

(II) or (III), respectively: R2 denotes H, C1-C7-alkyl, C3-C6-cycloalkyl or -(CH2)m,-Ra; R3 denotes aryl or heteroaryl, which can be substituted with CN, Cl, F, Br, CF3, CHF2, C3-C6-cycloalkyl or denotes heteroaryl which can be possibly substituted with C1-C7-alkyl; R4 denotes H, -OH, Cl, F, Br, CN, -CHF2, CF3, C1-C7-alkyl, C3-C6-cycloalkyl or -(CH2)m-Re; R5 denotes C1-C7-alkyl, -(CH2)n-O-Rf, or -(CH2)n-Re; Ra denotes -OH; Re denotes -OH; Rf denotes C1-C7-alkyl; m equals 1-4; n equals 2-6; and pharmaceutically acceptable salts thereof. The invention also relates to a medicinal agent containing said derivatives, use thereof in preparing medicinal agents suitable for treating diseases of the central nervous system.

EFFECT: novel compounds suitable for treating diseases of the central nervous system are obtained and described.

29 cl, 111 ex

Aromatic compound // 2416608

FIELD: chemistry.

SUBSTANCE: invention describes a novel compound of general formula (1), where radicals R1, R2, X1, Y and A are as described in claim 1 of the invention. The invention also describes a method of obtaining compounds of formula (1), as well as a pharmaceutical composition based on said compounds, for treating fibrosis.

EFFECT: novel compounds with excellent collagen formation suppression, cause fewer side-effects and which are safer are obtained.

62 cl, 2717 ex, 432 tbl

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