Kinase inhibitors

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula : a pharmaceutically acceptable salt or solvate thereof, having Syk kinase inhibiting properties. The invention also relates to a pharmaceutical composition containing said compound, methods of treating diseases whose development is aided by c-kit receptor activity, such as arthritis, rheumatoid arthritis, tumours, mantle cell lymphoma, as well as a method of inhibiting angiogenesis.

EFFECT: improved method.

13 cl, 4 tbl, 10 dwg

 

The SCOPE of the INVENTION

The present invention relates to substituted azaindole, their receipt, containing these compounds, pharmaceutical compositions, and their use in the treatment of pathological conditions that can be controlled by the inhibition of protein kinases.

The LEVEL of TECHNOLOGY

Protein kinases are involved in signaling processes that control the activation, growth and differentiation of cells in response to signals transmitted extracellular mediators, as well as to changes in the environment. In General, these kinases can be classified into several groups: those which preferentially phosphorylate serine and/or threonine residues, and those that selectively phosphorylate the tyrosine residues [S.K.Hanks and T.Hunter, FASEB. J., 1995, 9, str-596]. Serine/trionychinae include, for example, isoforms of protein kinase C [A.C.Newton, J. Biol. Chem., 1995, 270, str-28498] and cyclin-dependent kinases such as cdc2 [J.Pines, Trends in Biochemical Sciences, 1995, 18, page 195-l97]. Tyrosine kinase include membrane receptors growth factor, such as the receptor for epidermal growth factor [S.Iwashita and M.Kobayashi, Cellular Signalling, 1992, 4, p.123-132], and not containing the cytosolic receptor kinase like kinase p56tck, p59fYn, ZAP-70 and csk [C.Chanet. al., Ann. Rev. Immunol., 1994, l2, str-592].

The unusually high activity of protein kinases was observed in the clinical pictures of many diseases, you are the bathrooms impaired cellular functions. Directly or indirectly this could be caused, for example, a violation of the relevant control mechanism kinase related, for example, by mutation, overproductive or impaired activation of the enzyme, either excessive or insufficient production of cytokines or growth factors are also involved in the transduction of signals before or after synthesis of the kinase. In all these cases, you can expect a positive effect of selective inhibition of the action of the kinase.

Syk is a cytoplasmic proteincontaining with a molecular mass of 72 kDa, the expression of which is provided in a variety of hematopoietic cells, and is an important element in several stages, linking receptors antigens from cell response. Therefore, Syk plays a critical role in the signal transmission of the high-affinity IgE receptor, FcεR1 in the fat cells and signaling receptor-antigen in T - and b-lymphocytes. The transmission channels of the signal in the fat, T - and b-cells have common characteristics. The ligand-binding domain of the receptor is inactive, the inherent tyrosinekinase. However, they interact with the transmitting signal subunits that contain motifs activation immunoreceptor-based tyrosine (ITAM) [M.Reth, Nature, 1989, 338, str-384]. Such motifs are present in the β - and γ-subunits FcεR1, ξ-subunit receptor of T-cells (TCR) and β-is jedinica IgGα and IgG receptor B cells (BCR). [N.S.van Oers and A.Weiss, Seminars in Immunology, 1995, 7, str-236]. After binding of antigen and multimerization ITAM residues fosfauriliruyutza proteinkinase Src family. Syk refers to a unique class tyrosinekinase, having two tandem domains, a Src homology 2 (SH2) and the C-terminal catalytic domain. These SH2 domains with high affinity associated with ITAM, and such SH2 mediated the Association of Syk with the activated receptor stimulates activity of Syk kinase and localizes Syk in the plasma membrane.

In Syk-deficient mice inhibited the degranulation of mast cells, pointing to it as a significant object for the development of stabilizers mast cells [P.S.Costello, Oncogene, 1996, 13, str-2605]. Similar studies have demonstrated the important role of Syk in signal transmission in the BCR and TCR [A.M.Cheng, Nature, 1995, 378, str-306, (1995) and D.H.Chuet. al., Immunological Reviews, 1998, 165, str-180]. Apparently, Syk is also involved in ensuring the survival of eosinophils in response to IL-5 and GM-CSF [S.Yousefiet. al., J. Exp. Med., 1996, 183, SCR-1414]. Despite the key role of Syk in signal in the fat cells, BCR and T-cells, in the literature there is little information about the mechanism by which Syk transmits a signal subsequent to the effectors. It was shown that two adaptronic protein BLNK (linker protein in B-cells, SLP-65) and SLP-76 are substrates of Syk in B cells and mast cells, respectively, and who were studioflash their intermediate role in the chain between Syk and downstream effectors [M.Ishiai et. al., Immunity, 1999, 10, str-125 and L.R.Hendricks-Tayloret. al., J.Biol. Chem, 1997, 272, str-1367]. In addition, it is obvious that Syk plays an important role in the transmission channel signal CD40, which is essential for proliferation of b cells [M.Fariset. al., J.Exp. Med., 1994, 179, str-1931].

Syk is also involved in platelet activation stimulated by low-affinity receptor for IgG (Fc gamma RIIA) or collagen [F.Yanagaet. al., Biochem. J., 1995, 311, (Pt. 2) str-478].

The focal adhesion kinase (FAK) refers to preceptory the tyrosine kinases involved in signal channel-mediated integrin. FAK associates with integrins at sites of focal contacts, and it was shown that FAK activation and phosphorylation of tyrosine in many types of cells depends on integrins associated with their extracellular ligands. The results of several studies argue in favor of the hypothesis that FAK inhibitors can be used to treat cancer. For example, FAK-deficient cells weakly migrate in response to chemotactic signals, and overexpression of the C-terminal domain of FAK inhibits the proliferation of cells and chemotactic migration (Sieget. al., J. Cell Science, 1999, 112, 2677-2691; Richardson A. and Parsons, T., Cell, 1997, 97, 221-231); in addition, tumor cells treated antisense oligonucleotide-FAK, lose the ability to bind and undergo apoptosis (Xuet. al., Cel Growth Differ. 1996, 4, 413-418). It was reported overexpression of FAK in the case of prostate cancer, breast cancer, thyroid, colon and lung. The level of FAK expression directly correlates with tumors exhibiting the most aggressive phenotype.

Angiogenesis, or the formation of new blood vessels due to the proliferation of pre-existing vascular network plays a Central role in embryonic development and organogenesis. Anomalous enhanced neovascularization observed in rheumatoid arthritis, diabetic retinopathy and tumor growth (Folkman, Nat. Med., 1995, 1, 27-31.). Angiogenesis is a complex multistep process, which involves the activation, migration, proliferation and survival of endothelial cells. Detailed research in the field of angiogenesis of tumors in the last two decades has identified a number of therapeutic targets, including kinases, proteases and integrins, which led to the discovery of many new antiangiogenic agents, including inhibitors of KDR; some of them are currently undergoing clinical trials (Jekunen,et. al.Cancer Treatment Rev. 1997, 23, 263-286.). Inhibitors of angiogenesis can be used as main, auxiliary or even prophylactic drugs against the formation or re-growth of malignant tumors.

In yeast and Drosophila was what Yavlena several proteins, involved in the separation of chromosomes and the formation of the spindle. Disturbances in the functions of these proteins lead to incorrect separation of chromosomes and the formation of monopolar or destroyed spindles. These kinases are Ipl1 and euroracing from S.cerevisiae and Drosophila, respectively, which is necessary for the separation of centrosomes and segregation of chromosomes. In several laboratories has recently been cloned and characterized one of the human homolog of the yeast Ipl1. This kinase, called Aurora2, or STK15 BTAK, belongs to the family of serine/trainingin. In Bischof (Bischoff) with staff, it was shown that Aurora2 is oncogenic and amplificates for colorectal cancer man (EMBO J, 1998, 17, 3052-3065). She was also described in the case of cancer with the formation of epithelial tumors, such as breast cancer.

Currently, we found a new substituted azaindole having valuable pharmaceutical properties, in particular the ability to inhibit protein kinases, namely, the ability to selectively inhibit Syk kinase. This isoindoline derived like compounds disclosed in U.S. patent 6770643, however, in this patent there is no specific information about it.

BRIEF description of the INVENTION

The present invention relates to a compound of the formula I:

its pharmaceutically acceptable salt or prodrug or MES of such a compound, its salt or its prodrug.

The invention also relates to pharmaceutical compositions containing a compound of formula I, and the method of using the compounds of formula I for the treatment or prevention of a physiological condition associated with the presence of Syk to the patient.

The invention also relates to a method for obtaining compounds, which is an intermediate compound and is useful for obtaining the compounds of formula I.

In another aspect the present invention relates to pharmaceutical compositions containing compounds of General formula (I):

the corresponding N-oxides and prodrug; and a pharmaceutically acceptable salt and MES (e.g., hydrates) of such compounds; and N-oxides and prodrug; together with one or more pharmaceutically acceptable carriers or excipients.

BRIEF DESCRIPTION of FIGURES

FIGURE 1. The reaction scheme for obtaining compounds of formula I.

FIGURE 2. The average diameter of the hock joint of rats after injection of collagen type II from bovine nasal cartilage in incomplete Freund Freund and introduction A003397769, the compounds of formula I (3,0; 10 or 30 mg/kg 2 R/d) from day 6 to day 21. The average diameter of the hock female LEW rats sensitized by subcutaneous inye is the function of collagen in incomplete Freund Freund (400 μg/400 μl/rat) on day 0 and 7. Animals received a dose between 6 and 21 days.

FIGURE 3. Analysis of bone erosion (the ratio of the area of the bone to its volume) in the calcaneus CIA rats treated with A003397769 (3,0; 10 or 30 mg/kg).

FIGURE 4. The average diameter of the hock joint of rats after injection of collagen type II from bovine nasal cartilage in incomplete Freund Freund and introduction only A003397769 (3.0 mg/kg) or in combination with methotrexate (0.1 or 0.2 mg/kg) from day 6 to day 20 or 21 and in comparison with the animals treated with the carrier. The impact of the SYK inhibitor, A003397769, (3.0 mg/kg 2 R/d) in rat CIA in terms monotherapy or in combination with methotrexate.

FIGURE 5. The average diameter of the hock joint of rats after injection of collagen type II from bovine nasal cartilage in incomplete Freund Freund and introduction only A003397769 (10 mg/kg) or in combination with methotrexate (0.1 or 0.2 mg/kg) from day 6 to day 20 or 21 and in comparison with the animals treated with the carrier. The impact of the SYK inhibitor, A003397769, (10 mg/kg 2 R/d) in rat CIA in terms monotherapy or in combination with methotrexate.

6. Analysis of bone erosion (the ratio of the area of the bone to its volume) in the calcaneus CIA rats treated only A003397769 (3.0 or 10 mg/kg) or in combination with methotrexate (0.1 or 0.2 mg/kg).

7. The average diameter of the hock joint of rats after injection of collagen type II from bovine nasal cartilage in incomplete Freund's Froin the and and the introduction of A003397769 A (10 or 30 mg/kg 2 R/d) from day 12 to day 21. The average diameter of the hock female Lewis rats sensitized by subcutaneous injection of collagen in incomplete Freund Freund (400 μg/400 μl/rat) on day 0 and 7. The average for both legs.

FIG. Analysis of bone erosion (the ratio of the area of the bone to its volume) in the calcaneus CIA rats treated with A003397769 A (10 or 30 mg/kg 2 R/d).

FIG.9. The effect of the compounds of formula I on the weight of the rats.

FIGURE 10. The effect of the compounds of formula I at a concentration of hemoglobin.

A DETAILED DESCRIPTION of the INVENTION

Thus, in one aspect the present invention relates to pharmaceutical compositions containing compounds of General formula (I):

also known as: 2-[4-(7-ethyl-5H-pyrrolo[2,3-b]pyrazin-6-yl)-phenyl]-propan-2-ol.

In the present description assumes that the term "compounds of the present invention" and similar expressions include the above-described herein, the compound of General formula (I), which expression includes the prodrugs, pharmaceutically acceptable salt and solvate, e.g. hydrates, where permitted by the context. Similarly, the reference to intermediate compounds, regardless of whether they are themselves the subject of the present invention extends to their salts and solvate, where permitted by the context. To clarify specific cases, Engl is permitted by the context, sometimes noted in the text, however, these cases are only illustrative and do not exclude other cases, if permitted by the context.

Abbreviations:

ATP - adenosine triphosphate;

DTT - dithiothreitol;

PBS - saline solution in phosphate buffer.

Used above and throughout the description of the invention, the following terms, unless otherwise indicated, have the following meanings:

"Patient" means humans and other mammals.

"Prodrug" means a compound thatin vivoconverted by metabolic means (e.g. by hydrolysis) to a compound with the formula (I), including its N-oxides. For example, the ester compounds of formula (I)containing a hydroxyl group can be converted by hydrolysisin vivoin the original molecule. Alternatively, the ester compounds of formula (I)containing a carboxy group can be converted by hydrolysisin vivoin the original molecule.

Suitable esters of compounds of formula (I)containing a hydroxyl group, are for example acetates, citrates, lactates, tartratami, malonate, oxalates, salicylates, propionate, succinate, fumarate, maleate, methylene-bis-β-hydroxynaphthoate, gentisate, isethionate, di-p-toluoyltartaric, methansulfonate, econsultancy, bansilalpet, p-toluensulfonate, cyclohexylsulfamate and hinata.

A particularly useful class of esters of compounds of formula (I)containing a hydroxyl group can be obtained from the acid compounds selected from those described in the work Bundgaardet. al., J. Med. Chem., 1989, 32, str-2507, and includes substituted (aminomethyl)-benzoate, such as dialkylaminomethyl, in which the two alkyl groups may be combined together (or separated by an oxygen atom or a possibly substituted by a nitrogen atom, for example alkilirovanny a nitrogen atom, more precisely (morpholinomethyl)benzoate, for example 3 - or 4-(morpholinomethyl)benzoate, and (4-alkylpiperazine-1-yl)benzoate, for example 3 - or 4-(4-alkylpiperazine-1-yl)benzoate.

Some compounds of the present invention are basic, and such compounds are useful in the free base form or in the form of pharmaceutically acceptable salts obtained by addition of such compounds.

Salts formed when joining acids, more convenient for use; and in practice, use of salt, in essence, is to use the forms of the free base. Acids which can be used to prepare salts include preferably those which, due to their reaction with the free base form of pharmaceutically acceptable salts, i.e. salts, the anions of which are not toxic to the patient in pharmaceutical doses of the salts, and put the AUX inhibitory effect inherent in the free base is not broken side effects attributed to the anions. Although pharmaceutically acceptable salts of these basic compounds are preferred, all salts formed by addition of acids, useful as sources of the free base forms, even if the particular salt, per se, are only required as intermediate product as, for example, when the salt is formed only for purposes of purification and identification, or when it is used as intermediate compounds in the preparation of pharmaceutically acceptable salt by ion exchange processes. Pharmaceutically acceptable salt, covered by the present invention include salts derived from mineral acids and organic acids, and include hydrohalide, such as hydrochloride and hydrobromide, sulfates, phosphates, nitrates, sulfamate, acetates, citrates, lactates, tartratami, malonate, oxalates, salicylates, propionate, succinate, fumarate, maleate, methylene-bis-b-hydroxynaphthoate, gentisate, isethionate, di-p-malwarecity, methansulfonate, econsultancy, bansilalpet, p-toluensulfonate, cyclohexylsulfamate and hinata.

In addition to the use as active compounds, salts of the compounds according to the invention can be used for purification of the compounds, on the example on the basis of the difference in solubilities between the salts and the parent compounds, by-products and / or source materials, methods, well known to specialists in this field.

Compounds of the present invention have useful pharmacological activity and accordingly are entered into pharmaceutical compositions and used in the treatment of patients suffering from certain medical disorders. Thus, the present invention provides, in accordance with a further aspect, the compounds of the invention and compositions containing compounds of the invention for use in therapy.

Compounds covered by the present invention, inhibit or block the catalytic activity of the kinase, in accordance with the tests described in the literature, and proceduresin vitroas described below, the results of which should correlate to pharmacological activity in humans and other mammals. Thus, in one implementation of the present invention represented by the compounds forming the subject of the present invention and compositions containing them, which can be used in the treatment of patients suffering from disease or susceptible to diseases, which can facilitate the introduction of inhibitors of protein kinase (e.g., Syk, FAK, KDR or Aurora2). For example, the compounds of the present invention are useful in the treatment of inflammatory diseases is any, such as asthma; inflammatory dermatoses (e.g., psoriasis, dermatitis herpetiformis, eczema, and necrotic cutaneous vasculitis, bullous disorders); allergic rhinitis and allergic conjunctivitis and inflammation of the joints, including arthritis, rheumatoid arthritis and other arthritic conditions such as rheumatoid spondylitis, gouty arthritis, traumatic arthritis, rubella arthritis, psoriatic arthritis and osteoarthritis. These compounds are also useful in the treatment of chronic obstructive pulmonary disease (COPD), acute of invoice, autoimmune diabetes, autoimmune encephalomyelitis, colitis, atherosclerosis, peripheral vascular disease, cardiovascular disease, multiple sclerosis, restenosis, myocarditis, B-cell lymphoma, systemic lupus erythematosus, reactions, transplant rejection and other cases of transplant rejection, various forms of cancer and tumors, such as colorectal cancer, prostate cancer, breast cancer, thyroid cancer, colorectal cancer and lung cancer) and inflammatory bowel disease. These compounds are also useful as antiangiogenic drugs for the treatment of tumors. In addition, the compounds of the present invention are useful as drugs for the control of tumor cells.

The special is the first implementation of therapeutic methods, are the subject of the present invention is the treatment of joint inflammation.

Additional special implementation of therapeutic methods that are the subject of the present invention is the treatment of rheumatoid arthritis.

A special implementation of therapeutic methods that are the subject of the present invention is the treatment of cancers, tumors and other proliferative disorders.

Another specific application of therapeutic methods that are the subject of the present invention is the treatment of cancer, accompanied by the formation of liquid tumors.

Additional special implementation of therapeutic methods that are the subject of the present invention is the treatment of lymphoma cells in the mantle zone.

Additional special implementation of therapeutic methods that are the subject of the present invention is the treatment of disorders by inhibiting angiogenesis.

In accordance with another item of the present invention proposes a method of treatment of a patient (human or animal)suffering from disease or susceptible to diseases that may be alleviated by the introduction of an inhibitor of protein kinase (e.g., Syk, FAK, KDR or Aurora2), for example, conditions described above, including the introduction of the patient is the effective number of connections, leaving the subject of the present invention, or preparation containing the compound of the present invention. By "effective amount" refers to the number of connections that constitute the subject of the present invention, which is effective as an inhibitor of the catalytic activity of protein kinases such as Syk, FAK, KDR or Aurora2, and thus able to produce the desired therapeutic effect.

Included in this description reference to the treatment applicable to preventive therapy and treatment diagnosed pathological conditions.

The present invention also extends to pharmaceutical compositions comprising the compound forming the subject of the present invention, in combination with a pharmaceutically acceptable carrier or excipient.

The compound of the present invention may be administered in any suitable way. In practice, the connection of the present invention may, in General, be administered parenterally, topically, rectally, orally or by inhalation; in particular, orally or by inhalation.

In accordance with the present invention, the preparations can be prepared by conventional methods using one or more pharmaceutically acceptable excipients or fillers. Helper is idestam, among other things, include diluents, sterile aqueous medium and various non-toxic organic solvents. The compositions can be prepared in the form of tablets, pills, granules, powders, aqueous solutions or suspensions, injectable solutions, elixirs or syrups, and may contain one or more agents selected from the group consisting of sweeteners, flavors, colorants and stabilizers to obtain pharmaceutically acceptable preparations. The choice of media and the content of the active substance in the carrier is usually determined in accordance with the solubility and chemical properties of the active compounds, the specific route of administration and the provisions to be observed in pharmaceutical practice. For example, for the preparation of tablets can be used fillers such as lactose, sodium citrate, calcium carbonate, dicalcium phosphate and dezintegriruetsja agents, such as starch, alginic acid and certain complex silicates in combination with sliding agents such as magnesium stearate, sodium lauryl sulfate and talc. For the preparation of capsules is convenient to use the lactose and polyethylene glycols of high molecular weight. When using aqueous suspensions they may contain emulsifying agents or substances that contribute to the formation of suspensions. You can also use razbam the teli, for example, sucrose, ethanol, polyethylene glycol, propylene glycol, glycerin and chloroform, or a mixture thereof.

For parenteral applications, the use of emulsions, suspensions or solutions of the compounds of the invention in vegetable oils, such as sesame, peanut, or olive, or aqueous-organic solutions, such as water and propylene glycol, injectable organic esters, such as, for example, etiloleat, along with a sterile aqueous solution of the pharmacologically acceptable salts. Saline solutions of the compounds used in the present invention are particularly suitable for administration by intramuscular or subcutaneous injection. Aqueous solutions, including solutions of salts in pure distilled water can be used intravenously provided that their pH is brought to an appropriate level that they are buffered appropriately and made isotone adding a sufficient amount of glucose or sodium chloride, and that they are sterilized by heat treatment, irradiation or filtration.

For local application can be used gels (water or alcohol based), creams or ointments containing compounds of the invention. Compounds of the invention can also be included in the gel or matrix base for application in a patch, which will provide controlled wisweb the Denia connection through the transdermal barrier.

When used in inhalation the compounds of the invention can be dissolved or are in suspension in a suitable carrier for use in spray or aerosol spray can, or may be absorbed or adsorbed on a suitable solid carrier for use in a dry powder inhaler.

Solid compositions for rectal injection include suppositories prepared in accordance with known methods and containing at least one component of the invention.

The percentage of active ingredient in the compositions of the invention can vary, it is necessary to contain such a proportion that would ensure the appropriate dosage.

Obviously, several standard dosage forms may be administered at about the same time. The dose administered will be determined by the physician and depends on the desired therapeutic effect, the route of administration and duration of treatment, and the patient's condition. For adults doses typically range from about 0.001 to about 50, preferably from about 0.001 to about 5 mg/kg of body weight when introduced by inhalation, from about 0.01 to about 100, preferably from 0.1 to 70, more precisely from 0.5 to 10 mg/kg of body weight per day in oral administration, and from priblizitel is but about 0.001 to about 10, preferably from 0.01 to 1 mg/kg of body weight per day intravenously. In each case, the dose will be determined in accordance with factors for treatment of the patient, such as age, weight, General health, and other factors that may affect the efficacy of the pharmaceutical composition.

In accordance with the invention compounds can be administered as often as needed to achieve the desired therapeutic effect. Some patients may show a rapid response to a higher or lower dose, and these may be sufficient much weaker dose. For other patients may need long-term treatment with a frequency of from 1 to 4 doses per day, in accordance with the physiological needs of each individual patient. In General, the active substance can be applied orally 1 to 4 times a day. Of course, some patients will need to register no more than one or two doses per day.

Compounds of the present invention can be obtained by application or adaptation of known methods, by which is meant methods used above or described in the literature, for example, described in the paper by R.C. Larock, Comprehensive Organic Transformations, VCH publishers, 1989.

In the following R what actions may be necessary to protect reactive functional groups, for example, hydroxyl group, amino group, aminogroup, tigroup or carboxy groups, where these groups are required in the final product, to avoid their unwanted participation in the reactions. Can be used conventional protective groups in accordance with the accepted practice examples, see T.W. Greene and P. G. M. Wuts in "Protective Groups in Organic Chemistry," John Wiley and Sons, 1991.

It is obvious that the compounds forming the subject of the present invention may contain asymmetric centers. These asymmetric centers can independently from each other to be in the R - or S-configuration. For professionals in the field it is obvious that certain compounds forming the subject of the present invention may also exhibit geometric isomerism. It should be understood that the present invention applies to the individual geometrical isomers and stereoisomers and mixtures thereof, including racemic mixtures of the above compounds with formula (I). Such isomers can be distinguished from their mixtures by known methods or their modifications, e.g. chromatographic methods or methods of recrystallization, or to obtain separately from the corresponding isomers of the intermediate compounds.

According to another point of the invention formed by adding acid salt compounds that are the subject of the present invention, which can be obtained by reaction of the free base with the appropriate acid using known methods or their modifications. For example formed by adding acid salt compounds that are the subject of the present invention, can be obtained either by dissolving the free base in water, aqueous solution of ethanol or other suitable solvents containing the appropriate acid and the release of salt by evaporating the solution, or the reaction of the free base and acid in an organic solvent, where the salt precipitates or evaporated from the solution.

The compounds forming the subject of the present invention can be regenerated from the salts formed when joining acids by known methods or their modifications. For example, the parent compound forming the subject of the present invention can be regenerated from the salts obtained in accession, acid, alkali, for example aqueous sodium bicarbonate solution or aqueous solution of ammonia.

The compounds forming the subject of the present invention can be regenerated from the salts, obtained by joining the Foundation, using known methods or their modifications. For example, the parent compound forming the subject of the present invention can be regenerated from the salts, obtained by joining the Foundation, by treatment with an acid, for example hydrochloric acid.

The compounds forming the subject on the present invention, can be easily obtained or formed in the process constituting the subject matter of the present invention, as the solvate (e.g. hydrate). Hydrates of the compounds that are the subject of the present invention can be easily obtained by recrystallization from a mixture of water/organic solvent such organic solvents as dioxane, tetrahydrofuran or methanol.

According to another distinctive feature of the invention formed by adding the base salts of the compounds constituting the subject matter of this invention, can be obtained by the reaction of the free acid with the appropriate base by means of known methods or their modifications. For example formed by adding the base salts of the compounds constituting the subject matter of this invention, can be obtained either by dissolving the free acid in water or an aqueous solution of ethanol or other suitable solvents containing the appropriate base, and the release of salt by evaporating the solution, or the reaction of the free acids and bases in an organic solvent, where the salt precipitates or evaporated from the solution.

Raw materials and intermediate compounds can be obtained by application or modification of known methods, for example methods described in PR the least or their obvious chemical equivalents.

The present invention is further illustrated by, among other things, the following illustrative example.

The spectra of nuclear magnetic resonance (NMR) 300 MHZ1H were recorded on instruments Varian Mercury. In the spectra of nuclear magnetic resonance (NMR) magnitude of chemical shift (δ) are quoted in ppm (MD) on tetramethylsilane. Abbreviations have the following meanings: "C" = singlet, d = doublet; t = triplet; kV = Quartet; m = multiplet, "DD" = doublet of doublets, DDD = doublet of double doublets.

Experiments on liquid chromatography high pressure and mass spectrometry (GHMC) to determine retention time (RT) and associated mass ions was carried out using one of the following methods. Mass spectra (MS) are recorded on a time-of-flight mass spectrometer Micromass LCT. The method includes the ionization elektrorazpredelenie positive ions and scan mass m/z 100 to 1000. Liquid chromatography is performed using a binary pump and degasser Agilent™ series 1100; stationary phase: column Phenomenex Synergi™ 2µ Hydro-RP 20X4,0 mm, mobile phase: A=0.1% of formic acid (FA) in water, B=0,1% FA in acetonitrile. Volume of 5 μl using system CTC Analytical PAL. The flow rate is 1 ml/min Gradient ranging from 5% B to 90% B over 3 minutes, and from 90% B to 100% B C is 2 minutes. Auxiliary detector: UV detector Agilent 1100 series, wavelength =220 nm and evaporative light scattering detector (ELS) Sedere SEDEX™ 75, detector temperature =46°C, the nitrogen pressure= 4 bar.

The retention time thin-layer chromatography (TLC) RF was determined using tablets with silica gel Merck™.

EXAMPLE 1

2-[4-(7-ethyl-5H-pyrrolo[2,3-b]pyrazin-6-yl)-phenyl]-propan-2-ol

Just got 6.0 g of 2-[4-(7-ethyl-5H-pyrrolo[2,3-b]pyrazin-6-yl)phenyl]propan-2-ol (compound 1) in two steps from n-propylpyrazine (compound 2) and 4-acetylbenzoate (compound 3).

Synthesis of compound 1 was carried out as follows: the combination ofn-propylpyrazine (compound 2) and 4-acetylbenzoate (compound 3) with bis(trimethylsilyl)-amidon sodium in tetrahydrofuran at 40°C gave the intermediate compound 4 with 30%yield. Reaction of compound 4 with chloride Metalmania in tetrahydrofuran at 0°C gave the desired compound 1 with a yield of 74% after recrystallization from 2-propanol. The synthesis is shown in figure 1.

EXPERIMENTAL PART

2-[4-(7-ethyl-5H-pyrrolo[2,3-b]pyrazin-6-yl)phenyl]alanon (compound 4). Solutionn-propylpyrazine (compound 2, 912 mg, 7,46 mmol) in tetrahydrofuran (5 ml) was added by pipette for seven minutes to a solution of bis(trimethylsilyl)amide, sodium (2M solution in THF; 13 ml, 26 mmol, 3.5 equiv.) at 20°C. the Obtained solution is dark purple-red is the main color, and the temperature dropped to 16.4°C. Solution of 4-acetylbenzoate (compound 3, 1.08 g, 7.4 mmol) in tetrahydrofuran (5 ml) was added over 18 minutes of 13.3°C. the Temperature dropped to 12.6°C, to obtain the solution brown. The mixture was stirred at room temperature for 1 hour, heated to approximately 35°C for 6 hours and then left to mix at room temperature for about 60 hours. The mixture was poured into saturated aqueous sodium bicarbonate solution (200 ml) and was extracted with ethyl acetate (2×150 ml). The ethyl acetate was washed with water (100 ml) and concentrated on the device Bbchi with a bath temperature of 40°C and at a pressure of from 80 to 10 Torr to obtain a solid dark yellow color. The solid was used in the powder in diethyl ether (25 ml), was filtered and was washed in ether (25 ml). The solid was air-dried to obtain 600 mg (30,3%) of compound 4 in the form of a solid yellow:

1H NMR (300 MHz, CDCl3, figure 1) δ 8,5 (1H, d, J=2 Hz), 8,15 (2H, d, J=8 Hz), and 8.1 (1H, d, J=3 Hz), and 7.9 (2H, d, J=8 Hz), 3,1 (2H, square, J=9 Hz), 2,7 (3H, s), 1,4 (3H, t, J=9 Hz).

1-[4-(7-ethyl-5H-pyrrolo[2,3-b]pyrazin-6-yl)phenyl]propan-2-ol (compound 1). To a chilled (~5°C) solution of chloride Metalmania (3M in THF; 81,3 ml, 244 mmol, 10 equiv.) in tetrahydrofuran (116 ml) was added a solution of compound 4 (6.5 g, 24.4 mmol) in tetrahydrofuran (348 ml) Pipa is coy over 90 minutes, maintaining the temperature around 0°C as adding solution. In the process of adding observed the formation of a bright yellow solution. After one hour, TLC (ethyl acetate/n-heptane 1/1) showed the presence of starting material, and a new point is moved at a lower Rf. The party has repaid by careful addition of saturated aqueous sodium bicarbonate solution (about 660 ml). To the resulting thick mass was added ethyl acetate (250 ml) and water (250 ml). The aqueous phase was removed and extragonadal with ethyl acetate (2×250 ml). Fractions of ethyl acetate were combined, washed with water (2×200 ml) and concentrated on the device Bbchi with a bath temperature of 40°C and at a pressure of from 80 to 10 Torr to obtain 7.5 g (109%) 1 in the form of a solid light beige color:1H NMR (DMSO-d6, figure 2) δ 12,0 (1H, s), 8,35 (1H, d, J=3.5 Hz), and 8.2 (1H, d, J=3.5 Hz), and 7.6 (4H, s) to 5.1 (1H, s), 2,9 (2H, square, J=8 Hz), 1,5 (6H, s), 1,3 (3H, t, J=8 Hz).

This material was combined with material from a previous experiment. The combined material (8.5 g) was heated under reflux with 2-propanol (150 ml) to obtain a clear solution is light brown in color, which was filtered while hot through a heated Buchner funnel under vacuum. In the flask to filter formed precipitate, and the mixture was heated to the boiling temperature to obtain a transparent solution. The material of the cooling gap is Ali to room temperature while stirring to obtain a solid of light yellow color, which was filtered, washed with cold 2-propanol and dried in a vacuum oven at 50°C to obtain 6.0 g (71%) of compound 1 in the form of solid light yellow: LC/MS:RT=2,55 min, MS: 282 (M+H);1H NMR (DMSO-d6, figure 3) δ 12,0 (1H, s), 8,35 (1H, d, J=3 Hz), and 8.2 (1H, d, J=3 Hz), and 7.6 (4H, s) to 5.1 (1H, s), 2,9 (2H, square, J=9 Hz), 1,5 (6H, s), 1,3 (3H, t, J=9 Hz).

Elemental analysis of compound 1 shown in Table 1.

THEORETICAL INTEREST

SHARE ELEMENTS: C 72,57%, H for 6.81%, N 14,93%, O 5,69%

EXPERIMENT 1: C 72,46%, H 7,05%, N 15,07%

Table 1
The PROCEDURE of IN VITRO TESTING FOR SYK
The object of the analysis:Tyrosine(Y)kinase spleen
Short name:Syk
The format of the analysis:Phosphorylation of the substrate
Format detectionFlashPlate with streptavidin
ModulationInhibition

FlashPlate microplates Plus coated with streptavidin (PerkinElmer Life Sciences™) are designed to perform radiometric analysis within the Board is and. The inner surface of each hole sustainable covered with a thin layer of scintillator on the basis of polystyrene, which caused covalently-linked layer of streptavidin. These tablets are suitable for a wide range of analytical applications that use biotinylated molecules invaders. Poly (Glu,Tyr) 4:1 (PGT) is a statistical copolymer, which can act as a substrate for the tyrosine-specific protein kinases. The analysis assesses the degree of phosphorylation of the substrate PGT-Biotin by Syk. The enzyme transfers [γ33P]-phosphate from [γ33P]-ATP on the polymeric substrate. The analysis was carried out in solution in neisvaziuosiu 384-well pad. After stopping the reaction with phosphoric acid, the reaction mixture was transferred to 384-well streptavidin tablet FlashPlate. Biotinylated substrate was recorded on the tablet, and other reagents, including hot ATP, washed with water. Then he measured the radioactivity in each well.

The enzymatic reaction was carried out in neisvaziuosiu 384-well pad. The final concentration of reagents in each well were: to 7.77 nm Syk, 15,5 nm substrate PGT-Biotin, 0.1 MCI33P-ATP, 50 mm Tris, pH 7.5, 10 mm MgCl2, 3 mm MnCl2, 1 mm DTT, 0.1 mg/ml γ-globulin. The reaction volume was 22 mm. Reaction time: 120 minutes. Temperature is RA: room temperature. The reaction was stopped by adding 20 μl of 9%phosphoric acid, and 30 μl of reaction mixture was transferred into a 384-well streptavidin tablet FlashPlate. After 90 minutes incubation at room temperature the tablet was washed with 0.02% Tween-20 in 50 mm Tris, pH 7.5. The expense of radioactivity led to the scintillation counter Topp Count™.

Diluted solutions of the enzyme prior to use prepared and kept on ice.

Freshly prepared solutions of MnCl2and DTT was added to the analytical buffer immediately prior to use.

Materials for analysis are listed in table 2.

td align="center"> for 125.8
Table 2
MaterialsProviderCatalog numberMol. weightPurpose
1 M Tris, pH 7.5FisherBP1757-500The buffer
1 M MgCl2SigmaM-1028for 95.2Cofactor of the enzyme
1 MnCl2SigmaM-1787Cofactor of the enzyme
DTTSigmaD-5545154,2Antioxidant
γ-GlobulinsSigmaG-5009The stabilizer protein
DMSOSigma-Aldrich15 493-9Solvent
StaurosporinSigmaS-4400466,5Standard inhibitor
Phosphoric acid (85%)SigmaP-656098,0Stop solution
1 MCI [γ is 33P]-ATPPerkinElmerNEG6xxSubstrate
10× PBS, pH 7,4FisherBP399-1Wash buffer
Tween 20FisherBP337-5001227,54Detergent
384-well polypropylene tabletCorning3657Tablet for connection
Nswazwi 384-well plateCorning3652The reaction tablet
384-well streptavidin tablet FlashplatePerkinElmerSMP-410 (A)The tablet takeover
Sealant tape Top Seal APackard6005185Sealing tablet
Washing machine Elx405Bio-TekWashing tablet
Top CountPackardAccounts the chick
Station FXBeckmanA dispenser for liquids
Beckman 2000BeckmanA dispenser for liquids

Enzyme:

Syk with FLAG-epitope (0,184 mg/ml, MW=35531,81 Amu) was obtained and purified using known in the art methods.

Substrate:

Biotin-conjugated Poly (Glu,Tyr) 4:1 purchased from Cisbio international™ (catalog No. 61GT0BLB, batch No. 16).

Used solutions for the analysis are given in table 3.

Table 3
ReagentChemicalsSolventConcentration
Analytical bufferTris, pH 7.5H2O50 mm
Newly added MnCl2and DTTMnCl23 mm
MgCl2 10 mm
DTT1 mm
γ-globulins0,1%
The solution of enzyme and substrateSykAnalytical bufferto 7.77 nm
Keep in nesvezhih tabletsThe substrate PGT-Biotin15,5 nm
A solution of ATP/33P-ATPATP/33P-ATPAnalytical bufferof 0.1 µci/well
Stop solutionPhosphoric acidH2O9%
Wash bufferTween -201xPBS, pH 7,40,02%

Dilution connection:

1. Compounds were received as 10 mm/well of a solution of 10 μl/well in 100% DMSO in 96-well polypropylene plate with U-image of the second bottom with blank near H. Added 90 μl/well of 100% DMSO to obtain 100 μl of 1 mm solution of the compound was sealed and kept the plate at room temperature in the dark.

2. Preparing the final tablet: in 384-well polypropylene plate with a round bottom (the tablet to store Corning) was added to 23 μl/well H2O to columns 3 and 13, 20 ál/well of 30% DMSO to columns 4 through 12 and columns 14 through 22 (leaving blank rows O and P).

3. Was prepared by dilution of the compounds (10 dilutions, 300 μm, 100 μm, 30 μm, and so on) on a Biomek 2000™ analysis software kinase: using a multichannel pipette with spouts 20 µl removed 8th (number N) transferred 10 μl/well of 1 mm connections from column 1 in the original tablet in column 3 in the final tablet to prepare the first dilution to 300 μm. Then in the final tablet was mixed and transferred 10 μl/well of a dilution of 300 μm from column 3 to column 4 for the preparation of dilution of 100 microns. Mixed and transferred 10 μl/well of a dilution of 100 μm from column 4 to column 5, to obtain a dilution of 30 μm etc. Prepared duplicate dilutions for each connection, for example, endured 10 μl/well connections from A1 original tablet in A3 and B3 in the final tablet. Mixing and transfer were repeated. Then transferred to 10 μl/well of 1 mm connections from column 2 in the original tablet in column 13 in the final tablet for sentence on what the service dilution 300 μm. Then in the final tablet was mixed and transferred 10 μl/well of a dilution of 300 μm from the column 13 column 14 for the preparation of dilution 100 microns, etc. From one full 96-meadow original tablet with a blank next N you can cook up to six of the final tablets.

4. Prepared standard maternal tablet to connect: added 5 μl/well of 10 mm solution roscovitine in 100% DMSO into the ranks of H1 and H2 in 96-well polypropylene plate with a U-shaped bottom, then diluted to 1 mm solution of 45 μl/well of 100% DMSO.

5. Prepared standard tablet for drawing connections: in 384-well polypropylene plate with a round bottom (the tablet to store Corning™) was added to 23 μl/well H2O to columns 3 and 13, 20 ál/well of 30% DMSO to columns 4 through 12 and columns 14 through 22 (leaving blank rows O and P).

6. Prepared standard dilution of compounds (10 dilutions, 300 μm, 100 μm, 30 μm, and so on) on a Biomek 2000™ using standard analysis software: single 20 ál pipette bore 10 μl/well of 1 mm standard connection from the column N1 in maternal tablet in column O1 in the final tablet to prepare the first dilution to 300 μm. Then in the final tablet was mixed and transferred 10 μl/well of a dilution of 300 μm from the column O3 column O4 for the preparation of dilution of 100 microns. Mixed and transferred 10 ál/Lou is ka dilution of 100 μm from the column O4 in column A5, to obtain a dilution of 30 μm etc. Prepared duplicate dilutions for each standard compound, for example, endured 10 µl/hole connection from N1 maternal tablet in O3 and P3 in the final tablet. Mixing and transfer were repeated. Then transferred to 10 μl/well of 1 mm compound from the column of H2 in maternal tablet A13 column end tablet for preparation of dilution 300 μm. Then in the final tablet was mixed and transferred 10 μl/well of a dilution of 300 μm from the column A13 column A for preparation of dilution 100 μm etc.

7. In the final tablet standard compounds were added 20 μl/well of 30% DMSO (upper control) from A to H and 20 µl/hole 45% H3PO4(lower control) from I to J in column 23.

Order analysis:

1. In the analysis tablet (nswazwi 384-well plate, Corning™) was added 10 μl of an enzyme solution and substrate, 2 μl of test compounds were incubated at room temperature for 30 min (stage pre-incubation of the enzyme/compound).

2. The reaction was started by adding 10 μl of ATP solution/33P-ATP.

3. Incubated at room temperature for 120 minutes.

4. The reaction was stopped by adding 20 μl of stop buffer.

5. Bore 30 μl of the reaction mixture in 384-well streptavidin tablet Flashplate.

8. Incubated PR the room temperature for 90 minutes.

9. Double-washed streptavidin tablet Flashplate 100 μl/well proryvnym buffer using an automatic dispenser Elx405.

10. Sealed and read the tablet (40 sec/well) on a scintillation counter Top Count™.

*Analysis point 1 to point 5 was carried out using the station Biomek™ FX.

The equation for the processing of the curve when determining the IC50:

Y=lower limit+(upper limit - lower limit)/(1+10^((LogIC50-X)*Slope)).

X is the logarithm of concentration.

Y is the response.

Y begins to change from the lower limit and reaches the upper limit; the curve has a sigmoidal shape.

This approach is identical to the "four-parameter logistic equation.

According to the results of this analysis, the concentration of the IC50in connection with formula I totaled 1.7 nanomoles.

ANALYSIS of the viability of HEMATOLOGICAL MALIGNANT CELL LINES USING MTS REAGENT

1. PURPOSE

This method is used to determine the viability of the cell line liquid tumors after treatment with the test compound. Tumor cells contain in suspension in the logarithmic growth phase. To-day use cells resuspended to a density of from 0.05 to 0.1 million/ml and incubated in 96-Lucena tablet with a test compound for 96 hours. Cell viability was measured by incubating cells with PE who Ghent MTS company Promega. Cell viability is proportional to the change in absorption at a wavelength of 490 nm. The effect of test compounds on cell viability is determined by comparing the absorbance of the control and treated by the connection of the cells as a percentage of the viability of control cells.

2. PROCEDURE

A. Materials

1. Cells:

Cell line liquid tumors were obtained either from the set of American Tissue Culture Collection or from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH).

2. The cellular environment:

Complete RPMI: medium RPMI-1640 with 25 mm HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) and L-glutamine (Gibco/Invitrogen™, no cat. 22400-089)+10% thermoinactivation fetal bovine serum (FBS) (Gibco/Invitrogen™, no cat. 16140-071)+1x penicillin/streptomycin (Gibco/Invitrogen, No. cat. 15070-063) + 50 μg/ml plasmion (Invivogen™, no cat. ant-mpt)

cRPMI without phenol red: medium RPMI-1640 without phenol red, L-glutamine (Gibco/Invitrogen™, no cat. Gibco/Invitrogen, 11835-030)+10% thermoinactivation fetal bovine serum (FBS) (Gibco/Invitrogen, No. cat. 16140-071)+1x penicillin/streptomycin (Gibco/Invitrogen™, no cat. 15070-063)

3. Other liquid reagents:

Reagent Promega™ MTS (cell water titer 96 No. cat. G358B)

Dimethyl sulfoxide (DMSO) (Sigma™, no cat. D2650)

4. Supplies:

Sterile 96-well polystyrene transparent obrabotan the e tablets for tissue cultures with lids (Falcon™, No. cat. 3072)

5. Equipment:

The reader 96-well plates (SpectraMAX GeminiEM™, Molecular Devices, USA)

B. Method

Day 1. Preparation of test compounds

Dissolved and serially diluted test compounds in DMSO at 1000-fold concentration.

Dilute test compound 1:100 in cRPMI medium without phenol red in sterile 96-well tablets for tissue culture.

12-channel pipette, transfer 10 μl of diluted compound in a clean 96-well plate to be used for incubation of the cells.

After adding 100 μl of cell cultures at final concentrations of the test compounds will be equal to 1.

Day 1. Preparation of liquid cells of the tumor to treatment with compounds

Collect the cells in complete RPMI from the culture with logarithmic growth phase at a density of about 0.3 to 0.7 million/ml to quantify cells and adjust their number to 0.1 million/ml in cRPMI without phenol red (for the fast-growing cells with a doubling time that is equal to or less than 24 hours, such as K562, use of 0.05 million/ml, to avoid excessive growth).

To move three times in 100 μl of cells in wells of 96-well transparent tablet to a total of 10 000 cells/well. Note that each well contains 10 μl of 10-fold concentrated under test the connection is to be placed, prepared, as shown above.

Incubation of cells

Incubate cells for 72-96 hours at 370°C in the tissue culture incubator in an atmosphere of 5% CO2.

C. Measurement: to determine the viability of cells

1. Thaw the reagent MTS company Promega and add 20 µl of each cell pipet dosing.

2. Mix the reagents in the wells by shaking the tablet and place the tablet in a tissue culture incubator at a temperature of 370°C.

3. Prepare a blank control without cells", adding the MTS reagent to the number of wells containing 100 μl of cRPMI without phenol red.

4. Incubate at 370°C until the absorbance at 490 nm for control cells will not be >1,5.

5. Stir cell culture shaking, in order to ensure uniformity of color in the holes, make sure there are no air bubbles. If they are present, the tablet centrifuged in a tabletop centrifuge at 1000 g to remove air bubbles.

6. To consider the absorption on the reader 96-well plates Molecular Devices.

D. Analysis of results:

1. Copy and paste the text data in the Excel spreadsheet.

2. Average idle data without cells and subtract the resulting value from the absorbance of each well containing cells.

3. To average the data triple holes, adjusted idle experience is and, and calculate the standard deviation of the fluctuations of reproducibility.

4. To average the data for control cells:

5. To calculate the average data for cells treated with the compound in percent from the averaged data for control cells as follows:

(The value for the treated cells/value of control)* 100=% of control values

The results:

We have established antiproliferative activity of the compounds of formula I. As shown in table 4, the compound with the formula I exhibits a stable activity against two cell lines LMB Jeko-1 and Granta-519. In addition, the connection with formula I exhibits a suppressive action on a relatively broad range of hematological cell lines (6 of 15).

Table 4
The impact of the connection Syk on the viability
IC50(µm)% inhibition at Max. conc. (10 μm)
DiseaseCell lineA003397769
N=4
A003397769
N=4
AMLHL-603,7; 8,; >10; >1035
AMLKG-11,991
AMLML-2>1028
In-ALLNalm-65,584
B-CLLJVM-23,5; 3,6; 7,7; >1060
B-CLLJVM-26,8; 7,0; 8,4; >1059
B-NHLDLCL-21,4; >10; >10; >100
B-NHLDOHH-24,575
CMLJurl-MK11,2; >10; >10; >1028
CMLK5623,9; >10; >10; >1021
LMBJeko-1 3,197
LMBGranta-5193,467
MML-3635,561
MMRPMI8226>1036
T-ALLJurkat6,8; >10; >10; >1043

INHIBITION of COLLAGEN-INDUCED ARTHRITIS IN RATS

Introduction

Collagen-induced arthritis (CIA) is a well-described model of rheumatoid arthritis (RA) of a person who can be induced in genetically susceptible rodents after immunization with collagen type II (cII) in Freund. Both CIA and RA lead to severe swelling/inflammation of the joints, synovial hyperplasia and erosion of cartilage and bone. This chronic inflammatory arthritis induced by immunization with cII, includes as a component of T-cells, as demonstrated in a weakened form CIA in T-deficient mice, and the component b-cells. For b-deficient mice, mice with X-linked immunodeficiency or mice with a null mutation in CXCR5 not develop CIA.

Methods:

And moisala and promotion: female Lewis rats were immunotherapies on day 0 and was stimulated at day 7 the introduction of cII from bovine nasal cartilage mixed with incomplete Freund adjuvant with the final concentration of collagen 1.0 mg/ml Animals in the base of the tail was administered to 400 μg cII.

The scheme of prevention of the introduction: the connection with formula I (3,0; 10 and 30 mg/kg) was administered orally (p/o) twice a day (2 R/d)from day 6 to day 21, inclusive.

Scheme combined introduction: rats were administered compound with the formula I (3.0 and 10 mg/kg, p/o 2 p/d) or methotrexate (MTX, 0.1 and 0.2 mg/kg, p/o 1 p/d) as monotherapy or in various combinations, a single dose of the compounds of formula I with a single dose MTX, starting from day 6 to day 21, inclusive, simultaneously or sequentially.

therapeutic scheme introduction: the connection with formula I (10 and 30 mg/kg, p/o 2 p/d) was administered orally from day 12 to day 21, inclusive.

Pathology of the joint: swelling of the joint was measured with rounding to the nearest 0.01 mm using an electronic compass. The measurement results recorded 7 times during the study, starting with day 6 and ending at day 21. Body weight was recorded on the same days. Day 21 hind legs were removed at the border of the hairline just above the hock and were fixed in 10%neutral buffer formalin solution.

Microct analysis: hock joints was investigated using microct scanner with a conical beam focusing. Initially managed the overall scanning to select the volume of samples for analysis, and then was made the fix is tion provisions measurement and computer reconstruction. In hocks, analyzed the bone surface to its volume, which determines the difficulty level surface for a given volume.

Statistical analysis: to determine the swelling/inflammation of a joint data were analyzed using the program Everstat v.5 and two-factor repeated ANOVA analysis with postexperimental test Dannetta. Data microct analyzed using Everstat using one way ANOVA and multiple criteria comparison Newman-Coils. The data presented in the form of mean ± SOS, and the values of p<0,05 was considered significant.

The results:

Prophylactic administration:

Measurement of swelling/inflammation of a joint using a digital compass (figure 2).

The compound of formula I (3.0 mg/kg) significantly reduced the swelling/inflammation compared with rats treated with media only at day 12.

The compound of formula I (10 mg/kg) significantly reduced the swelling/inflammation compared with rats receiving media in the period from day 12 to day 19.

The compound of formula I (30 mg/kg) significantly reduced the swelling/inflammation compared with rats receiving media in the period from day 12 to day 21.

Microct analysis (figure 3).

The compound of formula I (3, 10 or 30 mg/kg, 2 R/d) showed a significant reduction in erosion of the coast is in comparison with rats receiving media (measured by the ratio of bone surface to its volume).

The combined introduction:

Measurement of swelling/inflammation of a joint using a digital compass (figure 4, 5).

The compound of formula I (10 mg/kg) significantly reduced the swelling/inflammation compared with rats receiving media in the period from day 15 to day 21.

- The combined introduction of the compounds of formula I (10 mg/kg) with MTX (or of 0.2 or 0.1 mg/kg) showed a significant reduction in the swelling/inflammation of the hock in the period from day 15 to day 21 compared with the swelling/inflammation observed in rats treated with either compound of formula I (10 mg/kg), or MTX (0.2 or 0.1 mg/kg) as monotherapy.

The compound of formula I (3.0 mg/kg) as monotherapy or as combination therapy with MTX (0.1 mg/kg) did not show a significant impact on the severity of the disease, determined by the swelling/inflammation of a joint.

The compound of formula I (3.0 mg/kg) as a combination therapy with MTX (0.2 mg/kg) showed a significant decrease in the swelling/inflammation of the joint is greater than observed with the introduction of any of the drugs as monotherapy according to the results of measurements performed in the period from day 18 to day 21.

Microct analysis (Fig.6):

- Three-dimensional images showed significant erosion/razrusheny the bones in the joints of rats, receiving media.

The rats of the compounds of formula I (10 mg/kg, p/o 2 p/d) resulted in a significant reduction in bone erosion compared with rats treated with the carrier. In rats treated with 3 mg/kg of the compounds of formula I, a significant reduction was not observed.

- Monotherapy MTX (0.2 mg/kg, p/o 1 p/d) resulted in a significant reduction in bone erosion. This effect was not observed in rats receiving 0.1 mg/kg MTX.

- With the introduction of combinations of the compounds of formula I and MTX all groups showed a significant decrease of the ratio of bone surface to its volume in comparison with rats treated with the media.

- Combination therapy with the compounds of formula I (10 mg/kg, p/o 2 p/d) with MTX (0.2 mg/kg, 1 R/d) resulted in additive protection against erosion of the bone, which was observed in rats treated with monotherapy or compound of formula I, or MTX.

Therapeutic introduction

Measurement of swelling/inflammation of a joint using a digital compass (Fig.7).

Even with the introduction of therapeutic scheme, in which the administration was Acrocephalus until the arthritis became visually evident, the compound of formula I (10 and 30 mg/kg, p/o 2 p/d) significantly reduced the swelling/inflammation over a period of 15-21 days.

Microct analysis (Fig):

Compared with rats receiving the carrier, the rat CIA, which therapeutically injected with a compound of formula I(10 or 30 mg/kg), demonstrated a significant reduction in bone erosion, which was determined by the ratio of bone surface to its volume.

Conclusion:

Studies show that inhibition of Syk kinase with the compound of formula I significantly delayed the beginning and the development of CIA in rats that were determined to reduce swelling/inflammation of the joint and bone erosion. It is important that a significant inhibition of the development of the disease and its severity was observed in rats, infusion of medication which was Acrocephalus before showing visible signs of arthritis. New clinical regimens RA usually held in combination with MTX. Our data, obtained on the model of arthritis rodents demonstrate additive effects when combined introduction of the compounds of formula I with MTX, and therefore, the combined introduction of the compounds of formula I and MTX is able to provide a synergistic clinical effects in patients with RA.

ANALYSIS of ANGIOGENESIS

Female Lewis rats (5 weeks of age, 150-175 g) received anesthesia with xylazine (4 mg/kg) and ketamine (80 mg/kg). Then subcutaneously in the back of the animals implanted cellulose sponge (diameter 10 mm, Vivoxid Ltd.™, Turku, Finland)containing 50 μl of a solution containing 400 ng FGF-2) and FGF-2 (fibroblast growth factor 2) or vehicle (physiological saline solution, bovine serum albumin 0,08%). Over the next two days on lineuse angiogenesis induced by daily introduction under the skin and in the sponge 50 μl of a solution of FGF-2 or medium (basal conditions). One week after implantation of the sponge animals were killed with excessive doses of pentobarbital and sponge analyzed. Then the lips were crushed and homogenized in Lisina buffer (NaCl 150 mm, EDTA 1 mm, Triton X100 1%, deoxycholate sodium 0,5%, NaF, 10 mm Tris/HCl 30 mm pH 7.8, containing a mixture of protease inhibitors (P8340, Sigma-Aldrich™, St Louis, U.S.A.)) in test tubes Lysing Matrix D (MP Biomedicals™, Illkirch, France) in a Fastprep homogenizer (Qbiogene™, Illkirch, France). hemoglobin, reflecting the volume of the vessels were determined using analysis Drabkin (Pierce Biotechnology™, Rockford, Illinois, U.S.A.). Compounds were administered via the oral probe in the form of a suspension in the aqueous methylcellulose of 0.6%solution of Tween-80 0,5%.

The effect of the compounds of formula I on the weight of the rats is reflected in figure 9. The effect of the compounds of formula I at a concentration of hemoglobin (mg/ml) is given in figure 10.

1. The compound of formula (I):

or a pharmaceutically acceptable salt, or MES of such compounds.

2. Pharmaceutical composition having the properties of an inhibitor of Syk kinase, containing a compound according to claim 1 in an effective amount, together with at least one pharmaceutically acceptable carrier or excipient.

3. A method of treating inflammation of the joints, mediated by the activity of Syk kinase, in a patient, comprising an introduction to the needy in this patient an effective amount of a compound according to claim 1.

4. A method of treating rheumatoid arthritis, mediated by the activity of Syk kinase, in a patient, comprising an introduction to the needy in this patient an effective amount of a compound according to claim 1.

5. A method of treating inflammation of the joints, mediated by the activity of Syk kinase, in a patient, comprising an introduction to the needy in this patient an effective amount of a compound according to claim 1, in combination with methotrexate.

6. A method of treating rheumatoid arthritis, mediated by the activity of Syk kinase, in a patient, comprising an introduction to the needy in this patient an effective amount of a compound according to claim 1, in combination with methotrexate.

7. A method of treating tumor-mediated activity of Syk kinase, in a patient, comprising an introduction to the needy in this patient an effective amount of a compound according to claim 1.

8. A method of treating lymphoma cells, mantle zone, mediated by the activity of Syk kinase, in a patient, comprising an introduction to the needy in this patient an effective amount of a compound according to claim 1.

9. Method of inhibiting angiogenesis, mediated by the activity of Syk kinase, in a patient, comprising an introduction to the needy in this patient an effective amount of a compound according to claim 1.

10. The compound according to claim 1, where the salt is selected from hydrochloride, hydrobromide, sulfate, phosphate, nitrate, sulpham, acetate, citrate, lactate, who Arteta, malonate, oxalate, salicylate, propionate, succinate, fumarata, maleate, methylene-bis-b-hydroxynaphthoate, gentisate, isetionate, di-n-tamatert, methansulfonate, aconsultant, bansilalpet, n-toluensulfonate, cyclohexylsulfamate and hinata.

11. The compound according to claim 1, where the salt is a hydrochloride.

12. The compound according to claim 1, where the salt is an acetate.

13. The compound according to claim 1, where the salt is a citrate.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention describes novel compounds of general formula in which is or (values of radicals are given in the claim), a method of producing said compounds, a pharmaceutical composition containing said compounds and therapeutic application thereof.

EFFECT: compounds are cysteine protease inhibitors and can be used in medicine.

25 cl, 1 tbl, 41 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing 2,4,6,8-tetraazabicyclo[3.3.0]octane-3,7-dione (glycoluril). The reaction takes place at 80°C for 60 minutes, where concentrated sulphuric acid is used in an aqueous medium and reagents are taken in the following molar ratios: glyoxal 2.0; urea 4.0; sulphuric acid 0.4; water 12, and the freshly prepared glyoxal solution is added while stirring for 20 minutes, after which the mixture is stirred for 40 more minutes.

EFFECT: novel method of producing glycoluril, which increases output of the end product and is simpler.

1 cl, 1 tbl, 1 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula where R1 is selected from H, F, CI, Br, CF3, C1-C6 alkoxy and OH; R2 is selected from H and C1-C6 alkyl; n equals 1-5; m equals 0 or 1; and Y is selected from CH2, NR3, (NR3R4)+X-, O and S; R3 and R4 are independently selected from H and C1-C4 alkyl; and X- is selected from pharmaceutically acceptable anions. The invention also relates to a method of producing said compound and to an antiviral pharmaceutical composition based on said compound of formula (I).

EFFECT: obtaining novel compounds and a composition based on said compounds, which can be used in medicine to treat a viral diseases such as herpes.

19 cl, 2 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: invention describes novel macrocyclic compounds of formulae pharmaceutically acceptable salts or stereoisomers thereof, where R1 = -OR5, -NH-SO2R6; R2 = hydrogen; R3 = C1-6-alkyl; R4 = isoquinolinyl, possibly substituted; n equals 4 or 5; R5 = hydrogen; R6 = C3-7-cycloalkyl, and a pharmaceutical composition containing said compounds.

EFFECT: novel compounds have hepatitis C virus replication inhibitory action and can be used in medicine.

6 cl, 32 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention describes a compound of general formula where A1 is selected from the following formula R1c denotes a hydrogen atom, a lower alkenyl group or a -Q3-A3(R1d)R1e group; A3 denotes a methane or lower alkyl group; Q3 denotes a single bond; R1d and R1e independently denote a hydrogen atom, hydroxyl group, lower alkyl group or hydroxyl-containing lower alkyl group, or together form a lower alkylene group in which one or two or more methylene groups constituting the lower alkylene group can be independently substituted with an oxygen atom; R1 denotes a lower alkenyl group or a lower alkynyl group; R2 denotes a phenyl, pyridyl or thienyl group, which can contain a -Q4-A4(R1g)R1h group; A4 denotes a nitrogen atom, a lower alkyl group optionally substituted with a hydroxy-lower alkyl group, or a methane group optionally substituted with a halogen atom, a hydroxyl group, a lower alkyl group or a hydroxy-lower alkyl group; Q denotes a single bond or a lower alkylene group in which one or two or more methylene groups constituting the lower alkylene group can be independently substituted with an oxygen atom; R1g and R1h independently denote a hydrogen atom, a lower alkyl group or a lower alkylsulphonyl group; R5 and R6 independently denote a hydrogen atom, a lower alkyl group or a hydroxyl-containing lower alkyl group, or a pharmaceutically acceptable salt thereof. The invention also describes a pharmaceutical composition based on compounds of formula I, having anti-cancer activity, an anticancer agent, a codrug, as well as an exposure sensitising agent containing the pharmaceutical composition.

EFFECT: novel compounds are obtained and described, having excellent Well-kinase inhibitory action and can therefore be used in medicine, especially when treating different malignant tumours.

13 cl, 21 ex

FIELD: chemistry.

SUBSTANCE: invention describes novel macrocyclic compounds of general formulae (I-c) (I-d), pharmaceutically acceptable salt or stereoisomer thereof, where R1 = -OR11 or -NH-SO2R12; R2 = hydrogen and R3 =C1-6-alkyl; n = 3-6; W is a radical of formula , where R5 = phenyl, possibly substituted with C1-6alkyl or alkoxy; thiazolyl, possibly substituted with C1-6alkyl; or pyridyl; R11 denotes hydrogen; R12 = C3-7-cycloalkyl, and a pharmaceutical composition containing said compounds.

EFFECT: said compounds are hepatitis C virus inhibitors and can be used in medicine.

3 cl, 6 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel antiviral active components - substituted indoles of general formula 1 and pharmaceutically acceptable salts thereof, which can be used to treat and/or prevent viral diseases caused by hepatitis C virus (HCV). In general formula , R1 denotes a hydrogen atom, optionally substituted C1-C4alkyl, C6cycloalkyl, phenyl, ethoxycarbonyl, nitro group; R2 denotes a hydrogen atom; R3 denotes N-mono- or N,N-disubstituted 1-methylene-piperidine-3-carboxamide of general formula 1a or N-mono- or N,N-disubstituted 1-methylene-piperdine-4-carboxamide of general formula 1b; R4 denotes a hydrogen atom, optionally substituted C2-C3alkyl, a -CH2-R12 group, where R12 denotes a hydrogen atom or phenyl which is optionally substituted with halogen or C1-C4alkyl; or R2, R3, and R4 together with atoms with which they are bonded form a substituted azaheterocycle of general formula 1.2; or R2 and R3 together with carbon atoms with which they are bonded form a substituted 2,3,4,9-tetrahydro-1H-carbazole of general formula 1.1, in which R1 denotes methyl, ethoxycarbonyl, nitro group; R4 denotes a hydrogen atom, methyl, C2-C3alkyl substituted with N-benzylamine; R7 and R8 denote hydrogen atoms or R7 and R8 together with a carbon atom with which they are bonded form a C=O group; R5 and R6, which are optionally identical, denote a hydrogen atom, optionally substituted C1-C3alkyl or C3-C6cycloalkyl; or R5 and R6 together with a nitrogen atom with which they are bonded form an optionally substituted 5- or 6-member azaheterocyclyl containing one or two nitrogen atoms, etc.

EFFECT: improved properties of compounds.

11 cl, 1 tbl, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to a compound of formula (I): or its pharmaceutically acceptable salt where Q is 2,6-pyrimidyl; where Q is optionally substituted by 1-5 substitutes JQ; Z is a link or NH; R1 is H; R2 is H; R3 is halogen or -(U)m-X where m is equal to 0; X is H or halogen; JQ is halogen, OCF3, -(Vn)-R", -(Vn)-CN or -(Vn)-(C1-4 halogenaliphatic group) where JQ is not H; V is C1-10aliphatic group where up to three methylene groups are substituted by GV where Gv is selected from -NH-, -NR-, -O-, -S-, -CO2-, -C(O)CO-, -C(O), -C(O)NH-, -C(O)NR-, -C(=N-CN)-, -NHCO-, -NRCO-, -NHSO2-, -NRSO2-, -NHC(O)NH-, -NRC(O)NH-, -NHC(O)NR-, -NRC(O)NR or -SO2-; and where V is optionally substituted by 1-6 substitutes JV; R" is H or an optionally substituted group selected from C1-6aliphatic group, C3-10cycloaliphatic group, C6-10aryl, 5-10-member heteroaryl or 5-10-member heterocyclyl; or two R" groups on the same substitute or various substitutes together with atom (s) whereto each group R" is attached, form optionally substituted 3-8-member heterocyclyl; where each optionally substituted R" group is independently and optionally substituted by 1-6 substitutes JR; R is an optionally substituted group selected from C1-6aliphatic group and C6-10aryl where each group R is independently and optionally substituted by 1-4 substitutes JR; each Jv and JR are independently selected from halogen, L, - (Ln)-R', - (Ln)-N(R')2, -(Ln)-OR', C1-4haloalkyl, -(Ln)-CN, - (Ln)-OH, -CO2R', -CO2H or -COR'; or two Jv, JR groups on the same substitute or various substitutes together with atom (s) whereto each group JV and JR is attached, form a 5-7-member saturated, unsaturated or partially saturated ring; R' is H or C1-6aliphatic group; L is C1-6aliphatic group where up to three methylene units are substituted by -C(O)-; each n is independently equal to 0 or 1. Besides, an invention refers to of a pharmaceutical composition for ROCK or JAK kinase inhibition on the basis of the given compounds, to a method of ROCK or JAK kinase activity inhibition, and also to application of the compounds of formula I, for preparing a drug where Q, Z, R1, R2 and R3 are those as described in cl. 1 of the patent claim, effective as protein kinase inhibitors, especially JAK and ROCK families kinase inhibitors.

EFFECT: there are prepared and described new compounds which can find the application in medicine.

42 cl, 6 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel pyrrolopyrimidines of general formula (I) or pharmaceutically acceptable salts thereof, having JANUS: JAK2, JAK3 protein kinase, protein kinase A (PKA), serine/threonine protein kinase ROCK inhibiting properties. The compounds can be used to treat such diseases as allergy, asthma, atopic dermatitis and others. In structural formula (I): R1 denotes H; R2 denotes H; Z1 denotes C1-6aliphatic group or C5-7cycloaliphatic group, optionally substituted with 0-1 groups Jz; if the bond between Z1 and C is a double bond, then Z1 can also denote =O or =C(R)2; Z2 denotes H; or C1-10halogenalkyl, -(Vn)-CN, (Vn)-(heterocyclyl), where the heterocyclyl is a 6-member ring containing a nitrogen atom or two oxygen atoms as heteroatoms, -(Vn)-(phenyl) or -(Vn)-(C3-10cycloaliphatic group), optionally substituted with 0-1 groups Jz; or Z1 and Z2 together with the carbon atom with which they are bonded form a ring Q; Z3 denotes H or C1-6alkyl, optionally substituted with 0-1 groups Jz; or Z1, Z2 and Z3 together with the carbon atom with which they are bonded form a 6-8-member saturated bicyclic ring Q; where if the bond between Z1 and C is a triple bond, then Z2 and Z3 are absent; if the bond between Z1 and C is a double or triple bond, then Z3 is absent or Z2 and Z3 are absent; Q denotes a 3-8-member saturated or partially saturated monocyclic ring containing 0-2 heteroatoms selected from nitrogen, oxygen or sulphur, where said Q is optionally and independently condensed with Q1; where said Q is optionally substituted with 0-4 groups JQ, where said Q is optionally substituted with 0-4 groups JQ. Values of other radicals are given in the claim.

EFFECT: high efficiency of using the compositions.

35 cl, 5 dwg, 5 tbl, 14 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel conformationally stable compounds of general formula (I), which imitate the secondary structure of reverse-configuration regions of biologically active peptides and proteins which are reverse-configuration mimetics. The compounds can be used to inhibit or treat disorders modulated by Wnt-signalling pathway, such as cancer, especially colorectal cancer. The invention also relates to a library containing the disclosed compound. In general formula (I), A denotes -(C=O)-, B denotes -(CHR4)-, D denotes -(C=O)-, E denotes -(ZR6)-, G denotes -(XR7)-, Z denotes CH, X denotes a nitrogen atom, W denotes -(C=O)NH-, R1 denotes benzyl; R2 denotes a heterocyclylC1-6alkyl group, including a 9-member condensed bicyclic ring having 2-3 heteroatoms selected from nitrogen, oxygen or sulphur atoms; a substituted hetercyclylC1-6alkyl group, including a 9-member condensed bicyclic ring having 2-3 heteroatoms selected from nitrogen, oxygen or sulphur atoms, an the ring has 1-3 substitutes independently selected from a group comprising halogen, piperidinyl, morpholinyl, C2-6alkenyl, phenyl, hydroxyphenyl, C1-6alkoxycarbonyl, dialkylamino, hydroxypiperidinyl, C1-6alkyl, hydroxyC1-6alkylpiperazinyl, amino, piperidinyl carbonyl; heterocyclyl-C1-6alkyl group having a 9-member condensed bicyclic ring which has one or two nitrogen atoms; and other values given in the claim. R4 denotes a substituted benzyl, having a substitute selected from disodium phosphate, monosodium phosphate, phosphate; R6 denotes hydrogen; R7 denotes: C1-6alkyl; C1-6alkynyl; C2-6alkenyl; substituted benzyl, having one or more substitutes independently selected from halogen and C1-6alkyl.

EFFECT: high efficiency of the compounds.

8 cl, 34 dwg, 19 tbl, 25 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula or pharmaceutically acceptable salt thereof, synthesis methods thereof, pharmaceutical compositions containing said compounds, and use thereof to prepare a medicinal agent having mTOR kinase and/or PI3K kinase inhibiting action.

EFFECT: improved properties of the derivatives.

15 cl, 72 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology, and can be used for treatment of ovarian cancer recurrences. For this purpose combined courses of subtotal body irradiation (STBI) and chemical therapy (CT) are carried out. STBI is performed with field of diaphragm dome to feet in single dose 0.1 Gy to course dose 1 Gy. CT is carried out immediately after STBI. After each combined course of STBI and CT additionally, after 1-1.5 months course of CT is carried out. After that, after 1-1.5 months entire course of treatment (STBI+CT+CT) is repeated until remission is achieved or total STBI dose not higher than 4 Gy. If after achievement of total STBI dose - 4 Gy, there is no remission, only CT is continued with 1-2 month intervals between courses. All CT courses, performed in the process of treatment are performed with medications from group of taxanes and platinum derivatives.

EFFECT: method ensures increase of remission cases and duration of recurrence-free period in patients due to enhance of cytostatic action onto tumour cells as a result of introduction into treatment scheme of additional courses of chemical therapy between combined courses of STBI and CT, as well as due to reduction of intervals between therapy courses.

1 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, photodynamic therapy of tumours. Photosensitiser (PS), in particular radachlorine is added to nanoparticles of emulsion of perfluorocarbons (PFC), consisting of perfluorodecalin and perfluoromethylcyclohexylpiperidine, stabilised with proxanol 268 solution. Emulsion of PFC with PS is added to stem cells of bone marrow (SCBM), in particular, autologic. Their joined cultivation is carried out, after which SCBM is introduced to subject with malignant tumour, in particular breast tumour. Zone of tumour growth is subjected to impact by light irradiation in dose sufficient for complete or partial tumour destruction. Subject can be represented by a mammal, in particular, human being.

EFFECT: method ensures targeted impact, complete or partial tumour regress, excludes systemic phototoxic injuries.

6 cl, 3 ex, 3 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical composition for injection targeted local application which involves a sterile suspension of platinum complex (OC-6-43)-bis(acetato)-(1-adamantilamino)ammine-dichloroplatinum (IV) (LA-12) in a pharmaceutically acceptable hydrophilic or hydrophobic injection liquid phase with 100% of platinum complex particles sizing less than 250 mcm.

EFFECT: invention provides preparing the sterile platinum complex suspension LA-12 suitable for injection application and containing a sufficient therapeutically effective amount of the complex.

13 cl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, oncology and concerns early diagnosis of postradiation myocardial damage in patients suffering lung cancer at the stages of combination treatment by examining a morphofunctional myocardial status. It involves gated myocardial 99mTc-MIBI SPECT described by two diagnostic criteria: myocardial perfusion values and left ventricular contractility. The criteria are evaluated fivefold: prior to treatment, after 2 courses of neoadjuvant chemotherapy and after 1, 6 and 12 months following a radical surgery with intraoperative radiation therapy. The derived values are compared with the reference values prior to treatment, and if observing a persistent decrease, postradiation myocardial damage is diagnosed.

EFFECT: technique provides adequate assessment of the progression myocardial status, higher diagnostic accuracy and reliability of early postradiation myocardial damages - up to one year prior to treatment in the given group of patients with lung of cancer having an initial considerable risk of development of a cardiac pathology.

1 ex, 1 app, 2 dwg

FIELD: chemistry.

SUBSTANCE: method is realised by treating a compound of formula

with boronic acid or ether thereof of formula

,

in which two OR15 groups together with the boron atom with which they are bonded form a pinacolato boronate ester group in the presence of a Pd catalyst. The invention relates to a method of producing a pharmaceutically acceptable salt of thieno[3,2-d]pyrimidine of formula

.

The invention also relates to a pharmaceutical composition, having phosphatidyl inositol-3-kinase inhibitor activity, containing thieno[3,2-d]pyrimidine of formula (I) as an active ingredient, a method of preparing said composition and use of thieno[3,2-d]pyrimidine of formula (I) or pharmaceutically acceptable salt thereof in producing a medicinal agent for inhibiting phosphatidyl inositol-3-kinase.

EFFECT: use of the derivative as a phosphatidyl inositol-3-kinase inhibitor.

11 cl, 13 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology, and can be used for treatment of patients with non0resectable signet ring cell adenocarcinoma. For this purpose laparotomy is performed. Into left gastric artery installed and fixed is catheter, through which 50-70 ml of ozonised physiological solution are introduced. After 30 minutes, 1000 mg of 5-fluoracyl and 50 mg leukovorin are introduced in the same way. After that, catheter is connected with port, installed in recess, specially created in subcutaneous adipose cellular tissue and brought under skin of anterior abdominal wall, wound is sewn layer-by layer. 7 days after operation into port, installed in anterior abdominal wall, connected with catheter, 50-70 ml of ozonised physiological solution are introduced. After 30 minutes, 1000 mg of 5-fluoracyl and 50 mg leukovorin are introduced in the same way. Procedure is repeated 5 more times with 1 week interval.

EFFECT: method ensures increase of anti-tumour activity of chemical preparations at the background of non-specific stimulation of patients' immunity, achieved by introduction of ozonised physiological solution.

1 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology and can be used in treatment of soft tissue sarcomas as one of complex treatment components. For this purpose sampling of 400 ml of blood into vial with hemopreservative is carried out with its further centrifugation at 1500 rotations per minute during 30 minutes. After that, plasma is divided into two equal parts into sterile vials: into one of them introduced are 600 mg/m2 of cyclophpsphane, into the other - methotrexate 40 mg/m2, and blood cells are connected with doxorubicin - 40 mg/m2. All mixtures are incubated for 30 minutes at 37°C. After that on the first and seventh day of treatment intravenously simultaneously introduced are blood cells with doxorubicin and mixtures of autoplasm with cyclophosphane and then with methotrexane are injected around tumour on its circumference.

EFFECT: method makes it possible to increase efficiency of said pathology treatment due to ensuring considerable reduction of tumour size before surgery.

1 ex

FIELD: medicine.

SUBSTANCE: group of inventions relates to medicine and is intended for treatment of malignant tumor in patient. At least one fixed dose of pertuzumab is introduced. Fixed dose is selected from group of approximately 420 mg, 525 mg, 840 m, 1050 mg.

EFFECT: group of inventions makes it possible to simplify dose selection for patient, reduce costs for individual selection of pertuzumab dose.

9 cl, 4 tbl, 14 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine and deals with composition, which contains non-covalent complex of pig alpha-fetoprotein (PAFP), obtained from blood and amniotic liquid of pig embryos by butanol extraction, and at least one agent, which induces apoptosis. Invention also relates to application of claimed composition for inhibition of proliferation of patient's cancer cell, which has alpha-fetoprotein receptor on its surface; application of claimed composition for treatment of multiple drug resistance of refractory malignant neoplasms in patient.

EFFECT: invention ensures reliable and cheap source of product.

25 cl, 8 ex, 7 dwg, 4 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine. A method involves application of IL-31 antagonist for IL-31 induced signal transduction inhibition in dorsal root ganglion cells. The IL-31 antagonist inhibits IL-31 polypeptide binding containing amino acid residues 27-164 SEQ ID NO:2 with its heterodimeric receptor containing IL-31RA and OSMR-beta. The antagonist represents a humanized monoclonal antibody or a chimeric antibody.

EFFECT: use of the method effectively relieves inflammation, pain and itching.

8 ex

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