Cyclopamine analogues and methods of their application

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to novel compounds of formula 1: or its pharmaceutically acceptable salt; in which each R1 and R8 independently represents H or hydroxyl; each R2 and R9 independently represents H or hydroxyl; R5 represents H; each R3, R4, R6, R7, R13 and R14 independently represents H; or R1 and R2, taken together, form =O; or R4 and R5, taken together form double bond; or R5 and R6, taken together form double bond; R10 and R11, taken together form double bond; R12 represents H, alkyl, hydroxyl, aralkyl, halogenalkyl, alcoxyl,- -[(W)- N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-O]qR21 or -[(W)-N(R21)]qR21; where each W independently represent bivalent alkyl or aralkyl radical, and q is equal 1, 2, 3 or 4; each R15, R16 and R17 independently represent H; each R18 and R19 independently represents H; and each R21 independently represents H, alkyl, aryl or aralkyl. Invention also relates to pharmaceutical composition. Claimed invention provides cycloamine analogues which can be applied to counteract phenotypic effects of undesirable activation of Hedgehog pathway, such as acquiring function from Hedgehog, Ptc loss of function or mitigated acquired from function mutations.

EFFECT: compounds by claimed invention are especially applicable in cancer treatment.

19 cl, 1 tbl, 28 ex

 

A related application

This application is related to the following applications USA: serial number 60/605020, filed August 27, 2004, serial number 60/617170, filed October 8, 2004, serial number 60/625676, filed November 5, 2004, and serial number 60/683169, filed may 19, 2005, each of which is incorporated here by reference in their entirety.

The level of technology

The Hedgehog signaling pathway is an essential part of many processes as part of embryonic development. Members of the secreted proteins of the Hedgehog family control cell proliferation, differentiation and tissue formation. The path was first deciphered in the body of the fruit fly Drosophila, but has since shown that he largely remains in invertebrates and vertebrates, including humans. Total activity of the Hedgehog signaling pathway decreases after embryogenesis in most cells, but the path remains active in some types of adult cells. It was recently shown that unregulated activation of the Hedgehog signaling pathway leads to certain types of cancer, as described below.

Hedgehog polypeptide is a secretory protein that functions as a signaling ligand in the Hedgehog pathway. Typical genes and Hedgehog proteins are described in PCT publications WO 95/18856 and WO 96/17924. Three different forms of Hedgehog protein found in humans; Sonic Hedgehog (Shh), Desert Hedgehog (Dhh) and Indin Hedgehog (Ihh). Sonic the Hedgehog is the most common representative of the Hedgehog in mammals and also the most well-described ligand Hedgehog family. Before secretion of Shh undergoes intramolecular cleavage and reaction of lipid modification. Lipid-modified peptide is responsible for all activity signaling.

Two transmembrane protein involved in the transduction of the Hedgehog signal path; twelve-transmembrane Patched receptor (Ptc) and the seven-transmembrane Smoothened protein (Smo).

Research in technology suggest that Hedgehog acts by binding Ptc, thus deblocare inhibitory effect of Ptc on Smo. As Ptc and Smo both are transmembrane proteins, the proposed scenario is that they are physically linked, forming a receptor complex, although indirect mechanisms of action are also likely. Depression from Smo inhibition of Ptc is most likely involves a conformational change in Smo. Ptc, however, is not necessary for the activity of Smo, as Smo becomes constitutively activated in the complete absence Pathed protein (Alcedo et al., above; Quirk et al. (1997) Cold Spring Harbor Symp. Quant. Biol. 62: 217-226). As soon as Smo is initiated, it is fast and highly fosfauriliruetsa and converts the signal, which activates transcription through the transcription factors Gli (the homologue of the protein CiDrosophila) (Alexandre et al. (196) Genes Dev. 10: 2003-13). The transcription factor Gli1 polysoude regulates many of the genes included in the growth and development of (Alexandre et al., above).Hedgehog signaling is a necessary part of many stages of development, especially in the formation of left-right symmetry. Loss or reduction of Hedgehog signaling leads to the development of many deficiencies and deformities, of which the most serious is cyclopia (Belloni et al. (1996)Nature Genetics, 14: 353-6).

Recently it was reported that activating mutations of forming Hedgehog pathways occurs in sporadic basal cell carcinoma (Xie et al. (1998)Nature391: 90-2) and primary neuroectodermal tumors of the Central nervous system (Reifenberger et al. (1998)Cancer Res.58: 1798-803). Unregulated activation of the Hedgehog pathway has also been identified in many types of cancer such as cancer of the gastrointestinal tract, including cancer of the pancreas, esophagus cancer, stomach cancer (Berman et al. (2003)Nature425: 846-51, Thayer et al. (2003)Nature425: 851-56), lung cancer (Watkins et al. (2003)Nature422: 313-317cancer of the prostate (Karhadkar et al. (2004)Nature431: 707-12, Sheng et al. (2004)Molecular Cancer3: 29-42, Fan et al. (2004)Endocrinology145:3961-70), breast cancer (Kubo et al. (2004)Cancer Research64: 6071-74, Lewis et al. (2004)Journal of Mammary Gland Biology and Neoplasia2:165-181) and hepatocellular cancer (Sicklick et al. (2005) ASCO conference, Mohini et al. (2005) AACR conference).

Shows steingebaude low molecular weight compounds activity of the Hedgehog pathway leads to cell death in several types of cancer, with unregulated activation of the Hedgehog pathway (see, for example,, Berman et al., 2003 Nature425: 846-51).

Antagonists of the Hedgehog pathway are currently being explored in a large number of clinical conditions, where therapeutic effect for condition or disorder can be obtained through the inhibition of one or more aspects of the activity of the Hedgehog pathway. Although the main focus is on cancer, the researchers found that the inhibition by low molecular weight compounds Hedgehog path showed improvement of symptoms of psoriasis (Tas, et al., 2004 Dermatology209: 126-131, published US patent application 20040072913(incorporated herein by reference)). Psoriasis is a very common chronic disorder of the skin, usually characterized by skin lesions, including erythematous papules and patches with silvery scale, although there are changes on the skin and in other parts of the body. Psoriasis, as currently believed, is an autoimmune disease, but its etiology is still poorly understood.

Inhibitor of the Hedgehog pathway, which attracted considerable interest, is a natural product cyclopamine. Cyclopamine was first isolated from LilyVeratrum californicumin 1966 after it was found that the offspring of sheep, which contained feeding in the pasture, was born with severe congenital deformities. To identify and apply the o(a), causes congenital deformation data, the FDA investigated possible sources of teratogens and identified jermenovci family of steroid alkaloids, including connection cyclopamine as teratogens responsible for congenital deformities.

Much later it was found that the mechanism of action cyclopamine was a direct inhibition of the activity of the Hedgehog pathway (Cooper et al. (1998)Science280: 1603-7, Chen et al. (2002)Genes and Development16: 2743-8). It is shown that cyclopamine and related compounds have anticancer activity through effects on Hedgehog pathway. Despite the initial term, any members of this family of compounds, or their analogs has not led to successful development of anti-cancer tool. The present invention fulfills this need and has other benefits.

The invention

The present invention provides analogues of steroidal alkaloids family cyclopamine that are useful for inhibiting cell proliferation and/or maintenance of apoptosis in the cell, including for the treatment of proliferative disorders such as cancer. Antagonists of the Hedgehog pathway of the present invention can be used to suppress proliferation (or other biological consequences) cells or tissues, such as in patients characterized as having a phenotype with irennoj functions Ptc, the phenotype of increased Smo function or phenotype of increased function Hedgehog.

In certain areas these methods are used to counteract the phenotypic effects of unwanted activation of the Hedgehog pathway, such as resulting from mutations increase the function Hedgehog, an underactive Ptc or increase the function of Smo. For example, the present methods can include bringing the cells into contact (in vitroorin vivowith an antagonist of the Hedgehog pathway of the present invention (as defined below) in an amount sufficient to exert an antagonistic effect on Smo-dependent activation pathway. Such antagonism will stop or slow down unwanted proliferation of cells and can cause cell death.

In some embodiments, the implementation of the methods and compounds of the present invention can be used for the regulation of cell proliferation and/or cell deathin vitroand/orin vivosuch as the treatment of cancer of the head, neck, nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx, hypopharynx, salivary glands, paraganglia, pancreas, stomach, skin, esophagus, liver and biliary tree, bone, small intestine, colon, rectum, ovary, prostate, lung, breast, lymphatic system, blood, Central nervous system, ostogo brain or brain.

In some embodiments, the implementation of the methods and compounds of the present invention can be used to treat the symptoms of psoriasis patients. Compounds of the present invention can be used for the treatment of psoriasis as a single agent or in combination with one or more means for the treatment of psoriasis. In preferred embodiments of the compounds of the present invention are applied topically in a patient in need thereof.

Compounds of the present invention can be further included in the composition as a pharmaceutical composition, comprising a pharmaceutically acceptable excipient, for administration to a patient as a means of cancer treatment. Antagonists of the Hedgehog pathway of the present invention and/or compositions comprising them, can be administered to a patient to treat conditions involving unwanted proliferation of cells, such as cancer and/or tumors of the head, neck, nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx, hypopharynx, salivary glands, paraganglia, pancreas, stomach, skin, esophagus, liver and biliary tree, bone, small intestine, colon, rectum, ovary, prostate, lung, breast, lymphatic system, blood, Central nervous system, bone marrow or brain. In some embodiments, the implementation of such compounds of recomposition applied systematically, for example, parenteral and/or locally, such as local.

Detailed description of the invention

Definitions

Definitions of terms used herein are intended to enable these modern definitions are recognized for each term in the chemical and pharmaceutical sectors. Where appropriate, examples are provided. The definitions apply to terms as they are used throughout this specification, unless otherwise limited in specific cases or individually, or as part of a larger group.

The term "heteroatom" is known in the art and refers to an atom of any element other than carbon atoms or hydrogen. Illustrative heteroatoms include boron, nitrogen, oxygen, phosphorus, sulfur and selenium.

The term "alkyl" is known in the art and includes saturated aliphatic groups, including alkyl groups with non-branched chain alkyl groups, branched-chain, cycloalkyl (polycyclic) groups, alkyl substituted cycloalkyl group and cycloalkylation alkyl groups. In some embodiments, the implementation of alkyl with non-branched chain or branched-chain has about 30 or fewer carbon atoms in the main chain (for example, C1-C30for unbranched chain, C3-C30for branched CE and), and alternative, approximately 20 or less. Similarly, cycloalkyl have from about 3 to about 10 carbon atoms in their ring structure, and alternative about 5, 6 or 7 carbon atoms in the ring structure. Alkyl groups, unless otherwise specified, may optionally be substituted by suitable substituents. The number of substituents is usually limited by the number of available valences on the alkyl group; thus, the alkyl group may be substituted by one or more of the hydrogen atoms that could be present in unsubstituted group. Suitable substituents for the alkyl groups include halogen, =O, =N-CN, =N-OR', =NR', OR', NR'2, SR', SO2R', SO2NR'2, NR'r SO2R', NR'r CONR'2, NR'r COOR', NR'r COR', CN, COOR', CONR'2, OOCR', COR', and NO2where each R' is independently H, C1-C6the alkyl, C2-C6heteroalkyl, C1-C6, acyl, C2-C6heteroatom, C6-C10the aryl, C6-C10heteroaryl, C7-C12arylalkyl or C6-C12heteroallyl, each of which is optionally substituted by one or more group selected from halogen, C1-C4of alkyl, C1-C4heteroalkyl, C1-C6aryl, C1-C6heteroaryl, hydroxy, amino, and =O; and where two R' in which cave one Deputy or on adjacent atoms, can be linked to form a 3-7-membered rings, optionally containing up to three heteroatoms selected from N, O and S.

If the number of carbon atoms is not defined otherwise, lower alkyl refers to an alkyl group as defined above, but having from one to about ten carbon atoms, alternatively from one to about six carbon atoms in the structure of the main chain. Similarly, lower alkenyl and lower quinil have similar chain length.

The term "aralkyl" is known in the art and refers to an alkyl group, substituted aryl group (for example, an aromatic or heteroaromatic group).

The terms "alkenyl" and "quinil" is known in the art and refer to unsaturated aliphatic groups analogous in length and possible substitution to alcelam described above, but they contain at least one double or triple bond, respectively, and may contain a mixture of both double and triple bonds. Alkeneamine and alkyline group optionally substituted, unless otherwise specified, the same substituents described above for alkyl groups.

Heteroalkyl, heteroalkyl and heteroalkyl and the like are defined similarly to the corresponding hydrocarbonyl (alkyl, alkenyl and alkynylaryl) groups, but the term "hetero" refers to groups is m, containing 1-3 O, S or N heteroatoms or combinations thereof within the balance of the main chain; thus at least one carbon atom, respectively, alkyl, alkenylphenol or alkenylphenol groups replaced by one of these heteroatoms, forming heteroalkyl, heteroalkyl or heteroalkyl group. Typical and preferred dimensions for waterform alkyl, alkenyl and etkinlik groups are usually the same as for the corresponding hydrocarbonrich groups, and the substituents that may be present on heterograph are the same as described above for hydrocarbonrich groups. It is also understood that unless otherwise specified, for reasons of chemical stability of these groups do not include more than two adjacent heteroatom, excluding those where oxoprop is present on the N or S, as in sulfonyloxy group.

The term "aryl" is known in the art and refers to 5-, 6 - and 7-membered monocyclic aromatic radicals, which may include from zero to four heteroatoms, for example benzene, naphthalene, anthracene, pyrene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazin, pyridazine and pyrimidine, and the like. Aryl groups having heteroatoms in the ring structure may also be referred to as aryl heterocycles or gets auromatically. The aromatic ring can be substituted at one or more positions of the ring with such substituents as described above, for example, halogen, azide, alkyl, aralkyl, alkenyl, quinil, cycloalkyl, hydroxyl, CNS group, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl group, silicom, simple ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ether complex, heterocyclyl, aromatic or heteroaromatic residues, -CF3, -CN, or the like. The term "aryl" also includes polycyclic system having two or more cyclic rings in which two or more carbons are common to two adjoining rings (the rings are "condensed ring"), where at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkenyl, arinami and/or heterocycles.

The terms ofortho,metaandpairknown in the art and refers to 1,2-, 1,3 - and 1,4 - twice replaced the benzenes, respectively. For example, the name "1,2-xylene and ortho-xylene" are synonymous.

The terms "heterocyclyl", "heteroaryl" or "heterocyclic group" are known in the art and refer to from 3 to PR is approximately 10-membered ring structures, alternative 3 - to about 7-membered rings, ring structures which comprise 1-4 heteroatoms. Heterocycles may also be politiclly. Heterocyclic groups include, for example, thiophene, tianren, furan, Piran, isobenzofuran, chrome, Xanten, pinoxaden, pyrrole, imidazole, pyrazole, isothiazol, isoxazol, pyridine, pyrazin, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, hemolysin, isoquinoline, quinoline, phthalazine, naphthiridine, cinoxacin, hinzelin, cinnolin, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthrolin, fenesin, phenarsazine, fentazin, furazan, phenoxazine, pyrrolidin, oxolan, tolan, oxazole, piperidine, piperazine, morpholine, lactones, lactams, such as azetidinone and pyrrolidinone, solitary, Sultani etc. Heterocyclic nucleus may be substituted in one or more positions with such substituents as described above, for example, halogen, alkyl, aralkyl, alkenyl, quinil, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl group, silyl, simple ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic residues, -CF3, -CN, or the like.

The terms "politikil" or "polycyclic group" are known in the art and Rel is relevant to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkenyl, Ariely and/or heterocyclyl), in which two or more carbons are common to two attached rings, such as rings are condensed ring. Rings attached through non-adjacent atoms are called bridge rings. Each of the rings polycycle can be substituted with such substituents as described above, for example, halogen, alkyl, aralkyl, alkenyl, quinil, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl group, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic residue, -CF3, -CN, or the like.

The term "carbocycle" is known in the art and refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

The term "nitro" is known in the art and refers to-NO2the term "halogen" is known in the art and refers to-F, -Cl, -Br or-I; the term "sulfhydryl" is known in the art and refers to-SH; the term "hydroxyl" means -- HE; and the term "sulfonyl" is known in the art and refers to-SO2. The halide defines the corresponding anion of halogen, and pseudohalogen connection" formulated the definition on str from "Advanced Inorganic Chemistry"by Cotton and Wilkinson.

The terms "amine"and "amino" and "ammonium" is known in the art and refers to unsubstituted and substituted amines, for example the balance, which may be represented by the General formula:

where R50, R51 and R52 each independently represents hydrogen, alkyl, alkenyl, -(CH2)m-R61, or R50 and R51, taken together with the N atom to which they are attached, form a heterocycle of 4 to 8 atoms in the ring structure; R61 represents an aryl, cycloalkyl, cycloalkenyl, heterocycle or politics; and m represents zero or an integer in the range 1-8. In other embodiments R50 and R51 (and optionally R52)each independently represent hydrogen, alkyl, alkenyl, or -(CH2)m-R61. Thus, the term "alkylamino" includes an amine group, as defined above, having attached a substituted or unsubstituted alkyl, i.e. at least one of R50 and R51 is an alkyl group.

The term "acylamino" is known in the art and refers to the residue, which may be represented by the General formula:

where R50 is as defined above, and R54 represent hydrogen, alkyl, alkenyl or (CH2)m-R61, where m and R61 are defined above.

The term "amido" is known in the art as aminosilanes carbon is l and includes the remainder, which may be represented by the General formula:

where R50 and R51, as defined above. Certain embodiments of the amide in the present invention will not include imides which may be unstable.

The term "alkylthio" refers to an alkyl group as defined above, attached via a sulfur radical. In some embodiments, the implementation of the remainder of alkylthio represented by one of-S-alkyl, -S-alkenyl, -S-quinil and-S-(CH2)m-R61, where m and R61 are defined above. Typical ancilliary include methylthio, ethylthio and the like.

The term "carboxyl group" is known in the art and includes such residues, which can be represented by the General formula:

where X50 is a bond or oxygen, or sulfur, and R55 and R56 represents a hydrogen, alkyl, alkenyl, -(CH2)m-R61 or a pharmaceutically acceptable salt, R56 represents a hydrogen, alkyl, alkenyl or -(CH2)m-R61, where m and R61 are defined above. In cases where X50 represents oxygen and R55 or R56 are not hydrogen, the formula represents an ester. In cases where X50 represents oxygen and R55, as specified above, the remainder is referred to here as a carboxyl group, and, preferably, when R55 is bodoro is, the formula is a carboxylic acid. In cases where X50 represents oxygen and R56 represents a hydrogen, the formula represents a formate. Usually in cases where the oxygen atom of the above formula is replaced by sulfur, the formula represents a thiocarbonyl group. In cases where X50 is a sulfur, and R55 or R56 represents a hydrogen, the formula represents a complex tiefer. In cases where X50 is a sulfur, and R55 represents a hydrogen, the formula represents thiocarbonic acid. In cases where X50 is a sulfur and R56 represents a hydrogen, the formula represents toformat. On the other hand, in cases where X50 is a bond and R55 is hydrogen, the above formula represents a ketone group. In cases where X50 is a bond and R55 represents a hydrogen, the above formula represents an aldehyde group.

The term "carbarnoyl" refers to-O(C=O)NRR', where R and R' are independently H, aliphatic group, aryl group or heteroaryl group.

The term "oxo" refers to the carbonyl oxygen (=O).

The terms "alkoxyl" or "alkoxy" are known in the art and refers to an alkyl group as defined is prohibited above, attached via radical oxygen. Typical CNS groups include methoxy, ethoxy, propyloxy, tert-butoxy and the like. Simple ether is a two hydrocarbon, United covalent bond through oxygen. Accordingly, the Deputy of alkyl, which forms alkilany broadcast or included in the CNS group, which may be represented by one of-O-alkyl, -O-alkenyl, -O-quinil, -O-(CH2)m-R61, where m and R61 described above.

The term "sulfonate" is known in the art and refers to the residue, which may be represented by the General formula:

where R57 is an electron pair, hydrogen, alkyl, cycloalkyl or aryl.

The term "sulfate" is known in the art and includes the remainder, which may be represented by the General formula:

where R57, as defined above.

The term "sulfonamide" is known in the art and

includes the remainder, which may be represented by the General formula:

in which R50 and R56, as defined above.

The term "sulfamoyl" is known in the art and refers to the residue, which may be represented by the General formula:

where R50 and R51, as is designated above.

The term "sulfonyl" is known in the art and refers to the residue, which may be represented by the General formula:

where R58 is one of the following: hydrogen, alkyl, alkenyl, quinil, cycloalkyl, heterocyclyl, aryl or heteroaryl.

The term "sulfoxide" is known in the art and refers to the residue, which may be represented by the General formula:

where R58 is defined above.

The term "phosphoryl" is known in the art and may be represented by the General formula:

where Q50 represents S or O, and R59 represents a hydrogen, lower alkyl or aryl. When there is substitution, for example, alkyl, phosphoryl group phosphorylative can be represented by the General formula:

where Q50 and R59 each independently as defined above, and Q51 represents O, S or N. When Q50 is an S, the phosphoryl residue is a "Fosforit".

The term "phosphoramidite" is known in the art and may be represented by the General formula:

where Q51, R50, R51 and R59, as defined above.

The term "phosphoramidite" is known in the art and may be represented by the General formula:

where Q51, R50, R51 and R59, as defined above, and R60 represents a lower alkyl or aryl.

Can be replacement alkenyl and etkinlik groups similar to the substituents of receiving, for example, aminoalkyl, aminoalkyl, aminoalkyl, aminoalkyl, aminoalkyl, aminoalkyl, tolchenov, tyulkanov, carbonization of alkenyl or alkinyl.

The definition of each expression, for example, "alkyl", "m", "n" and the like are intended to be independent from the definition elsewhere in the same structure when it is repeatedly found in this structure.

The term "selenology" is known in the art and refers to an alkyl group attached through selenography. Typical "sleeeper can be a Deputy of alkyl, which is chosen from-Se-alkyl, -Se-alkenyl, -Se-quinil and-Se-(CH2)m-R61, m and R61 defined above.

The terms "trifles", "toil", "methylsulphonyl" and "neatly" is known in the art and refer to trifloromethyl, p-toluensulfonyl, methanesulfonyl and nonoperatively groups, respectively. The terms "triplet", "toilet", "mesilate and nonflat" is known in the art and refer to triftormetilfullerenov, p-toluensulfonate, methanesulfonate and nonattributable, fu is clonally groups and molecules that contain these groups.

The abbreviations Me, Et, Ph, Tf, Nf, Ts and Ms denote methyl, ethyl, phenyl, trifloromethyl, nonattributable, p-toluensulfonyl and methanesulfonyl respectively. A more complete list of abbreviations used in this technical field chemists-organically conventional qualification appears in the first issue of each volumeJournal of Organic Chemistry;this list is typically presented in the table entitledStandard List of Abbreviations.

Some compounds included in the compositions of the present invention may exist in the preferred geometric or stereoisomeric forms. The present invention considers all such compounds, including CIS - and TRANS-isomers, R - and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures and other mixtures thereof, falling within the range of the invention. Additional asymmetric carbon atoms may be present in the Deputy, such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in the invention.

If, for example, desirable preferred enantiomer of the compounds of the present invention, it can be obtained by asymmetric synthesis or by modification with chiral auxiliary compound, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide a chemically pure desired enantiomers. Alternatively, in cases where the molecule contains a basic functional group, such as amino, or an acidic functional group such as carboxyl group, diastereomeric salts are formed with an appropriate optically active acid or base, with subsequent separation of diastereoisomers, which is carried out by fractional crystallization or chromatographic method known in the art, and subsequent isolation of pure enantiomers. Similarly, the preferred enantiomer can be isolated from the racemic mixture of its enantiomers using chiral chromatographic methods known in the art.

It is understood that the Deputy or substituted implies that such substitution is in accordance with a permitted valence of the substituted atom and the substituent, and that the substitution results in a stable connection, for example, which does not undergo spontaneous transformation, such as rearrangement, cyclization, elimination, or other reaction.

The term "substituted" is also considered to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic the mandated the prevalence of organic compounds. Illustrative substituents include, for example, described herein above. Permissible substituents can be one or more and same or different suitable for organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. Do not imply that the invention is limited to any permissible substituents of organic compounds.

As used here, the phrase "protective group" means a temporary deputies, which protect a potentially reactive functional group from undesired chemical transformations. Examples of such protective groups include esters of carboxylic acids, Silovye esters of alcohols and acetals and ketals of aldehydes and ketones, respectively. The field of chemical protective groups was considered (Green, T.W.; Wuts, P.G.M. Protective Groups in Organic Synthesis, 2nded.; Wiley: New York, 1991). Protected forms of the compounds of the invention are included in the scope of this invention.

For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics,67th Ed., 1986-87, on the cover.

The phrase "aberrant modification or mutation of a gene refers to a genetic time is eniam, as, for example, deletion, substitution or insertion of nucleotides in the gene, as well as significant chromosomal rearrangements of the gene and/or abnormal methylation of the gene. Similarly, abnormal expression of a gene refers to aberrant levels of transcription of the gene relative to such levels in a normal cell under these conditions, as well as splicing unstable type of mRNA transcribed from the gene.

Basal cell carcinoma exist in different clinical and histological forms, such as nodular-ulcerative, superficial, pigmented, annular, fibroepithelioma and Sevodnya syndrome. Basal cell cancers are the most common skin tumors found in humans. Most new cases of skin cancer non-melanoma, belong to this category.

The term "carcinoma" refers to a malignant neoplasm composed of epithelial cells tending to infilterate into the surrounding tissues and give rise to metastases. Typical cancer include basal cell carcinoma, which is an epithelial tumor of the skin, which rarely metastasize, has the potential for local invasion and destruction; squamous cell carcinoma, which refers to carcinomas, coming from squamous epithelium and having a cubic cell; ka is conocerla, which includes a malignant tumor made up of cancerous and sarcomatous tissues; adenomatosnuu carcinoma, carcinoma, characterized by cylinders or strips or mucinous stroma, separated or surrounded by nests or cords of small epithelial cells found in breast and salivary glands and mucous glands of the respiratory tract; epidermoid carcinoma, which refers to malignant cells, which tend to differentiate in the same way as similar from the epidermis; i.e. they have a tendency to the formation of thorn cells and transfer the keratinization; nasopharyngeal carcinoma, which refers to a malignant tumor arising in the epithelial cover the space behind the nose; and renal cell cancer, which refers to cancer of the renal parenchyma, composed of canalave cells in the changed location.

Other cancerous epithelial growths are papillomas, which are benign tumors that proceedeth out of the epithelium, and are caused by the human papilloma virus; and epidermoid, which are tumors of the brain or meninges, forming the inclusion of ectodermal elements during the laying groove of the neural tube.

The term "ED50" means a dosage environments is TBA, which produces 50% of its maximum response or effect.

Effective number of pending connections over existing methods of treatment, refers to the amount of antagonist in the composition which, when applied as part of a desired dosage regimen, causing, for example, a change in the level of cell proliferation and/or level of cell survival according to clinically acceptable standards for diseases for which treatment will be applied.

The term "epithelial" and "epithelium" refers to the cellular covering of internal and external body surfaces (skin, mucous and serous membranes), including the glands and other structures that emanate from them, such as corneal, esophageal, epidermal and hair epithelial cells of the follicle. Another typical epithelial tissue includes the olfactory epithelium, which is pseudolegally epithelium lining the olfactory region of the nasal cavity and contains the olfactory receptors; glandular epithelium, which refers to the epithelium, composed of secreting cells; squamous epithelium, which refers to the epithelium composed of flattened plate cells. The term "epithelial" can also refer to the transitional epithelium, which typically is in the lining of hollow organs, which was subjected to what are the big mechanical change, such as the contraction and stretching, such as cloth, which represents a transition between the multilayered squamous and columnar epithelium.

The state of cell growth refers to the rate of cell proliferation and/or differentiation of cells. Altered state growth is a state of growth, characterized by abnormal levels of cell proliferation, such as cell showing increased or decreased the proliferation rate relative to normal cells.

The term "antagonist of the Hedgehog pathway" refers to a tool that slows down the function of the Hedgehog pathway, for example, suppresses the transcription of target genes (Gli1 and Ptc genes), which in normal cells is induced by contact of the cells with a Hedgehog. In addition to changing Smoothened-dependent pathway in some embodiments, the implementation of the Hedgehog antagonists of the path of the present invention can be used to overcome the reduction in the Ptc function, Smoothened enhancing the function and/or improves the function of the Hedgehog. The terms "reduced function" and "increasing function" respectively refer to an aberrant modification or mutation, for example, gene Ptc, Hedgehog Smoothened gene or gene, or a decrease or increase of the expression level of this gene, which results in a phenotype which resembles the contact of the cell with a Hedgehog protein, such as aberrant activation of the Hedgehog pathway Il is reminiscent of an underactive Smo. The mutation may include a reduction in the ability of the products of genes Ptc or Smo for the regulation of the activity level of proteins Gli/Ci, such as Gli1, Gli2 and Gli3.

As used here, an immortalized cells are cells that have been chemically modified, and/or recombinant cells, and indicate that these cells have the ability to grow in an unlimited number of divisions in culture.

The term "LD50" means the dose of the drug that is lethal for 50% of test patients.

"Patient" or "subject" means the body is subjected to treatment by the method of the present invention, and may imply or person, or animal.

The phrase "pharmaceutically acceptable" is used here to refer to those compounds, materials, compositions and/or dosage forms which are, within the investigated medical judgments are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable ratio of benefit/risk.

The phrase "pharmaceutically acceptable carrier", as used here, means a pharmaceutically acceptable material, composition or binder, such as a liquid or solid filler, solvent, inert of napolnitel is, excipient (e.g., lubricant, powder magnesium, calcium or zinc stearate or stearic acid) or soluble encapsulating material included in the transfer or transportation of the considered compound from one organ or body part to another body or body part. Each carrier must be acceptable in the sense to be compatible with other ingredients of the composition and is not harmful for the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include : (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; (4) powdered tragakant; (5) malt; (6) gelatin; (7) talc; (8) inert fillers, such as cocoa butter and wax for suppositories; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as etiloleat and tillaart; (13) agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid is; (16) pyrogen-free water; (17) isotonic saline; (18) ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and/or polyanhydrides and (22) other non-toxic compatible substances used in pharmaceutical compositions.

The term "prodrug" is intended to encompass compounds which, under physiological conditions, are converted into therapeutically active tool of the present invention. A common method for the production of prodrugs should include selected residues, which are hydrolyzed under physiological conditions to form the desired molecule. In other embodiments, the prodrug is converted by enzymatic activity (or other physiological activity) of an animal host.

As used here, "proliferating" and "proliferation" refers to cells in mitosis.

Throughout this application the term "proliferative disorder of the skin" refers to any disease/disorder of the skin, marked by unwanted or aberrant proliferation of skin tissue. These States are usually characterized by the proliferation of epidermal cells or incomplete differentiation of cells and include, for example, X-linked ichthyosis, psoriasis, atopic dermatitis, allergic contact dermatitis, epidermolizei wipe the keratosis and seborrheic dermatitis. For example, epidermodysplasia is a form of defective development of the epidermis. Another example is bullosa, which refers to the weakened condition of the epidermis with the formation of a bull or spontaneously, or at the site of injury.

The term "therapeutic index" refers to therapeutic index of a drug defined as LD50/ED50.

The term "transgenic cell" refers to cells that are spontaneously transformed into a state of unlimited growth, i.e. they have acquired the ability to grow through an indefinite number of divisions in culture. Transformed cells can be characterized by such terms as "neoplastic", "anaplastic" and/or "hyperplastic", in relation to their lower growth control.

The term "subject"as used here, refers to an animal, typically a mammal, or the person who was the target of treatment, observation and/or experiment. When this term is used in conjunction with the introduction of the compound or drug, the subject is subjected to treatment, observation and/or the introduction of compounds or drugs. The phrase "therapeutically effective amount", as used here, means the number of connections, material, or composition comprising the compound of the present invention that I have is effective for in order to produce some desired therapeutic effect in at least a subpopulation of the cells of the animal in an acceptable ratio of benefit/risk, applicable to any treatment.

The phrases "parenteral administration" and "applied parenterally"as used here means the routes of administration other than enteral and local introduction, usually by injection, and include, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, lower eyelid, intracardiac, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, pagkapanalo, subarachnoid, spinal and vnutrigrudne injection and infusion.

The phrases "systemic impact", "applied systemically", "peripheral effect and applicable peripheral", as used here, means introducing the compound, drug or other material other than directly to the rest of the Hedgehog pathway associated with the disorder, such that arrives in the system of the patient and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.

The term "sugar"as used here, refers to a natural or non-natural monosaccharide, disaccharide or oligosaccharide, comprising one or more of the pyranose rings or fu is anosy. Sugar can be covalently linked to steroid alkaloid present invention through an ether linkage or through an alkyl linkage. In certain embodiments of the implementation charigny the balance can be covalently linked to steroid alkaloid present invention in the anomeric center Zaharenko rings.

The term "biradical", as used herein, refers to any number of divalent groups such as alkyl, alkeline, alkyline, aryl, cycloalkyl, heterocytolysine, kalkilya, heteroaryl and heteroalkyl group. For example,is divalent alkyl radical;is divalent alkyl radical;is Uralkaliy divalent radical; andis (alkyl)heteroalkyl divalent radical. Typical examples include alkylene General structure (CH2)xwhere X is 1-6 and the corresponding alkanolamine and akinleye linkers having 2-6 carbon atoms and one or more double or triple links; cycloalkene group having 3-8 ring members; and kalkilya group, where one open valence is on the aryl ring, and each of the alkyl fragments, such as and its isomers.

Compounds of the invention

The present invention provides analogues cyclopamine, as well as their isolated and purified form, including synthetic and semi-synthetic analogs and pharmaceutical compositions containing such analogs. In one embodiment, the present invention provides compounds represented by the compound of the formula1:

(1)

or its pharmaceutically acceptable salt;

in which each R1and R8independently represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl, heteroalkyl, halide, sulfhydryl, alkylthio, aaltio, Uralkali, hydroxyl, alkoxyl, aryloxy, acyloxy, amino, alkylamino, arylamino, acylamino, aralkylamines, nitro, atillio, carboxamide, sulfonamide, carboxyl, nitrile, sulfate, -OP(L)(OR20)2, -X-C(L)-R21or-X-C(L)-X-R21;

where Rlmay also be a sugar;

X represents O or NR, where R represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl or aralkyl;

L represents O or S;

R2and R9independently represent H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, alkoxyl, aryloxy, acyloxy, halide, sulfhydryl, alkylthio, arieti is, Uralkali, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamines, heteroaryl or heteroalkyl;

each R5and R11independently represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamines, alkylsilane, aralkylamines, Arellano, alkylthio, Uralkali, aaltio, heteroaryl or heteroalkyl;

each R3, R4, R6, R7, R13and R14independently represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamines, heteroaryl or heteroalkyl;

or R1and R2and/or R8and R9taken together with the carbon to which they are bound, form a -(C=O)-, -(C=S)-, -(C=N(OR20))-, -(C=N(R20))-, -(C=N(N(R20)(R20))) or form an optionally substituted 3-8-membered cycle; or

R4and R5taken together, and/or R5and R6taken together, and/or R10and R11taken together form a double bond or form a group represented by1b

where Z represents NR21, O or C(R23)(R23);

R12represents H, alkyl, who lceil, quinil, aryl, cycloalkyl, heteroseksualci, hydroxyl, aralkyl, heteroaryl, heteroalkyl, halogenated, alkoxy, -C(O)R21, -CO2R21, -SO2R21, -C(O)N(R21)(R21), -[C(R21)2]q-R21, -[(W)-N(R21)C(O)]qR21, -[(W)-C(O)]qR21, -[(W)-C(O)O]qR21, -[(W)-(O)C(O)]qR21, -[(W)-SO2]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-N(R21)]qR21or -[(W)-S]qR21;

where each W is biradical, and q is 1, 2, 3, 4, 5 or 6;

R15, R16and R17independently represent H, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamines; or R15and R16taken together with the carbon to which they are attached, form-C(O)- or-C(S)-;

R18and R19independently represent H, alkyl, aralkyl, halide, amido, or ester;

R20represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl or heteroaryl; or any two of R20in a single substituent can be taken together to form a 4-8 membered optionally substituted cycle;

R21represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl is, heteroseksualci, aralkyl, heteroaryl, heteroalkyl or -[C(R20)2]p-R25where p is 0-6; or any two of R21in a single substituent can be taken together to form a 4-8 membered optionally substituted cycle;

R23represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl, heteroalkyl, halide, alkoxyl, aryloxy, acyloxy, silyloxy, nitrile, -C(O)R21, -CO2R21, -SO2R21and-C(O)N(R21)2;

R25represents hydroxyl, acylamino, -N(R20)COR20, -N(R20)C(O)OR20, -N(R20)SO2(R20), -COR20N(R20)2, -OC(O)R20N(R20)(R20), -SO2N(R20)(R20), -N(R20)(R20), -COOR20, -C(O)N(HE)(R21), -OS(O)2OR20, -S(O)2OR20, -OP(L)(OR20)(OR20), -NP(O)(OR20)(OR20) or-P(O)(OR20)(OR20).

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R13, R14, R15, R16and R17represent hydrogen.

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R1represents hydroxyl, sugar, -OP(L)(OR20 )2, -X-C(L)-R21or-X-C(L)-X-R21; or R1and R2taken together with the carbon to which they are attached, form-C(O)-.

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R4and R5taken together form a double bond.

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R1and R2taken together with the carbon to which they are attached, form-C(O)-.

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R1represents hydroxyl, and R2represents N.

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R1represents hydroxyl, R2represents H; and R5and R6taken together form a double bond; or R5and R6taken together form a group represented by1b;

where

Z represents C(R23)(R23).

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R10and Rsup> 11taken together form a double bond; or R10and Rlltaken together form a group represented by1b;

where

Z represents C(R23)(R23).

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R5and R6taken together form a double bond and R10and Rlltaken together form a double bond.

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R1and R2taken together with the carbon to which they are attached, form-C(O)-; R4and R5taken together form a double bond; and R10and R11taken together form a double bond; or R10and R11taken together form a group represented by1b;

where

Z represents C(R23)(R23).

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R1represents hydroxyl, and R2represents H; R10and Rl1taken together form a double bond; or R10and R11taken together form a group represented by1b;

img src="https://img.russianpatents.com/1063/10637109-s.jpg" height="22" width="40" />

where

Z represents C(R23)(R23).

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R8and R9represent hydrogen; or R8and R9taken together with the carbon to which they relate, are-C(O)-.

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R12represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroalkyl, halogenated, hydroxyl, alkoxyl, -[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-C(O)]qR21or -[(W)-C(O)O]qR21.

In some embodiments, the communication of the present invention presents1and the attendant definitions wherein R13, R14, R15, R16and R17represent hydrogen; and R12represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroalkyl, halogenated, hydroxyl, alkoxyl, -[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-C(O)]qR21or -[(W)-C(O)O]qR21.

In some embodiments, osushestvlyaetsya the present invention presents 1and the attendant definitions wherein R4and R5taken together form a double bond; R1and R2taken together with the carbon to which they are attached, form-C(O)-; and R12represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroalkyl, halogenated, hydroxyl, alkoxyl, -[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-C(O)]qR21or -[(W)-C(O)O]qR21.

In some embodiments, the communication of the present invention represented by 1 and the attendant definitions wherein R1represents hydroxyl and R2represents H; and R12represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroalkyl, halogenated, hydroxyl, alkoxyl, -[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-C(O)]qR21or -[(W)-C(O)O]qR21.

In some embodiments, the communication of the present invention compound represented by the formula:

where

or

R12represents H, alkyl, aryl, cycloalkyl, heteroseksualci, hydroxyl, aralkyl, heteroaryl, heteroalkyl, halogenated, alkoxyl, -C(O)R21, -CO2R21, -SO2R21, -C(O)N(R21)(R21), -[C(R21)2]p-R21, -[(W)-N(R21)C(O)]qR21, -[(W)-C(O)]qR21, -[(W)-C(O)O]qR21, -[(W)-OC(O)]qR21, -[(W)-SO2]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-N(R21)]qR21or -[(W)-S]qR21; where each W independently represents biradical;

q is 1, 2, 3, 4, 5 or 6;

R20represent H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl or heteroaryl; or any two of R20in a single substituent can be taken together to form a 4-8 membered optionally substituted cycle;

R21represents H, alkyl, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl, heteroalkyl or -[C(R20)2]p-R25; or any two of R21can be taken together to form a 4-8 membered optionally substituted cycle;

R25represents hydroxyl, acylamino, -N(R20)COR20, -N(R20)C(O)OR20, -N(R20)SO2(R20), -COR20N(R20)2, -OC(O)R20N(R20)(R20), -SO2N(R20)(R20), -N(R20)(R20), -COOR20, -C(O)N(HE)(R21), -OS(O)2OR19 , -S(O)2OR20, -OP(L)(OR20)(OR20), -NP(O)(OR20)(OR20) or-P(O)(OR20)(OR20).

The present invention specifically provides compounds represented by the group consisting of

In some embodiments, the communication of the present invention represented by any of the aforementioned compound and the attendant definitions, where the compound represented by the formula:

In one embodiment, the present invention provides compounds represented by the compound of the formula2:

or its pharmaceutically acceptable salt;

in which each R1and R8independently represent H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl, heteroalkyl, halide, sulfhydryl, alkylthio, aaltio, Uralkali, hydroxyl, alkoxyl, aryloxy, acyloxy, amino, alkylamino, arylamino, acylamino, aralkylamines, nitro, atillio, carboxamide, sulfonamide, carboxyl, nitrile, sulfate, -OP(L)(OR20)2, -X-C(L)-R21or-X-C(L)-X-R21;

where Rlmay also be a sugar;

X presented yet a O or NR, where R represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl or aralkyl;

L represents O or S;

R2and R9independently represent H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, alkoxyl, aryloxy, acyloxy, carboxyl, halide, sulfhydryl, alkylthio, aaltio, Uralkali, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamines, heteroaryl or heteroalkyl;

each R5and R11independently represent H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamines, alkylsilane, aralkylamines, Arellano, alkylthio, Uralkali, aaltio, heteroaryl or heteroalkyl;

each R3, R4, R6, R7, R13and R14independently represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamines, heteroaryl or heteroalkyl;

where R1and R2and/or R8and R9taken together with the carbon to which they are bound, form a -(C=O)-, -(C=S)-, -(C=N(OR20))-, -(C=N(R20))-, -(C=N(N(R20)(R20))) or form an optionally substituted 3-8-membered cycle; Il the

R4and R5taken together, and/or R5and R6taken together, and/or R10and R11taken together form a double bond or form a group represented by1b

where Z represents NR21, O or C(R23)(R23);

R12represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, hydroxyl, aralkyl, heteroaryl, heteroalkyl, halogenated, alkoxy, -C(O)R21, -CO2R21, -SO2R21, -C(O)N(R21)(R21), -[C(R21)2]q-R21, -[(W)-N(R21)C(O)]qR21, -[(W)-C(O)]qR21, -[(W)-C(O)O]qR21, -[(W)-OC(O)]qR21, -[(W)-SO2]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-N(R21)]qR21or -[(W)-S]qR21;

where W represents biradical, and q is 1, 2, 3, 4, 5 or 6;

R15, R16and R17independently represent H, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamines; or R15and R16taken together with the carbon to which they are attached, form-C(O)- or-C(S)-;

R18and R19independently represent H, alkyl, aralkyl, halide, amido, or ester;

R20independently before the hat is N, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl or heteroaryl; or any two of R20in a single substituent can be taken together to form a 4-8 membered optionally substituted cycle;

R21independently represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl, heteroalkyl or -[C(R20)2]p-R25where p is 0-6; or any two of R21in a single substituent can be taken together to form a 4-8 membered optionally substituted cycle;

R23independently represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl, heteroalkyl, halide, alkoxyl, aryloxy, acyloxy, silyloxy, nitrile, -C(O)R21, -CO2R21, -SO2R21and-C(O)N(R21)2; and

R25represents hydroxyl, acylamino, -N(R20)COR20, -N(R20)C(O)R20, -N(R20)SO2(R20), -COR20N(R20)2, -OC(O)R20N(R20)(R20), -SO2N(R20)(R20), -N(R20)(R20), -COOR20, -C(O)N(HE)(R21), -OS(O)2OR20, -S(O)2OR20, -OP(L)(OR20)(OR20), -NP(O)(OR20)(OR20) or-P(O)(OR20)(OR20),

provided that there is at least one group, is expressed by formula 1b, on the specified connection formula 2.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions, where in this description R13, R14, R15, R16and R17represent hydrogen.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R1represents hydroxyl, sugar, OP(L)(OR20)2, -X-C(L)-R21or-X-C(L)-X-R21; or R1and R2taken together with the carbon to which they are attached, form-C(O)-.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R4and R5taken together form a double bond.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R1and R2taken together with the carbon to which they are attached, form-C(O)-.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R1represents a hydroxyl, a R2represents N.

In some embodiments, the communication of the present invention represented by f is rmulas 2 and the attendant definitions, where R1represents hydroxyl, and R2represents H; R5and R6taken together form a double bond; or R5and R6taken together form a group represented by 1b;

where

Z represents C(R23)(R23).

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R10and R11taken together form a double bond; or R10and R11taken together form a group represented by 1b;

where

Z represents C(R23)(R23).

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R5and R6taken together form a double bond, and R10and R11taken together form a double bond.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R1and R2taken together with the carbon to which they are attached, form-C(O)-; R4and R5taken together form a double bond; and R10and R11taken together form a double bond; or R10and R11taken together, form a group, before the purposes of the 1b;

where

Z represents C(R23)(R23).

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R1represents hydroxyl and R2represents H; R10and R11taken together form a double bond; or R10and R11taken together form a group represented by 1b;

where

Z represents C(R23)(R23).

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R8and R9represent hydrogen; or R8and R9taken together d carbon, with which they are associated, are-C(O)-.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R12represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroalkyl, halogenated, hydroxyl, alkoxyl, -[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-C(O)]qR21or -[(W)-C(O)O]qR21.

In some embodiments, the communication of the present invention to depict what aulani formula 2 and the attendant definitions, where R13, R14, R15, R16and R17represent hydrogen, and R12represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroalkyl, halogenated, hydroxyl, alkoxyl, -[(W)-N(R21)C(O)]q21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)]qR21, -[(W)-C(O)]qR21or - [(W)-C(O)Oh]qR21.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R4and R5taken together form a double bond; R1and R2taken together with the carbon to which they are attached, form-C(O)-; and R12represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroalkyl, halogenated, hydroxyl, alkoxyl, -[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-C(O)]qR21or -[(W)-C(O)O]qR21.

In some embodiments, the communication of the present invention represented by formula 2 and the attendant definitions wherein R1represents hydroxyl and R2represents H; and R12represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroalkyl, halogenated, hydroxyl, alkoxyl, -[(W)-N(R 21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-C(O)]qR21or -[(W)-C(O)O]qR21.

In some embodiments, the communication of the present invention compound represented by the formula:

or

where

R12represents H, alkyl, aryl, cycloalkyl, heteroseksualci, hydroxyl, aralkyl, heteroaryl, heteroalkyl, halogenated, alkoxy, -C(O)R21, -C(O)N(R21)(R21), -[C(R21)2]p-R21, -[(W)-N(R21)C(O)]qR21, -[(W)-C(O)]qR21, -[(W)-C(O)O]qR21, -[(W)-OC(O)]qR21, -[(W)-SO2]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-C(O)N(R21)]qR21, -[(W)-O]qR21, -[(W)-N(R21)]qR21or -[(W)-S]qR21;

q is 1, 2, 3, 4, 5 or 6;

R20represents H, alkyl, alkenyl, quinil, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl, or heteroalkyl; or any two of R20in a single substituent can be taken together to form a 4-8 membered optionally substituted cycle;

R21represents H, alkyl, aryl, cycloalkyl, heteroseksualci, aralkyl, heteroaryl, heteroalkyl or -[C(R20)2]p 25; or any two of R21can be taken together to form a 4-8 membered optionally substituted cycle;

R25represents hydroxyl, acylamino, -N(R20)COR20, -N(R20)C(O)OR20, -N(R20)SO2(R20), -COR20N(R20)2, -OC(O)R20N(R20)(R20), -SO2N(R20)(R20), -N(R20)(R20), -COOR20-C(O)N(HE)(R21), -OS(O)2OR19, -S(O)2OR20, -OP(L)(OR20)(OR20), -NP(O)(OR20)(OR20) or-P(O)(OR20)(OR20).

The present invention specifically provides compounds represented by the group consisting of;

,,

,,,,

,,,

In some embodiments, the communication of the present invention represented by any of the aforementioned compounds and attendant op what edelenyi, where the compound represented by the formula:

In some embodiments implementing the present invention relates to pharmaceutical compositions containing any one or more of the aforementioned compounds; and pharmaceutically acceptable filler.

Synthesis of steroidal alkaloid compounds

Cyclopropylidene steroid alkaloid derivatives of the present invention can be obtained directly from the steroid alkaloid, isolated as a natural product (or synthesized) or N-protected forms of these compounds. Suitable protective groups for the nitrogen include, but are not limited to, Fmoc, Alloc, Boc, Troc, trifurcation, Totila, Cbz, atellana and Bn.

For cyclopropylamine steroid alkaloid can be applied to a variety of cyclopropylamine funds. 1,1-Halogenallylacetic complexes, including reactive substances known as carbenoid commonly used for cyclopropylamine olefins. These reagents standard way get using diadakan or diazoalkane and metal or ORGANOMETALLIC compounds such as Et2Zn,iBu3Al, samarium, copper, rhodium or palladium. In some embodiments, the implementation of Et2Zn and deadmetal use to influence cyclopropylamine. Can the e applied to other known methods of cyclopropylamine, such as methods with the use of sulfur ylides to interact with the olefin, paired with the carbonyl to join groups CH2or CH-alkyl or CH-aryl, and catalyzed by metal decomposition diazoalkane and α-diazocarbonyl compounds such as diazomethane and ethyl diazo acetate: these methods can easily give cyclopropanes with alkyl, aryl, alkoxycarbonyl(-COOR) or acyl substituents. For example, the accession of ethyldiazoacetate (EtO2C-C(N2)-Me) to the olefinic compound in an organic solvent containing a metal catalyst, such as copper or palladium, leads to the formation of cyclopropane containing the group represented by the formula1bwhere Z is C(R23)2in which one of R23represents Me and the other R23represents COOEt.

Through the careful selection cyclopropenones funds selectivity with respect to position can be achieved when cyclopropylamine steroid alkaloids more than one olefin. For example, if diameter and Et2Zn is used for cyclopropylamine of such as iervin under certain conditions, only the olefin, more rich in electrons will interact.

The diastereoselectivity of cyclopropylamine can be controlled in different ways. For example, lowering the temperature of reaction is AI cyclopropylamine can lead to higher diastereoselectivity. The alternative, can be used chiral cyclopropane tool that can distinguish each Deserializer side steroid alkaloid. Enantioselectivity when cyclopropylamine can also be achieved by using a substrate-directed reactions (i.e. cyclopropylamine allylic alcohols using Et2Zn/CH2I2reagents).

Reaction cyclopropylamine can be carried out in an aprotic solvent, preferably that in which the ingredients of the reaction are essentially soluble. Suitable solvents include ethers, such as diethyl ether, 1,2-dimethoxyethane, diglyme, tert-butyl methyl ether, tetrahydrofuran and the like; halogenated solvents such as chloroform, dichloromethane, dichloroethane and the like; aliphatic or aromatic hydrocarbon solvents such as benzene, xylene, toluene, hexane, pentane and the like; esters and ketones such as ethyl acetate, acetone and 2-butanone; polar aprotic solvents such as acetonitrile, dimethylsulfoxide, dimethylformamide and the like; or combinations of two or more solvents. In a preferred embodiment, dichloromethane is the solvent used to cyclopropylamine when using dialkylzinc and diameter.

After synthesis cyclopropanemethanol with rodnoy alkaloid Central part of the connection can be derivatization using a variety of functionalization reactions, known in this field. Representative examples include reactions combination with palladium to alcoholgeodon or aryl halides, oxidation, recovery, reactions with nucleophiles, reaction with electrophiles, the reaction of penicillinase, the introduction of a protective group, removing the protective groups, etc.

In the presence of Lewis acids or Bronsted cyclopropylamino analogues of the present invention undergo a previously unknown realignment and expansion cycle, which allows to obtain new cyclopamine analogues in which the ring D is increased by one carbon atom. Cyclopropane ring can be substituted or unsubstituted. In cases where cyclopropane ring is substituted, the group attached to the methylene cyclopropane will be incorporated in the D ring after rearrangement and ring extensions. Suitable acids include, but are not limited to, ZnI2BF3, methansulfonate acid, variatsionnye acid and HCl. In a preferred embodiment of the invention used the Lewis acid is a BF3. These homologisation analogs can be further functionalized using a variety of functionalization reactions known in this field. Representative examples include reactions combination with palladium to alkoholen the Dov or aryl halides, oxidation, recovery, reactions with nucleophiles, reaction with electrophiles, the reaction of penicillinase, the introduction of a protective group, removing the protective groups, etc.

The methods of the invention

The present invention further provides methods of treating, improving one or more symptoms and reduce the severity of hyperproliferative disorders such as cancer, as well as other disorders and conditions mediated by the Hedgehog pathway.

Currently, the antagonists of the Hedgehog pathway are investigated in a large number of clinical conditions, where therapeutic effect can be obtained for the condition or disorder, slowing one or more aspects of the activity of the Hedgehog pathway. Although the main focus of research has been directed to cancer, the researchers found that inhibition of Hedgehog pathway of low molecular weight compounds improves the symptoms of psoriasis (Tas, et al., 2004 Dermatology209: 126-131,published asUS 20040072913(included here as a reference)). Psoriasis is a very common, chronic skin disorder that is usually characterized by skin lesions, usually consisting of erythematous papules and patches with silvery scale, although there are changes on the skin and in other parts of the body. At the present time believe that psoriasis is an autoimmune disease, but its cause all asalaha studied. In the first stage, the local application cyclopamine on psoriatic lesions led to complete or partial regression of the lesions with a decrease in inflammatory cells (Tasand others,above).

Antagonists of the Hedgehog pathway of the present invention can be used for treatment or prevention of psoriasis when used as a single agent or when used in combination with one or more other protivopsoriaticescoe means, including, but not limited to data, corticosteroids, tar, calcipotriene, tazarotene, calcineurin inhibitors, irradiation with ultraviolet rays, methotrexate, retinoids, cyclosporine, immunomodulating drugs, etanercept, alefacept, efalizumab and infliximab.

It is shown that many tumors and proliferative state depend on the way the Hedgehog. On the growth and survival of these cells can be affected by treatment with the compounds of the present invention. For example, inhibition of the Hedgehog pathway of low molecular weight compounds to slow the growth of basal cell carcinoma (Williams,and others, 2003PNAS100: 4616-21),Protocol (Berman et al., 2002 Science297: 1559-61),pancreatic cancer (Bermanet al., 2003Nature425:846-51),cancer of the stomach (Bermanet al., 2003Nature425:846-51,published PCT applicationWO 05/013800),esophageal cancer (Bermanet al., 2003Nature425: 846-51),sakalagraha (Watkins et al., 2003.Nature422:313-7)and prostate cancer (Karhadkaret al., 2004.Nature431:707-12).

In addition, it has been shown that many types of cancer have uncontrolled activation of the Hedgehog pathway, such as breast cancer (Kuboet al., 2004.Cancer Research64: 6071-4),hepatocellular cancer (Patilet al., 2005. 96thAnnual AACR conference, abstract#2942Sicklicket al., 2005.ASCO annual meeting, abstract#9610),hematological malignant diseases (Watkins and Matsui, unpublished results), basal cell carcinoma (Bale & Yu,2001.Human Molec. Genet.10: 757-762Xieet al., 1998Nature391:90-92),medulloblastoma (Pietschet al., 1997.Cancer Res.57: 2085-88) andstomach cancer (Maet al., 2005Carcinogenesis May19, 2005(Epub)).

Cancer or neoplastic diseases and related disorders that can be treated by the introduction of the compounds and compositions of the present invention include, but are not limited to data, cancer of the adrenal cortex, cancer of the anal canal, aplastic anemia, bile duct cancer, bladder cancer, bone cancer, tumors of the brain/CNS, breast cancer, cervical cancer, non-Jackinsky lymphoma, colon cancer, rectal cancer, endometrial cancer, esophageal cancer, tumors of the Ewing family, cancer, eye, gallbladder cancer, gastrointestinal carcinoid tumor, gastrointestinal gastrointestinal stromal tumors, gestational USSR disease, the disease is Ognina, Kaposi's sarcoma, kidney cancer, larynx cancer and podletochnyh cancer, acute lymphocytic leukemia, acute myeloid leukemia, children's leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, liver cancer, lung cancer, carcinoid lung tumor, non-Hodgskins lymphoma, male breast cancer, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, cancer of the nasal cavity and paranasal region, nasopharynx cancer, neuroblastoma, oral cancer, cancer of the oropharynx carcinoma, osteosarcoma, ovarian cancer, pancreatic cancer, cancer of the penis, pituitary tumor, prostate cancer, retinoblastoma, rhabdomyosarcoma cancer , salivary gland cancer, sarcoma, melanoma, skin cancer non-melanoma, stomach cancer, cancer of the testes, cancer of the thymus, thyroid cancer, sarcoma cancer, cancer of the vagina, vulvar cancer, macroglobulinemia waldenstrom and tumor Wilms ' tumor.

The methods and compositions of the present invention can be used in the treatment of cancer in humans, such as basal cell cancers and other tumors of epithelial tissues such as skin. Additionally, the compounds of the present invention can be used as part of treatment for basal cell carcinoma, pancreatic cancer, prostate cancer, sarcoma, lymphoma, leukemia, stomach cancer, cancer of the gastrointestinal tract, multiple myeloma, m is coletocna lung cancer, gliomas, breast cancer, hepatocellular cancer or Protocol through the application of a therapeutically effective amount of at least one of the compounds of the present invention as the sole means or in combination with another anticancer agent.

The methods and compositions of the present invention can be used in the treatment of neoplastic or hyperplastic transformation, such as those that can occur in the Central nervous system. For example, the compounds of the present invention can be used to cause such transformed cells to become or post mitotic or apoptotic. The provided method can therefore be used as part of treatment, for example, malignant gliomas, meningiomas, medulloblastoma, neuroectodermal tumors, and ependymomas.

In one embodiment, an existing method may be used as part of treatment of malignant Protocol and other primary malignant neuroectodermal tumors of the Central nervous system.

In some embodiments implementing the present invention relates to a method of treatment of cancer, comprising the administration to the patient, if he needs it, a therapeutically effective amount of one or more of any of the above compounds.

In some embodiments implementing the present invention relates to a method of treatment of cancer, includes introduction to the needy in this patient a therapeutically effective amount of a compound of any one or more of the above compounds, in cases when the cancer is localized in the patients head, neck, nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx, hypopharynx, salivary glands, paraganglioma, pancreas, stomach, skin, esophagus, liver and biliary tree, bone, small intestine, colon, rectum, ovaries, prostate, lung, breast, lymphatic system, blood, Central nervous system, bone marrow or the brain.

In some embodiments implementing the present invention relates to a method of treating cancer, comprising an introduction to the needy in this patient a therapeutically effective amount of a compound of any one or more of the above compounds, in cases when the cancer is a basal cell cancer, pancreatic cancer, prostate cancer, sarcoma, lymphoma, leukemia, stomach cancer, esophagus cancer, cancer of the bile ducts, colon cancer, multiple myeloma, small cell lung cancer, glioma, breast cancer, hepatocellular cancer, or medulloblastoma.

In some embodiments implementing the present invention relates to the aforementioned method, where the connection is s used in combination with radiation therapy or other anti-cancer chemotherapeutic agent.

In some embodiments implementing the present invention relates to any of the above-mentioned method where the compound is applied locally to the tumor or systemically to the patient's body.

In some embodiments implementing the present invention relates to any of the above-mentioned method, where the method of administration specified connection represents the inhalation, oral, intravenous, sublingual, ocular, transdermal, rectal, vaginal, local, intramuscular, intraarterial, intrathecal, subcutaneous, buccal or intranasal.

In some embodiments implementing the present invention relates to any of the above-mentioned method, where the route of administration is oral, intravenous or local.

In some embodiments implementing the present invention relates to a method antagonistic effects on Hedgehog path in a cell, comprising bringing into contact with cells expressing Smoothened, with an effective amount of any one or more of the aforementioned compounds.

In some embodiments implementing the present invention relates to a method antagonistic effects on Hedgehog path in a cell, comprising bringing into contact with cells expressing Smoothened, with an effective amount of any one or more of the above-mentioned connection is to be made where the specified cell expressing Smoothened, in contact with the specified connectionin vitro.

In some embodiments implementing the present invention relates to a method antagonistic effects on Hedgehog path in a cell, comprising bringing into contact with cells expressing Smoothened, with an effective amount of any one or more of the above compounds, where the specified cell expressing Smoothened, in contact with the specified connectionin vivo.

In some embodiments implementing the present invention relates to a method antagonistic effects on Hedgehog path in a cell, comprising bringing into contact with cells expressing Smoothened, with an effective amount of any one or more of the above compounds, where the specified cell expressing Smo, is in contact with the specified connection within the patient's body.

In some embodiments implementing the present invention relates to a method of treatment or prevention of psoriasis in a patient, including an introduction to the needy in this patient a therapeutically effective amount of any one or more of the aforementioned compounds.

In some embodiments implementing the present invention relates to the aforementioned method of treatment or prevention of psoriasis, where the route of administration specified what about the composition of a local call.

In some embodiments, the implementation of one or more compounds of the present invention are used for treatment and prevention of psoriasis in combination with one or more protivopsoriaticescoe means, including, but not limited to data, corticosteroids, tar, calcipotriene, tazarotene, calcineurin inhibitors, irradiation with ultraviolet rays, methotrexate, retinoids, cyclosporine, immunomodulating drugs, etanercept, alefacept, efalizumab and infliximab.

Treatment of cancer in combination with chemotherapy or radiation therapy

In some embodiments, the implementation of one or more compounds of the present invention are used to treat or prevent cancer or neoplastic disease in combination with one or more anticancer chemotherapeutics, including, but not limited to data, gemcitabine, methotrexate, Taxol, mercaptopurine, tioguanin, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosamine, cisplatin, carboplatin, mitomycin, dacarbazine, procarbazine, etoposide, prednisolone, dexamethasone, citarum, computacin, bleomycin, doxorubicin, idarubitsin, daunorubicin, dactinomycin, plicamycin, mitoxantrone, asparaginase, vinblastine, vincristine, vinorelbine, paclitaxel, and docetaxel. In a preferred embodiment ar is about or more compounds of the present invention are used for treatment and prevention of cancer or neoplastic disease in combination with one or more chemotherapeutic or other anti-cancer means, including, but not limited to data, radiation (e.g. gamma radiation), nitrogen mustard (for example, cyclophosphamide, ifosfamide, trofosfamide, chlorambucil, estramustin and melphalan), nitrosoanatabine (for example, carmustine (BCNU) and lomustin (CCNU)), alkyl sulphonates (e.g., busulfan and treosulfan), triazine (e.g., dacarbazine and temozolomide), platinum compounds (e.g., cisplatin, carboplatin and oxaliplatin), Vinca alkaloids (e.g. vincristine, vinblastine, vindesine and vinorelbine), taxoid (e.g., paclitaxel and docetaxel), appendfiles (e.g., etoposide, teniposide, topotecan 9-aminocamptothecin, containinaton, Kristol, mitomycin C and mitomycin C), antimetabolites, inhibitors of DHFR (for example, methotrexate and trimetrexate), inhibitors of IMP dehydrogenase (e.g., mycophenolic, tianfuan, ribavirin and EICAR)inhibitors ribonucleotides (e.g., hydroxyurea and deferoxamine), analogs of uracil (e.g., fluorouracil, floxuridine, doxifluridine, redirected and capecitabine), analogs of cytosine (e.g., cytarabine (Ara C), cytosine arabinoside and fludarabine), purine analogues (for example, mercaptopurine and tioguanin), antiestrogens (for example tamoxifen, raloxifene and megestrol), LHRH agonists (for example, goserelin and acetate leuprolide), antiandrogens (for example flutamide, and bikalutamid), analogs of vitamin D3 (e.g. the R, EB 1089, CB 1093 and KH 1060), photodynamic therapy (for example, verteporfin (BPD-MA), phthalocyanine, photosensitizer Pc4 and dimethoxyphenethyl AND (2BA-2-DMHA)), cytokines (for example, α-interferon, γ-interferon and tumor necrosis factor)inhibitors isoprenaline (e.g., lovastatin), dopaminergic neurotoxins (e.g., ion 1-methyl-4-phenylpyridine)inhibitors of the cell cycle (for example, staurosporin), aktinomitinov (for example, actinomycin D, and dactinomycin), bleomycin (for example, bleomycin A2, bleomycin B2 and peplomycin), anthracyclines (e.g. daunorubicin, doxorubicin (adriamycin), idarubitsin, epirubicin, pirarubicin, zorubicin and mitoxantrone), MDR inhibitors (e.g. verapamil), inhibitors of Ca2+-ATP-ases (e.g., thapsigargin), antibodies (e.g., Avastin, Erbitux, Rituxan, bexxar), corticosteroids (e.g., prednisone, predisone etc), imatinib, thalidomide, lenalidomide, bortezomib, gemcitabine, erlotinib, gefitinib, sorafenib and sutinen.

Chemotherapeutic agent and/or radiation therapy can be administered according to therapeutic protocols well known in the art. Specialists in the art it will be obvious that the introduction of the chemotherapeutic and/or radiation therapy may vary differently depending on the disease, against which is applied the treatment, and the known effects of chemotherapeutic drugs and/or radiation therapy for this disease. In addition, in accordance with the knowledge of the skilled Clinician, therapeutic protocols (e.g., dosage and time of administration) may be different due to the observed patient effects from the applied therapeutics (i.e. antineoplastics funds or radiation) and in view of the observed responses of the disease to introduce a therapeutic agent and the observed adverse effects.

In addition, in General, the compounds of the present invention and a chemotherapeutic agent should not be administered in the same pharmaceutical composition, and may, because the physical and chemical characteristics of different, must be administered through various routes. For example, the compounds of the present invention can be administered intravenously to generate and maintain good blood levels, while the chemotherapeutic agent may be administered orally. The definition of the method of introduction and feasibility of the introduction, if possible, in the same pharmaceutical composition is within the knowledge of a qualified Clinician. The initial introduction may be performed according to established protocols known in the art, and then by a qualified Clinician may change the dosage, route of administration and time of administration, the basis of IVaS on observed effects.

Preferred chemotherapeutic agents or radiation will depend upon the diagnosis of the doctors and their judgments about the patient's condition and appropriate Protocol introduction.

The compound of the present invention and a chemotherapeutic agent and/or radiation may be administered simultaneously (e.g., simultaneously, essentially simultaneously or within the same treatment Protocol) or sequentially, depending on the nature of the proliferative disease, the condition of the patient and the actual choice of chemotherapeutic and/or radiation, which are administered together (i.e. within a single Protocol introduction) with the compound of the present invention.

If the connection of the present invention and a chemotherapeutic agent and/or radiation are not introduced simultaneously or essentially simultaneously, then the optimal order of introduction of the compounds of the present invention and a chemotherapeutic and/or radiation may be different for different tumors. Thus, in certain situations, the connection of the present invention may be administered first, followed by the appointment of chemotherapeutic and/or radiation; and in other situations chemotherapeutic agent and/or radiation may be administered first, followed by the appointment of preprogram the Oia of the present invention. This additional assignment may be repeated during a single treatment Protocol. Determination of the order of introduction and the number of repetitions of the introduction of each therapeutic agent for the treatment Protocol is fully within the knowledge of a qualified physician after evaluation of the disease being treated, and the condition of the patient. For example, the chemotherapeutic agent and/or radiation may be administered first, especially if it is a cytotoxic agent and then the treatment continued with the introduction of the compounds of the present invention, accompanied, when filled usefulness, the introduction of the chemotherapeutic and/or radiation, and so on until the treatment Protocol is completed.

Thus, in accordance with experience and knowledge of the practitioner may continue the treatment to modify each Protocol for the introduction of a component (therapeutic agent, for example, the compound of the present invention, chemotherapeutic agent or radiation) for treatment according to the individual needs of the patient.

The pharmaceutical composition

In yet another embodiment, the present invention provides pharmaceutically acceptable compositions, which include therapeutically effective amount of one or more compounds described above (formula1and ), formed together with one or more pharmaceutically acceptable carriers and/or solvents. The pharmaceutical compositions of the present invention may be specially formulated for administration in solid or liquid form, including adapted for the following: (1) oral administration, for example, medicine (aqueous or non-aqueous solutions or suspensions), tablets, e.g., intended for buccal, sublingual, and systemic use, pills, powders, granules, pastes for adhesion to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or composition for the continuous introduction; (3) topical application, for example, as a cream, ointment or plaster controlled release or spray solution applied to the skin; (4) vnutrivlagalischno or rectally, for example, as a Royal ring, cream or foam; (5) sublingually; (6) eye; (7) transdermally; or (8) through the lungs or (9) through the nose. As set forth above, certain embodiments of the existing connections may contain a basic functional group, such as amino or alkylamino, and, thus, capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable acids. The term "pharmaceutically who ielemia salts" in this respect refers to the relatively non-toxic salts of the compounds of the present invention with inorganic and organic acids. These salts can be preparedin situin the binder substance for injection or in the form of dosage forms in the production process, or by separately reacting a purified compound of the invention in its free base form with a suitable organic or inorganic acid, and excretion of salt, which is formed during subsequent purification. Typical salts include bromohydrin, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naftilan, mesilate, glucoheptonate, lactobionate and laurylsulphate, and the like (see, for example, Bergeet al.(1977) "Pharmaceutical Salts", J.Pharm. Sci.66:1-19).

Pharmaceutically acceptable salts of the compounds of the present invention include the conventional nontoxic salts or Quaternary ammonium salt compounds, for example, non-toxic organic or inorganic acids. For example, such conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric, Hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and salts derived from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, tartaric, citric, ascorbic, palmitin the traveler, maleic, hydroxymaleimide, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluensulfonate, methanesulfonate, ethicality, oxalic acid, setinova etc.

In other cases, the compounds of the present invention may contain one or more acidic functional groups and, thus, possess the ability to form pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term “pharmaceutically acceptable salts” in these instances refers to the relatively non-toxic, inorganic and organic basic additive salts of the compounds of the present invention. These salts can probably be obtainedin situin the environment for the introduction or methods, dosage forms, or via a specific interaction of the purified compound in its free acid form with a suitable base such as the hydroxide, carbonate or bicarbonate pharmaceutically acceptable metal cation, with ammonia or pharmaceutically acceptable oranicheskih primary, secondary or tertiary amine. Representative alkali or alkaline earth salts include salts of lithium, sodium, potassium, calcium, magnesium and aluminum and other Representative organic amines, applicable to basic education-additive sole is, include ethylamine, diethylamine, Ethylenediamine, ethanolamine, diethanolamine, piperazine and the like (see, for example, Berge et al.,supra).

The compositions may also be present moisturizers, emulsifiers and lubricants such as sodium lauryl sulfate and magnesium stearate, and colouring agents, funds release, covering tools, sweeteners, fragrances and flavors, preservatives and antioxidants.

Examples of pharmaceutically acceptable antioxidants include : (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, bottled hydroxyanisol (BHA), bottled hydroxytoluene (OSH), lecithin, propylgallate, alpha-tocopherol and the like; and (3) metalhalide tools, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

Methods of obtaining data ready-made forms or compositions include the stage of bringing into Association the compound of the present invention with the carrier and, optionally, one or more accessory ingredients. In General, the finished form get through homogeneous and directly bringing into Association the compound of the present invention with liquid carries the representatives or finely divided solid carriers or both and then, if necessary, shaping the product.

The pharmaceutical compositions of the present invention suitable for parenteral administration contain one or more compounds of the invention in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be introduced into sterile injectable solutions or dispersions just prior to use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostatic, soluble substances, which give the finished form isotonic properties in relation to the blood of the intended recipient or suspendida, or condensing means.

Examples of suitable aqueous and nonaqueous carriers which may be used in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), and suitable mixtures of vegetable oils, such as olive oil, and injectable organic esters, such as etiloleat. The desired fluidity can be installed, for example, through the application of covering materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of the project for a surface-active particles.

These compositions may also contain adjuvants such as preservatives, humectants, emulsifying and dispersing funds. Prevention of the action of microorganisms on the compounds of the present invention can be achieved by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, fenolcarbonove acid, etc. may Also be desirable to include in the composition isotonic agents such as sugars, sodium chloride, etc. in Addition, prolonged absorption of the injectable pharmaceutical form can be given by the inclusion of means, which delay absorption such as aluminum monostearate and gelatin.

In some cases, to rollover effects medicines it is desirable to slow the absorption of medicines in the subcutaneous or intramuscular injection. This can be achieved by applying a liquid suspension of crystalline or amorphous material having poor water solubility. The degree of absorption of drug then depends upon its degree of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of the dosage form for parenteral administration is achieved n the means of dissolution or suspension of the drug in an oil environment.

Injectable deposited forms receive through education microencapsulating matrix compounds of the present invention in biodegradable polymers such as polylactide-polyglycolide. Depending on the relationship of drug to polymer and the nature of the particular polymer used, the rate of release of drug can be monitored. Examples of other biodegradable polymers include poly(orthoclone esters) and poly(anhydrides). Deposited injectable forms also get the introduction of the drug in liposomes or microemulsions which are compatible with body tissues.

The finished compositions of the invention suitable for oral administration may be in the form of capsules, porridge, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragakant), powders, granules or in the form of a solution or suspension in aqueous or non-aqueous liquid, or in the form of a liquid emulsion oil-in-water or water-in-oil, or as an elixir or syrup, or in the form of hearts (using an inert base, such as gelatin and glycerin or sucrose and acacia)and/or as a gargle for mouth and the like, each of which contains a predetermined quantity of compounds of the present invention in the form of active ingredi the NTA. The compound of the present invention may also be administered in the form of a bolus, electuary or paste.

When the compounds of the present invention is administered in the form of pharmaceutical agents to humans and animals, they can enter as such or in the form of a pharmaceutical composition containing, for example, from 0.1 to 99% (more preferably, from 10 to 30%) of active ingredient in combination with pharmaceutically acceptable carrier.

These compounds can be administered to humans or other animals for therapy by any suitable route of administration, including oral, nasal, or through, for example, a spray, rectally, intrawaginalno, parenteral, intracisternal or topically, or by powders, ointments or drops, including buccal and sublingual route.

Regardless of the chosen route of administration of the compounds of the present invention, which can be used in a suitable hydrated form, and/or pharmaceutical compositions of the present invention receive in the form of pharmaceutically acceptable dosage forms by using common methods known to experts in this field.

Actual dosage levels of active ingredients in the pharmaceutical compositions of the present invention can be modified in such a way as to obtain an amount of active ingredient that is effective is output to achieve the desired therapeutic effect for a particular patient, composition and method of administration, without being toxic to the patient.

The selected dosage level will depend on many factors, including the activity of the specific compound of the present invention or a complex ester, salt or amide, route of administration, time of administration, rate of excretion or metabolism of specific compounds, the rate and extent of absorption, the duration of the treatment, other drugs, compounds and/or substances used in combination with the specific compound, the age, sex, weight, condition, General health and prior medical history of the patient to be treated, and like factors well known in medicine.

The physician or veterinarian, which is the ordinary specialists in this field can easily determine and prescribe the required effective amount of the pharmaceutical composition. For example, the physician or veterinarian could start doses of the compounds of the invention used in the pharmaceutical composition at levels lower than required to achieve the desired therapeutic effect and gradually increase the dosage until you reach the desired effect.

In General, a suitable daily dose of a compound of the invention will be this number of connections, which is the lowest dose, effective for a therapeutic effect. Such an effective dose will generally depend on the factors described above. Generally, oral, intravenous, intracerebroventricular and subcutaneous doses of the compounds of the present invention for a patient, when used for the indicated analgesic effects will be in the range from about 0.0001 to 100 mg per kilogram of body weight per day.

As desired, the effective daily dose of the active compounds may be administered as two, three, four, five, six or more subdot, administered separately at appropriate intervals throughout the day, optionally, in single dosage forms. The preferred dosage is one introduction to day.

While it is possible to introduce only one of the compounds of the present invention, preferably compounds as pharmaceutical forms (compositions).

The patient receiving this treatment is any animal in need this treatment, including primates, particularly humans and other mammals, such as horses, cattle, pigs and sheep; in General, poultry and Pets.

Compounds of the invention can be administered as such or in admixtures with pharmaceutically acceptable carriers and can also be entered in connected and with antimicrobial means, such as penicillins, cephalosporins, aminoglycosides and glycopeptides. Concomitant therapy thus includes sequential, simultaneous and separate introduction of active compound in such a way that therapeutic effects of the first input connection does not disappear completely, when you enter the next one.

Illustrating examples

Now, when the invention is generally described, it will be more easily understood with reference to the following examples which are included merely to illustrate certain aspects of the present invention and imply that they do not limit the invention.

Example 1

Derivatization cyclopamine

Stage And

To a solution of cyclopamine2(250 mg, 0.6 mmol, 1 EQ.) in DHM at room temperature add Fmoc-OSu (205 mg, 0.6 mmol, 1 EQ.) and the resulting mixture was stirred at room temperature overnight. The resulting solution of the crude Fmoc-cyclopamine then cooled to 0°C and treated with 15% diethylzinc in toluene (0.5 ml, 0.6 mmol, 1 EQ.) and stirred for 30 min (flask A).

Diameter (0.4 ml, 6 mmol, 10 EQ.) in DHM (20 ml) at 0°C is treated with 15% diethylzinc in toluene (3 ml, 3 mmol, 5 EQ.) and the resulting solution is stirred those who tell 5 min (the bulb).

The contents of the flask In transferred into the flask And through the cannula and the resulting suspension is stirred for 5 h at room temperature. The reaction is quenched with HCl (1 M), stirred for 10 min (until then, until you dissolve all white substance) and extracted with DHM (5×). The organic extracts are dried (MgSO4), filtered through celite and concentrated in vacuo. The residue is purified flash chromatography (1:1 hexane/AcOEt). Target 11,12-monocytotropic receive in the form of a 9:1 mixture of diastereoisomers, along with 20% diastereomeric bis-cyclopropylalanine products (80% of total extract). This mixture is separated using preparative SFC chromatography.

Stage

Fmoc-monosyllabically3(100 mg, 0.15 mmol, 1 EQ.) in DHM (2 ml) at room temperature is treated with diethylamine (0.5 ml, 4.8 mmol, 32 EQ.) overnight, the resulting solution was concentrated in vacuo and the residue adsorb on silica gel and purified flash chromatography (2:1→1:1 hexane/AcOEt, and then 95:5→90:10→20:80 DHM/Meon). The desired product is obtained in the form of a white solid (95% yield). MS (ESI(+)) m/e 426,31 (M+H)+.

Example 2

Derivatization cyclopamine

Stage And

To a solution of hydracarina acid5(3,g, 20 mmol, 1 EQ.) in anhydrous chloroform (30 ml) at 75°C, add thionyl chloride (1.75 ml, 24,1 mmol, 1.2 EQ.) dropwise over a period of 3 minutes the Mixture is refluxed for 3.5 hours, the Solvent is distilled off to obtain crude carboxylic acid as a pale yellow viscous liquid. The crude substance is used without further purification.

Stage

To a biphasic mixture of7(3,16 g, 24,1 mmol, 1.2 EQ.) in DHM (30 ml) and aqueous NaOH solution (2.0 M, 30 ml, 3 EQ.) at 25°C. add a solution of carboxylic acid6(to 3.38 g, 20 mmol, 1 EQ.) in DHM (10 ml) and the resulting mixture was stirred at 25°C for 3 hours the Mixture is then neutralized with an aqueous solution of HCl (2 M, 30 ml). The organic layer is then separated and the aqueous layer was extracted with DHM (3×50 ml). The combined organic layers washed with HCl (2.0 M, 25 ml), water (3×50 ml), saturated brine (50 ml), dried over magnesium sulfate and the solution evaporated under reduced pressure. The crude substance chromatographic on silica gel using 5% Meon:DHM as eluent and the column next elute 10% Meon:DHM obtaining 1,141 g connection8.

Stage

To a mixture of acid8(264 mg, 1 mmol, 1 EQ.), EDCI (231 mg, 1.2 mmol, 1.2 equiv.) triethylamine (168 μl, 1.2 mmol, 1.2 equiv.) in DHM (2 ml) at 0°C. add Lyamin (90,3 μl, 1.2 mmol, 1.2 equiv.) the resulting mixture was stirred at 0°C and allow it to warm to 25°C. over a period of 2 hours, the Reaction mixture was added to water (50 ml), extracted with DHM (4×25 ml), the combined organic layers washed with 1M HCl (2×25 ml), water (3×25 ml), saturated brine (25 ml), dried over magnesium sulfate and the solvent evaporated under reduced pressure to get 287,5 mg of the desired product. This substance is used without further purification.

Stage D

To a solution of allylamine9(70 mg, 0.23 mmol, 1 EQ.) in acetone (1 ml) and water (0.3 ml) add a solution of osmium tetroxide (0.35 ml, 0.035 mmol, 0.15 in EQ., 2.5 wt./wt. in tert-butanol). The reaction mixture was immediately cooled on ice soon after adding solution OsO4and the resulting dark brown solution was stirred at 0°C for 15 min Periodate sodium (110 mg, 0.51 mmol, 2.2 EQ.) add 5 portions to the above mixture and stirring is continued for 1 h at 0°C., and the mixture allowed to warm to 25°C over a period of 2 hours, the Reaction mixture was diluted with DHM (3 ml), filtered through a small layer of magnesium sulfate and the precipitate on the filter is washed DHM (4×3 ml). The filtrate is concentrated and the residue (67,9 mg) was filtered through a small layer of RP silica gel using 5% Meon:DHM with the receipt of 38.9 mg treb is imago product.

Stage E

To a suspension of1(10 mg, is 0.023 mmol, 1 EQ.) in acetonitrile (2 ml) add a solution of aldehyde10(17 mg, 0,056 mmol, 2.4 EQ.) in acetonitrile (0.3 ml) followed by the addition of triacetoxyborohydride sodium (6.5 mg, 0,031 mmol, 1.3 EQ.) and the reaction mixture was stirred at 25°C for 16 hours the Solvent is then evaporated under reduced pressure and the residue chromatographic on silica gel (7 cm × 10 mm)using 3% methanol:DHM with getting to 24.6 mg of the crude substance. This substance is again subjected to column chromatography on silica gel using 2% methanol:DHM and extract of 18.2 mg of the crude product, which was further purified preparative TLC using 3% methanol:DHM as a developing solvent (2-pass) with the receipt of 6.3 mg of the desired product. MS (ESI(+)) m/e 714,4 (M+H)+.

Example 3

Derivatization cyclopamine

Stage And

Cyclopamine2(20 mg, 0,049 mmol, 1 EQ.) suspended in dry toluene (0.6 ml) and added cyclohexanone (150 μl, about 1.47 mmol, 30 EQ.) with the subsequent addition of isopropoxide aluminum (79 mg, 0,392 mmol, 8 EQ.). The resulting mixture is heated at the boil under reflux for 2 h, cooled to room temperature, diluted with AcOEt is quenched with a salt solution of Rochela. The bi-phase mixture is stirred overnight, the layers separated, the aqueous layer was extracted with AcOEt and the combined organic extracts dried (MgSO4), filtered and concentrated in vacuo. The residue is purified flash chromatography (DHM, DHM/Meon 98:2 and 95:5). The target substance is obtained as a white crystalline solid (70% yield).

Stage

Diameter (40 μl, 0.5 mmol, 25 EQ.) in DHM (0,52 ml) at 0°C is treated with 15% diethylzinc in toluene (0.2 ml, 0.2 mmol, 10 EQ.) and the resulting solution stirred for 5 minutes Add connection12(10 mg, 0.02 mmol, 1 equiv.) in DHM (0.35 ml) and the resulting mixture was stirred at room temperature (removing the bath with ice) for 3 h, quenched with 2 N. NaOH and stirred for 10 min, the layers separated and the aqueous layer was extracted with DHM (2×). The organic extracts are dried (MgSO4), filtered and concentrated in vacuo. The residue is purified flash chromatography (DHM/Meon 92:8). Cyclopropylamino substance get in the form of a white solid. MS (ESI(+)) m/e 424,5 (M+H)+.

Example 4

Derivatization cyclopamine

Stage And

To a solution of cyclopamine2(250 mg, 0.6 mmol, 1 EQ.) in DHM (10 ml) at room temperature add Fmoc-OSu (mg, 0.6 mmol, 1 EQ.), the resulting mixture was stirred at room temperature overnight and concentrated in vacuo. The crude Fmoc-cyclopamine receive in the form of not-quite-white foam.

Stage

A solution of the crude Fmoc-cyclopamine3(15 mg, 0,024 mmol, 1 EQ.) in DHM (0.5 ml) cooled to -78°C and treated with sodium bicarbonate (4 mg, 0,047 mmol, 1,96 EQ.) then add msrv (4 mg, 0,024 mmol, 1 EQ.). The reaction mixture was stirred at -78°C for 1 h, diluted with H2O and extracted with DHM (3×). The organic extracts are washed with 10% NaHCO3and saturated saline solution, dried (MgSO4), filtered and concentrated in vacuo. The crude substance is purified preparative TLC (hexane/AcOEt 1:2) to give the epoxide as a white foam (70% yield).

Stage

A solution of compound14(11 mg, of 0.017 mmol, 1 EQ.) in DHM (0.5 ml) is treated at room temperature Et2NH (0.5 ml, 4.8 mmol, 282 equiv.) the resulting solution was stirred at room temperature overnight and concentrated in vacuo. The residue is purified preparative TLC (DHM/Meon 9:1). Connection13receive in the form of a white solid (90% yield). MS (ESI(+)) m/e 428,5 (M+H)+.

Example 5

Derivatization cyclopamine

Stage And

Connection2(of 1.30 g, 3.2 mmol, 1 EQ.) weighed and loaded into a reaction chamber. Potassium carbonate (of 0.91 g, 6.6 mmol, 2.1 EQ.) weighed and loaded into a reaction chamber followed by the addition of dichloroethane (6.0 ml, 76 mmol, 23,8 EQ.) and anhydrous DMSO (5 ml). The reaction mixture is heated to 70°C for 36 h in nitrogen atmosphere. The reaction mixture is cooled to room temperature, diluted with DHM (15 ml) and washed twice with water (2×15 ml). The organic layer is dried over sodium sulfate, filtered (washing DHM as needed), and concentrated to dryness to obtain a pale yellow solid. Flash chromatography (DHM/EtOAc) to obtain the target substance in the form of a white crystalline solid.

Stage

Connection16(0,111 g, 0,233 mmol, 1 EQ.) transferred into the reaction flask, placed in a nitrogen atmosphere and dissolved in anhydrous DHM (2 ml). Add chloridometer (0,238 ml of 3.27 mmol, 14 EQ.). The solution is cooled to -15°C. Diethylzinc (1M in heptane, and 1.63 ml, and 1.63 mmol, 6.5 EQ.) is added dropwise within 30 min, carefully controlling exothermic effect. The reaction mixture was kept between -10°C and -14°C for several hours up until TLC does not indicate the flow of the original substance. The reaction is then quenched careful on what deliverance THF (6 ml) and then aqueous citrate buffer (pH 4.5, 10 ml). The layers allowed to warm to room temperature. Add a saturated solution of sodium sulfate. Layers are well mixed, transfer to a connecting funnel with excess DHM and collect the organic layer. The organic layer was washed with aqueous sodium hydroxide (1 N., 10 ml) and saturated sodium sulfate (10 ml), dried over sodium sulfate before concentrating to dryness. The crude substance is purified flash chromatography to obtain the desired product with 55% yield.

Example 6

Derivatization cyclopamine

To a solution of compound11(5 mg, 0.01 mmol, 1 EQ.) and the connection10(10 mg, 0.04 mmol, 3 EQ.) in anhydrous DHM (5 ml) is added solid triacetoxyborohydride (8 mg, of 0.004 mmol, 3 EQ.) and the resulting suspension stirred at 25°C for 2 h, the Reaction mixture was quenched with sodium bicarbonate, extracted with DHM (4×10 ml), the organic layer is collected and washed with saturated salt solution (1×20 ml), dried over magnesium sulfate and concentrate under reduced pressure. The crude substance cleanse PTSH (DHM/Meon 95:5) to obtain 8 mg of the desired product.

Example 7

Derivatization cyclopamine

Stage And

Cyclopamine2(20 mg, 0.04 mmol) is suspended in dry toluene (0.6 ml) and added cyclohexanone (150 μl, of 1.47 mmol, 30 EQ.) with the subsequent addition of isopropoxide aluminum (79 mg, 0,392 mmol, 8 EQ.). The resulting mixture is heated at the boil under reflux for 2 h, cooled to room temperature, diluted with ethyl acetate and quenched with a salt solution of Rochela. The bi-phase mixture is stirred overnight, the layers are combined, the aqueous layer was extracted with ethyl acetate and the combined organic extracts are dried over MgSO4), filtered and concentrated in vacuo. The residue is purified flash chromatography (DHM, DHM/methanol 98:2 and 95:5). The target substance12receive in the form of a white crystalline solid (70% yield).

Stage

Diameter (40 μl, 0.5 mmol, 2.5 EQ.) in DHM (0,52 ml) at 0°C is treated with 15% diethylzinc in toluene (0.2 ml, 0.2 mmol, 1 equiv.) and the resulting solution stirred for 5 min (see where white precipitate). Add northward12(10 mg, 0.02 mmol, 1 equiv.) in DHM (0.35 ml) and the resulting mixture was stirred at room temperature (removing the bath with ice) for 3 h, quenched with NaOH (2N) and stirred for 10 min, the layers separated and the aqueous layer was extracted with DHM (2×). The organic extracts dried over MgSO4, filtered and concentrated in vacuo. The residue is purified flash chromatography (DHM/Meon 92:8). Cyclopropylamino wishes is in 11receive in the form of a white solid.

Stage

To a solution of cyclopropylamine11(10 mg, 24 μmol, 1 EQ.) in DHM (0.5 ml) at 0°C in an argon atmosphere add BF3·Et2O (30 μl, 0.24 mmol, 10 EQ.) and the resulting solution stirred at 0°C for 1.5 h, diluted with DHM and quenched with a saturated solution of sodium bicarbonate. The organic phase is washed with saturated sodium bicarbonate and the combined aqueous layers extracted with DHM. The combined organic layers washed with saturated saline solution, dried over MgSO4, filtered and concentrated in vacuo. The residue is purified preparative TLC (DHM/Meon 9:1). The target connection18receive in the form of a white solid (90% yield). MS (ESI(+)) m/e 424,62 (M+H)+.

Example 8

Derivatization cyclopamine

Stage And

To a solution of cyclopamine2(250 mg, 0.6 mmol, 1 EQ.) in DHM (10 ml) at room temperature add Fmoc-OSu (205 mg, 0.6 mmol, 1 EQ.) and the resulting mixture was stirred at room temperature overnight. The resulting solution of the crude Fmoc-cyclopamine then cooled to 0°C and treated with 15% diethylzinc in toluene (0.5 ml, 0.6 mmol, 1 EQ.) and stirred for 30 min (the number is a a, yellowish solution).

Diameter (0.4 ml, 6 mmol, 10 EQ.) in DHM (20 ml) at 0°C is treated with 15% diethylzinc in toluene (3 ml, 3 mmol, 5 EQ.) and the resulting solution stirred for 5 min (the bulb In, a white precipitate).

The contents of the flask In transferred into the flask And through the cannula and the resulting suspension is stirred for 5 h at room temperature. The reaction is quenched with 1 N. hydrochloric acid, stirred for 10 min (until then, until you dissolve all white substance) and extracted with DHM (5×). The organic extracts dried over MgSO4, filtered through celite and concentrated in vacuo. The residue is purified with flash chromatography (hexane/ethyl acetate 1:1). Target 11,12-monocytotropic5receive in the form of a 9:1 mixture of diastereoisomers, along with 20% diastereomeric bis-cyclopropylalanine products (80% of total extract). This mixture is separated using preparative SFC chromatography.

Stage

Fmoc-cyclopropylalanine3(14 mg, 22 μmol, 1 EQ.) dissolve in DHM (0.5 ml), cooled to 0°C and treated with BF3.Et2O (27 μl, 0.22 mmol, 10 EQ.) within 1 h, quenched with saturated sodium bicarbonate solution, the layers separated and the aqueous layer was extracted with DHM. The combined organic extracts dried over MgSO4filter and concentrate vacuume. The residue is purified preparative thin-layer chromatography (hexane/ethyl acetate 2:1). The target substance, Fmoc-protected cyclopamine receive in the form of a clear oil.

A solution of the crude Fmoc-protected cyclopamine (20 mg, 31 μmol, 1 EQ.) in DHM (0.5 ml) is treated with Et2NH (0.5 ml, 4.8 mmol, 154 EQ.) overnight, concentrated in vacuo and the residue purified flash chromatography (DHM, DHM/methanol 98:2 and 95:5). Connection required to receive in the form of an oil which crystallizes upon standing. MS (ESI(+)) m/e 426,29 (M+H)+.

Example 9

Derivatization cyclopamine

Stage And

Connection6(23 mg, 54 μmol, 1 EQ.) dissolve in DHM (1 ml) and to it add methyliodide (0.17 ml, 0.54 mmol, 10 EQ.). The reaction mixture is allowed to mix at room temperature in a nitrogen atmosphere over night. The next morning TLC/LC-MS indicates that there is still a little WILLOWS (original substance). Add a spatula Na2CO3and the mixture is stirred for another one hour. The crude substance is loaded into the system Biotage 25 Si+M and elute DHM/EtOAc/MeOH (82,5/10/7,5). Receive an amorphous substance: 16 mg MS (ESI(+)) m/e 440,32 (M+H)+.

Example 10

Derivatization cyclopamine

Stage And

Hydrocinnamaldehyde22(1.13 g, 6.7 mmol, 1 EQ.) and allylamine (of 0.77 ml, 10 mmol, 1.5 EQ.) solubilizer in THF (20 ml) and the reaction mixture was stirred at room temperature for 24 hours. A white precipitate is formed. The reaction mixture is filtered. The filtrate is dried over MgSO4, filtered and concentrated in vacuo. Allocate a colorless oil, which turns into a wax-like solid (1.1 g).

To a solution of allylamine (0,81 mg, 0.27 mmol, 1 EQ.) in a mixture of acetone:water (9 ml; 3:1) at 0°C. add a solution of OsO4(0,55 ml, 2.5 wt./wt. in tert-BuOH) and the resulting brown mixture is stirred for 10 minutes Solid periodate sodium (of 0.13 g, 0.59 mmol, 2.2 EQ.) add three portions and the mixture is stirred at 0°C and allowed to warm to 25°C over a period of 2 hours Light off-white Crampton mixture is diluted with DHM (25 ml), dried over magnesium sulfate, the solids filtered through a layer of celite, the filtrate is concentrated under reduced pressure. The crude substance slowly becomes yellow-black color. The crude substance is loaded into the system Biotage 25+S and clean, elwira a mixture of hexane/EtOAc (1:1 to 1:2) to give a colorless oil which solidifies upon drying (250 mg).

Stage

To a solution of18(108 mg, 0.25 mmol, 1 EQ.) and aldehyde21(100 mg, 0,52 mm is l, 2.1 EQ.) in DHM (5 ml) add triacetoxyborohydride sodium (100 mg, 0.47 mmol, 1.9 EQ.) one portion and the suspension was stirred at room temperature for 7 hours the Reaction is quenched by adding Meon and filtered through celite. By evaporation to dryness obtain 230 mg of oil. The substance is purified by chromatography (SiO2, column 3 cm × 4 cm), elwira a mixture of hexane/EtOAc (4:6 to 2:8) to give 38 mg of the desired product. MS (ESI(+)) m/e 599,74 (M+H)+.

Example 11

Derivatization cyclopamine

Stage And

2-Phenylethanolamine25(1.13 g, 5.5 mmol, 1 EQ. and allylamine high (0.56 ml, 7,3 mmol, 1.3 EQ.) solubilizer in THF (20 ml) and allowed to interact at room temperature for 24 hours of a white precipitate is Formed. The reaction mixture is filtered. The filtrate is dried over MgSO4, filtered and concentrated in vacuo. Allocate a slightly yellow oil (1.1 g). The crude substance is used directly in the next stage.

To a solution of arylsulfonamides (0.15 g, 0.66 mmol, 1 EQ.) in a mixture of acetone:water (4 ml; 3:1) at 0°C. add a solution of OsO4(of 0.13 ml, 2.5 wt./wt. in tert-BuOH) and the resulting brown mixture is stirred for 10 minutes Solid periodate sodium (0.31 g, of 1.46 mmol, 2.2 EQ.) add three portions and the mixture is stirred at 0°C and give her regrets is up to 25°C during the period, equal to 2 hours Light off-white Crampton mixture is diluted with DHM (25 ml), dried over magnesium sulfate, the solids filtered through a layer of celite, the filtrate is concentrated under reduced pressure. The crude substance is purified on SiO2(column 2 cm × 12 cm), elwira a mixture of hexane/EtOAc (7:3) to obtain the desired substance (16 mg).

Stage

To a solution of18(15 mg, of 35.4 mmol, 1 EQ.) and aldehyde26(16 mg, 70 μmol, 2 EQ.) in DHM (3 ml) add triacetoxyborohydride sodium (20 mg, 94 mmol, 2.6 EQ.) one portion at room temperature. After 24 h the reaction is quenched by adding a few drops of the Meon and filtered through celite. The crude substance is purified preparative TLC (1 mm (first elution: toluene/acetone (9:1), the second elution: toluene/acetone (4:1)) to obtain 4 mg of colorless oil. MS (ESI(+)) m/e 635,43 (M+H)+.

Example 12

Derivatization cyclopamine

Stage And

Connection2(of 1.30 g, 3.2 mmol, 1 EQ.) weighed and loaded into a reaction chamber. Potassium carbonate (of 0.91 g, 6.6 mmol, 2.1 EQ.) weighed and loaded into a reaction chamber followed by the addition of dichloroethane (6.0 ml, 76 mmol, 23,8 EQ.) and anhydrous DMSO (5 ml). The reaction mixture is heated to 70°C for 36 h in the atmosphere of nitrogen. The reaction mixture is cooled to room temperature, diluted with DHM (15 ml) and washed twice with water (2×15 ml). The organic layer is dried over sodium sulfate, filtered (washing DHM as needed) and concentrated to dryness to obtain a pale yellow solid. Flash chromatography (DHM/EtOAc) to obtain the target substance in the form of a white crystalline solid.

Stage

Connection16(0,111 g, 0,233 mmol, 1 EQ.) transferred into the reaction flask, placed in a nitrogen atmosphere and dissolved in anhydrous DHM (2 ml). Add chloridometer (0,238 ml of 3.27 mmol, 14 EQ.). The solution is cooled to -15°C. Diethylzinc (1M in heptane, and 1.63 ml, and 1.63 mmol, 6.5 EQ.) is added dropwise within 30 minutes, carefully controlling exothermic effect. The reaction mixture was kept between -10°C and -14°C for several hours up until TLC does not indicate the flow of the original substance. The reaction is then quenched by careful addition of THF (6 ml) and then aqueous citrate buffer (pH 4.5, 10 ml). The layers allowed to warm to room temperature. Add a saturated solution of sodium sulfate (10 ml). Layers are well mixed, transferred into a separation funnel with excess DHM and collect the organic layer. The organic layer was washed with aqueous sodium hydroxide (1M, 10 ml) and a saturated solution of sodium sulfate (0 ml), dried over sodium sulfate before concentrating to dryness. The crude substance is purified flash chromatography to obtain the desired product with 55% yield.

Stage

Connection15(1.25 g, 2.56 mmol, 1 equiv.) dissolve in DHM (22 ml) under nitrogen atmosphere and the solution cooled to an internal temperature of 0.9°C. Undiluted BF3-OEt2(1.6 ml, 12.8 mmol, 5 EQ.) add portions within a few hours, at the same time monitoring the reaction using IHMS. The reaction mixture is allowed slowly to warm to 10°C. to complete the reaction. The reaction is quenched Meon (5 ml) at 0°C, diluted with KOH (2 M, 30 ml) and stirred at room temperature for 2 hours. The layers are separated and the organic layer washed with water, dried over Na2SO4filter and concentrate to dryness. Chromatography with DHM/EtOAc get the desired product.

Example 13

Derivatization cyclopamine

Stage And

Connection27(29 mg, 60 μmol, 1 EQ.) placed in 5-ml round bottom flask. Add butanone (2 ml) and Al(OiPr)3(12.3 mg, 60 μmol, 1 EQ.). The contents of the round bottom flask heated to boiling under reflux in an argon atmosphere for 7 hours, the Reaction mixture was then quenched with a solution (2 ml), formed by mixing citric acid (500 g), NaOH (15.7 g) and water (500 ml). The resulting mixture is stirred at high speed until the dispersion of the emulsion. The mixture is then extracted with EtOAc (3×10 ml). The organic layers collected, dried over Na2SO4and concentrate. The crude substance is purified column chromatography. MS (ESI(+)) m/e 486,26 (M+H)+.

Example 14

Derivatization cyclopamine

Stage And

Connection28(25 mg, 0,051 mmol, 1 EQ.) dissolved in anhydrous 2-methoxyethanol (1 ml, 12.7 mmol, 234 EQ.). Add potassium carbonate (7,1 mg, 0,051 mmol, 1 EQ.) and the reaction mixture is heated to 120°C. reaction Monitoring performed by TLC. When TLC indicates stop the reaction, the reaction mixture is cooled to room temperature. The reaction mixture was then diluted with ethyl acetate and washed with water. The organic layer is dried over sodium sulfate and concentrated to dryness. Chromatography with DHM/EtOAc get the desired product. MS (ESI(+)) m/e 526,66 (M+H)+.

Example 15

Derivatization cyclopamine

Stage And

To a solution of hydracarina acid5(3,01 g, 20 mmol, 1 EQ.) in 30 ml of anhydrous chloroform at 75°C add thionyl the reed (1.75 ml, 24,1 mmol, 1.2 EQ.) dropwise over a period of 3 minutes the Mixture is refluxed for 3.5 hours, the Solvent is distilled off to obtain crude carboxylic acid as a pale yellow viscous liquid. The crude substance is used without further purification.

Stage

To a biphasic mixture of7(3,16 g, 24,1 mmol, 1.2 EQ.) in DHM (30 ml) and aqueous NaOH solution (2.0 M, 30 ml, 3 EQ.) at 25°C. add a solution of carboxylic acid6(to 3.38 g, 20 mmol, 1 EQ.) in DHM (10 ml) and the resulting mixture was stirred at 25°C for 3 hours the Mixture is then neutralized with an aqueous solution of HCl (2 M, 30 ml). The organic layer is then separated and the aqueous layer was extracted with DHM (3×50 ml). The combined organic layers washed with HCl (2.0 M, 25 ml), water (3×50 ml), saturated brine (50 ml), dried over magnesium sulfate and the solution evaporated under reduced pressure. The crude substance chromatographic on silica gel using 5% Meon:DHM as eluent and the column next elute 10% Meon:DHM obtaining 1,141 g connection8.

Stage

To a mixture of acid8(264 mg, 1 mmol, 1 EQ.), EDCI (231 mg, 1.2 mmol, 1.2 equiv.) triethylamine (168 μl, 1.2 mmol, 1.2 equiv.) in DHM (2 ml) at 0°C. add allylamine (90,3 μl, 1.2 mmol, 1.2 equiv.) and the resulting mixture is stirred PR is 0°C and allow it to warm to 25°C during the period, equal to 2 hours, the Reaction mixture was added to water (50 ml), extracted with DHM (4×25 ml), the combined organic layers washed with 1M HCl (2×25 ml), water (3×25 ml), saturated brine (25 ml), dried over magnesium sulfate and the solvent evaporated under reduced pressure to get 287,5 mg of the desired product. This substance is used without further purification.

Stage D

To a solution of allylamine9(70 mg, 0.23 mmol, 1 EQ.) in acetone (1 ml) and water (0.3 ml) add a solution of osmium tetroxide (0.35 ml, 0.035 mmol, 2.5 wt./wt. in tert-butanol) and the reaction mixture is immediately cooled in a bath with ice after adding a solution of OsO4. The resulting dark brown solution was stirred at 0°C for 15 min Periodate sodium (110 mg, 0.51 mmol, 2.2 EQ.) add 5 portions to the above mixture and stirring is continued for 1 h at 0°C., and the mixture allowed to warm to 25°C over a period of 2 hours, the Reaction mixture was diluted with DHM (3 ml), filtered through a small layer of magnesium sulfate and the precipitate on the filter is washed DHM (several times). The filtrate is concentrated and the residue (67,9 mg) was filtered through a small layer of RP silica gel using 5% Meon:DHM with the receipt of 38.9 mg of the desired product.

Stage E

To a solution of19(0,0242 g, 0,0569 mmol, 1 EQ) and aldehyde 10(0,0346 g, 0,114 mmol) in 3.0 ml DHM at 23°C add triacetoxyborohydride sodium (24,1 mg, 0,114 mmol, 2 EQ.) one portion and the resulting mixture is stirred for 16 hours. After complete conversion of the original substance in the desired product, which is obvious according to GHMC and TLC, the mixture is taken in 2.5 ml of methanol and purified by reversed-phase preparative chromatography (acetonitrile-buffer 20 mm ammonium carbonate, the main method). Fraction concentrate and take in a minimum volume of acetonitrile, the solution is frozen and lyophilizers obtaining 0,007 g (0,0098 mmol, 17%) as a white solid. MS (ESI(+)) m/e 714,6 (M+H)+.

Example 16

Derivatization cyclopamine

Stage And

To a solution of18(35 mg, 0.08 mmol, 1 EQ.) and aldehyde32(34 mg, 0,17 mmol, 2.0 EQ.) in THF (2.0 ml) add triacetoxyborohydride sodium (35 mg, 0,17 mmol, 2.0 EQ.) one portion. The solution is allowed to mix at 23°C for 12 hours the Mixture is then concentrated and purified by chromatography on silica gel using a gradient of 1:1 hexane:ethyl acetate with subsequent 1:2, 1:4 and ethyl acetate. Some of the material still eluted, so the column is then washed in a flash mode, a mixture of 9:1 ethyl acetate:methanol. The desired product eluted together with n is that the amount of aldehyde, therefore, the substance is further purified preparative HPLC. (Native method 50_100). The required fraction is frozen and lyophilizers with getting oily residue (12 mg, 0.02 mmol, 24% yield). MS (ESI(+)) m/e 614,44 (M+H)+.

Example 17

Derivatization cyclopamine

Stage And

To a solution of18(8.0 mg, 0.02 mmol, 1 equiv.) and aldehyde34(6.0 mg, 0.04 mmol, 2.0 EQ.) in CH2Cl2(1.0 ml) add triacetoxyborohydride sodium (8.0 mg, 0,17 mmol, 2.0 EQ.) one portion. The solution is allowed to mix at 23°C for 12 hours the Mixture is then concentrated and purified by chromatography on silica gel using a gradient of 1:1 hexane:ethyl acetate with subsequent 1:2, 1:4, to select the desired product, collisuem with a certain amount of aldehyde. This substance is further purified preparative HPLC. The required fraction is frozen and lyophilizers obtaining a white powder (4,9 mg, 0,009 mmol, 46% yield). MS (ESI(+)) m/e 570,41 (M+H)+.

Example 18

Derivatization cyclopamine

Stage And

To a solution of18(6.0 mg, 0.01 mmol, 1 EQ.) and benzaldehyde36(3.0 mg, 0.02 mmol, 2.0 EQ.) in CH2Cl2(0.5 ml) add triacetoxyborohydride sodium (6.0 mg, 0.02 mmol, 2.0 EQ.) one for the tution. The solution is allowed to mix at 23°C for 12 hours the Mixture is then concentrated and purified by chromatography on silica gel using a gradient of 4:1 hexane:ethyl acetate with subsequent 1:1 to scroll to the desired product, collisuem with a certain amount of aldehyde. This substance is further purified preparative HPLC. The required fraction is frozen and lyophilizers obtaining a white powder (0.6 mg, 0.001 mmol, 8% yield).

Example 19

Derivatization cyclopamine

Stage And

To a solution of18(6.0 mg, 0.01 mmol, 1 EQ.) and 4-phenoxybenzaldehyde38(6.0 mg, 0.02 mmol, 2.0 EQ.) in CH2Cl2(0.5 ml) add triacetoxyborohydride sodium (6.0 mg, 0.02 mmol, 2.0 EQ.) one portion. The solution is allowed to mix at 23°C for 12 hours the Mixture is then concentrated and purified by chromatography on silica gel using a gradient of 4:1 hexane:ethyl acetate with subsequent 1:1 to scroll to the desired product, collisuem with a certain amount of aldehyde. This substance is further purified preparative HPLC. The required fraction is frozen and lyophilizers obtaining a white powder (1.8 mg, of 0.003 mmol, 21% yield). MS (ESI(+)) m/e 606,4 (M+H)+.

Example 20

Derivatization cyclopamine

Stage And

To a mixture of18(40 mg, 0.09 mmol, 1 EQ.) and sodium bicarbonate (15 mg, 0.18 mmol, 2.0 EQ.) in CH2Cl2(0.5 ml) add bromoethanol40(33 μl, 0.47 mmol, 5.0 EQ.). The solution is heated to boiling under reflux for 4 hours the Mixture is then concentrated and purified by chromatography on silica gel using a gradient DHM, with subsequent 38:1:1 dichloromethane:ethyl acetate:methanol, then 36:3:1, then 17:2:1) to highlight the desired product as oil (12 mg, was 0.026 mmol, 27% yield). MS (ESI(+)) m/e 468,24 (M+H)+.

Example 21

Derivatization cyclopamine

Stage And

To a solution of18(100 mg, 0.24 mmol, 1 EQ.) and aldehyde42(42 mg, 0.35 mmol, 1.5 EQ.) in CH2Cl2(2.5 ml) add triacetoxyborohydride sodium (100 mg, 0.47 mmol, 2.0 EQ.) one portion. The solution is allowed to mix at 23°C for 12 h, and IHMS shows only 50% conversion. To the mixture add a further equivalent of aldehyde36(26 mg, 0.24 mmol, 1.0 EQ.) and triacetoxyborohydride sodium (48 mg, 0.24 mmol, 1.0 EQ.) and allow to mix for 2 hours the Mixture is then concentrated and purified by chromatography on silica gel using a gradient of 1:1 hexane:ethyl acetate with subsequent 1:2 and 1:4, to select the desired product, collisuem with some quantities of the m aldehyde. This substance is further purified preparative HPLC. The required fraction is frozen and lyophilizers obtaining a white powder (53 mg, 0.10 mmol, 43% yield). MS (ESI(+)) m/e 526,66 (M+H)+.

Example 22

Derivatization cyclopamine

Stage And

To a solution of18(15 mg, 0.04 mmol, 1 equiv.) and aldehyde44(6.9 mg, 0.04 mmol, 1.0 EQ.) in CH2Cl2(0.6 ml) add triacetoxyborohydride sodium (15 mg, 0.07 mmol, 2.0 EQ.) one portion. The solution is allowed to mix at 23°C for 12 h, and IHMS shows only 50% conversion. To the mixture add a further equivalent of aldehyde44(6.9 mg, 0.04 mmol, 1.0 EQ.) and triacetoxyborohydride sodium (7.5 mg, 0.04 mmol, 1.0 EQ.) and allow to mix for 12 hours the Mixture is then concentrated and purified by chromatography on silica gel using a gradient of 1:1 hexane:ethyl acetate with subsequent 1:2 and 1:4, to select the desired product as oil (12 mg, 0,19 mmol, 54% yield). MS (ESI(+)) m/e 635,43 (M+H)+.

Example 23

Derivatization cyclopamine

Stage And

To a mixture of18(12 mg, 0.03 mmol, 1 EQ.) and potassium carbonate (40 mg, 0.28 mmol, 10 EQ.) in DMF (0.5 ml) is added hydrochloride 2-(dimethylamino)ethylchloride46(20 mg, 0.14 mmol, 5.0 equiv.) The solution is stirred for 2 h at 23°C and the reaction does not occur. The solution is further heated at 65°C for 12 h, quenched with water (2 ml) and then extracted with diethyl ether (2×10 ml). The combined organic extracts washed with saturated saline and dried with MgSO4. The mixture is then concentrated and purified preparative HPLC. The required fraction is frozen and lyophilizers obtaining a white powder (17, 1.8 mg, of 0.004 mmol, 13% yield). MS (ESI(+)) m/e 495,71 (M+H)+.

Example 24

Derivatization cyclopamine

Stage And

To a solution of18(100 mg, 0.24 mmol, 1 EQ.) and chloracetamide48(250 mg, 1.2 mmol, 5.0 equiv.) in CH2Cl2(1.0 ml) is added triethylamine (160 μl, 1.2 mmol, 5.0 EQ.). The solution is heated to boiling under reflux and stirred for 72 h the Mixture was then concentrated and purified by chromatography on silica gel using a gradient of 4:1 hexane:ethyl acetate followed by 2:1, 1:1 and 1:2, to select the desired product as a mixture of two spots. The same conditions of the column and repeat the desired product is isolated in the form of an oil (17 mg, 0.14 mmol, 12% yield).

Example 25

Derivatization cyclopamine

Connection18(103 mg, 0.24 mmol, 1 EQ.) dissolve in DHM (3.0 ml) and cooled to -78°C. To this solution add msrv (77 wt.%, 54 mg, 0.24 m is ol, 1.0 EQ.) and then the solution is allowed to warm to 22°C for 12 hours the Reaction takes place with a 50% conversion (IHMS). The solution is quenched with sodium bicarbonate and extracted with DHM. The combined organic phases are dried with magnesium sulfate, filtered and concentrated. The substance is further purified by chromatography on silica gel using a gradient DHM:EtOAc:MeOH, equal to 95:2,5:2,5, next 92,5:5,0:2,5, then 85:10:5, to scroll to the desired product, collisuem with a small amount of impurities. Combined fractions are concentrated and purified preparative HPLC to obtain 3.4 mg of the desired product. MS (ESI(+)) m/e 440,63 (M+H)+.

Example 26

Inhibition of the Hedgehog pathway in cell culture, using analogues cyclopamine

Effects to destroy specific cancer cells Hedgehog paths can be identified using the following analysis. Cells SNT/2 differentiate into osteoblasts in contact with voiced Hedgehog peptide (Shh-N). During differentiation of these osteoblasts produce high levels of alkaline phosphatase (AP), which can be measured in enzymatic analysis (Nakamura et al., 1997BBRC237: 465). Compounds that block differentiation SNT/2 in osteoblasts (Shh dependent event), can therefore be identified through recovery when designing AR (van der Horst et al., 2003 Bone 33: 899). Details of the analysis are described below. The results, appr is xemeraldia (EC 50for inhibition) analysis of differentiation shown in the table below.

Protocol analysis

Cell culture

SNT/2 cells fibroblasts mesoderm of mouse embryos (obtained from ATSS) were cultured in basal MEM medium (Gibco/Invitrogen), supplemented with 10% heat inactivated FBS (Hyclone), 50 units/ml penicillin and 50 µg/ml streptomycin (Gibco/Invitrogen) at 37°C With 5% CO2in an atmosphere of air.

Analysis of alkaline phosphatase

SNT/2 cells are placed in 96-well tablets at a density of 8×103cells/well. Cells are grown to confluence (72 hours). After adding voiced Hedgehog (250 ng/ml) (R&D Systems) and/or processing by the connection of the cells are lysed in 110 µl lisanova buffer (50 mm Tris pH 7.4, with 0.1% TritonX100), dies voiced and lysates push through dies 0.2 μm PVDF (Corning). 40 μl of the lysates analyzed for the activity of AR in the solution alkaline buffer (Sigma)containing 1 mg/ml p-nitrophenylphosphate. After incubation for 30 min at 37°C. the plate read using a reading device for spots Envision at 405 nm. Total protein was quantitatively evaluated using the test kit protein ICA from Pierce according to the manufacturer's instructions. The activity of AR normalize with respect to the total protein. Note that "A" indicates that the IC50is less than 200 nm, "B" indicates that the IC50extending t is 200-500 nm, "C" indicates that the IC50>500 nm.

Approximated EU50
for inhibition
ConnectionEU50analysis of differentiation
1
4And
11
17A
18A
19B
20C
21A
24B
27C
28C
29A
30A
31A
33A
35A
37C
39C
41A
43B
45B
47B

Example 27

The way preparative supercritical fluid chromatographic purification (SFC)

The described method preparative supercritical fluid chromatography for the purification of the compounds of the present invention.

The software used:

SFC: Berger PrepSFC System

UV detector: Knauer Model K-2501

Column: Berger 5 microns silica, 20 mm 250 nm

The SFC conditions:

Mobile phase: CO2- 95%methanol - 5%

Flow rate: 50,00 ml/min

The column temperature: 35°C

Isocratic for 40 min with 5% methanol in supercritical CO2

Amount: 1000 ál

The concentration of the specimen are usually at 5.0 mg/ml

Get sample: the sample was dissolved in 20% DHM/80% methanol

Products elute between 25 and 40 min

Options UV detector

Wavelength = 210 nm; resolution = 1.0 nm

Example 28

The method of liquid chromatography - mass spectrometry (IHMS)

The described method of liquid chromatography - mass spectrometry for the compounds of the present invention.[A] report on the input method

The mobile phase Waters Alliance 2795 LC

Solvents
And%0,0
In%10.0 Acetonitrile
With%90,0 water 20 mm NH4HCO3
D%0,0 water 0,1A
The stream mode1,00
Flow (ml/min)1,500
Stop time (min)4,00
Min. pressure (bar)0
Max. pressure (bar)300
DegasserThe amount of delay100,0 ál
Column Waters Alliance 2795 LC
The position of the columnColumn 3
Time trim (min)0,00
The column temperature (°C)35
The temperature limit of the column(°C)5
Quick trim Waters Alliance 2795 LC
Track systemOff.
The flow system (ml/min)0,00
System time (min)0,00
While re-balancing (min)0,00
Volume predalone (ál)0,00
Waters Alliance 2795 I/O
Switch 1Unchanged
Switch 2Unchanged
Switch 3Unchanged
Switch 4Unchanged
Setting the analog outputThe flow velocity
Gradient temporary table Waters Alliance 2795 LC
Gradient the temporary table has 5 inputs, which are:
TimeAnd%In%With%D%StreamCurve
0,000,010,090,00,0 1,5001
3,000,085,015,00,01,5006
3,200,0100,00,00,01,5006
3,500,0100,00,00,01,5006
3,600,010,090,00,01,5006

Temporary table external events Waters Alliance 2795 LC

The inputs of the temporary table external events pump no

The input parameters Waters Alliance 2795

Input typeSerial
The entry modePartial loop
Air border pre-treatment of the sample (ál)4,0
Air border subsequent processing of the sample (ál)4,0
Pre-boot time (min)0,00

The parameters of the sampler Waters Alliance 2795

The sample temperature (ºc)20
Limit sample temperature (ºc)20
The depth of grooves (mm)0
Speed groovesNormal
The depth of the wells searchFailure
Height check cardFailure
The parameters of the washing Waters Alliance 2795
The frequency of flushingInput
Fast flush ()3
Washing time (s)10
Wash cycles2
The volume of secondary rinse (ál)600,0

Waters996 PDA

The initial wavelength (nm)220,00
End wavelength (nm)400,00
Resolution (nm)1,2
The rate of sampling (range/s)1,000
Response filter1
The exposure time (MS)Automatically
Interpolation 656Yes
The stop time of data collection (min)4,000
Save to disk:Yes
Analog channel 1 Waters996 PDA
Output modeOff.
Analog channel 2 Waters996 PDA
Output modeOff
End of report

[B]Experimental report

NameExperiment by default
Creation timeFriday June 04, 2004 10:25:33
The instrument ID
The version number1,0
Duration (min)4,0
The name of the calibration fileC:MassLynxInfinity_2002.PROACQUDB AICS_040408.cal
The delay of the solvent provided a check valve0
The number of functions1

Function 1: MS scan time is 0.00 to 3.50, mass 200,00 to 1000,00 ES+

TypeMS scan
Ion modeES+
Data formatThe centroid
FileC:MasslynxInfinity_2002.PROACQUDBdefault.ipr
Starting weight200,0
The final weight1000,0
The scan time (sec)1,0
The time between scans in seconds)0,2
Start time (min)0,0
End time (min)3,5
[S] Settings ZQ
Source (ES+)InstallReading
Capilar (kV)3,20with 3.27
Cone (V)35,0037,24
Extractor (V)3,002,81
The RF lens (V)0,00,0
The source temperature (°C)130129
Temperature desolvatation (°C)350350
The gas flow cone (l/h)2033
The desolvatation gas flow (l/h)450466
AnalyzerInstallReading
LM 1 resolution15,0
NM 1 resolution15,0
The ion energy 10,5
Improver (V)550-547
Valve pressure
Pirani pressure (mbar)<1e-4 mbar
MUX configuration
ProbeStandard

The incorporation by reference

All of the U.S. patents and published patent applications U.S. cited in this description are incorporated into it by reference.

Equivalents

Specialists in this field will recognize, or be able to identify, using no more than routine experimentation, many equivalents of the specific embodiments of the invention described in this document. Imply that such equivalents are covered by the following claims.

1. The compound of formula 1

or its pharmaceutically acceptable salt,
in which each R1and R8independently represents H or hydroxyl;
each R2and R9independently represents H or hydroxyl;
R5represents H;
each R3, R4, R6, R7, R13and R14independently represents H;
Il is R 1and R2taken together form =O; or
R4and R5taken together form a double bond; or
R5and R6taken together form a double bond;
R10and R11taken together form a double bond;
R12represents H, alkyl, hydroxyl, aralkyl,
halogenated, alkoxy, -[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, [(W)-O]qR21or -[(W)-N(R21)]q21;
where each W independently represents a divalent alkyl or Uralkaliy radical, and q is 1, 2, 3 or 4;
each R15, R16and R17independently represents H;
each R18and R19independently represents H and
each R21independently represents H, alkyl, aryl or aralkyl.

2. The compound according to claim 1, which has the formula
or
where R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17, R18and R19such as defined in claim 1, or its pharmaceutically acceptable salt.

3. The compound according to claim 1, where
R1represents a hydroxyl;
or R1and R2taken together form =O, and/or
R4and R5taken together, brisout double bond.

4. The compound according to claim 1, where R1and R2taken together form =O or R1represents hydroxyl, and R2represents N.

5. The compound according to claim 1, where R5and R6taken together form a double bond.

6. The compound according to claim 1, where R8and R9represent hydrogen.

7. The compound according to claim 1, where R12represents H, alkyl, aralkyl, halogenated, hydroxyl or alkoxyl.

8. The compound according to claim 1 of the formula
or
or its pharmaceutically acceptable salt,
where R12represents H, alkyl, hydroxyl, aralkyl, halogenated, alkoxy, -[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-O]qR21or
-[(W)-N(R)]qR21where each W independently represents a divalent alkyl or Uralkaliy radical;
q is 1, 2, 3 or 4;
each R21independently represents H, alkyl, aryl or aralkyl.

9. The compound according to claim 1, selected from the group consisting of






br/>



and

and their pharmaceutically acceptable salts.

10. The compound of formula 2

or its pharmaceutically acceptable salt,
in which each R1and R8independently represent H or hydroxyl;
each R2and R9independently represents H or hydroxyl;
R5represents H;
each R3, R4, R6, R7, R13and R14independently represents H;
or R1and R2taken together form =O; or
R4and R5taken together form a double bond; or
R5and R6taken together form a double bond;
R10and R11taken together form a group represented by 1b

where Z is O or C(R23)(R23);
R12represents H, alkyl, hydroxyl, aralkyl, halogenated, alkoxy,
-[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR21, -[(W)-O]qR21or -[(W)-N(R21)]qR21;
where each W independently represents a divalent alkyl or Kalki the capacity radical, a q equal to 1, 2, 3 or 4;
each R15, R16and R17independently represents H;
each R21independently represents H, alkyl, aryl or aralkyl and
each R23independently represents N.

11. The connection of claim 10, which has the formula

or

where R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, R15, R16and R17such as defined in claim 10, or its pharmaceutically acceptable salt.

12. The connection of claim 10, where
R1represents a hydroxyl; or R1and R2taken together form =O; and/or
R4and R5taken together form a double bond.

13. The connection of claim 10, where R5and R6taken together form a double bond.

14. The connection of claim 10, where R8and R9represent hydrogen.

15. The connection of claim 10, where R12represents H, alkyl, aralkyl or halogenated.

16. The compound of claim 10 of the formula
or
or its pharmaceutically acceptable salt,
where R12represents H, alkyl, hydroxyl, aralkyl, halogenated, alkoxy,
-[(W)-N(R21)C(O)]qR21, -[(W)-N(R21)SO2]qR 21, -[(W)-O]qR21or -[(W)-N(R21)]qR, where each W independently represents a divalent alkyl or Uralkaliy radical; and
q is 1, 2, 3 or 4; and
each R21independently represents H, alkyl, aryl or aralkyl.

17. The compound of claim 10 selected from the group consisting of

and
and their pharmaceutically acceptable salts.

18. Pharmaceutical composition having anti-cancer activity containing a compound according to any one of claims 1 to 17; and pharmaceutically acceptable filler.

19. The compound according to any one of claims 1 to 17 for use as a drug for cancer treatment.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds, specifically to 4-substituted-3-(1-alkyl-2-chloro-1H-indol-3-yl)furan-2,5-diones of general formula I , where R1=H, C1-C6 alkyl; R2=H, C1-C6 alkyl, C1-C6 alkoxy; R3=phenyl, naphthyl, 2-phenyl-1-ethenyl, thienyl, furyl, pyrrolyl, benzothiophenyl, benzofuranyl, indolyl, synthesis method thereof and use as compounds capable of photochemical generation of stable fluorophores of formula II, which can be used, for instance in information storage systems, particularly as photosensitive components of material for three-dimensional recording and storage of information. The invention also relates to novel 4,5-substituted-6-alkyl-1H-furo[3,4-c]carbazole-1,3(6H)diones of general formula II , where R1=H, C1-C6 alkyl; R2=H, C1-C6 alkyl, C1-C6 alkoxy; R4=H, R5=phenyl, R4, R5=benzo, naphtho, thieno, furo, pyrrolo, benzothieno, benzofuro, indolo, method for synthesis of said compounds and use as fluorophores.

EFFECT: obtaining novel compounds and possibility of using said compounds as fluorophores.

14 cl, 2 tbl, 8 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel cyclic amine compounds of formula (1) or pharmaceutically acceptable salts thereof: . In formula (1), X is O, S, NR2 (where R2 is H, C1-C12 alkyl); when X is O, S, then R1 is H, CN, COOH, C2-C13 alkoxycarbonyl, carbamoyl group; and when X is NR2 (where R2 assumes values given above), R1 is CN; Ar1 and Ar2 are identical or different and each represents an aryl which can be substituted with 1-3 halogens; or Ar1 and Ar2 together with neighbouring carbon atoms to which they are bonded form a group with formula (b): (where ring S and ring T are identical and each is a benzene ring; Y is O); ring B is a benzene ring which can be substituted with 1-3 substitutes independently selected from a group comprising halogen, C1-C12 alkyl, C1-C8 halogenalkyl, C1-C12 alkoxy, C1-C8 halogenalkoxy group; n is an integer from 1 to 10; p, q are identical or different and each is an integer equal to 1 or 2. Formula (1) compounds are bonding inhibitors of the α2C-adrenoreceptor.

EFFECT: possibility of using compounds in pharmaceutical compositions.

7 cl, 1 tbl, 8 ex

FIELD: chemistry.

SUBSTANCE: in formula compounds, each of R1, R2, R3, R4 is a substitute for a cyclic system, chosen from hydrogen, halogen, C1-C6-alkyl; C1-C6-alkoxy group; X is a heteroatom, chosen from oxygen or sulphur; R5 and R6 independently represent amino group substitutes, chosen from hydrogen, possibly substituted C1-C6-alkyl; possibly substituted C3-C6-cycloalkyl, which can be annealed with a benzene ring; possibly substituted phenyl, which can be annealed with dioxole, dioxine, -(CH2)n group, where n=4 to 6, or with a 5 or 6-member possibly substituted and possibly condensed azaheterocyclyl; possibly substituted saturated or unsaturated 5-6-member heterocyclyl, containing 1-2 heteroatoms, chosen form nitrogen, oxygen, sulphur and possibly condensed with a benzene ring, or R5 and R6 together with the nitrogen atom to which they are bonded, form an optionally substituted 5 or 6-member azahetero ring, possibly containing an additional heteroatom, chosen from nitrogen, and possibly annealed with a benzene ring or spiro-condensed with dioxole, where substitutes in the said alkyl, cycloalkyl, phenyl and heterocyclyl are chosen from halogen atoms, possibly substituted C1-C6-alkyl, CF3, possibly substituted C3-C6-cycloalkyl, possibly substituted phenyl, 5 or 6-member heterocyclyl, nitro group, substituted amino group, alkyloxycarbonyl, substituted carbonyl, aminocarbonyl, alkylsulphanyl.

EFFECT: design of an efficient method of producing new substituted furo[2,3-b]quinoline-2-carboxamides and substituted thieno[2,3-b]quinoline-2-carboxamides or their racemates, or their optical isomers, as well as their pharmaceutically acceptable salts and/or hydrates of general formula (I), which have antituberculous activity.

9 cl, 1 dwg, 7 tbl, 5 ex

FIELD: pharmacology.

SUBSTANCE: invention relates to novel compounds - tetrahydronaphthyridine derivatives of formula (I) or their pharmaceutically acceptable salts, where R1 represents C1-6alkoxycarbonyl group optionally substituted with 1-5 substituents, etc; R2 represents C1-6alkyl group; R3 represents hydrogen or and all; R4 represents C1-4alkylene group; R5 represents optionally substituted unsaturated 5-8-member heterocyclic group containing 1-4 heteroatoms independently selected from oxygen and nitrogen atoms; R6, R7 and R8 represent independently hydrogen atom, hydroxygroup, cyanogroup, C1-6alkyl group, C1-6alkoxygroup, mono- or di- C1-6alkylcarbamoyl group or mono- or di- C1-6alkylaminogroup, optionally substituted with 1-6 substituents independently selected from halogen atom, C1-6alkoxygroup and aminogroup; R10 represents optionally substituted with 1-2 substituents phenyl group; which possess inhibiting activity with respect to cholesteryl ester transfer protein (CETP).

EFFECT: novel tetrahydronaphthyridine derivatives and method of obtaining them.

12 cl, 408 ex, 38 tbl

FIELD: chemistry.

SUBSTANCE: present invention relates to new annelated azaheterocyclic amides, including a pyrimidine fragment, with the general formula 1, method of obtaining them and their application in the form of free bases or their pharmaceutically accepted salts as inhibitors of P13K kinase, in compounds with the general formula 1: , where: X represents an oxygen atom, sulphur atom or not necessarily substituted at the nitrogen NH group, where the substitute is selected from lower alkyls and possibly a substituted aryl; Y represents an atom of nitrogen or substituted at the carbon atom CH group, where the substitute is selected from lower alkyls; Z represents an oxygen atom; R1 represents a hydrogen atom or not necessarily substituted C1-C6alkyl, or Z represents a nitrogen atom, which is together with a carbon atom, with which it is joined, form through Z and R1 annelated imidazole cycle; R2 and R3 independently from each other represent hydrogen, not necessarily substituted with C1-C6alkyl, C3-C6cycloalkyl, not necessarily substituted with phenyl, not necessarily substituted with 6-member aza-heteroaryl, under the condition, when Y represents a nitrogen atom, or R2 and R3 independently from each other represent not necessarily substituted C1-C6alkyl, not necessarily substituted with phenyl, not necessarily substituted with 5-7-member heterocycle with 1-2 heteroatoms, selected from nitrogen and oxygen, and possibly annelated with a phenyl ring, under the condition, when Y does not necessarily represent a substituted carbon atom at the CH group, and X represents an oxygen atom, sulphur atom, or R2 represents hydrogen, and R3 represents a substituted aminoC1-C6alkyl and not necessarily substituted 5-6-member aza-heterocycloalkyl, under the condition, when Y represents a group which is substituted at the CH atom, and X represents an oxygen atom, sulphur atom, or R2 represents hydrogen, and R3 represents phenyl which is not necessarily substituted, pyridyl which is not necessarily substituted, pyrimidinyl which is not necessarily substituted, under the conditions, when R1 represents a substituted aminoC1-C6alkyl, substituted C2-C3hydroxyalkyl and aza-heterocycloalkyl not necessarily substituted, Y represents a group with CH substituted, and X represents an oxygen atom, sulphur, and the substitute of the above indicated substituted alkyl, phenyl, heterocycle, pyridyl, pyrimidyl are selected from groups of hydroxyl-, cyano-groups, hydrogen, lower alkyls, possibly mono- or di-substituted lower alkyl sulfamoyl, carbamoyl, C1-C6alkoxycarbonyl, amino, mono- or di-lower alkyl-amine, N-(lower alkyl), N-(phenylC1-C6alkyl)amine, phenyl, possibly substituted with a halogen atom, C1-C6alkyl, haloid-C1-C6alkyl; phenylC1-C6alkyl, saturated or non-saturated 5-6-member heterocycle containing 1-2-heteroatoms, selected from nitrogen, oxygen and sulphur, and possible condensation with a benzene ring R4 represents hydrogen or a lower alkyl.

EFFECT: obtaining new annelated aza-heterocyclic amides, including a pyrimidine fragment, with the general formula with the possibility of their application in the form of free bases or their pharmaceutically accepted salts as inhibitors of PI3K kinase.

16 cl, 5 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: in general formula (I) , R1 represents similar or different 2 groups, each of which is selected from group consisting of C1-3alkyl, or when R1 are two adjacent groups, two groups R1, taken together, can form saturated or unsaturated 5- or 6-member cyclic group, which can have 1 or 2 oxygens as heteroatom; X represents oxygen or sulphur; values of other radicals are given in invention formula.

EFFECT: increase of composition efficiency.

16 cl, 11 tbl, 31 ex

FIELD: medicine; pharmacology.

SUBSTANCE: new annelated asaheterocycles include pyrimidine fragment of general formula I in the form of free bases or pharmaceutically acceptable salts. Compounds of this invention possess properties of PI3 kinase inhibitors. In general formula I X represents oxygen atom or sulphur atom; Z represents oxygen atom, R1 represents hydrogen atom or optionally substituted C1-C6alkyl, or Z represents nitrogen atom together with bound carbon atom forming through Z and R1 optionally substituted annelated imidazoline cycle; R2 represents optionally substituted C1-C6alkyl, optionally substituted C3-C8cycloalkyl, optionally substituted phenyl, possibly annelated with 5-6-term heterocyclyl containing heteroatoms chosen from oxygen and nitrogen, optionally substituted 5-6-term heterocyclyl containing heteroatoms chosen from nitrogen, oxygen and/or sulphur, possibly annelated with phenyl ring. Invention also concerns method of production of compounds, pharmaceutical compositions and medical products.

EFFECT: effective application for preparation of medical products for oncologic therapy.

14 cl, 3 dwg, 1 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: claimed are novel pyrazole derivatives of formula II or its pharmaceutically acceptable salts, where C ring is selected from phenyl or pyridinyl ring and R2, R2', Rx and Ry are such as said in given description. C ring has ortho-substituent and is optionally substituted in non-ortho positions. R2 and R2' , optionally taken with their intermediate atoms, form condensed ring system, such s indazole ring, and Rx and Ry, optionally taken together with their intermediate atoms, form condensed ring system, such a quinazoline ring.

EFFECT: possibility to use compositions as inhibitors of protein kinases as inhibitors GSK-3 and other kinases and apply them for protein kinase-mediated diseases.

41 cl, 8 tbl, 423 ex

FIELD: chemistry.

SUBSTANCE: invention relates to the bonds consistent with the general formula (I) or the general formula (II) where: R1 = H; Z = O or S; P1 = CR5R6, P2 = CR7R8, Q = CR9R10; each of R5, R6, R7, R8, R9 and R10 denotes H; Y = CR12R13-CO, where R12, R13 is selected from C0-7-alkyl; C3-6-cycloalkyl or phenyl-C0-7-alkyl; and where the phenyl ring doesn't have to be substituted R19, specified below; in the group (X)0, X = CRI4R15, where R14 and R15 are independently selected from C0-7-alkyl, and o represents a number from zero to three; (W)n, W = O, S, C(O), S(O) or S(O)2 or NR16, where R16 denotes H, and n equals zero or one; (V)m, V = C(O), NHC(O), C(O)NH or CR17R18, where R17 and R18 denotes H, and m represents a number from zero to three, on condition that, when m is more than one, (V)m contains a maximum of one carbonyl group; U = a stable 5-7-member monocyclic or 8-11-member dicyclic ring, which is saturated or non-saturated and which has from zero to four heteroatoms selected from: , where R19 represents: C0-7-alkyl, C3-6-cycloclkyl, Ar-C0-7-alkyl, O-C0-7-aklyl, NH-C0-7-alkyl, N(C0-7-alkyl)2, O-phenyl, S-phenyl; or, as a part of CHR19 or CR19 group, R19 can represent a halogen; where Ar represents a stable 5- or 6- member monocyclic or stable 9- or 10- member dicyclic ring, which is unsaturated as determined earlier for U, and where Ar doesn't have to be substituted R19, which is of importance specified above; C0-7-alkyl represents a stable linear or a branched aliphatic carbon chain, which contains from 0 to 7 carbon atoms, which doesn't have to be substituted with one, two or three halogen atoms and doesn't have to contain one or few heteroatoms selected from O, N and S, where the heteroatom is present only when C0-7-alkyl contains as a minimum one carbon atom; C3-6-cycloalkyl relates to C0-7-alkyl, certainly higher than the additionally contained carboxyl ring, which doesn't have to be substituted with one or more halogens, selected from F, Cl, Br and I or heteroatoms, selected from N, O, S; A represents O; B, D and G are independently selected from: CR19, where R19 is as specified above or N; E represents O or S; J, L, M, R, T, T2, T3 and T4 which are independently selected from: CR19 and N, where R19 is as specified above; T5 represents N; q represents a number from one to three, determining in this way a 5-, 6- or 7- member ring or its salt, hydrate or solvate. The bonds of the general formula (I) or the general formula (II), represent cruzipain inhibitors and inhibitors of other cisteinproteases and can be used as therapeutic agents, for example, in cases of Chagas disease or for confirmation of target oriented therapeutic bonding.

EFFECT: new bonds which posses helpful biological properties have been discovered.

27 cl, 156 ex, 2 tbl

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to heterocycle-substituted tricyclic compounds of the formula (I): or their pharmaceutically acceptable salts wherein R means hydrogen atom; R1 and R2 are chosen independently from group comprising hydrogen atom or alkyl comprising 1-6 carbon atoms; R3 means hydrogen atom; n1 and n2 = 0-3 independently under condition that they both do not mean 0; Het means pyridyl wherein pyridyl is added to B through cyclic carbon atom and it comprises from 1 to 4 substitutes (W) chosen independently from group comprising -NR4R5, -NHCOR26, -NHSO2R16; R21 means aryl and R21 means heteroaryl wherein heteroaryl represents furyl, thienyl, pyridyl, thiazolyl, pyrrolidinyl, azethidinyl; R4 and R5 mean hydrogen atom or alkyl comprising 1-6 carbon atoms, or R4 and R5 mean in common -(CH2)3-, -(CH2)4-, -(CH2)5- or -(CH2)2NR7-(CH2)2- wherein R7 means hydrogen atom or alkyl comprising 1-6 carbon atoms; R8, R, R10 and R11 mean hydrogen atom; B means -(CH2)n4CR12=CR12a(CH2)n5 wherein n4 and n5 = 0-2 independently; R12 and R12a are chosen independently from group comprising hydrogen atom or alkyl comprising 1-6 carbon atoms; R21 means from 1 to 3 substitutes chosen independently from group comprising hydrogen atom, trifluoromethyl, trifluoromethoxy, halogen atom, cyano, alkyl comprising 1-6 carbon atoms, alkoxy group comprising 1-6 carbon atoms, or -CR29(=NOR28); R22 means -COR23, -S(O)R31, -S(O)2R31 or -COOR27; R23 means cycloalkyl comprising 3-7 carbon atoms, (C3-C7)-cycloalkyl-(C1-C6)-alkyl, cycloalkyl comprising 3-7 carbon atoms containing from 1 to 3 substitutes chosen from group comprising halogen atom, (C1-C3)-alkoxy-(C1-C3)-alkyl, hydroxy group and alkoxy group comprising 1-6 carbon atoms, aryl, aryl-(C2-C6)-alkyl; R27 means alkyl comprising 1-6 carbon atoms, phenyl or benzyl; R28 and R29 are chosen independently from group comprising hydrogen atom or alkyl comprising 1-6 carbon atoms; R31 means alkyl comprising 1-6 carbon atoms, halogenalkyl comprising 1-6 carbon atoms, aryl, aryl-(C1-C6)-alkyl. Also, invention relates to pharmaceutical compositions containing these substances and their using for preparing a drug used in treatment of thrombosis, atherosclerosis, restenosis, hypertension, stenocardia, arrhythmia, heart failure and cancer.

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.

10 cl, 11 tbl, 9 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to compounds of formula IB , where radicals R1-R5 have values, given in invention formula. In range of claimed invention also described are pharmaceutical compositions, which include compounds of IB formula, and methods of application of such compounds and compositions for treatment of different malfunctions, mainly selected from immune response reactions.

EFFECT: compounds by claimed invention have inhibiting action with respect to proteinkinases and, in particular with respect to JAK-3, ROCK or Aurora kinases.

55 cl, 6 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to novel derivatives of 2,4-di(hetero)arylaminopyrimidine of general formula I or its pharmaceutically acceptable salt, possessing properties of ZAP-70 inhibitors. In compounds of formula I: Z stands for =CR2-; each of radicals R0 and R1 represents hydrogen; R2 represents (C1-C4)alkoxy; R3 represents -SO2NH2; or R1 and R2 form together with C-atoms, to which they are bound, 5-7-member non-aromatic carbocyclic or heterocyclic residue, where said heterocyclic residue includes 1 or 2 heteroatoms, selected from N and O, and heterocyclic residue, containing 1-2 atoms of oxygen can be substituted with fluorine atoms; R4 and R6 represent hydrogen, R5 represents hudrogen, halogen, (C1-C4)alkyl or CF3; one of R7, R8 and R9 represents NR10R11, and one or two others represent hydrogen, halogen, COOH, CF3 or (C1-C4)alkyl; R10 and R11 independently represents hydrogen or (C1-C4)alkyl. Invention also relates to methods of obtaining compounds.

EFFECT: compounds can be applied, for instance in case of acute or chronic rejection of organ or tissue, in treatment of atherosclerosis and other diseases, when inhibition of ZAP-70 is of importance.

9 cl, 7 tbl, 150 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology and can be applied for treatment of malignant tumors, resistant to platinum-based medications. Method by invention includes introduction to patient of humanised antibody against 2C4 HER2 or its fragment, and chemical therapeutic medication of antimetabolite, each in amount, effective for treatment of malignant tumor.

EFFECT: application of invention allows to treat such tumors as malignant tumor of ovary, peritoneal carcinoma and carcinoma of Fallopian tubes, resistant to platinum-based medications, due to more active suppression by antibody 2C4 HER2 of HER dimerisation.

30 cl, 4 tbl, 6 dwg, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine and pharmacology and represents stable pharmaceutical composition for parenteral application, which in one reservoir contains fixed doses of recombinant gamma and alpha 2b interferons for treatment of pathologic phenomena, characterised by benign non-physiological or malignant cell growth of tissues or organs, and which additionally contains pharmaceutically acceptable filling agents or carriers.

EFFECT: invention ensures higher tumour treatment efficiency due to synergic impact of composition components, as well as composition stability.

18 cl, 14 ex, 19 tbl, 9 dwg

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely, to oncology, and can be used in treatment of locally advanced and disseminated renal cell carcinoma. Method includes determination of absolute concentration of cells with phenotypes CD57+ in mm (X5) and CD4+CD25+Treg in mm3 (X4), as well as levels of spontaneous production of interleukine-6 in pg/ml (X3), interleukine-8 in pg/ml (X2) and interferon-γ in pg/ml (XI). Obtained data are used to calculate discriminant functions and the largest value of the function is used to predict patient's life duration and to administer treatment depending on predicted life duration.

EFFECT: application of invention allows to carry out adequate treatment of patients with different life duration, to optimise regimens of medication introduction, to improve remote results of treatment of patients with disseminated and locally advanced renal cell carcinoma.

12 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, oncology, and can be used for treating locally advanced breast cancer. For this purpose, the method starts with chemotherapy. 15-30 minutes later, it is followed with hyperthermia at temperature 41.0-42.0°C on the periphery of a tumour at exposure time 90-60 min, respectively. If observing involution of the tumour more than by 30% by RECIST criteria, radical mastectomy followed by radiation therapy is performed. A less evident involution requires preoperative radiation therapy followed by radical mastectomy.

EFFECT: method allows improving a local therapeutic effect due to the fact that on the periphery of the tumour, total vascular dilatation and blood flow increase are observed that allows for maximum penetration of medicinal agents into the tumour, and in the centre of the tumour, high temperature is generated because of decreased blood flow and therefore heat extraction thereupon by the end of the procedure in the tumour region, blood flow is slowed in vessels and stops almost, the cumulated medicinal agents are kept in the tumour for a long time that allow prolonging the cytotoxic action of chemopreparations on malignant cells.

3 cl, 6 dwg, 2 tbl, 5 ex

FIELD: medicine.

SUBSTANCE: there is claimed isolated human antibody or its fragment, which binds to human EGFR. Antibody contains corresponding CDR areas of light and heavy chain. Its conjugate with anti-neoplastic means or marker is described. Also described are: coding nucleic acid, expression vector, recombinant cell-host for obtaining antibodies and method of inhibiting growth of tumor, expressing EGFR on the basis of antibody.

EFFECT: application of invention provides antibodies with affinity comparable or higher, than in IMC-C225, which neutralises EGFR activation, what can be applied in medicine for treatment of tumours.

36 cl, 14 dwg, 6 tbl, 13 ex

FIELD: chemistry.

SUBSTANCE: claimed is method of obtaining N-pyrazinoyl-(L)-phenylalanyl-(L)-leucine boronic acid or its anhydride with application of reaction of (1S,2S,3R,5S)-pinandiol-(S)-1-clor-3-methylbutane-I-boronate with lithium bis(trimethylsilyl)-amide with formation of (1S,2S,3R,5S)-pinandiol-(R)-1-bis(trimethylsilyl)amino-3-methylbutane-1-boronate, which is subjected to extractive purification with water and is isolated in pure form and with further interaction with trifluoroacetic acid with formation of corresponding acid-additive salt, which is condensed with N-pyrazinoil-(L)-phenylalanine in presence of condensing agent TBTU and diisopropylethylamine in methylene chloride, N-pyrazinoil-(L)-phenylalanine being obtained by condensation of N,O-bis-silyl derivative of (L)-phenylalanine, formed from (L)-phenylalanine under impact of N,O-bis(trimethylsilyl)acetamide, and imidazolyl of pyrazine carbonic acid, with formation of (1S,2S,3R,5S)- pinandiol-N-pyrazinoil-(L)-phenylalanyl-(L)-leucine boronate, which is subjected to acid hydrolysis with formation of N-pyrazinoyl-(L)-phenylalanyl-(L)-leucine boronic acid or its anhydride.

EFFECT: method allows top obtain target product with high output, high purity using simpler and safer technology.

8 ex

FIELD: chemistry.

SUBSTANCE: medicine-ligand conjugates are powerful cytotoxins in which the medicine is bonded to the ligand through a peptide, hydrazine or disulphide linker.

EFFECT: agents are highly effective.

63 cl, 8 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds which have inhibiting properties on activity of at least one kinase selected from veGFR2, EphB4, PDGFRβ and c-Kit, having general formula I , where R denotes 0-1 substitutes independently selected from halogen, (C1-C6)-alkyl, (C1-C6) -alkoxy; R7 and R8 together with carbon atoms with which they are bonded form a condensed 5-member heteroaryl ring substituted with a -(Z1)mR1 group, where the heteroaryl ring contains at least one nitrogen atom and optionally an extra nitrogen atom; R1 is pyridinyl which can be substituted, where the substitutes are independently selected from hydroxy, cyano, halogen, (C1-C6)-alkyl, (C1-C6) - alkoxy, morpholinyl, imidazolyl, tetrazolyl, aminocarbonyl and -NHR10, where R10 is selected from hydrogen; (C1-C6)-alkyl; allyl, phenyl or pyrimidinyl, which may be substituted with CN; thiazolyl, pyridinyl, sulphonyl which is substituted with (C1-C6)-alkyl, an acyl which may be substituted with (C1-C6)-alkoxy, imidazolyl, cyclopropyl, (C1-C6)-alkyl, possibly substituted with NH2, NH(C1-C4-alkyl), N((C1-C4)-alkyl (C1-C4-alkyl)), hydroxy, (C1-C4)alkoxy, morpholinyl, pyrrolidinyl; Z1 is - CR5R6-, where each of R5 and R6 is selected from hydrogen; and m equals 1; R2 is benzo[1,3]dioxo-5-lyl or phenyl, which may be substituted with (C1-C6)-alkyl, halogen(C1-C6)-alkyl, halogen, phenyl, (C1-C6)-alkylsulphonyl or (C1-C6)-alkoxy, whose alkyl part may be substituted with hydroxyl, halogen, dioxanyl; and each of R3 and R4 is selected from hydrogen; provided that the formula 1 compound is not 5-(phenylcarbamoylamino)-3-(2-(4-pyridyl)ethyl)indole, 1-(4-chloro-3-(trifluoromethyl)phenyl)-3-(1-(pyridin-4-yl)-1H-indol-5-yl)urea and 1(2-methoxy-5-(trifluoromethyl)phenyl)-3-(1-pyridin-4-yl)-1H-indol-5-yl)urea, as well as their pharmaceutically acceptable salts. The invention also relates to a pharmaceutical composition, an inhibiting method and a method of preparing a pharmaceutical composition.

EFFECT: more effective use of the compounds.

29 cl, 1 dwg, 6 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel cyclic amine compounds of formula (1) or pharmaceutically acceptable salts thereof: . In formula (1), X is O, S, NR2 (where R2 is H, C1-C12 alkyl); when X is O, S, then R1 is H, CN, COOH, C2-C13 alkoxycarbonyl, carbamoyl group; and when X is NR2 (where R2 assumes values given above), R1 is CN; Ar1 and Ar2 are identical or different and each represents an aryl which can be substituted with 1-3 halogens; or Ar1 and Ar2 together with neighbouring carbon atoms to which they are bonded form a group with formula (b): (where ring S and ring T are identical and each is a benzene ring; Y is O); ring B is a benzene ring which can be substituted with 1-3 substitutes independently selected from a group comprising halogen, C1-C12 alkyl, C1-C8 halogenalkyl, C1-C12 alkoxy, C1-C8 halogenalkoxy group; n is an integer from 1 to 10; p, q are identical or different and each is an integer equal to 1 or 2. Formula (1) compounds are bonding inhibitors of the α2C-adrenoreceptor.

EFFECT: possibility of using compounds in pharmaceutical compositions.

7 cl, 1 tbl, 8 ex

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