RU2403256C2 - Cyclopamine analogues and methods of their application - Google Patents

Abstract

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

Description

Related Application

This application is associated with the following US applications: 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 herein by reference in their entirety.

State of the art

The hedgehog signaling pathway is an essential part of numerous processes in embryonic development. Members of the secreted proteins of the Hedgehog family control cell proliferation, differentiation, and tissue formation. The pathway was initially deciphered in the body of the fruit fly Drosophila, but since then it has been shown that it is largely preserved in invertebrates and vertebrates, including humans. The full activity of the Hedgehog signaling pathway decreases after embryogenesis in most cells, but the pathway remains active in some types of adult cells. Recently, unregulated activation of the Hedgehog signaling pathway has been shown to lead to certain types of cancer, as described in detail below.

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

Two transmembrane proteins are involved in signal transduction along the Hedgehog pathway; the twelve-transmembrane Patched receptor (Ptc) and the seven-transmembrane Smoothened protein (Smo).

Engineering studies suggest that Hedgehog acts by binding to Ptc, thereby releasing the inhibitory effect of Ptc on Smo. Since Ptc and Smo, both, are transmembrane proteins, the proposed scenario is that they physically bind to form a receptor complex, although indirect mechanisms of action are also likely. Depression of Smo from Ptc inhibition most likely includes a conformational change in Smo. Ptc, however, is not necessary for Smo activity, since Smo becomes constitutively activated in the complete absence of Pathed protein ( Alcedo et al., Supra ; Quirk et al. (1997) Cold Spring Harbor Symp. Quant. Biol. 62: 217-226 ) Once Smo is initiated, it quickly and highly phosphorylates and converts a signal that activates transcription via Gli transcription factors (protein homologue Ci Drosophila ) ( Alexandre et al. (1996) Genes Dev. 10: 2003-13 ). The Gli1 transcription factor up-regulates many genes involved in growth and development ( Alexandre et al., Supra). 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 many deficiencies and deformities, of which cyclopia is the most serious ( Belloni et al. (1996) Nature Genetics , 14: 353-6 ).

Recently, mutations in the formation of the Hedgehog pathway have been reported to occur with sporadic basal cell carcinoma ( Xie et al. (1998 ) Nature 391: 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 detected in numerous types of cancer, such as cancer of the gastrointestinal tract, including pancreatic cancer, esophageal cancer, and cancer of the stomach ( Berman et al. (2003) Nature 425: 846-51, Thayer et al. (2003 ) Nature 425: 851-56 ), lung cancer ( Watkins et al. (2003) Nature 422: 313-317 , prostate cancer ( Karhadkar et al. (2004) Nature 431: 707-12, Sheng et al. (2004) Molecular Cancer 3: 29-42, Fan et al. (2004) Endocrinology 145: 3961-70), breast cancer ( Kubo et al. (2004) Cancer Research 64: 6071-74, Lewis et al. (2004) Journal of Mammary Gland Biology and Neoplasia 2: 165-181 ) and hepatocellular cancer ( Sicklick et al. (2005) ASCO conference, Mohini et al. (2005) AACR conference ).

Inhibition of the activity of the Hedgehog pathway by low molecular weight compounds has been shown to result in cell death in a number of different types of cancer having unregulated activation of the Hedgehog pathway (see, for example , Berman et al., 2003 Nature 425 : 846-51 ).

Hedgehog pathway antagonists are currently being investigated in a large number of clinical settings where a therapeutic effect for a condition or disorder can be obtained by inhibiting one or more aspects of the Hedgehog pathway activity. Although the focus is on cancer, the researchers found that inhibition of the low molecular weight Hedgehog compounds showed an improvement in the symptoms of psoriasis ( Tas, et al., 2004 Dermatology 209 : 126-131, published US patent application 20040072913 (incorporated herein by reference)). Psoriasis is a very common chronic skin disorder, usually characterized by skin lesions, including erythematous papules and spots with silver scales, although there are changes on the skin and in other parts of the body. Psoriasis is currently believed to be an autoimmune disease, but its etiology is still poorly understood.

The Hedgehog pathway inhibitor, which has attracted considerable interest, is the natural cyclopamine product. Cyclopamine was first isolated from Veratrum californicum lily in 1966 after it was found that the offspring of sheep kept on pasture were born with severe congenital deformities. In order to identify the agent (a) that causes these birth defects, the FDA examined the possible sources of teratogens and identified the iervine family of steroid alkaloids, including the cyclopamine compound, as teratogens responsible for birth defects.

Much later it was found that the mechanism of action of cyclopamine was a direct inhibition of the activity of the Hedgehog pathway ( Cooper et al. (1998) Science 280: 1603-7, Chen et al. (2002) Genes and Development 16: 2743-8 ). Cyclopamine and related compounds have been shown to have anticancer activity through action on the Hedgehog pathway. Despite the initial prospect, no members of this family of compounds or their analogues have led to the successful development of an anti-cancer agent. The present invention fulfills this need and has other related advantages.

SUMMARY OF THE INVENTION

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

In certain areas, the present methods are used to counteract the phenotypic effects of unwanted activation of the Hedgehog pathway, such as mutations resulting in increased Hedgehog function, decreased Ptc function, or increased Smo function. For example, the present methods may include contacting the cell ( in vitro or in vivo) with the Hedgehog pathway antagonist of the present invention (as defined below) in an amount sufficient to antagonize the Smo-dependent pathway activation. Such antagonism will stop or slow down unwanted cell proliferation and can lead to cell death.

In some embodiments, the methods and compounds of the present invention can be used to regulate cell proliferation and / or cell death in vitro and / or in vivo , such as the treatment of cancer of the head, neck, nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx, larynopharynx, salivary glands, paragangliomas, pancreas, stomach, skin, esophagus, liver and gall tree, bone, small intestine, colon, rectum, ovaries, prostate, lung, mammary gland, lymphatic system , blood, central nervous system, bone marrow, or brain.

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

The compounds of the present invention can be further formulated as a pharmaceutical composition comprising a pharmaceutically acceptable excipient, for administration to a patient as a treatment for cancer. Hedgehog antagonists of the pathway of the present invention and / or compositions comprising them can be administered to a patient for the treatment of conditions including unwanted cell proliferation, for example, cancer and / or tumors of the head, neck, nose, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx , larynopharynx, salivary glands, paragangliomas, pancreas, stomach, skin, esophagus, liver and gall tree, bone, small intestine, colon, rectum, ovaries, prostate, lung, chest, lymphatic system, blood, central nervous system system, bone marrow, or brain. In some embodiments, the implementation of such compounds or compositions are applied systemically, for example parenterally and / or locally, for example topically.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

The definitions of the terms used here are intended to include these current definitions recognized for each term in the chemical and pharmaceutical fields. Where necessary, examples are provided. Definitions apply to terms, as they are used everywhere in this specification, unless otherwise limited in specific cases, either 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 or hydrogen atoms. Illustrative heteroatoms include boron, nitrogen, oxygen, phosphorus, sulfur, and selenium.

The term “alkyl” is known in the art and includes saturated aliphatic groups, including straight chain alkyl groups, branched chain alkyl groups, cycloalkyl (polycyclic) groups, alkyl substituted cycloalkyl groups and cycloalkyl substituted alkyl groups. In some embodiments, a straight chain or branched chain alkyl has about 30 or fewer carbon atoms in the main chain (e.g., C ₁ -C ₃₀ for a straight chain, C ₃ -C ₃₀ for a branched chain), and alternatively, about 20 or less . Similarly, cycloalkyls have from about 3 to about 10 carbon atoms in their ring structure, and alternatively, about 5, 6 or 7 carbon atoms in the ring structure. Alkyl groups, unless otherwise specified, may optionally be substituted with suitable substituents. The number of substituents is usually limited by the number of available valencies on the alkyl group; thus, the alkyl group may be substituted with one or more hydrogen atoms that might be present on the unsubstituted group. Suitable substituents for alkyl groups include halogen, = O, = N-CN, = N-OR ', = NR', OR ', NR' ₂ , SR ', SO ₂ R', SO ₂ NR ' ₂ , NR'SO ₂ R ', NR'CONR' ₂ , NR'COOR ', NR'COR', CN, COOR ', CONR' ₂ , OOCR ', COR' and NO ₂ , where each R 'is independent H, C ₁ -C ₆ alkyl, C ₂ -C ₆ heteroalkyl, C ₁ -C ₆ , acyl, C ₂ -C ₆ heteroacyl, C ₆ -C ₁₀ aryl, C ₆ -C ₁₀ heteroaryl, C ₇ -C ₁₂ arylalkyl or C ₆ -C ₁₂ heteroarylalkyl, each of which is optionally substituted with one or more groups selected from halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ heteroalkyl, C ₁ -C ₆ aryl, C ₁ -C ₆ heteroacyl, hydroxy, amino and = ABOUT; and where two R 'in the same substituent or on adjacent atoms, can be connected in the form of 3-7-membered rings, optionally containing up to three heteroatoms selected from N, O and S.

Unless otherwise specified, the number of carbon atoms 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 backbone structure. Similarly, lower alkenyl and lower alkynyl have similar chain lengths.

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

The terms “alkenyl” and “alkynyl” are known in the art and refer to unsaturated aliphatic groups similar in length and possible substitution to alkyls described above, but they contain at least one double or triple bond, respectively, and may contain a mixture of double, and triple bonds. Alkenyl and alkynyl groups are also optionally substituted, unless otherwise specified, with the same substituents described above for alkyl groups.

Heteroalkyl, heteroalkenyl and heteroalkynyl and the like are defined similarly to the corresponding hydrocarbyl (alkyl, alkenyl and alkynyl) groups, but the terms “hetero” refer to groups that contain 1-3 O, S or N heteroatoms or combinations thereof within the remainder of the main chain; thus, at least one carbon atom of the corresponding alkyl, alkenyl or alkynyl groups is replaced by one of these heteroatoms, forming a heteroalkyl, heteroalkenyl or heteroalkynyl group. Typical and preferred sizes for heteroforms of alkyl, alkenyl and alkynyl groups are usually the same as for the corresponding hydrocarbyl groups, and the substituents that may be present on the hetero groups are the same as those described above for the hydrocarbyl groups. It is also understood that unless otherwise specified, for reasons of chemical stability, such groups do not include more than two adjacent heteroatoms, excluding those where the oxo group is present on N or S, as in the sulfonyl 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, pyrazine, pyridazine and pyrimidine and the like. Aryl groups having heteroatoms in the ring structure may also be referred to as aryl heterocycles or heteroaromatic. The aromatic ring may be substituted at one or more ring positions with such substituents as described above, for example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino, nitro, sulfhydryl, imino, amido, phosphonate , phosphinate, carbonyl, carboxyl group, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic residues, —CF ₃ , —CN or the like. The term “aryl” also includes polycyclic systems having two or more cyclic rings in which two or more carbon atoms are common to two adjacent rings (the rings are “fused rings”), where at least one of the rings is aromatic for example, other cyclic rings may be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and / or heterocycles.

The terms ortho, meta and steam are known in the art and refer to 1,2-, 1,3- and 1,4- doubly substituted benzenes, respectively. For example, the names "1,2-dimethylbenzene" and "ortho-dimethylbenzene" are synonymous.

The terms “heterocyclyl”, “heteroaryl” or “heterocyclic group” are known in the art and refer to from 3 to about 10 membered ring structures, alternatively from 3 to about 7 membered rings, ring structures of which include 1 4 heteroatoms. Heterocycles may also be polycycles. Heterocyclic groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxanthene, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, fenarsazin, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, varnish tams, such as azetidinones and pyrrolidinones, sultams, sultones, etc. The heterocyclic ring may be substituted in one or more positions by such substituents as described above, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl a group, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic residues, —CF ₃ , —CN or the like.

The terms “polycyclyl” or “polycyclic group” are known in the art and refer to two or more rings (eg, cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and / or heterocyclyls) in which two or more carbon atoms are common to two attached rings, for example rings are condensed rings. Rings connected through non-adjacent atoms are called bridged rings. Each of the polycyclic rings may be substituted with such substituents as described above, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl group, silyl , ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic residue, —CF ₃ , —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 —NO ₂ , the 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 —OH; and the term “sulfonyl” is known in the art and refers to —SO ₂ . The halide defines the corresponding halogen anion, and the “pseudo-halide compound” is defined on page 560 of Advanced Inorganic Chemistry by Cotton and Wilkinson.

The terms "amine", "amino" and "ammonium" are known in the art and refer to unsubstituted and substituted amines, for example, a residue that can be represented by general formulas:

where R50, R51 and R52 each independently represents hydrogen, alkyl, alkenyl, - (CH ₂ ) _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 ring structure; R61 represents aryl, cycloalkyl, cycloalkenyl, heterocycle or polycyclic; and m is zero or an integer in the range of 1-8. In other embodiments, R50 and R51 (and optionally R52) each independently represent hydrogen, alkyl, alkenyl, or - (CH ₂ ) _m —R61. Thus, the term “alkylamine” includes an amine group, as defined above, having attached substituted or unsubstituted alkyl, i.e., at least one of R50 and R51 represents an alkyl group.

The term “acylamino” is known in the art and refers to a residue that can be represented by the general formula:

where R50, as defined above, and R54 are hydrogen, alkyl, alkenyl or (CH ₂ ) _m —R61, where m and R61 are as defined above.

The term "amido" is known in the art as an amino-substituted carbonyl and includes a residue that can 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 that may be unstable.

The term “alkylthio” refers to an alkyl group, as defined above, attached via a sulfur radical. In some embodiments, the alkylthio residue is represented by one of —S-alkyl, —S-alkenyl, —S-alkynyl and —S- (CH ₂ ) _m —R61, where m and R61 are as defined above. Typical alkylthio groups include methylthio, ethylthio and the like.

The term “carboxyl group” is known in the art and includes those moieties that can be represented by general formulas:

where X50 is a bond or oxygen or sulfur, and R55 and R56 are hydrogen, alkyl, alkenyl, - (CH ₂ ) _m -R61 or a pharmaceutically acceptable salt, R56 is hydrogen, alkyl, alkenyl or - (CH ₂ ) _{m is} R61, where m and R61 are as defined above. In cases where X50 is oxygen and R55 or R56 are not hydrogen, the formula is an ester. In cases where X50 is oxygen and R55, as defined above, the residue is referred to herein as a carboxyl group, and preferably when R55 is hydrogen, the formula is carboxylic acid. In those cases where X50 is oxygen and R56 is hydrogen, the formula is formate. Typically, in cases where the oxygen atom of the above formula is replaced by sulfur, the formula is a thiocarbonyl group. In those cases where X50 is sulfur and R55 or R56 is not hydrogen, the formula is a thioester. In cases where X50 is sulfur and R55 is hydrogen, the formula is thiocarboxylic acid. In those cases where X50 is sulfur and R56 is hydrogen, the formula is thioformate. On the other hand, in cases where X50 is a bond and R55 is not hydrogen, the above formula represents a ketone group. In cases where X50 is a bond and R55 is hydrogen, the above formula represents an aldehyde group.

The term “carbamoyl” refers to —O (C = O) NRR ′, where R and R ′ are independently H, aliphatic groups, aryl groups or heteroaryl groups.

The term “oxo” refers to carbonyl oxygen (= O).

The terms “alkoxyl” or “alkoxy” are known in the art and refer to an alkyl group, as defined above, attached via an oxygen radical. Typical alkoxy groups include methoxy, ethoxy, propyloxy, tert-butoxy and the like. The ether is two hydrocarbons connected by a covalent bond through oxygen. Accordingly, an alkyl substituent which forms an alkyl ester or is part of an alkoxyl group which may be represented by one of —O-alkyl, —O-alkenyl, —O-alkynyl, —O- (CH ₂ ) _m —R61, where m and R61 are described above.

The term "sulfonate" is known in the art and refers to a residue that can be represented by the general formula:

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

The term "sulfate" is known in the art and includes a residue that can be represented by the general formula:

in which R57, as defined above.

The term "sulfonamido" is known in the art and

includes a residue that can 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 a residue that can be represented by the general formula:

in which R50 and R51, as defined above.

The term "sulfonyl" is known in the art and refers to a residue that can be represented by the general formula:

in which R58 is one of the following: hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl.

The term "sulfoxido" is known in the art and refers to a residue that can be represented by the general formula:

in which R58 is defined above.

The term "phosphoryl" is known in the art and can be represented by the general formula:

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

where Q50 and R59 are each independently defined above, and Q51 is O, S or N. When Q50 is S, the phosphoryl residue is “phosphorothioate”.

The term "phosphoramidite" is known in the art and can be represented by general formulas:

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

The term "phosphonamidite" is known in the art and can be represented by general formulas:

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

Alkenyl and alkynyl groups can be replaced with similar substituents to obtain, for example, aminoalkenyls, aminoalkynyls, amidoalkenyls, amidoalkynyls, iminoalkenyls, iminoalkynyls, thioalkenyls, thioalkynyls, carbonyl-substituted alkenyls or alkynyls.

The definition of each expression, for example, “alkyl”, “m”, “n” and the like, is intended to be independent of the definition in another place of the same structure when it is repeatedly found in this structure.

The term "selenoalkyl" is known in the art and refers to an alkyl group attached through a seleno group. A typical “selenoester” may be an alkyl substituent selected from —Se-alkyl, -Se-alkenyl, -Se-alkynyl, and -Se- (CH ₂ ) _m —R61, m and R61, as defined above.

The terms "triflil", "tosyl", "methylsulfonyl" and "nonaflil" are known in the art and refer to trifluoromethanesulfonyl, p-toluenesulfonyl, methanesulfonyl and nonafluorobutanesulfonyl groups, respectively. The terms "triflate", "tosylate", "mesylate" and "nonaflate" are known in the art and refer to trifluoromethanesulfonate, p-toluenesulfonate, methanesulfonate and nonafluorobutanesulfonate, the functional groups and molecules of which contain these groups.

The abbreviations Me, Et, Ph, Tf, Nf, Ts and Ms are methyl, ethyl, phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl, p-toluenesulfonyl and methanesulfonyl, respectively. A more complete list of abbreviations used in the art by organic chemists of ordinary skill appears in the first issue of each volume of the Journal of Organic Chemistry; this list is usually presented in the table entitled Standard List of Abbreviations.

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

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

It is understood that the substituent or substituted implies that such substitution is in accordance with the permissible valency of the substituted atom and substituent and that the substitution leads to a stable compound, 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 of the acceptable substituents of organic compounds. In a broad aspect, acceptable substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Illustrative substituents include, for example, those described hereinabove. Permissible substituents may be one or more of the same or different for suitable organic compounds. For the purposes of this invention, heteroatoms, such as nitrogen, may have hydrogen substituents and / or any permissible substituents of the organic compounds described herein that satisfy the valency of the heteroatoms. This invention is not intended to be limited by any acceptable substituents of organic compounds.

As used herein, the phrase “protecting group” means temporary substituents that protect a potentially reactive functional group from undesired chemical reactions. Examples of such protective groups include esters of carboxylic acids, silyl esters of alcohols and acetals, and ketals of aldehydes and ketones, respectively. The field of chemistry of protective groups has been examined (Green, TW; Wuts, PGM Protective Groups in Organic Synthesis, 2 ^nd ed .; Wiley: New York, 1991). Protected forms of the compounds of the invention are included within the scope of this invention.

For the purposes of this invention, 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 such genetic lesions as, for example, deletion, substitution or insertion of nucleotides in a gene, as well as significant chromosomal gene rearrangements and / or pathological gene methylation. Similarly, improper gene expression refers to aberrant levels of gene transcription relative to such levels in a normal cell under similar conditions, as well as splicing of an unstable type of mRNA transcribed from a gene.

Basal cell carcinomas exist in various clinical and histological forms, such as nodular-ulcerative, superficial, pigmented, annular, fibroepithelial and non-nevoid syndrome. Basal cell cancers are the most common skin tumors found in humans. Most new cases of non-melanoma skin cancer fall into this category.

The term "carcinoma" refers to a malignant neoplasm composed of epithelial cells, which tend to infiltrate into surrounding tissues and give rise to metastases. Typical carcinomas include basal cell carcinoma, which is an epithelial tumor of the skin that, rarely metastasizing, has the potential for local invasion and destruction; squamous cell carcinoma, which refers to carcinomas originating from squamous epithelium and having cubic cells; carcinosarcoma, which includes malignant tumors composed of carcinomatous and sarcomatous tissues; adenocystic carcinoma, a carcinoma characterized by cylinders or stripes or a mucinous stroma, separated or surrounded by nests or cords of small epithelial cells found in the mammary and salivary glands and mucous glands of the respiratory tract; epidermoid carcinoma, which refers to malignant cells that tend to differentiate in the same way as the like from the epidermis; those. they tend to form spike cells and tolerate keratinization; nasopharyngeal carcinoma, which refers to a malignant tumor that occurs in the epithelial lining of the space behind the nose; and renal cell carcinoma, which refers to cancer of the renal parenchyma composed of tubule cells in a changed location.

Other carcinomatous epithelial growths are papillomas, which are benign tumors originating from the epithelium, and have a causative agent - the papilloma virus; and epidermoidomas, which refer to a tumor of the brain or meninges that forms the inclusion of ectodermal elements during the laying of the groove of the neural tube.

The term "ED50" means a dose of a drug that produces 50% of its maximum response or effect.

An effective amount of a compound of interest relative to existing methods of treatment refers to the amount of antagonist in a composition that, when used as part of a desired dosage regimen, causes, for example, a change in cell proliferation level and / or cell survival rate according to clinically acceptable standards for disease, to who will be treated.

The terms “epithelial” and “epithelium” refer to the cell coating of the internal and external surfaces of the body (skin, mucous membranes and serous membranes), including glands and other structures emanating from them, for example corneal, esophageal, epidermal and hair epithelial follicle cells. Another typical epithelial tissue includes the olfactory epithelium, which is a pseudo-layered epithelium lining the olfactory region of the nasal cavity and containing olfactory receptors; glandular epithelium, which refers to an epithelium consisting of secreting cells; squamous epithelium, which refers to an epithelium made up of smoothed plate cells. The term "epithelium" may also refer to a transitional epithelium, which is typically located in the lining of hollow organs, which undergo a large mechanical change, such as contraction and stretching, for example, tissue, which is a transition between a multilayer squamous and columnar epithelium.

A cell growth state refers to a cell proliferation rate and / or a cell differentiation state. An altered growth state is a growth state characterized by a pathological level of proliferation, for example, a cell showing an increased or decreased level of proliferation relative to a normal cell.

The term “Hedgehog pathway antagonist” refers to an agent that slows down the function of the Hedgehog pathway, for example, inhibits the transcription of target genes (Gli1 and Ptc genes), which is induced in normal cells by contacting a cell with a Hedgehog. In addition to changing the Smoothened-dependent pathway, in some embodiments, the Hedgehog antagonists of the pathway of the present invention can be used to overcome decreased Ptc function, Smoothened increased function and / or increased Hedgehog function. The terms "decrease in function" and "increase in function" respectively refer to an aberrant modification or mutation, for example, of a Ptc gene, Hedgehog gene, or Smoothened gene, or a decrease or increase in the expression level of such a gene that leads to a phenotype that, for example, resembles cell contact with a Hedgehog protein, such as aberrant activation of the Hedgehog pathway or resembles a decrease in Smo function. Mutation may include a decrease in the ability of Ptc or Smo gene production to regulate the activity level of Gli / Ci proteins, such as Gli1, Gli2 and Gli3.

As used here, immortalized cells refer to cells that have been chemically altered and / or recombinant cells, and mean that such cells have the ability to grow through an unlimited number of divisions in culture.

The term "LD50" means a dose of a drug that is lethal to 50% of the tested patients.

“Patient” or “subject” means an organism being treated by the method of the present invention, and may mean either a human or an animal.

The phrase “pharmaceutically acceptable” is used herein to refer to those compounds, materials, compositions and / or dosage forms which, within the limits of the medical judgment examined, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reaction or other problem or complications commensurate with an acceptable benefit / risk ratio.

The phrase “pharmaceutically acceptable carrier,” as used herein, means a pharmaceutically acceptable material, composition or binder, such as a liquid or solid excipient, solvent, inert excipient, excipient (eg, a lubricant, magnesia powder, calcium or zinc stearate or stearic acid ) or soluble encapsulating material included in the transfer or transportation of the subject compound from one organ or body part to another organ or body part. Each carrier must be acceptable in the sense of being compatible with the other ingredients of the composition and not harmful to the patient. Some examples of materials that 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 carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) inert excipients, such as cocoa butter and suppository wax; (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) polyhydric alcohols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar-agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (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 formulations.

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

As used herein, “proliferating” and “proliferating” refers to cells in a state of mitosis.

Throughout this application, the term "proliferative skin disorder" refers to any disease / skin disorder marked by unwanted or aberrant proliferation of skin tissue. These conditions are usually characterized by epidermal cell proliferation or incomplete cell differentiation and include, for example, X-linked ichthyosis, psoriasis, atopic dermatitis, allergic contact dermatitis, epidermolytic hyperkeratosis and seborrheic dermatitis. For example, epidermodysplasia is a form of defective development of the epidermis. Another example is epidermolysis, which refers to a weakened state of the epidermis with the formation of bullae either spontaneously or at the site of injury.

The term “therapeutic index” refers to the therapeutic index of a drug, defined as LD50 / ED50.

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

The term "subject", as used here, refers to an animal, usually a mammal, or a person who was the goal of treatment, observation and / or experiment. When this term is used in combination with the administration of a compound or drug, the subject is treated, monitored and / or administered with the compound or drug. The phrase “therapeutically effective amount” as used herein means an amount of a compound, material or composition comprising a compound of the present invention that is effective to produce some desired therapeutic effect in at least a subpopulation of animal cells in an acceptable benefit / ratio risk applicable to any treatment.

The phrases “parenteral administration” and “applied parenterally” as used herein mean methods of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac , intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, spinal and sternal injection and infusion.

The phrases "systemic exposure", "applied systemically", "peripheral exposure" and "applied peripheral", as used here, mean the introduction of a compound, drug or other material, other than directly to the rest of the Hedgehog pathway associated with the disorder, such what enters the patient’s system and, thus, is subject to metabolism and other similar processes, for example, subcutaneous administration.

The term “sugar,” as used herein, refers to a natural or non-natural monosaccharide, disaccharide or oligosaccharide comprising one or more pyranose or furanose rings. Sugar can be covalently linked to the steroidal alkaloid of the present invention via an ether linkage or via an alkyl linkage. In certain embodiments, the saccharide residue may be covalently linked to the steroid alkaloid of the present invention at the anomeric center of the saccharide ring.

является алкильным двухвалентным радикалом;

является также алкильным двухвалентным радикалом;

является аралкильным двухвалентным радикалом; и

является (алкил)гетероалкильным двухвалентным радикалом. Типичные примеры включают алкилены общей структуры (СН₂)_х, где Х равно 1-6 и соответствующие алкениленовые и алкиниленовые линкеры, имеющие 2-6 углеродных атомов и одну или более двойных или тройных связей; циклоалкиленовые группы, имеющие 3-8 кольцевых элементов; и аралкильные группы, где одна открытая валентность находится на кольце арила, и каждый из алкильных фрагментов, таких как

, и его изомеры.The term “biradical”, as used herein, refers to any number of divalent groups, for example, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl and heteroalkyl groups. For example,

is an alkyl divalent radical;

is also an alkyl divalent radical;

is an aralkyl divalent radical; and

is a (alkyl) heteroalkyl divalent radical. Typical examples include alkylenes of the general structure (CH ₂ ) _x , where X is 1-6 and the corresponding alkenylene and alkynylene linkers having 2-6 carbon atoms and one or more double or triple bonds; cycloalkylene groups having 3-8 ring elements; and aralkyl groups, where one open valency is on the aryl ring, and each of alkyl fragments, such as

, and its isomers.

Compounds of the invention

The present invention provides cyclopamine analogues, as well as their isolated and purified forms, including synthetic and semi-synthetic analogues, as well as pharmaceutical compositions containing such analogues. In one embodiment, the present invention provides compounds represented by a compound of formula 1 :

(1)

(one)

or a pharmaceutically acceptable salt thereof;

in which each R ¹ and R ⁸ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, heteroaralkyl, halide, sulfhydryl, alkylthio, arylthio, aralkylthio, hydroxyl, alkoxy, aryloxy, acylo , alkylamino, arylamino, acylamino, aralkylamino, nitro, acylthio, carboxamide, sulfonamide, carboxyl, nitrile, sulfate, -OP (L) (OR ²⁰ ) ₂ , -XC (L) -R ²¹ or -XC (L) -X -R ²¹ ;

where R ^l may also be sugar;

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

L represents O or S;

R ² and R ⁹ independently represent H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, alkoxyl, aryloxy, acyloxy, halide, sulfhydryl, alkylthio, arylthio, aralkylthio, hydroxyl, amino, alkylamino, arylamino, aralkylamino, heteroaryl or heteroaralkyl;

each R ⁵ and R ¹¹ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamino, alkylseleno, arylalkyl alkylthio, aralkylthio, arylthio, heteroaryl or heteroaralkyl;

each R ³ , R ⁴ , R ⁶ , R ⁷ , R ¹³ and R ¹⁴ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino , arylamino, acylamino, aralkylamino, heteroaryl or heteroaralkyl;

or R ¹ and R ² and / or R ⁸ and R ⁹ , taken together with the carbon to which they are bonded, form - (C = O) -, - (C = S) -, - (C = N (OR ²⁰ )) -, - (C = N (R ²⁰ )) -, - (C = N (N (R ²⁰ ) (R ²⁰ ))) or form an optionally substituted 3-8 membered ring; or

R ⁴ and R ⁵ taken together and / or R ⁵ and R ⁶ taken together and / or R ¹⁰ and R ¹¹ taken together form a double bond or form a group represented by 1b

where Z represents NR ²¹ , O or C (R ²³ ) (R ²³ );

R ¹² represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, hydroxyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, alkoxyl, —C (O) R ²¹ , —CO ₂ R ²¹ , —SO ₂ R ²¹ , -C (O) N (R ²¹ ) (R ²¹ ), - [C (R ²¹ ) ₂ ] _q -R ²¹ , - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -C (O)] _q R ²¹ , - [(W) -C (O) O] _q R ²¹ , - [(W) - (O) C (O)] _q R ²¹ , - [ (W) -SO ₂ ] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -N (R ²¹ )] _q R ²¹ or - [(W) -S] _q R ²¹ ;

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

R ¹⁵ , R ¹⁶ and R ¹⁷ independently represent H, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamino; or R ¹⁵ and R ¹⁶ taken together with the carbon to which they are attached form —C (O) - or —C (S) -;

R ¹⁸ and R ¹⁹ independently represent H, alkyl, aralkyl, halide, amido or ester;

R ²⁰ represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl or heteroaralkyl; or any two R ²⁰ in one substituent may be taken together to form a 4-8 membered optionally substituted ring;

R ²¹ represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, heteroaralkyl or - [C (R ²⁰ ) ₂ ] _p -R ²⁵ where p is 0-6; or any two R ²¹ in one substituent may be taken together to form a 4-8 membered optionally substituted ring;

R ²³ represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, heteroaralkyl, halide, alkoxy, aryloxy, acyloxy, silyloxy, nitrile, —C (O) R ²¹ , —CO ₂ R ²¹ , -SO ₂ R ²¹ and -C (O) N (R ²¹ ) ₂ ;

R ²⁵ represents hydroxyl, acylamino, —N (R ²⁰ ) COR ²⁰ , —N (R ²⁰ ) C (O) OR ²⁰ , —N (R ²⁰ ) SO ₂ (R ²⁰ ), —COR ²⁰ N (R ²⁰ ) ₂ , -OC (O) R ²⁰ N (R ²⁰ ) (R ²⁰ ), -SO ₂ N (R ²⁰ ) (R ²⁰ ), -N (R ²⁰ ) (R ²⁰ ), -COOR ²⁰ , -C (O) N (OH) (R ²¹ ), -OS (O) ₂ OR ²⁰ , -S (O) ₂ OR ²⁰ , -OP (L) (OR ²⁰ ) (OR ²⁰ ), -NP (O) ( OR ²⁰ ) (OR ²⁰ ) or -P (O) (OR ²⁰ ) (OR ²⁰ ).

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ and R ¹⁷ are hydrogen.

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹ is hydroxyl, sugar, —OP (L) (OR ²⁰ ) ₂ , —XC (L) —R ^21, or —XC (L) —X— R ²¹ ; or R ¹ and R ² taken together with the carbon to which they are bonded form —C (O) -.

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, where R ⁴ and R ⁵ taken together form a double bond.

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, where R ¹ and R ² , taken together with the carbon to which they are attached, form —C (O) -.

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹ is hydroxyl and R ² is H.

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹ is hydroxyl, R ² is H; and R ⁵ and R ⁶ taken together form a double bond; or R ⁵ and R ⁶ taken together form a group represented by 1b ;

Where

Z represents C (R ²³ ) (R ²³ ).

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹⁰ and R ¹¹ taken together form a double bond; or R ¹⁰ and R ^ll taken together form a group represented by 1b ;

Where

Z represents C (R ²³ ) (R ²³ ).

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, where R ⁵ and R ⁶ taken together form a double bond and R ¹⁰ and R ^ll taken together form a double bond.

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹ and R ² , taken together with the carbon to which they are attached, form —C (O) -; R ⁴ and R ⁵ taken together form a double bond; and R ¹⁰ and R ¹¹ taken together form a double bond; or R ¹⁰ and R ¹¹ taken together form a group represented by 1b ;

Where

Z represents C (R ²³ ) (R ²³ ).

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹ is hydroxyl and R ² is H; R ¹⁰ and R ^l1 taken together form a double bond; or R ¹⁰ and R ¹¹ taken together form a group represented by 1b ;

Where

Z represents C (R ²³ ) (R ²³ ).

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ⁸ and R ⁹ are hydrogen; or R ⁸ and R ⁹ taken together with the carbon to which they are attached are —C (O) -.

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹² is H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, hydroxyl, alkoxyl, - [(W) -N (R ²¹ ) C (O )] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) - O] _q R ²¹ , - [(W) -C (O)] _q R ²¹ or - [(W) -C (O) O] _q R ²¹ .

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ and R ¹⁷ are hydrogen; and R ¹² represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, hydroxyl, alkoxyl, - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -C (O)] _q R ²¹ or - [(W) -C (O) O] _q R ²¹ .

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ⁴ and R ⁵ taken together form a double bond; R ¹ and R ² taken together with the carbon to which they are bonded form —C (O) -; and R ¹² represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, hydroxyl, alkoxyl, - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -C (O)] _q R ²¹ or - [(W) -C (O) O] _q R ²¹ .

In some embodiments, the compounds of the present invention are represented by 1 and the accompanying definitions, wherein R ¹ is hydroxyl and R ² is H; and R ¹² represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, hydroxyl, alkoxyl, - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -C (O)] _q R ²¹ or - [(W) -C (O) O] _q R ²¹ .

In some embodiments, the compounds of the present invention are represented by a compound of the formula:

Where

или

or

R ¹² represents H, alkyl, aryl, cycloalkyl, heterocycloalkyl, hydroxyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, alkoxyl, —C (O) R ²¹ , —CO ₂ R ²¹ , —SO ₂ R ²¹ , —C (O ) N (R ²¹ ) (R ²¹ ), - [C (R ²¹ ) ₂ ] _p -R ²¹ , - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -C (O)] _q R ²¹ , - [(W) -C (O) O] _q R ²¹ , - [(W) -OC (O)] _q R ²¹ , - [(W) -SO ₂ ] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -N (R ²¹ )] _q R ²¹ or - [(W) -S] _q R ²¹ ; where each W independently represents a biradical;

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

R ²⁰ represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl or heteroaralkyl; or any two R ²⁰ in one substituent may be taken together to form a 4-8 membered optionally substituted ring;

R ²¹ represents H, alkyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, heteroaralkyl or - [C (R ²⁰ ) ₂ ] _p -R ²⁵ ; or any two R ²¹ can be taken together to form a 4-8 membered optionally substituted ring;

R ²⁵ represents hydroxyl, acylamino, —N (R ²⁰ ) COR ²⁰ , —N (R ²⁰ ) C (O) OR ²⁰ , —N (R ²⁰ ) SO ₂ (R ²⁰ ), —COR ²⁰ N (R ²⁰ ) ₂ , -OC (O) R ²⁰ N (R ²⁰ ) (R ²⁰ ), -SO ₂ N (R ²⁰ ) (R ²⁰ ), -N (R ²⁰ ) (R ²⁰ ), -COOR ²⁰ , -C (O) N (OH) (R ²¹ ), -OS (O) ₂ OR ¹⁹ , -S (O) ₂ OR ²⁰ , -OP (L) (OR ²⁰ ) (OR ²⁰ ), -NP (O) ( OR ²⁰ ) (OR ²⁰ ) or -P (O) (OR ²⁰ ) (OR ²⁰ ).

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

In some embodiments, the compounds of the present invention are represented by any of the aforementioned compounds and the accompanying definitions, wherein the compound is represented by the formula:

In one embodiment, the present invention provides compounds represented by a compound of formula 2 :

or a pharmaceutically acceptable salt thereof;

in which each R ¹ and R ⁸ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, heteroaralkyl, halide, sulfhydryl, alkylthio, arylthio, aralkylthio, hydroxyl, alkoxy, aryloxy, acylo , alkylamino, arylamino, acylamino, aralkylamino, nitro, acylthio, carboxamide, sulfonamide, carboxyl, nitrile, sulfate, -OP (L) (OR ²⁰ ) ₂ , -XC (L) -R ²¹ or -XC (L) -X -R ²¹ ;

where R ^l may also be sugar;

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

L represents O or S;

R ² and R ⁹ independently represent H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, alkoxyl, aryloxy, acyloxy, carboxyl, halide, sulfhydryl, alkylthio, arylthio, aralkylthio, hydroxyl, amino, alkylamino, a acylamino, aralkylamino, heteroaryl or heteroaralkyl;

each R ⁵ and R ¹¹ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamino, alkylseleno, arylalkyl alkylthio, aralkylthio, arylthio, heteroaryl or heteroaralkyl;

each R ³ , R ⁴ , R ⁶ , R ⁷ , R ¹³ and R ¹⁴ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino , arylamino, acylamino, aralkylamino, heteroaryl or heteroaralkyl;

where R ¹ and R ² and / or R ⁸ and R ⁹ , taken together with the carbon to which they are bonded, form - (C = O) -, - (C = S) -, - (C = N (OR ²⁰ )) -, - (C = N (R ²⁰ )) -, - (C = N (N (R ²⁰ ) (R ²⁰ ))) or form an optionally substituted 3-8 membered ring; or

R ⁴ and R ⁵ taken together and / or R ⁵ and R ⁶ taken together and / or R ¹⁰ and R ¹¹ taken together form a double bond or form a group represented by 1b

where Z represents NR ²¹ , O or C (R ²³ ) (R ²³ );

R ¹² represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, hydroxyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, alkoxyl, —C (O) R ²¹ , —CO ₂ R ²¹ , —SO ₂ R ²¹ , -C (O) N (R ²¹ ) (R ²¹ ), - [C (R ²¹ ) ₂ ] _q -R ²¹ , - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -C (O)] _q R ²¹ , - [(W) -C (O) O] _q R ²¹ , - [(W) -OC (O)] _q R ²¹ , - [(W) -SO ₂ ] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W ) -O] _q R ²¹ , - [(W) -N (R ²¹ )] _q R ²¹ or - [(W) -S] _q R ²¹ ;

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

R ¹⁵ , R ¹⁶ and R ¹⁷ independently represent H, alkoxyl, aryloxy, acyloxy, halide, hydroxyl, amino, alkylamino, arylamino, acylamino, aralkylamino; or R ¹⁵ and R ¹⁶ taken together with the carbon to which they are attached form —C (O) - or —C (S) -;

R ¹⁸ and R ¹⁹ independently represent H, alkyl, aralkyl, halide, amido or ester;

R ²⁰ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl or heteroaralkyl; or any two R ²⁰ in one substituent may be taken together to form a 4-8 membered optionally substituted ring;

R ²¹ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, heteroaralkyl or - [C (R ²⁰ ) ₂ ] _p -R ²⁵ where p is 0-6; or any two R ²¹ in one substituent may be taken together to form a 4-8 membered optionally substituted ring;

R ²³ independently represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, heteroaralkyl, halide, alkoxy, aryloxy, acyloxy, silyloxy, nitrile, —C (O) R ²¹ , —CO ₂ R ²¹ , -SO ₂ R ²¹ and -C (O) N (R ²¹ ) ₂ ; and

R ²⁵ represents hydroxyl, acylamino, —N (R ²⁰ ) COR ²⁰ , —N (R ²⁰ ) C (O) R ²⁰ , —N (R ²⁰ ) SO ₂ (R ²⁰ ), —COR ²⁰ N (R ²⁰ ) ₂ , -OC (O) R ²⁰ N (R ²⁰ ) (R ²⁰ ), -SO ₂ N (R ²⁰ ) (R ²⁰ ), -N (R ²⁰ ) (R ²⁰ ), -COOR ²⁰ , -C (O) N (OH) (R ²¹ ), -OS (O) ₂ OR ²⁰ , -S (O) ₂ OR ²⁰ , -OP (L) (OR ²⁰ ) (OR ²⁰ ), -NP (O) ( OR ²⁰ ) (OR ²⁰ ) or -P (O) (OR ²⁰ ) (OR ²⁰ ),

provided that at least one group represented by formula 1b exists on said compound of formula 2.

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein, in this specification, R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ and R ¹⁷ are hydrogen.

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ¹ is hydroxyl, sugar, OP (L) (OR ²⁰ ) ₂ , —XC (L) —R ²¹ or —XC (L) —XR ²¹ ; or R ¹ and R ² taken together with the carbon to which they are attached form —C (O) -.

In some embodiments, the compounds of the present invention are represented by formula 2 and the accompanying definitions, where R ⁴ and R ⁵ taken together form a double bond.

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ¹ and R ² , taken together with the carbon to which they are attached, form —C (O) -.

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ¹ is hydroxyl and R ² is H.

In some embodiments, the compounds of the present invention are represented by formula 2 and the accompanying definitions, wherein R ¹ is hydroxyl and R ² is H; R ⁵ and R ⁶ taken together form a double bond; or R ⁵ and R ⁶ taken together form a group represented by 1b;

Where

Z represents C (R ²³ ) (R ²³ ).

In some embodiments, the compounds of the present invention are represented by formula 2 and the accompanying definitions, wherein R ¹⁰ and R ¹¹ taken together form a double bond; or R ¹⁰ and R ¹¹ taken together form a group represented by 1b;

Where

Z represents C (R ²³ ) (R ²³ ).

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ⁵ and R ⁶ taken together form a double bond, and R ¹⁰ and R ¹¹ taken together form a double bond.

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ¹ and R ² , taken together with the carbon to which they are attached, form —C (O) -; R ⁴ and R ⁵ taken together form a double bond; and R ¹⁰ and R ¹¹ taken together form a double bond; or R ¹⁰ and R ¹¹ taken together form a group represented by 1b;

Where

Z represents C (R ²³ ) (R ²³ ).

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ¹ is hydroxyl and R ² is H; R ¹⁰ and R ¹¹ taken together form a double bond; or R ¹⁰ and R ¹¹ taken together form a group represented by 1b;

Where

Z represents C (R ²³ ) (R ²³ ).

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ⁸ and R ⁹ are hydrogen; or R ⁸ and R ⁹ taken together by the d carbon to which they are bonded are —C (O) -.

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ¹² is H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, hydroxyl, alkoxyl, - [(W) -N (R ²¹ ) C ( O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -C (O)] _q R ²¹ or - [(W) -C (O) O] _q R ²¹ .

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ and R ¹⁷ are hydrogen and R ¹² is H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, hydroxyl, alkoxyl, - [(W) -N (R ²¹ ) C (O)] _q ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -C (O)] _q R ²¹ or - [(W) -C (O) O] _q R ²¹ .

In some embodiments, the compounds of the present invention are represented by formula 2 and the accompanying definitions, wherein R ⁴ and R ⁵ taken together form a double bond; R ¹ and R ² taken together with the carbon to which they are bonded form —C (O) -; and R ¹² represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, hydroxyl, alkoxyl, - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -C (O)] _q R ²¹ or - [(W) -C (O) O] _q R ²¹ .

In some embodiments, the compounds of the present invention are represented by Formula 2 and the accompanying definitions, wherein R ¹ is hydroxyl and R ² is H; and R ¹² represents H, alkyl, cycloalkyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, hydroxyl, alkoxyl, - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -C (O)] _q R ²¹ or - [(W) -C (O) O] _q R ²¹ .

In some embodiments, the compounds of the present invention are represented by a compound of the formula:

или

or

Where

R ¹² represents H, alkyl, aryl, cycloalkyl, heterocycloalkyl, hydroxyl, aralkyl, heteroaryl, heteroaralkyl, haloalkyl, alkoxyl, —C (O) R ²¹ , —C (O) N (R ²¹ ) (R ²¹ ), - [C (R ²¹ ) ₂ ] _p -R ²¹ , - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -C (O)] _q R ²¹ , - [ (W) -C (O) O] _q R ²¹ , - [(W) -OC (O)] _q R ²¹ , - [(W) -SO ₂ ] _q R ²¹ , - [(W) -N ( R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -C (O) N (R ²¹ )] _q R ²¹ , - [(W) -O] _q R ²¹ , - [(W) -N ( R ²¹ )] _q R ²¹ or - [(W) -S] _q R ²¹ ;

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

R ²⁰ represents H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, or heteroaralkyl; or any two R ²⁰ in one substituent may be taken together to form a 4-8 membered optionally substituted ring;

R ²¹ represents H, alkyl, aryl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaryl, heteroaralkyl or - [C (R ²⁰ ) ₂ ] _p -R ²⁵ ; or any two R ²¹ can be taken together to form a 4-8 membered optionally substituted ring;

R ²⁵ represents hydroxyl, acylamino, —N (R ²⁰ ) COR ²⁰ , —N (R ²⁰ ) C (O) OR ²⁰ , —N (R ²⁰ ) SO ₂ (R ²⁰ ), —COR ²⁰ N (R ²⁰ ) ₂ , -OC (O) R ²⁰ N (R ²⁰ ) (R ²⁰ ), -SO ₂ N (R ²⁰ ) (R ²⁰ ), -N (R ²⁰ ) (R ²⁰ ), -COOR ²⁰ , -C (O) N (OH) (R ²¹ ), -OS (O) ₂ OR ¹⁹ , -S (O) ₂ OR ²⁰ , -OP (L) (OR ²⁰ ) (OR ²⁰ ), -NP (O) ( OR ²⁰ ) (OR ²⁰ ) or -P (O) (OR ²⁰ ) (OR ²⁰ ).

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

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

In some embodiments, the compounds of the present invention are represented by any of the aforementioned compounds and the accompanying definitions, wherein the compound is represented by the formula:

In some embodiments, the present invention relates to a pharmaceutical composition comprising any one or more of the above compounds; and a pharmaceutically acceptable excipient.

Synthesis of Steroid Alkaloid Compounds

The cyclopropyl steroid alkaloid derivatives of the present invention can be obtained directly from a steroid alkaloid isolated in the form of a natural product (or synthesized) or N-protected forms of these compounds. Suitable nitrogen protecting groups include, but are not limited to, Fmoc, Alloc, Boc, Troc, trifluoroacetate, tosyl, Cbz, ethyl cyanide and Bn.

A variety of cyclopropanizing agents can be used to cyclopropane steroid alkaloid. 1,1-Haloalkyl metal complexes, including reactive substances known as carbenoids, commonly used for cyclopropanation of olefins. These reagents are typically prepared using diiodalkane or diazoalkane and metal or organometallic compounds such as Et ₂ Zn, i Bu ₃ Al, samarium, copper, rhodium or palladium. In some embodiments, Et ₂ Zn and diiodomethane are used to effect cyclopropanation. Other known cyclopropanation processes may also be used, such as those using sulfur ylides to react with a carbonyl conjugated olefin to attach CH ₂ or CH-alkyl or CH-aryl groups, and metal-catalyzed decomposition of diazoalkyl and α-diazocarbonyl compounds, such as diazomethane and ethyl diazoacetate: these methods readily provide cyclopropanes having alkyl, aryl, alkoxycarbonyl (—COOR) or acyl substituents. For example, the addition of ethyldiazopropionate (EtO ₂ CC (N ₂ ) -Me) to an olefin 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 formula 1b , where Z represents C (R ²³ ) ₂ , in which one R ²³ represents Me and the other R ²³ represents COOEt.

By careful selection of the cyclopropane agent, positional selectivity can be achieved by cyclopropanation of steroid alkaloids with more than one olefin. For example, if diiodomethane and Et ₂ Zn are used for cyclopropanation of yervin under certain conditions, only an electron-rich olefin will interact.

The diastereoselectivity of cyclopropanation can be controlled in many ways. For example, lowering the temperature of the cyclopropanation reaction can lead to higher diastereoselectivity. Alternatively, a chiral cyclopropan means can be used that can distinguish between each diastereol side of the steroid alkaloid. Enantioselectivity during cyclopropanation can also be achieved using substrate-directed reactions (i.e., cyclopropanation of allyl alcohols using Et ₂ Zn / CH ₂ I ₂ reagents).

Cyclopropanation reactions can be carried out in an aprotic solvent, preferably one in which the reaction ingredients are substantially 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, dimethyl sulfoxide, dimethylformamide and the like; or a combination of two or more solvents. In a preferred embodiment, dichloromethane is the solvent used for cyclopropanation when dialkylzinc and diiodomethane are used.

Following the synthesis of the cyclopropane steroid alkaloid central portion, the compound can be derivatized using a variety of functionalization reactions known in the art. Representative examples include coupling reactions with palladium to alkene halides or aryl halides, oxidation, reduction, reaction with nucleophiles, reactions with electrophiles, pericyclization reactions, introduction of protecting groups, deprotection, and the like.

In the presence of Lewis or Bronsted acids, the cyclopropylcyclopamine analogues of the present invention undergo previously unknown rearrangement and extension of the cycle, which allows to obtain new cyclopamine analogues in which the D ring increases by one carbon atom. The cyclopropyl ring may be substituted or unsubstituted. In those cases where the cyclopropyl ring is substituted, groups attached to cyclopropane methylene will be inserted into ring D after rearrangement and expansion of the ring. Suitable acids include, but are not limited to, ZnI ₂ , BF ₃ , methanesulfonic acid, diaryloxyphosphoric acids and HCl. In a preferred embodiment, the Lewis acid used is BF ₃ . These homogenized analogues can be further functionalized using a variety of functionalization reactions known in the art. Representative examples include coupling reactions with palladium to alkene halides or aryl halides, oxidation, reduction, reaction with nucleophiles, reactions with electrophiles, pericyclization reactions, introduction of protecting groups, deprotection, and the like.

Methods of the invention

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

Currently, Hedgehog pathway antagonists are being studied in a large number of clinical settings where a therapeutic effect can be obtained for a condition or disorder, slowing down one or more aspects of the Hedgehog pathway activity. Although the focus of the studies was on cancer, the researchers found that inhibition of the Hedgehog pathway by low molecular weight compounds improves the symptoms of psoriasis ( Tas, et al., 2004 Dermatology 209 : 126-131, published as US 20040072913 (incorporated herein by reference). Psoriasis is a very common, chronic skin disorder, usually characterized by skin lesions, usually including erythematous papules and spots with silver scales, although there are changes on the skin and other parts of the body. Psoriasis is currently believed to be an autoimmune disease, but its etiology is still poorly understood. In the first stage, local application of cyclopamine to psoriatic lesions led to complete or partial regression of the lesion with a decrease in inflammatory cells (Tas et al., Supra).

The Hedgehog antagonists of the present invention can be used to treat or prevent psoriasis when they are used as a single agent or when used in combination with one or more other psoriasis agents, including, but not limited to, corticosteroids, tar, calcipotriene, tazarotene, calcineurin inhibitors, irradiation with ultraviolet rays, methotrexate, retinoids, cyclosporin, immunomodulatory drugs, etanercept, alefasept, efalizumab and infliximab.

Many tumors and proliferative conditions have been shown to depend on the Hedgehog pathway. The growth and survival of such cells can be affected by treatment with the compounds of the present invention. For example, inhibition of the Hedgehog pathway by low molecular weight compounds has been shown to slow the growth of basal cell carcinoma (Williams et al., 2003 PNAS 100: 4616-21), medulloblastoma ( Berman et al., 2002 Science 297 : 1559-61), pancreatic cancer (Berman et al, 2003 Nature 425:. 846-51), gastric cancer (Be rm an et al, 2003 Nature 425:. 846-51, published application PCT WO 05/013800), esophageal cancer (Berman et al,. 2003 Nature 425 : 846-51), lung cancer (Watkins et al., 2003. Nature 422: 313-7) and prostate cancer (Karhadkar et al., 2004. Nature 431: 707-12).

In addition, it has been shown that many types of cancer have uncontrolled activation of the Hedgehog pathway, such as breast cancer (Kubo et al., 2004. Cancer Research 64: 6071-4), hepatocellular cancer (Patil et al., 2005. 96 ^th Annual AACR conference, abstract # 2942 Sicklick et al., 2005. ASCO annual meeting, abstract # 9610), hematologic malignancies (Watkins and Matsui, unpublished results), basal cell carcinoma (Bale & Yu, 2001. Human Molec. Genet. 10 : 757-762 Xie et al., 1998 Nature 391: 90-92), medulloblastoma (Pietsch et al., 1997. Cancer Res. 57 : 2085-88) and gastric cancer (Ma et al., 2005 Carcinogenesis May 19 2005 (Epub)).

Cancer or neoplastic diseases and related disorders that can be treated by administering the compounds and compositions of the present invention include, but are not limited to, adrenal cortex cancer, anal canal cancer, aplastic anemia, bile duct cancer, bladder cancer, bone cancer, tumors brain / central nervous system, breast cancer, cervical cancer, non-Hodgkin’s lymphoma, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, Ewing tumors, eye cancer, gall bladder cancer, gastrointestinal carcinoids e tumors, gastrointestinal stromal tumors, gestational trophoblastic disease, Hodgkin's disease, Kaposi’s sarcoma, kidney cancer, laryngeal cancer and submaryngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, childhood leukemia, chronic lymphocytic leukemia, chronic myeloid liver leukemia lung cancer, carcinoid tumor of the lung, non-Hodgkin lymphoma, male breast cancer, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, cancer of the nasal cavity and paranasal region and, nasopharyngeal cancer, neuroblastoma, oral cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumor, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, melanoma, non-skin cancer melanomas, stomach cancer, testicular cancer, thymus cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulva cancer, Waldenstrom macroglobulinemia and Wilms tumor.

The methods and compositions of the present invention can be used in the treatment of human cancer, for example, 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 the treatment of basal cell carcinoma, pancreatic cancer, prostate cancer, sarcoma, lymphoma, leukemia, gastric cancer, gastrointestinal cancer, multiple myeloma, small cell lung cancer, glioma, breast cancer, hepatocellular cancer or medulloblastoma by using a therapeutically effective amount of at least one of the compounds of the present invention as a single agent or in combination with another otivorakovym means.

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

In one embodiment, the existing method can be used as part of a treatment regimen for malignant medulloblastoma and other primary malignant neuroectodermal tumors of the central nervous system.

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

In some embodiments, the present invention relates to a method for treating cancer, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one or more of the aforementioned compounds, in cases where the cancer is localized in the patient’s head, neck, nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx, larynx, pharyngeal salivary glands, paraganglionic, in the pancreas, stomach, skin, esophagus, liver and gall tree, bone, small intestine, thorax of the colon, rectum, ovary, prostate, lung, breast, lymphatic system, blood, central nervous system, bone marrow, or the brain.

In some embodiments, the present invention relates to a method of treating cancer, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one or more of the aforementioned compounds, in cases where the cancer is basal cell cancer, pancreatic cancer, prostate cancer , sarcoma, lymphoma, leukemia, stomach cancer, esophageal cancer, bile duct cancer, colon cancer, multiple myeloma, small cell lung cancer, glioma, breast cancer, hepatocell lar cancer or medulloblastoma.

In some embodiments, the present invention relates to the aforementioned method, wherein the compound is used in combination with radiation therapy or another anti-cancer chemotherapeutic agent.

In some embodiments, the present invention relates to any of the aforementioned methods, wherein the compound is applied locally to a tumor or systemically to a patient.

In some embodiments, the present invention relates to any of the aforementioned methods, wherein the method of administering said compound is inhalation, oral, intravenous, sublingual, ophthalmic, transdermal, rectal, vaginal, local, intramuscular, intra-arterial, intrathecal, subcutaneous, buccal or intranasal.

In some embodiments, the present invention relates to any of the aforementioned methods, wherein the route of administration is oral, intravenous or topical.

In some embodiments, the present invention provides a method for antagonizing a Hedgehog pathway in a cell, comprising contacting a cell expressing Smoothened with an effective amount of any one or more of the aforementioned compounds.

In some embodiments, the present invention provides a method for antagonizing a Hedgehog pathway in a cell, comprising contacting a Smoothened expressing cell with an effective amount of any one or more of the aforementioned compounds, wherein said Smoothened expressing cell is contacted with said in vitro compound .

In some embodiments, the present invention relates to a method for antagonizing a Hedgehog pathway in a cell, comprising contacting a cell expressing Smoothened with an effective amount of any one or more of the above compounds, wherein said Smoothened expressing cell is contacted with said compound in vivo.

In some embodiments, the present invention provides a method for antagonizing a Hedgehog pathway in a cell, comprising contacting a cell expressing Smoothened with an effective amount of any one or more of the aforementioned compounds, wherein said Smo expressing cell is contacted with said compound in within the patient’s body.

In some embodiments, the present invention relates to a method for treating or preventing psoriasis in a patient, including administering to a patient in need thereof a therapeutically effective amount of any one or more of the above compounds.

In some embodiments, the present invention relates to the aforementioned method of treating or preventing psoriasis, wherein the method of administering said composition is local.

In some embodiments, one or more of the compounds of the present invention are used for the treatment and prevention of psoriasis in combination with one or more antipsoriatics, including, but not limited to, corticosteroids, tar, calcipotriene, tazarotene, calcineurin inhibitors, ultraviolet radiation, methotrexate, retinoids , cyclosporine, immunomodulatory drugs, etanercept, alefasept, efalizumab and infliximab.

Cancer treatment combined with chemotherapy or radiation therapy

In some embodiments, one or more compounds of the present invention are used to treat or prevent cancer or a neoplastic disease in combination with one or more anti-cancer chemotherapeutic agents, including, but not limited to, gemcitabine, methotrexate, taxol, mercaptopurine, thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosourea, cisplatin, carboplatin, mitomycin, dacarbazine, procarbizin, etoposide, prednisolone, dexamethasone, cytarbin, campatekin, bleoma icin, doxorubicin, idarubicin, daunorubicin, dactinomycin, plikamycin, mitoxantrone, asparaginase, vinblastine, vincristine, vinorelbine, paclitaxel and docetaxel. In a preferred embodiment, one or more compounds of the present invention are used for the treatment and prevention of cancer or a neoplastic disease in combination with one or more chemotherapeutic or other anti-cancer agents, including but not limited to radiation (e.g., gamma radiation), nitrogen mustard (e.g. cyclophosphamide, ifosfamide, trophosphamide, chlorambucil, estramustine and melphalan), nitrosoureas (e.g. carmustine (BCNU) and lomustine (CCNU)), alkyl sulfonates (e.g. busulfan and threosulfan), riazenes (e.g., dacarbazine and temozolomide), platinum compounds (e.g., cisplatin, carboplatin and oxaliplatin), periwinkle alkaloids (e.g., vincristine, vinblastine, vindesine and vinorelbine), taxoids (e.g., paclitaxel and docetaxol), epinodophyllines (e.g. , teniposide, topotecan 9-aminocamptothecin, camptirothinocane, crisnatol, mitomycin C and mitomycin C), antimetabolites, DHFR inhibitors (e.g. methotrexate and trimethrexate), IMP dehydrogenase inhibitors (e.g. mycophenolic, thiaziribonitrin and riazofirinurin, ri riboferin, E, Ductases (e.g., hydroxyurea and deferoxamine), Uracil analogues (e.g., fluorouracil, phloxuridine, doxyfluridine, ratitrexed and capecitabine), cytosine analogs (e.g. cytarabine (Ara C), cytosine arabinoside and fludarabine), and tyropinopurin, e.g. ), antiestrogens (e.g. tamoxifen, raloxifene and megestrol), LHRH agonists (e.g. goserelin and leuprolide acetate), antiandrogens (e.g. flutamide and bicalutamide), vitamin D3 analogues (e.g. EB 1089, CB 1093 and KH 1060), photodynamic therapy (e.g., vertoporphyrin (BP D-MA), phthalocyanine, Pc4 photosensitizer and demethoxyhypocrellin A (2BA-2-DMHA)), cytokines (e.g. α-interferon, γ-interferon and tumor necrosis factor), isoprenylation inhibitors (e.g. lovastatin), dopaminergic neurotoxins (e.g. , 1-methyl-4-phenylpyridine ion), cell cycle inhibitors (e.g., staurosporin), actinomycin (e.g., actinomycin D and dactinomycin), bleomycin (e.g., bleomycin A2, bleomycin B2, and peplomycin), anthracyclines (e.g., daunorubicin, docorubicin, (adriamycin), idarubicin, epirubicin, pyrarubicin, zora itsin and mitoxantrone), inhibitors of MDR (e.g., verapamil), inhibitors of Ca ²⁺ -ATPase (e.g., thapsigargin), antibodies (e.g., Avastin, Erbitux, Rituxan and bekssar), corticosteroids (e.g., prednilon, predizon and t. e.), imatinib, thalidomide, lenalidomide, bortezomib, gemcitabine, erlotinib, gefitinib, sorafenib and intranib.

A chemotherapeutic agent and / or radiation therapy may be administered according to therapeutic protocols known in the art. It will be apparent to those skilled in the art that administration of a chemotherapeutic agent and / or radiation therapy may vary differently depending on the disease against which the treatment is being applied and the known effects of the chemotherapeutic agent and / or radiation therapy on the disease. In addition, according to the knowledge of a qualified clinician, therapeutic protocols (e.g., dosages and time of administration) may be different due to the effects observed by the patient on the therapeutic agents used (i.e. antineoplastic or radiation) and due to the observed disease response on the administered therapeutic agents and the observed adverse effects.

In addition, in general, the compounds of the present invention and the chemotherapeutic agent should not be administered in the same pharmaceutical composition, and it is possible, since the physical and chemical characteristics are different, they must be administered via different routes. For example, the compounds of the present invention can be administered intravenously to create and maintain good blood levels, while a chemotherapeutic agent can be administered orally. The determination of the route of administration and the feasibility of administration, if possible in the same pharmaceutical composition, is within the knowledge of a qualified clinician. Initial administration can be done according to established protocols known in the art, and then a qualified clinician can change the dosage, route of administration, and time of administration based on the observed effects.

The preferred choice of chemotherapeutic agent or type of radiation will depend on the diagnosis of the doctors and their judgment on the patient's condition and the appropriate protocol for administration.

The compound of the present invention and a chemotherapeutic agent and / or radiation can be administered simultaneously (for example, simultaneously, essentially simultaneously or within the same treatment protocol) or sequentially, depending on the nature of the proliferative disease; the patient’s condition and the actual selection of a chemotherapeutic agent and / or radiation that are administered together (i.e., within a single administration protocol) with the compound of the present invention.

If the compound of the present invention and the chemotherapeutic agent and / or radiation are not administered simultaneously or essentially simultaneously, then the optimal administration order of the compound of the present invention and the chemotherapeutic agent and / or radiation may be different for different tumors. Thus, in certain situations, a compound of the present invention may be administered initially, followed by the administration of a chemotherapeutic agent and / or radiation; and in other situations, a chemotherapeutic agent and / or radiation may be administered initially, followed by the administration of a compound of the present invention. This additional appointment can be repeated during one treatment protocol. The determination of the order of administration and the number of repetitions of the administration of each therapeutic agent during the treatment protocol is completely within the knowledge of a qualified physician after evaluating the disease being treated and the condition of the patient. For example, a chemotherapeutic agent and / or radiation may be administered first, especially if it is a cytotoxic agent and then the treatment is continued with the administration of a compound of the present invention, accompanied, if approved, by administration of a chemotherapeutic agent and / or radiation, and so on, until the treatment protocol is completed .

Thus, in accordance with experience and knowledge, the practitioner may, during treatment, change each protocol for the administration of the component (therapeutic agent, for example, a compound of the present invention, chemotherapeutic agent or radiation) for treatment according to the needs of an individual patient.

Pharmaceutical Compositions

In yet another embodiment, the present invention provides pharmaceutically acceptable compositions that comprise a therapeutically effective amount of one or more of the compounds described above (Formulas 1 and 2 ), formed together with one or more pharmaceutically acceptable carriers and / or solvents. The pharmaceutical compositions of the present invention can be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, medicines (aqueous or non-aqueous solutions or suspensions), tablets, for example, intended for buccal, sublingual and systemic use, pills, powders, granules, pastes for gluing to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection, such as, for example, a sterile solution or suspension, or a composition for continuous administration; (3) topical application, for example, as a cream, ointment or controlled-release patch or spray solution applied to the skin; (4) intravaginal or rectal, for example, as a uterine ring, cream or foam; (5) sublingual; (6) through the eyes; (7) transdermally; (8) through the lungs; or (9) through the nose. As described above, certain embodiments of existing compounds may contain a basic functional group, such as amino or alkylamino, and are thus capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable acids. The term "pharmaceutically acceptable salts" in this regard refers to the relatively non-toxic salts of the compounds of the present invention with inorganic and organic acids. These salts can be prepared in situ in a binder for administration, either as dosage forms in a manufacturing process, or by separately reacting the purified compound of the invention in its free base form with a suitable organic or inorganic acid, and isolating the salt that forms during subsequent purification. Typical salts include bromohydrate, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, mesylate , lactobionate and lauryl sulfonate, and the like (see, for example, Berge et al. (1977) Pharmaceutical Salts, J. Pharm. Sci. 66: 1-19).

Pharmaceutically acceptable salts of the compounds of the present invention include conventional non-toxic salts or quaternary ammonium salts of 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, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isethionic, etc.

In other cases, the compounds of the present invention may contain one or more acidic functional groups and, thus, have the ability to form pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term “pharmaceutically acceptable salts” in these cases refers to the relatively non-toxic, inorganic and organic base addition salts of the compounds of the present invention. These salts can probably be prepared in situ in an administration medium or method of manufacturing a dosage form or by separately reacting the purified compound in its free acid form with a suitable base such as a pharmaceutically acceptable metal cation hydroxide, carbonate or bicarbonate, with ammonia or with a pharmaceutically acceptable oranic primary, secondary or tertiary amine. Representative alkaline or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium and aluminum salts and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. (see, for example, Berge et al., supra ).

Moisturizing agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, perfumes and fragrances, preservatives and antioxidants may also be present in the compositions.

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, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and the like; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Methods for preparing these formulations or compositions include the step of associating the compound of the present invention with a carrier and, optionally, one or more accessory ingredients. In general, formulations are obtained by uniformly and directly associating the compounds of the present invention with liquid carriers or finely divided solid carriers, or both, and then, if necessary, molding the product.

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 non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be incorporated into sterile injectable solutions or dispersions immediately before use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostats, soluble substances Twa that give the finished shape isotonic with the blood of the intended recipient or suspending or thickening agents.

Examples of suitable aqueous and non-aqueous vehicles that can be used in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerin, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. The desired fluidity can be established, for example, by the use of coating materials, such as lecithin, by maintaining the required particle size in the case of dispersions and by using surfactant particles.

These compositions may also contain adjuvants, such as preservatives, humectants, emulsifying and dispersing agents. 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, phenolsorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride and the like in the compositions. Additionally, prolonged absorption of the injectable pharmaceutical form can be given by the inclusion of agents that delay absorption, such as aluminum monostearate and gelatin.

In some cases, to prolong the effect of the drug, it is desirable to slow the absorption of the drug as a result of subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous substance having poor solubility in water. The degree of absorption of the drug then depends on its degree of dissolution, which in turn may depend on the size of the crystals and crystalline form. Alternatively, delayed absorption of the parenteral dosage form is achieved by dissolving or suspending the drug in an oil medium.

Injectable depot forms are prepared by forming microencapsulated matrices of the compounds of the present invention in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular polymer used, the release rate of the drug can be controlled. Examples of other biodegradable polymers include poly (ortho-esters) and poly (anhydrides). Depotable injectable formulations are also prepared by incorporating the drug into liposomes or microemulsions that are compatible with body tissues.

Finished compositions of the invention suitable for oral administration may be in the form of capsules, porridge, pills, tablets, troches (using a flavored base, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or in the form of pastilles (using an inert base such as gelatin and glycerin or sucrose and acacia), and / or mouthwash, etc., each of which contains a predetermined amount of a compound of the present invention as an active ingredient. The compound of the present invention may also be administered as a bolus, electuary or paste.

When the compounds of the present invention are administered as pharmaceuticals to humans and animals, they can be administered as such or as a pharmaceutical composition containing, for example, from 0.1 to 99% (more preferably from 10 to 30%) of the active ingredient in combination with a 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 by, for example, spray, rectally, intravaginally, parenterally, intracisternally or topically, or by powders, ointments or drops, including buccal and sublingual the way.

Regardless of the chosen route of administration, the compounds of the present invention, which can be used in a suitable hydrated form, and / or the pharmaceutical compositions of the present invention are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those skilled in the art.

Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic effect for a particular patient, composition and route of administration, without toxicity to the patient.

The dosage level chosen will depend on many factors, including the activity of the particular compound of the present invention or its ester, salt or amide, route of administration, time of administration, rate of excretion or metabolism of the particular compound used, rate and extent of absorption, duration of treatment, other drugs, compounds and / or substances used in combination with the particular compound used, age, gender, weight, condition, general health and previous history and illness of the patient to be treated, and similar factors well known in the medical field.

A general practitioner or veterinarian in the art can easily determine and prescribe the required effective amount of a pharmaceutical composition. For example, a therapist or veterinarian can start with 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 the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will be such an amount of a compound that is the smallest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above. In general, 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 of about 0.0001 to 100 mg per kilogram of body weight per day.

Optionally, an effective daily dose of the active compound may be administered in the form of two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally in unit dosage forms. The preferred dosage is one administration per day.

While it is possible to administer only one compound of the present invention, it is preferable to administer the compounds in the form of a pharmaceutical formulation (composition).

A patient receiving this treatment is any animal in need of this treatment, including primates, especially humans, and other mammals such as horses, cattle, pigs, and sheep; in general, poultry and pets.

The compounds of the invention may be administered as such or in mixtures with pharmaceutically acceptable carriers and may also be administered in conjunction with antimicrobial agents such as penicillins, cephalosporins, aminoglycosides and glycopeptides. Combination therapy, therefore, includes sequential, simultaneous and separate administration of the active compound so that the therapeutic effects of the first compound administered do not completely disappear when the next one is administered.

Illustrative examples

Now that the invention has been generally described, it will be more readily understood with reference to the following examples, which are included merely to illustrate certain aspects of the present invention and are intended to not limit the invention.

Example 1

Obtaining a cyclopamine derivative

Stage A

To a solution of cyclopamine 2 (250 mg, 0.6 mmol, 1 equiv.) In DCM at room temperature, Fmoc-OSu (205 mg, 0.6 mmol, 1 equiv.) Was added and the resulting mixture was stirred at room temperature for nights. The resulting crude Fmoc-cyclopamine solution was then cooled to 0 ° C and treated with 15% diethylzinc in toluene (0.5 ml, 0.6 mmol, 1 equiv.) And stirred for 30 minutes (flask A).

Diiodomethane (0.4 ml, 6 mmol, 10 eq.) In DCM (20 ml) at 0 ° C is treated with 15% diethylzinc in toluene (3 ml, 3 mmol, 5 eq.) And the resulting solution is stirred for 5 min (flask B).

The contents of flask B are transferred to flask A through a cannula and the resulting suspension is stirred for 5 hours at room temperature. The reaction was quenched with HCl (1 M), stirred for 10 minutes (until all white matter had dissolved) and extracted with DCM (5 ×). The organic extracts were dried (MgSO ₄ ), filtered through celite and concentrated in vacuo. The residue was purified by flash chromatography (1: 1 hexane / AcOEt). The desired 11,12-monocyclopropane is obtained as a 9: 1 mixture of diastereoisomers, along with 20% diastereomeric bis-cyclopropane products (80% of the total recovery). This mixture was separated using preparative SFC chromatography.

Stage B

Fmoc-monocyclopropylcyclopamine 3 (100 mg, 0.15 mmol, 1 equiv.) In DCM (2 ml) is treated with diethylamine (0.5 ml, 4.8 mmol, 32 equiv.) At room temperature overnight, resulting in the solution was concentrated in vacuo and the residue was absorbed on silica gel and purified by flash chromatography (2: 1 → 1: 1 hexane / AcOEt and then 95: 5 → 90: 10 → 20: 80 DCM / MeOH). The desired product is obtained as a white solid (95% yield). MS (ESI (+)) m / e 426.31 (M + H) ⁺ .

Example 2

Obtaining a cyclopamine derivative

Stage A

To a solution of hydrocinnamic acid 5 (3.01 g, 20 mmol, 1 equiv.) In anhydrous chloroform (30 ml) at 75 ° C was added thionyl chloride (1.75 ml, 24.1 mmol, 1.2 equiv.) over a period of 3 minutes The mixture was refluxed for 3.5 hours. The solvent was distilled off to obtain the crude acid chloride as a pale yellow viscous liquid. The crude material is used without further purification.

Stage B

To a biphasic mixture of 7 (3.16 g, 24.1 mmol, 1.2 equiv.) In DCM (30 ml) and an aqueous solution of NaOH (2.0 M, 30 ml, 3 equiv.) At 25 ° C add a solution acid chloride 8 (3.38 g, 20 mmol, 1 equiv.) in DCM (10 ml) and the resulting mixture was stirred at 25 ° C for 3 hours. The mixture was further neutralized with an aqueous solution of HCl (2 M, 30 ml) . The organic layer was further separated and the aqueous layer was extracted with DCM (3 × 50 ml). The combined organic layers were washed with HCl (2.0 M, 25 ml), water (3 × 50 ml), brine (50 ml), dried over magnesium sulfate and the solution was evaporated under reduced pressure. The crude material is chromatographed on silica gel using 5% MeOH: DCM as an eluent and the column is further eluted with 10% MeOH: DCM to give 1.141 g of compound 8 .

Stage C

To a mixture of acid 8 (264 mg, 1 mmol, 1 equiv.), EDCI (231 mg, 1.2 mmol, 1.2 equiv.) And triethylamine (168 μl, 1.2 mmol, 1.2 equiv.) DCM (2 ml) at 0 ° C add allylamine (90.3 μl, 1.2 mmol, 1.2 eq.), The resulting mixture is stirred at 0 ° C and allowed to warm to 25 ° C over a period of 2 hours. The reaction mixture was added to water (50 ml), extracted with DCM (4 × 25 ml), the combined organic layers were washed with 1M HCl (2 × 25 ml), water (3 × 25 ml), saturated brine (25 ml) ), dried over magnesium sulfate and the solvent is evaporated off under reduced pressure to obtain 287.5 mg as required th product. This substance is used without further purification.

Stage D

To a solution of allylamide 9 (70 mg, 0.23 mmol, 1 equiv.) In acetone (1 ml) and water (0.3 ml) is added a solution of osmium tetroxide (0.35 ml, 0.035 mmol, 0.15 equiv., 2.5 w / w in tert-butanol). The reaction mixture was immediately cooled on ice shortly after the addition of OsO ₄ solution, and the resulting dark brown solution was stirred at 0 ° C for 15 minutes. Sodium periodate (110 mg, 0.51 mmol, 2.2 eq.) Was added 5 portions to the above mixture and stirring was continued for 1 h at 0 ° C, and the mixture was allowed to warm to 25 ° C for a period of 2 hours The reaction mixture was diluted with DCM (3 ml), filtered through a small pad of magnesium sulfate and the filter cake washed with DCM (4 × 3 ml). The filtrate was concentrated and the residue (67.9 mg) was filtered through a small pad of RP silica gel using 5% MeOH: DCM to give 38.9 mg of the desired product.

Stage E

To a suspension of 1 (10 mg, 0.023 mmol, 1 equiv.) In acetonitrile (2 ml) was added a solution of aldehyde 10 (17 mg, 0.056 mmol, 2.4 equiv.) In acetonitrile (0.3 ml) followed by sodium triacetoxyborohydride (6.5 mg, 0.031 mmol, 1.3 eq.) And the reaction mixture was stirred at 25 ° C. for 16 hours. The solvent was then evaporated under reduced pressure and the residue was chromatographed on silica gel (7 cm × 10 mm) using 3% methanol: DCM to give 24.6 mg of a crude material. This material was re-subjected to silica gel column chromatography using 2% methanol: DCM and 18.2 mg of crude product was recovered, which was further purified by preparative TLC using 3% methanol: DCM as a developing solvent (2 passes) to give 6.3 mg product required. MS (ESI (+)) m / e 714.4 (M + H) ⁺ .

Example 3

Obtaining a cyclopamine derivative

Stage A

Cyclopamine 2 (20 mg, 0.049 mmol, 1 eq.) Is suspended in dry toluene (0.6 ml) and cyclohexanone (150 μl, 1.47 mmol, 30 eq.) Is added followed by aluminum isopropoxide (79 mg, 0.392 mmol) , 8 equiv.). The resulting mixture was heated at reflux for 2 hours, cooled to room temperature, diluted with AcOEt, and quenched with Rochelle brine. The biphasic mixture was stirred overnight, the layers were separated, the aqueous layer was extracted with AcOEt and the combined organic extracts were dried (MgSO ₄ ), filtered and concentrated in vacuo. The residue was purified by flash chromatography (DCM, DCM / MeOH 98: 2 and 95: 5). The target substance is obtained as a white crystalline solid (70% yield).

Stage B

Diiodomethane (40 μl, 0.5 mmol, 25 equiv.) In DCM (0.52 ml) at 0 ° C is treated with 15% diethylzinc in toluene (0.2 ml, 0.2 mmol, 10 equiv.) And obtained in as a result, the solution is stirred for 5 minutes. Compound 12 (10 mg, 0.02 mmol, 1 equiv.) In DCM (0.35 ml) was added and the resulting mixture was stirred at room temperature (while removing the ice bath) for 3 hours, quenched with 2 N NaOH and stirred for 10 min, the layers were separated and the aqueous layer was extracted with DCM (2 ×). The organic extracts were dried (MgSO ₄ ), filtered and concentrated in vacuo. The residue was purified by flash chromatography (DCM / MeOH 92: 8). The cyclopropane is obtained as a white solid. MS (ESI (+)) m / e 424.5 (M + H) ⁺ .

Example 4

Obtaining a cyclopamine derivative

Stage A

To a solution of cyclopamine 2 (250 mg, 0.6 mmol, 1 equiv.) In DCM (10 ml) at room temperature Fmoc-OSu (205 mg, 0.6 mmol, 1 equiv.) Was added, the resulting mixture was stirred at room temperature overnight and concentrated in vacuo. Crude Fmoc-cyclopamine is obtained as an off-white foam.

Stage B

A solution of the crude Fmoc-cyclopamine 3 (15 mg, 0.024 mmol, 1 equiv.) In DCM (0.5 ml) was cooled to -78 ° C and treated with sodium bicarbonate (4 mg, 0.047 mmol, 1.96 equiv.) Followed by the addition of mCPBA (4 mg, 0.024 mmol, 1 equiv.). The reaction mixture was stirred at -78 ° C for 1 h, diluted with H ₂ O and extracted with DCM (3 ×). The organic extracts were washed with 10% NaHCO ₃ and brine, dried (MgSO ₄ ), filtered and concentrated in vacuo. The crude material is purified by preparative TLC (hexane / AcOEt 1: 2) to give the epoxide as a white foam (70% yield).

Stage C

A solution of compound 14 (11 mg, 0.017 mmol, 1 equiv.) In DCM (0.5 ml) was treated at room temperature with Et ₂ NH (0.5 ml, 4.8 mmol, 282 equiv.), The resulting solution was stirred at room temperature overnight and concentrated in vacuo. The residue was purified by preparative TLC (DCM / MeOH 9: 1). Compound 13 was obtained as a white solid (90% yield). MS (ESI (+)) m / e 428.5 (M + H) ⁺ .

Example 5

Obtaining a cyclopamine derivative

Stage A

Compound 2 (1.30 g, 3.2 mmol, 1 equiv.) Was weighed and loaded into a reaction vessel. Potassium carbonate (0.91 g, 6.6 mmol, 2.1 eq.) Is weighed and loaded into a reaction vessel, followed by dichloroethane (6.0 ml, 76 mmol, 23.8 eq.) And anhydrous DMSO (5 ml ) The reaction mixture is heated to 70 ° C. for 36 hours under nitrogen. The reaction mixture was cooled to room temperature, diluted with DCM (15 ml) and washed twice with water (2 × 15 ml). The organic layer was dried over sodium sulfate, filtered (washing with DCM, as needed), and concentrated to dryness to give a pale yellow solid. Flash chromatography (DCM / EtOAc) afforded the target material as a white crystalline solid.

Stage B

Compound 16 (0.111 g, 0.233 mmol, 1 eq.) Was transferred to a reaction flask, placed under a nitrogen atmosphere, and dissolved in anhydrous DCM (2 ml). Chloroiodomethane (0.238 ml, 3.27 mmol, 14 equiv.) Was added. The solution is cooled to -15 ° C. Diethyl zinc (1M in heptane, 1.63 ml, 1.63 mmol, 6.5 eq.) Was added dropwise over 30 minutes, carefully controlling the exothermic effect. The reaction mixture was kept between -10 ° C and -14 ° C for several hours until TLC indicated the consumption of the starting material. The reaction is then quenched by careful addition of THF (6 ml) and then aqueous citrate buffer (pH 4.5, 10 ml). The layers are allowed to warm to room temperature. Saturated sodium sulfate solution was added. The layers are well mixed, transferred to a connecting funnel with excess DCM, and the organic layer is collected. The organic layer was washed with aqueous sodium hydroxide (1N, 10 ml) and saturated sodium sulfate (10 ml), dried over sodium sulfate before concentration to dryness. The crude material is purified by flash chromatography to give the desired product in 55% yield.

Example 6

Obtaining a cyclopamine derivative

To a solution of compound 11 (5 mg, 0.01 mmol, 1 equiv.) And compound 10 (10 mg, 0.04 mmol, 3 equiv.) In anhydrous DCM (5 ml), solid triacetoxyborohydride (8 mg, 0.004 mmol, 3 equiv.) And the resulting suspension was stirred at 25 ° C for 2 hours. The reaction mixture was quenched with sodium bicarbonate, extracted with DCM (4 × 10 ml), the organic layer was collected and washed with brine (1 × 20 ml), dried over magnesium sulfate and concentrated under reduced pressure. The crude material is purified by PTLC (DCM / MeOH 95: 5) to give 8 mg of the desired product.

Example 7

Obtaining a cyclopamine derivative

Stage A

Cyclopamine 2 (20 mg, 0.049 mmol) is suspended in dry toluene (0.6 ml) and cyclohexanone (150 μl, 1.47 mmol, 30 eq.) Is added thereto, followed by the addition of aluminum isopropoxide (79 mg, 0.392 mmol, 8 equiv.). The resulting mixture was heated under reflux for 2 hours, cooled to room temperature, diluted with ethyl acetate, and quenched with Rochelle brine. The biphasic mixture was stirred overnight, the layers were combined, the aqueous layer was extracted with ethyl acetate, and the combined organic extracts were dried (MgSO ₄ ), filtered and concentrated in vacuo. The residue was purified by flash chromatography (DCM, DCM / methanol 98: 2 and 95: 5). Target material 12 was obtained as a white crystalline solid (70% yield).

Stage B

Diiodomethane (40 μl, 0.5 mmol, 2.5 equiv.) In DCM (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 was stirred for 5 minutes (where a white precipitate was observed). Enon 12 (10 mg, 0.02 mmol, 1 equiv.) In DCM (0.35 ml) was added and the resulting mixture was stirred at room temperature (while removing the ice bath) for 3 hours, quenched with NaOH (2N) and stirred for 10 min, the layers were separated and the aqueous layer was extracted with DCM (2 ×). The organic extracts were dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by flash chromatography (DCM / MeOH 92: 8). Cyclopropane 11 is obtained as a white solid.

Stage C

To a solution of cyclopropyleneone 11 (10 mg, 24 μmol, 1 equiv.) In DCM (0.5 ml) at 0 ° C in argon atmosphere add BF ₃ · Et ₂ O (30 μl, 0.24 mmol, 10 equiv.) and the resulting solution was stirred at 0 ° C. for 1.5 hours, diluted with DCM and quenched with saturated sodium bicarbonate. The organic phase was washed with saturated sodium bicarbonate and the combined aqueous layers were extracted with DCM. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by preparative TLC (DCM / MeOH 9: 1). Target compound 18 was obtained as a white solid (90% yield). MS (ESI (+)) m / e 424.62 (M + H) ⁺ .

Example 8

Obtaining a cyclopamine derivative

Stage A

To a solution of cyclopamine 2 (250 mg, 0.6 mmol, 1 equiv.) In DCM (10 ml) at room temperature, Fmoc-OSu (205 mg, 0.6 mmol, 1 equiv.) Was added and the resulting mixture was stirred at room temperature overnight. The resulting crude Fmoc-cyclopamine solution was further 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, yellowish solution).

Diiodomethane (0.4 ml, 6 mmol, 10 eq.) In DCM (20 ml) at 0 ° C is treated with 15% diethylzinc in toluene (3 ml, 3 mmol, 5 eq.) And the resulting solution is stirred for 5 min (flask B, white precipitate).

The contents of flask B are transferred to flask A through a cannula and the resulting suspension is stirred for 5 hours at room temperature. The reaction is quenched with 1 N. hydrochloric acid, stirred for 10 min (until all white matter has dissolved) and extracted with DCM (5 ×). The organic extracts were dried over MgSO ₄ , filtered through celite and concentrated in vacuo. The residue was purified by flash chromatography (hexane / ethyl acetate 1: 1). Target 11,12-monocyclopropane 5 was obtained as a 9: 1 mixture of diastereoisomers, along with 20% diastereomeric bis-cyclopropane products (80% of the total recovery). This mixture was separated using preparative SFC chromatography.

Stage B

Fmoc-cyclopropylcyclopamine 3 (14 mg, 22 μmol, 1 eq.) Was dissolved in DCM (0.5 ml), cooled to 0 ° C and treated with BF ₃ ^. Et ₂ O (27 μl, 0.22 mmol, 10 equiv.) For 1 h, quenched with saturated sodium bicarbonate solution, the layers were separated and the aqueous layer was extracted with DCM. The combined organic extracts were dried over MgSO ₄ , filtered and concentrated in vacuo. The residue was purified by preparative thin layer chromatography (hexane / ethyl acetate 2: 1). The target substance, Fmoc-protected cyclopamine, is obtained as a clear oil.

A solution of the crude Fmoc-protected cyclopamine (20 mg, 31 μmol, 1 equiv.) In DCM (0.5 ml) was treated with Et ₂ NH (0.5 ml, 4.8 mmol, 154 equiv.) Overnight, concentrated in vacuum and the residue was purified by flash chromatography (DCM, DCM / methanol 98: 2 and 95: 5). The desired compound is obtained in the form of an oil, which crystallizes upon standing. MS (ESI (+)) m / e 426.29 (M + H) ⁺ .

Example 9

Obtaining a cyclopamine derivative

Stage A

Compound 6 (23 mg, 54 μmol, 1 equiv.) Was dissolved in DCM (1 ml) and methyl iodide (0.17 ml, 0.54 mmol, 10 equiv.) Was added to it. The reaction mixture was allowed to stir at room temperature under nitrogen overnight. The next morning, TLC / LC-MS indicates that there is still some IV (starting material). A spatula of Na ₂ CO _{3 is} added and the mixture is stirred for one more hour. The crude material is loaded onto a Biotage 25 Si + M system and eluted with DCM / EtOAc / MeOH (82.5 / 10 / 7.5). An amorphous substance is obtained: 16 mg. MS (ESI (+)) m / e 440.32 (M + H) ⁺ .

Example 10

Obtaining a cyclopamine derivative

Stage A

Hydrocinnamoyl chloride 22 (1.13 g, 6.7 mmol, 1 equiv.) And allylamine (0.77 ml, 10 mmol, 1.5 equiv.) Are solubilized in THF (20 ml) and the reaction mixture is stirred at room temperature for 24 hours A white precipitate forms. The reaction mixture is filtered. The filtrate was dried over MgSO ₄ , filtered and concentrated in vacuo. A colorless oil is isolated, which turns into a wax-like solid (1.1 g).

To a solution of allylamide (0.81 mg, 0.27 mmol, 1 equiv.) In a mixture of acetone: water (9 ml; 3: 1) at 0 ° C, a solution of OsO ₄ (0.55 ml, 2.5 wt. (w / w in t-BuOH) and the resulting brownish mixture was stirred for 10 minutes. Solid sodium periodate (0.13 g, 0.59 mmol, 2.2 eq.) Was added in three portions and the mixture was stirred at 0 ° C and allowed to warm to 25 ° C for a period of 2 hours. Light off-white creamy the mixture was diluted with DCM (25 ml), dried over magnesium sulfate, the solids were filtered through a layer of celite, the filtrate was concentrated under reduced pressure. The crude material slowly turns yellow-black. The crude material is loaded into the Biotage 25 + S system and purified by eluting with hexane / EtOAc (1: 1 to 1: 2) to give a colorless oil that solidifies upon drying (250 mg).

Stage B

Sodium triacetoxyborohydride (100 mg, 0.47) was added to a solution of 18 (108 mg, 0.25 mmol, 1 equiv.) And aldehyde 21 (100 mg, 0.52 mmol, 2.1 equiv.) In DCM (5 ml). mmol, 1.9 equiv.) in one portion and the suspension was stirred at room temperature for 7 hours. The reaction was quenched by the addition of MeOH and filtered through celite. Evaporation to dryness gives 230 mg of oil. The material was purified by chromatography (SiO ₂ , column 3 cm × 4 cm), eluting with 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

Obtaining a cyclopamine derivative

Stage A

2-Phenylethanesulfonyl chloride 25 (1.13 g, 5.5 mmol, 1 equiv. And allylamine (0.56 ml, 7.3 mmol, 1.3 equiv.) Are solubilized in THF (20 ml) and allowed to interact at room temperature within 24 hours, a white precipitate formed, the reaction mixture was filtered, the filtrate was dried over MgSO ₄ , filtered and concentrated in vacuo. A slightly yellow oil (1.1 g) was isolated. The crude material was used directly in the next step.

To a solution of allylsulfonamide (0.15 g, 0.66 mmol, 1 equiv.) In a mixture of acetone: water (4 ml; 3: 1) at 0 ° C, a solution of OsO ₄ (0.13 ml, 2.5 wt. (w / w in t-BuOH) and the resulting brownish mixture was stirred for 10 minutes. Solid sodium periodate (0.31 g, 1.46 mmol, 2.2 eq.) Was added in three portions and the mixture was stirred at 0 ° C and allowed to warm to 25 ° C for a period of 2 hours. Light off-white the creamy mixture was diluted with DCM (25 ml), dried over magnesium sulfate, the solids were filtered through a pad of celite, and the filtrate was concentrated under reduced pressure. The crude material is purified on SiO ₂ (column 2 cm × 12 cm), eluting with hexane / EtOAc (7: 3) to give the desired substance (16 mg).

Stage B

To a solution of 18 (15 mg, 35.4 μmol, 1 equiv.) And aldehyde 26 (16 mg, 70 μmol, 2 equiv.) In DCM (3 ml), sodium triacetoxyborohydride (20 mg, 94 μmol, 2.6 equiv. .) in one portion at room temperature. After 24 hours, the reaction was quenched by the addition of a few drops of MeOH and filtered through celite. The crude material is purified by preparative TLC 1 mm (first elution: toluene / acetone (9: 1), second elution: toluene / acetone (4: 1)) to give 4 mg of a colorless oil. MS (ESI (+)) m / e 635.43 (M + H) ⁺ .

Example 12

Obtaining a cyclopamine derivative

Stage A

Compound 2 (1.30 g, 3.2 mmol, 1 equiv.) Was weighed and loaded into a reaction vessel. Potassium carbonate (0.91 g, 6.6 mmol, 2.1 eq.) Is weighed and loaded into a reaction vessel, followed by dichloroethane (6.0 ml, 76 mmol, 23.8 eq.) And anhydrous DMSO (5 ml ) The reaction mixture is heated to 70 ° C. for 36 hours under nitrogen. The reaction mixture was cooled to room temperature, diluted with DCM (15 ml) and washed twice with water (2 × 15 ml). The organic layer was dried over sodium sulfate, filtered (washing with DCM, as needed) and concentrated to dryness to give a pale yellow solid. Flash chromatography (DCM / EtOAc) afforded the target material as a white crystalline solid.

Stage B

Compound 16 (0.111 g, 0.233 mmol, 1 eq.) Was transferred to a reaction flask, placed under a nitrogen atmosphere, and dissolved in anhydrous DCM (2 ml). Chloroiodomethane (0.238 ml, 3.27 mmol, 14 equiv.) Was added. The solution is cooled to -15 ° C. Diethylzinc (1M in heptane, 1.63 ml, 1.63 mmol, 6.5 eq.) Was added dropwise over 30 minutes, carefully controlling the exothermic effect. The reaction mixture was kept between -10 ° C and -14 ° C for several hours until TLC indicated the consumption of the starting material. The reaction is then quenched by careful addition of THF (6 ml) and then aqueous citrate buffer (pH 4.5, 10 ml). The layers are allowed to warm to room temperature. Saturated sodium sulfate solution (10 ml) was added. The layers are well mixed, transferred to a separatory funnel with excess DCM, and the organic layer is collected. The organic layer was washed with aqueous sodium hydroxide (1M, 10 ml) and saturated sodium sulfate solution (10 ml), dried over sodium sulfate before concentration to dryness. The crude material is purified by flash chromatography to give the desired product in 55% yield.

Stage C

Compound 15 (1.25 g, 2.56 mmol, 1 equiv.) Was dissolved in DCM (22 ml) in a nitrogen atmosphere and the solution was cooled to an internal temperature of 0.9 ° C. Undiluted BF ₃ -OEt ₂ (1.6 ml, 12.8 mmol, 5 eq.) Was added portionwise over several hours, while monitoring the reaction with LCMS. The reaction mixture was allowed to slowly warm to 10 ° C until completion of the reaction. The reaction was quenched with MeOH (5 ml) at 0 ° C, diluted with KOH (2 M, 30 ml) and stirred at room temperature for 2 hours. The layers were separated and the organic layer was washed with water, dried over Na ₂ SO ₄ , filtered and concentrated to dryness. Chromatography with DCM / EtOAc gives the desired product.

Example 13

Obtaining a cyclopamine derivative

Stage A

Compound 27 (29 mg, 60 μmol, 1 equiv.) Was placed in a 5 ml round bottom flask. Butanone (2 ml) and Al (O i Pr) ₃ (12.3 mg, 60 μmol, 1 equiv.) Are added. The contents of the round bottom flask were heated at reflux under argon for 7 hours. The reaction mixture was then quenched with a solution (2 ml) formed by a mixture of citric acid (500 g), NaOH (15.7 g) and water (500 ml). The resulting mixture is stirred at high speed until the emulsion is dispersed. The mixture was further extracted with EtOAc (3 × 10 ml). The organic layers were collected, dried over Na ₂ SO ₄ and concentrated. The crude material is purified by column chromatography. MS (ESI (+)) m / e 486.26 (M + H) ⁺ .

Example 14

Obtaining a cyclopamine derivative

Stage A

Compound 28 (25 mg, 0.051 mmol, 1 equiv.) Was dissolved in anhydrous 2-methoxyethanol (1 ml, 12.7 mmol, 234 equiv.). Potassium carbonate (7.1 mg, 0.051 mmol, 1 equiv.) Was added and the reaction mixture was heated to 120 ° C. Monitoring the reaction is carried out using TLC. When TLC indicated the stop of the reaction, the reaction mixture was cooled to room temperature. The reaction mixture was further diluted with ethyl acetate and washed with water. The organic layer was dried over sodium sulfate and concentrated to dryness. Chromatography with DCM / EtOAc gives the desired product. MS (ESI (+)) m / e 526.66 (M + H) ⁺ .

Example 15

Obtaining a cyclopamine derivative

Stage A

To a solution of hydrocinnamic acid 5 (3.01 g, 20 mmol, 1 eq.) In 30 ml of anhydrous chloroform at 75 ° C was added thionyl chloride (1.75 ml, 24.1 mmol, 1.2 eq.) Dropwise over period of 3 minutes The mixture was refluxed for 3.5 hours. The solvent was distilled off to obtain the crude acid chloride as a pale yellow viscous liquid. The crude material is used without further purification.

Stage B

To a biphasic mixture of 7 (3.16 g, 24.1 mmol, 1.2 equiv.) In DCM (30 ml) and an aqueous solution of NaOH (2.0 M, 30 ml, 3 equiv.) At 25 ° C add a solution acid chloride 8 (3.38 g, 20 mmol, 1 equiv.) in DCM (10 ml) and the resulting mixture was stirred at 25 ° C for 3 hours. The mixture was further neutralized with an aqueous solution of HCl (2 M, 30 ml) . The organic layer was further separated and the aqueous layer was extracted with DCM (3 × 50 ml). The combined organic layers were washed with HCl (2.0 M, 25 ml), water (3 × 50 ml), brine (50 ml), dried over magnesium sulfate and the solution was evaporated under reduced pressure. The crude material is chromatographed on silica gel using 5% MeOH: DCM as an eluent and the column is further eluted with 10% MeOH: DCM to give 1.141 g of compound 8 .

Stage C

To a mixture of acid 8 (264 mg, 1 mmol, 1 equiv.), EDCI (231 mg, 1.2 mmol, 1.2 equiv.) And triethylamine (168 μl, 1.2 mmol, 1.2 equiv.) DCM (2 ml) at 0 ° C add allylamine (90.3 μl, 1.2 mmol, 1.2 eq.) And the resulting mixture was stirred at 0 ° C and allowed to warm to 25 ° C over a period of 2 hours. The reaction mixture was added to water (50 ml), extracted with DCM (4 × 25 ml), the combined organic layers were washed with 1M HCl (2 × 25 ml), water (3 × 25 ml), saturated brine (25 ml) ), dried over magnesium sulfate and the solvent is evaporated off under reduced pressure to obtain 287.5 mg; wow product. This substance is used without further purification.

Stage D

To a solution of allylamide 9 (70 mg, 0.23 mmol, 1 equiv.) In acetone (1 ml) and water (0.3 ml) was added 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 an ice bath after adding a solution of OsO ₄ . The resulting dark brown solution was stirred at 0 ° C for 15 minutes. Sodium periodate (110 mg, 0.51 mmol, 2.2 eq.) Was added 5 portions to the above mixture and stirring was continued for 1 h at 0 ° C, and the mixture was allowed to warm to 25 ° C for a period of 2 hours The reaction mixture was diluted with DCM (3 ml), filtered through a small pad of magnesium sulfate and the filter cake washed with DCM (several times). The filtrate was concentrated and the residue (67.9 mg) was filtered through a small pad of RP silica gel using 5% MeOH: DCM to give 38.9 mg of the desired product.

Stage E

Sodium triacetoxyborohydride (24.1 mg, 0.114) was added to a solution of 19 (0.0242 g, 0.0569 mmol, 1 equiv.) And aldehyde 10 (0.0346 g, 0.114 mmol) in 3.0 ml of DCM at 23 ° C. mmol, 2 equiv.) in one portion and the resulting mixture was stirred for 16 hours. After complete conversion of the starting material to the desired product, which is obvious according to LCMS and TLC, the mixture is taken up in 2.5 ml of methanol and purified by reverse phase preparative chromatography (acetonitrile buffer 20 mM ammonium carbonate, basic method). Fractions were concentrated and collected in a minimal volume of acetonitrile, the solution was frozen and lyophilized to obtain 0.007 g (0.0098 mmol, 17%) as a white solid. MS (ESI (+)) m / e 714.6 (M + H) ⁺ .

Example 16

Obtaining a cyclopamine derivative

Stage A

To a solution of 18 (35 mg, 0.08 mmol, 1 equiv.) And aldehyde 32 (34 mg, 0.17 mmol, 2.0 equiv.) In THF (2.0 ml), sodium triacetoxyborohydride (35 mg, 0 , 17 mmol, 2.0 equiv.) In one portion. The solution was allowed to stir at 23 ° C. for 12 hours. The mixture was further concentrated and purified by silica gel chromatography using a 1: 1 gradient of hexanes: ethyl acetate followed by 1: 2, 1: 4 and ethyl acetate directly. Some material was still eluting, so the column was then washed in flash mode with a 9: 1 ethyl acetate: methanol mixture. The desired product is co-eluted with a certain amount of aldehyde; therefore, the substance is further purified by preparative HPLC. (Basic method 50_100). The desired fractions are frozen and lyophilized to give an oily residue (12 mg, 0.02 mmol, 24% yield). MS (ESI (+)) m / e 614.44 (M + H) ⁺ .

Example 17

Obtaining a cyclopamine derivative

Stage A

To a solution of 18 (8.0 mg, 0.02 mmol, 1 equiv.) And aldehyde 34 (6.0 mg, 0.04 mmol, 2.0 equiv.) In CH ₂ Cl ₂ (1.0 ml) was added sodium triacetoxyborohydride (8.0 mg, 0.17 mmol, 2.0 eq.) in one portion. The solution was allowed to mix at 23 ° C for 12 hours. The mixture was further concentrated and purified by silica gel chromatography using a 1: 1 gradient of hexanes: ethyl acetate followed by 1: 2, 1: 4 to isolate the desired product, eluted with some aldehyde. This substance is further purified by preparative HPLC. The desired fractions were frozen and lyophilized to give a white powder (4.9 mg, 0.009 mmol, 46% yield). MS (ESI (+)) m / e 570.41 (M + H) ⁺ .

Example 18

Obtaining a cyclopamine derivative

Stage A

To a solution of 18 (6.0 mg, 0.01 mmol, 1 equiv.) And benzaldehyde 36 (3.0 mg, 0.02 mmol, 2.0 equiv.) In CH ₂ Cl ₂ (0.5 ml) was added sodium triacetoxyborohydride (6.0 mg, 0.02 mmol, 2.0 eq.) in one portion. The solution was allowed to stir at 23 ° C. for 12 hours. The mixture was further concentrated and purified by silica gel chromatography using a 4: 1 gradient of hexanes: ethyl acetate followed by 1: 1 to isolate the desired product, eluted with some aldehyde. This substance is further purified by preparative HPLC. The desired fractions are frozen and lyophilized to give a white powder (0.6 mg, 0.001 mmol, 8% yield).

Example 19

Obtaining a cyclopamine derivative

Stage A

To a solution of 18 (6.0 mg, 0.01 mmol, 1 equiv.) And 4-phenoxybenzaldehyde 38 (6.0 mg, 0.02 mmol, 2.0 equiv.) In CH ₂ Cl ₂ (0.5 ml ) sodium triacetoxyborohydride (6.0 mg, 0.02 mmol, 2.0 eq.) is added in one portion. The solution was allowed to stir at 23 ° C. for 12 hours. The mixture was further concentrated and purified by silica gel chromatography using a 4: 1 gradient of hexanes: ethyl acetate followed by 1: 1 to isolate the desired product, eluted with some aldehyde. This substance is further purified by preparative HPLC. The desired fractions were frozen and lyophilized to give a white powder (1.8 mg, 0.003 mmol, 21% yield). MS (ESI (+)) m / e 606.4 (M + H) ⁺ .

Example 20

Obtaining a cyclopamine derivative

Stage A

To a mixture of 18 (40 mg, 0.09 mmol, 1 equiv.) And sodium bicarbonate (15 mg, 0.18 mmol, 2.0 equiv.) In CH ₂ Cl ₂ (0.5 ml) was added bromoethanol 40 (33 μl, 0.47 mmol, 5.0 equiv.). The solution was heated under reflux for 4 hours. The mixture was further concentrated and purified by silica gel chromatography using a gradient of DCM, followed by 38: 1: 1 dichloromethane: ethyl acetate: methanol, then 36: 3: 1, then 17: 2: 1 to isolate the desired product as an oil (12 mg, 0.026 mmol, 27% yield). MS (ESI (+)) m / e 468.24 (M + H) ⁺ .

Example 21

Obtaining a cyclopamine derivative

Stage A

To a solution of 18 (100 mg, 0.24 mmol, 1 equiv.) And aldehyde 42 (42 mg, 0.35 mmol, 1.5 equiv.) In CH ₂ Cl ₂ (2.5 ml) is added sodium triacetoxyborohydride (100 mg, 0.47 mmol, 2.0 equiv.) in one portion. The solution was allowed to mix at 23 ° C. for 12 hours, and LCMS showed only 50% conversion. An additional equivalent of aldehyde 36 (26 mg, 0.24 mmol, 1.0 eq.) And sodium triacetoxyborohydride (48 mg, 0.24 mmol, 1.0 eq.) Are added to the mixture and allowed to mix for 2 hours. The mixture is further concentrated and purified by silica gel chromatography using a 1: 1 gradient of hexanes: ethyl acetate followed by 1: 2 and 1: 4 to isolate the desired product, co-eluting with some aldehyde. This substance is further purified by preparative HPLC. The desired fractions were frozen and lyophilized to give a white powder (53 mg, 0.10 mmol, 43% yield). MS (ESI (+)) m / e 526.66 (M + H) ⁺ .

Example 22

Obtaining a cyclopamine derivative

Stage A

Sodium triacetoxyborohydride was added to a solution of 18 (15 mg, 0.04 mmol, 1 equiv.) And aldehyde 44 (6.9 mg, 0.04 mmol, 1.0 equiv.) In CH ₂ Cl ₂ (0.6 ml). (15 mg, 0.07 mmol, 2.0 equiv.) In one portion. The solution was allowed to mix at 23 ° C. for 12 hours, and LCMS showed only 50% conversion. An additional equivalent of aldehyde 44 (6.9 mg, 0.04 mmol, 1.0 eq.) And sodium triacetoxyborohydride (7.5 mg, 0.04 mmol, 1.0 eq.) Are added to the mixture and allowed to mix for 12 h. The mixture is further concentrated and purified by silica gel chromatography using a gradient of 1: 1 hexanes: ethyl acetate followed by 1: 2 and 1: 4 to isolate the desired product as an oil (12 mg, 0.19 mmol, 54% yield). MS (ESI (+)) m / e 635.43 (M + H) ⁺ .

Example 23

Obtaining a cyclopamine derivative

Stage A

To a mixture of 18 (12 mg, 0.03 mmol, 1 equiv.) And potassium carbonate (40 mg, 0.28 mmol, 10 equiv.) In DMF (0.5 ml), 2- (dimethylamino) ethyl chloride hydrochloride 46 ( 20 mg, 0.14 mmol, 5.0 eq.). The solution was stirred for 2 hours at 23 ° C and the reaction did not occur. The solution was further heated at 65 ° C for 12 hours, quenched with water (2 ml) and then extracted with diethyl ether (2 × 10 ml). The combined organic extracts were washed with brine and dried with MgSO ₄ . The mixture was further concentrated and purified by preparative HPLC. The desired fractions are frozen and lyophilized to give a white powder (17, 1.8 mg, 0.004 mmol, 13% yield). MS (ESI (+)) m / e 495.71 (M + H) ⁺ .

Example 24

Obtaining a cyclopamine derivative

Stage A

To a solution of 18 (100 mg, 0.24 mmol, 1 equiv.) And chloroacetamide 48 (250 mg, 1.2 mmol, 5.0 equiv.) In CH ₂ Cl ₂ (1.0 ml) add triethylamine (160 μl 1.2 mmol, 5.0 equiv.). The solution was heated under reflux and stirred for 72 hours. The mixture was further concentrated and purified by silica gel chromatography using a 4: 1 gradient of hexanes: ethyl acetate followed by 2: 1, 1: 1 and 1: 2 to isolate the desired product as a mixture of two spots. The same column conditions are repeated and the desired product is isolated as an oil (17 mg, 0.14 mmol, 12% yield).

Example 25

Obtaining a cyclopamine derivative

Compound 18 (103 mg, 0.24 mmol, 1 equiv.) Was dissolved in DCM (3.0 ml) and cooled to -78 ° C. To this solution was added mCPBA (77 wt.%, 54 mg, 0.24 mmol, 1.0 eq.) And then the solution was allowed to warm to 22 ° C for 12 hours. The reaction proceeds with 50% conversion (LCMS). The solution was quenched with sodium bicarbonate and extracted with DCM. The combined organic phases are dried with magnesium sulfate, filtered and concentrated. The substance is further purified by silica gel chromatography using a gradient of DCM: EtOAc: MeOH equal to 95: 2.5: 2.5, then 92.5: 5.0: 2.5, then 85: 10: 5 to isolate the desired product co-eluted with a small amount of impurity. The combined fractions were concentrated and purified by preparative HPLC to give 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 Cyclopamine Analogs

Hedgehog pathway specific cancer cell killing effects can be identified using the following analysis. C3H10T1 / 2 cells differentiate into osteoblasts upon contact with the Hedgehog-voiced peptide (Shh-N). Upon differentiation, these osteoblasts produce high levels of alkaline phosphatase (AR), which can be measured in an enzymatic assay (Nakamura et al., 1997 BBRC 237: 465). Compounds that block the differentiation of C3H10T1 / 2 into osteoblasts (Shh-dependent event) can therefore be identified by reduction during production of AR (van der Horst et al., 2003 Bone 33: 899). Details of the analysis are described below. Results approximating (EC ₅₀ for inhibition) differentiation analysis are shown in the table below.

Analysis protocol

Cell culture

C3H10T1 / 2 murine embryo mesoderm fibroblast cells (obtained from ATCC) 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% CO ₂ in an air atmosphere.

Alkaline phosphatase assay

C3H10T1 / 2 cells were placed in 96-well plates at a density of 8 × 10 ³ cells / well. Cells are grown before fusion (72 hours). After adding sonic Hedgehog (250 ng / ml), (R&D Systems) and / or compound treatment, the cells are lysed in 110 μl of lysis buffer (50 mM Tris pH 7.4, 0.1% TritonX100), the plates are sonicated and the lysates are forced through the plates 0.2 μm PVDF (Corning). 40 μl of lysates are analyzed for AP activity in a solution of alkaline buffer (Sigma) containing 1 mg / ml p-nitrophenyl phosphate. After an incubation of 30 minutes at 37 ° C., the plates were read using an Envision plate reader at 405 nm. Total protein is quantified using Pierce BCA Protein Assay Kit according to the manufacturer's instructions. AR activity is normalized with respect to the total protein. It is noted that “A” indicates that the IC ₅₀ is less than 200 nM, “B” indicates that the IC ₅₀ is 200-500 nM, “C” indicates that the IC ₅₀ > 500 nM.

Approximate EU ₅₀
for inhibition Compound EU ₅₀ differentiation analysis one FROM four BUT eleven FROM 17 A eighteen A 19 B twenty C 21 A 24 B 27 C 28 C 29th A thirty A 31 A 33 A 35 A 37 C 39 C 41 A 43 B 45 B 47 B

Example 27

Preparative supercritical liquid chromatographic purification (SFC) method

Describes a method of preparative supercritical liquid chromatography for the purification of compounds of the present invention.

Software Used :

SFC: Berger PrepSFC System

UV Detector: Knauer Model K-2501

Column: Berger 5 micron silicon dioxide, 20 mm - 250 nm

SFC Terms:

Mobile phases: СО ₂ - 95%; methanol - 5%

Flow rate: 50.00 ml / min

Column temperature: 35 ° C

Isocratic for 40 min at 5% methanol in supercritical CO ₂

Injection volume: 1000 μl

Sample concentrations typically pass at 5.0 mg / ml

Sample preparation: Samples were dissolved in 20% DCM / 80% methanol.

Products elute between 25 and 40 min.

UV Detector Parameters

Wavelength = 210 nm; and resolution = 1.0 nm

Example 28

The method of liquid chromatography - mass spectrometry (LCMS)

Describes a method of liquid chromatography - mass spectrometry for the compounds of the present invention. [A] Report on the input method

Mobile phase Waters Alliance 2795 LC

Solvents BUT% 0,0 AT% 10.0 Acetonitrile FROM% 90.0 water 20 mM NH ₄ HCO ₃ D% 0.0 water 0.1A Stream mode 1.00 Flow (ml / min) 1,500 Stop time (min) 4.00 Min pressure (bar) 0 Max. pressure (bar) 300 Degasser Included Delay volume 100.0 μl Column Waters Alliance 2795 LC Column position Column 3 Equilibration time (min) 0.00 Column temperature (° C) 35 Column temperature limit (° C) 5 Quick Balance Waters Alliance 2795 LC Track system Off System flow (ml / min) 0.00 System time (min) 0.00 Re-equilibration time (min) 0.00 Pre-column volume (μl) 0.00 Waters Alliance 2795 I / O Switch 1 Without changes Switch 2 Without changes Switch 3 Without changes Switch 4 Without changes Setting analog output Flow rate Waters Alliance 2795 LC Gradient Temporary Table The gradient temporary table contains 5 inputs, which are: Time BUT% AT% FROM% D% Flow Curve 0.00 0,0 10.0 90.0 0,0 1,500 one 3.00 0,0 85.0 15.0 0,0 1,500 6 3.20 0,0 100.0 0,0 0,0 1,500 6 3,50 0,0 100.0 0,0 0,0 1,500 6 3.60 0,0 10.0 90.0 0,0 1,500 6

Temporary table of external events Waters Alliance 2795 LC

The inputs of the temporary table of external pump events are absent

Waters Alliance 2795 input options

Input type Consistent Input mode Partial loop Sample pretreatment air boundary (μl) 4.0 Post-processing air boundary (μl) 4.0 Preload Time (min) 0.00

Waters Alliance 2795 autosampler options

Sample temperature (єС) twenty Sample Temperature Limit (єС) twenty Groove depth (mm) 0 Groove speed Normal Search hole depth Crash Verification Board Height Crash Flushing parameters of the Waters Alliance 2795 Flushing frequency Enter Quick flush time (s) 3 Flushing time (s) 10 Flushing cycles 2 Secondary Wash Volume (μl) 600,0

Waters996 PDA

Initial wavelength (nm) 220.00 Final wavelength (nm) 400.00 Resolution (nm) 1,2 Sampling rate (spectrum / s) 1,000 Filter response one Exposure Time (ms) Automatically Interpolation 656 Yes Data collection stop time (min) 4,000 Saving to disk: Yes Analog Channel 1 Waters996 PDA Output mode Off Analog Channel 2 Waters996 PDA Output mode Off End of report

[B] Pilot report

Title Default experiment Time of creation Friday June 04, 2004 10:25:33 AM Tool id Version number 1,0 Duration (min) 4.0 Calibration File Name C: \ MassLynx \ Infinity_2002.PRO \ ACQUDB \ NAICS_040408.cal Solvent retention provided by check valve 0 Number of functions one

Function 1: MS scan, time 0.00 to 3.50, mass 200.00 to 1000.00 ES +

Type of MS scan Ion mode ES + Data format Centroid Parameter file C: \ Masslynx \ Infinity_2002.PRO \ ACQUDB \ default.ipr Starting weight 200,0 Final mass 1000,0 Scan Time (sec) 1,0 Time between scans (sec) 0.2 Start time (min) 0,0 End time (min) 3,5 [C] ZQ settings Source (ES +) Setting Read values Capilar (kV) 3.20 3.27 Cone (V) 35.00 37.24 Extractor (V) 3.00 2.81 Lens RF (V) 0,0 0,0 Source temperature (° C) 130 129 Desolvation Temperature (° C) 350 350 Cone gas flow (l / h) twenty 33 Desolvation gas flow (l / h) 450 466 Analyzer Setting Read values LM 1 Resolution 15.0 NM 1 resolution 15.0 Ion Energy 1 0.5 Improver (V) 550 -547 Pressure valve Pirani pressure (mbar) <1e-4 mbar MUX configuration Probe Standard

Link inclusion

All of the US patents and published US patent applications cited in this description are included in it by reference.

Equivalents

Specialists in this field will be able to recognize or be able to identify, using nothing more than routine experiments, many equivalents of the specific embodiments of the invention described herein. Assume that such equivalents are covered by the following claims.

Claims (19)

1. The compound of formula 1

or a pharmaceutically acceptable salt thereof,
in which each R ¹ and R ⁸ independently represents H or hydroxyl;
each R ² and R ⁹ independently represents H or hydroxyl;
R ⁵ represents H;
each R ³ , R ⁴ , R ⁶ , R ⁷ , R ¹³ and R ¹⁴ independently represents H;
or R ¹ and R ² taken together form = O; or
R ⁴ and R ⁵ taken together form a double bond; or
R ⁵ and R ⁶ taken together form a double bond;
R ¹⁰ and R ¹¹ taken together form a double bond;
R ¹² represents H, alkyl, hydroxyl, aralkyl,
haloalkyl, alkoxyl, - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , [(W) -O] _q R ²¹ or - [(W) —N (R ²¹ )] _q ²¹ ;
where each W independently represents a divalent alkyl or aralkyl radical, aq is 1, 2, 3 or 4;
each R ¹⁵ , R ¹⁶ and R ¹⁷ independently represents H;
each R ¹⁸ and R ¹⁹ independently represents H and
each R ²¹ independently represents H, alkyl, aryl or aralkyl. 2. The compound according to claim 1, which has the formula

or

where R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ , R ¹⁷ , R ¹⁸ and R ¹⁹ are as defined in claim 1, or a pharmaceutically acceptable salt thereof. 3. The compound according to claim 1, where
R ¹ represents hydroxy;
or R ¹ and R ² taken together form = O, and / or
R ⁴ and R ⁵ taken together form a double bond. 4. The compound according to claim 1, where R ¹ and R ² taken together form = O or R ¹ represents hydroxyl, and R ² represents N. 5. The compound according to claim 1, where R ⁵ and R ⁶ taken together form a double bond. 6. The compound according to claim 1, where R ⁸ and R ⁹ represent hydrogen. 7. The compound according to claim 1, where R ¹² represents H, alkyl, aralkyl, haloalkyl, hydroxyl or alkoxyl. 8. The compound according to claim 1 of the formula

or

or a pharmaceutically acceptable salt thereof,
where R ¹² represents H, alkyl, hydroxyl, aralkyl, haloalkyl, alkoxyl, - [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -O] _q R ²¹ or
- [(W) -N (R)] _q R ²¹ where each W independently represents a divalent alkyl or aralkyl radical;
q is 1, 2, 3 or 4;
each R ²¹ independently represents H, alkyl, aryl or aralkyl. 9. The compound according to claim 1, selected from the group consisting of

and

and their pharmaceutically acceptable salts. 10. The compound of formula 2

or a pharmaceutically acceptable salt thereof,
in which each R ¹ and R ⁸ independently represent H or hydroxyl;
each R ² and R ⁹ independently represents H or hydroxyl;
R ⁵ represents H;
each R ³ , R ⁴ , R ⁶ , R ⁷ , R ¹³ and R ¹⁴ independently represents H;
or R ¹ and R ² taken together form = O; or
R ⁴ and R ⁵ taken together form a double bond; or
R ⁵ and R ⁶ taken together form a double bond;
R ¹⁰ and R ¹¹ taken together form a group represented by 1b

where Z represents O or C (R ²³ ) (R ²³ );
R ¹² represents H, alkyl, hydroxyl, aralkyl, haloalkyl, alkoxyl,
- [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -O] _q R ²¹ or - [(W) -N (R ²¹ )] _q R ²¹ ;
where each W independently represents a divalent alkyl or aralkyl radical, aq is 1, 2, 3 or 4;
each R ¹⁵ , R ¹⁶ and R ¹⁷ independently represents H;
each R ²¹ independently represents H, alkyl, aryl or aralkyl and
each R ²³ independently represents N. 11. The compound of claim 10, which has the formula

or

where R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ and R ¹⁷ such as defined in claim 10, or a pharmaceutically acceptable salt thereof. 12. The compound of claim 10, where
R ¹ represents hydroxy; or R ¹ and R ² taken together form = O; and / or
R ⁴ and R ⁵ taken together form a double bond. 13. The compound of claim 10, where R ⁵ and R ⁶ taken together form a double bond. 14. The compound of claim 10, where R ⁸ and R ⁹ represent hydrogen. 15. The compound of claim 10, where R ¹² represents H, alkyl, aralkyl or halogenated. 16. The compound according to claim 10 of the formula

or

or a pharmaceutically acceptable salt thereof,
where R ¹² represents H, alkyl, hydroxyl, aralkyl, haloalkyl, alkoxyl,
- [(W) -N (R ²¹ ) C (O)] _q R ²¹ , - [(W) -N (R ²¹ ) SO ₂ ] _q R ²¹ , - [(W) -O] _q R ²¹ or - [(W) -N (R ²¹ )] _q R, where each W independently represents a divalent alkyl or aralkyl radical; and
q is 1, 2, 3 or 4; and
each R ²¹ independently 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. A pharmaceutical composition having anti-cancer activity, comprising a compound according to any one of claims 1-17; and a pharmaceutically acceptable excipient. 19. The compound according to any one of claims 1 to 17 for use as a medicine for the treatment of cancer.