Compounds and compositions as hedgehog pathway modulators

Biphenyl-3-carboxamides and compositions as modulators of the Hedgeog path

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a national phase application of the United States of international application number PCTIUS20071068292 filed on May 4, 2007, whose application claims priority of the provisional US patent application number 601797,949, filed on May 5 2006. The full disclosure of these applications is incorporated herein by reference in its entirety and for all purposes.

Field of the Invention

The invention provides a method for modulating the activity of the Hedgehog signaling path. In particular, the invention provides a method for inhibiting aberrant growth states resulting from phenotypes such as loss of Ptc function, Hedgehog function gain, Smoothened function gain, or Gli function gain, which comprises contacting a cell with a sufficient amount of a compound of Formula I.

Background of the Invention

During embryonic development, the Hedgehog signaling pathway is essential for numerous processes, such as the control of cell proliferation, differentiation, and the formation of tissue patterns. However, the aberrant activity of the Hedgehog signaling path, for example as a result of better activation, can have pathological consequences. In this regard, activation of the Hedgehog pathway in adult tissues may result in specific types of cancer that include, but are not limited to, cancers of the brain, muscle and skin, prostate, medulloblastoma, pancreatic adenocarcinomas, and carcinomas. small cell lungs. The best activation of the Hedgehog signaling pathway contributes to the pathology and / or symptomatology of a number of diseases. In accordance with the above, the molecules that modulate the activity of the Hedgehog signaling path are useful as therapeutic agents in the treatment of these diseases.

Brief Description of the Invention

In one aspect, the present invention provides compounds of Formula I:

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where:

Yi and Y₂ are independently selected from N and CR₁₀; where R₁₀ is selected from hydrogen, halogen, alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms substituted by halogen, alkoxy of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms substituted by halogen , and -OXNR_10thRiob; where Ri_Oa and Riob are independently selected from hydrogen and alkyl of 1 to 6 carbon atoms;

Rt is selected from cyano, halogen, alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms substituted by halogen, alkoxy of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms substituted by halogen, aryl of 6 to 10 carbon atoms , dimethyl-amino, alkyl of 1 to 6 carbonosulfanyl atoms, and hetero-chloroalkyl of 3 to 8 carbon atoms optionally substituted with up to 2 alkyl radicals of 1 to 6 atoms carbon;

R₂ and Rs are independently selected from hydrogen, cyano, halogen, alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms substituted by halogen, alkoxy of 1 to 6 carbon atoms, alkoxy of 1 to 6 atoms carbon substituted by halogen, and dimethyl amino;

R₃ and R₄ are independently selected from hydrogen, halogen, cyano, alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms substituted by halogen, alkoxy of 1 to 6 carbon atoms, and alkoxy of 1 to 6 atoms of carbon substituted by halogen; o Ri and R₂, or R! and R₅, together with the phenyl with which they are both attached, form heteroaryl of 5 to 10 carbon atoms;

R₆ and R₇ are independently selected from hydrogen, alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms substituted by halogen, alkoxy of 1 to 6 carbon atoms, and alkoxy of 1 to 6 carbon atoms substituted by halogen ; with the proviso that R₆ and R₇ they are not both hydrogen;

Ra is selected from hydrogen, halogen, alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms substituted by halogen, alkoxy of 1 to 6 carbon atoms, and alkoxy of 1 to 6 carbon atoms substituted by halogen;

R₉ is selected from -S (O)₂R_{1 B} -0 (0 ^^, -OR_{1 B}-NR_12thRi2b and -Rn; wherein Rn is selected from aryl, heteroaryl, cycloalkyl, and heterocycloalkyl; R_12th and Ri₂t are independently selected from alkyl of 1 to 6 carbon atoms and alkyl of 1 to 6 carbon atoms substituted by hydroxyl;

wherein the aryl, heteroaryl, cycloalkyl, and heterocycloalkyl of Rg may be optionally substituted with 1 to 3 radicals independently selected from alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms substituted by halogen, alkoxy of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms substituted by halogen, aryl of 6 to 10 carbon atoms-alkyl of 0 to 4 carbon atoms, hetero-aryl of 5 to 10 carbon atoms-alkyl of 0 to 4 carbon atoms, cycloalkyl of 3 to 12 carbon atoms, and hetero-cycloalkyl of 3 to 8 carbon atoms;

wherein the aryl alkyl substituent of R₉ it is optionally substituted with 1 to 3 radicals independently selected from halogen, alkyl of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms substituted by halogen, alkoxy of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms substituted by halogen, and metll-plperazmil; and N-oxide derivatives, pro-drug derivatives, protected derivatives, individual isomers and mixtures of Isomers thereof; and pharmaceutically acceptable salts and solvates (eg, hydrates) of these compounds.

In a second aspect, the present invention provides a pharmaceutical composition, which contains a compound of Formula I or an N-oxide derivative, individual isomers and mixtures of isomers thereof; or a pharmaceutically acceptable salt thereof, mixed with one or more suitable excipients.

In a third aspect, the present invention provides a method for the treatment of a disease in an animal where the modulation of the activity of the Hedgehog pathway can prevent, inhibit, or alleviate the pathology and / or symptomatology of the diseases, whose The method comprises administering to the animal a therapeutically effective amount of a compound of Formula I, or an N-oxide derivative, individual isomers and mixtures of Isomers thereof, or a pharmaceutically acceptable salt thereof.

In a fourth aspect, the present invention provides the use of a compound of Formula I in the manufacture of a medicament for the treatment of a disease in an animal where the activity of the Hedgehog path contributes to the pathology and / or symptomatology of the illness.

In a fifth aspect, the present invention provides a process for the preparation of compounds of Formula I, and the N-oxide derivatives, pro-drug derivatives, protected derivatives, individual isomers and mixtures of isomers thereof, and pharmaceutically acceptable salts thereof.

Definitions "Alkyl", as a group and as a structural element of other groups, for example alkyl and alkoxy substituted by halogen and alkoxy, may be straight or branched chain. Alkoxy of 1 to 4 carbon atoms includes methoxy, ethoxy, and the like. Halogen-substituted alkyl includes trifluoro-methyl, pentafluoro-ethyl, and the like.

"Aryl" means a fused monocyclic or bicyclic aromatic ring assembly containing from 6 to 10 ring carbon atoms. For example, aryl can be phenyl or naphthyl, preferably feni I or.

"Arylene" means a divalent radical derived from an aryl group.

"Heteroaryl" is as defined for aryl above, wherein one or more of the ring members is a heteroatom. For example, heteroaryl of 5 to 10 carbon atoms is a minimum of

Ί five members, as indicated by carbon atoms, but these carbon atoms can be replaced by a heteroatom. Accordingly, heteroaryl of 5 to 10 carbon atoms includes pyridyl, indolyl, indazolyl, quinoxalinyl, quinolinyl, 5 benzo-furanyl, benzo-pyranyl, benzo-thiopyranyl, benzo- [1,3] -dioxol, imidazolyl, benzo-imidazolyl , pyrimidinyl, furanyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, thienyl, etc.

"Cycloalkyl" means a monocyclic, fused bicyclic, or bridged, saturated or partially unsaturated polycyclic ring assembly, which contains the number of atoms in the indicated ring. For example, cycloalkyl of 3 to 10 carbon atoms includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

"Hetero-cycloalkyl" means cycloalkyl, as defined in this application, with the understanding that one or more of the indicated carbon atoms of the ring are replaced by a fraction selected from -O-, -N =, -NR -, -C (O) -, -S-, -S (O) - or -S (O)₂-, where R is hydrogen, alkyl of 1 to 4 carbon atoms, or a nitrogen protecting group. For example, heterocycloalkyl of 3 to 8 carbon atoms, as used in this application to describe the compounds of the invention, includes morpholino, pyrrolidinyl, pyrro I idini l-2-one, piperazinyl, piperidinyl, pi pe rid ini lo na, 1 , 4-dioxa8-aza-espiro- [4.5] -dec-8-¡, thiomorpholino, sulfano-morphol, sulfonomorpholino, etc.

"Halogen" (or halo), preferably represents chlorine or fluorine, but can also be bromine or iodine.

"Hedgehog function gain" refers to an aberrant modification or mutation of a Ptc gene, Hedgehog gene, or Smoothened gene, or a decrease (or loss) in the expression level of this gene, which results in a phenotype that It seems to make contact with a cell with a Hedgehog protein, for example an aberrant activation of a Hedgehog path. The gain in function may include a loss of the ability of the Ptc genetic product to regulate the level of expression of the Gil genes, for example Gl1, Gl2, and Gli3. The term "Hedgehog function gain", is also used herein to refer to any similar cellular phenotype (for example, exhibiting excess proliferation), which occurs due to an alteration in any part of the transduction path. Hedgehog signal, including, but not limited to, a modification or mutation of the Hedgehog itself. For example, a tumor cell with an abnormally high proliferation rate due to the activation of the Hedgehog signaling path would have a "Hedgehog function gain" phenotype, even when the Hedgehog is not mutated in that cell.

"Patched loss of function" refers to an aberrant modification or mutation of a Ptc gene, or a reduced level of gene expression, which results in a phenotype that appears to make contact with a cell with a Hedgehog protein, for example the aberrant activation of a Hedgehog path. Loss of function may include a loss of the ability of the Ptc genetic product to regulate the expression of Gil genes, for example

GII1, GII2, and Gl¡3.

"Gil function gain" refers to an aberrant modification or mutation of a Gli gene, or a higher level of gene expression 5, which results in a phenotype that appears to make contact with a cell with a Hedgehog protein, for example the aberrant activation of a Hedgehog path.

"Smoothened function gain" refers to an aberrant modification or mutation of a Smo gene, or to a higher level of gene expression, which results in a phenotype that appears to make contact with a cell with a Hedgehog protein, for example the aberrant activation of a Hedgehog path.

"Treat," treating, and "treatment," refer to a method to relieve or abate a disease and / or its combined symptoms.

The present invention relates to the discovery that the signal transduction pathways regulated by Hedgehog, (Ptc) Patched, Gli, and / or Smoothened, can be modulated by the compounds of Formula I.

Description of Preferred Modalities

In one embodiment, with respect to the compounds of the

Formula I, Y ·, and Y₂ are selected from N and CR₁₀; where R₁₀it is selected from hydrogen, methyl, fluorine, chlorine, bromine, dimethyl-amino-ethoxy, and trifluoro-methyl; R₆ and R₇ they are independently selected from hydrogen, methyl, chlorine, fluorine, bromine, trifluoro-methyl, and methoxy; with the condition of R₆ and R₇ they are not both hydrogen; and R_to It is selected from hydrogen, fluorine, chlorine, methyl, and trifluoro-methyl.

In another mode, R! is selected from cyano, chloro, fluorine, methyl, ethyl, tertiary butyl, propyl, isobutyio, isopropyl, sopropyloxy, butoxy, methoxy, dimethyl-amino, ethoxy, methylsulfanyl, phenyl, trifluoro-methyl, trifluoro-methoxy, and piperazinyl, optionally substituted with up to 2 methyl radicals; R₂ and Rs are independently selected from hydrogen, chlorine, fluorine, cyano, methyl, trifluoro-methyl, sopropyloxy, methoxy, ethoxy, trifluoro-methoxy, and dimethyl-amino; and R₃ and R₄ they are independently selected from hydrogen, chlorine, methyl, methoxy, and cyano; or R! and R₂, or R! and R₅, together with the phenyl with which they are both attached, form quinoxalinyl.

In another mode, R₉ is selected from -S (O) 2R1 1! -ORn, -C (O) Rn, -NR_12thRi2b and -Rui θη where Rn is selected from thiomorpholino, sulfono-morpholino, sulfano-morpholino, morpholino, cyclohexyl, phenyl, azepan-1-yl, 2-oxopiperazin-1-yl, 1,4-oxazepan4-yl, piperidin-1-yl, tetrahydro-2H-pran-4-yl, piperidin-3-yl, piperazinyl, pyrrolidinyl, and 1,4-diazepan-1-yl; R_12th and Ri2b are independently selected from isobutyio and hydroxy-etiIo; wherein this thiomorpholino, sulfono-morpholino, sulfano-morpholino, morpholino, cyclohexyl, phenyl, azepan-1-yl, 2-oxopiperazin-1-yl, 1,4oxazepan-4-yl, piperidin-1-yl, tetrahydro-2H -piran-4-yl, piperidin-3¡lo, piperazinyl, pyrrolidinyl, or 1,4-diazepan-1-yl of R_g they may be optionally substituted with 1 to 3 radicals independently selected from methyl, ethyl, methoxy, benzyl, thienyl methyl, pyridin Im et i I or, benzo- [d] - [1,3] -dioxol- 6-yl, and 2,3-dihydro-benzo- [b] [1,4] -dioxin-7-yl; wherein the phenyl or benzyl substituent of R_git is optionally substituted with 1 to 3 radicals independently selected from methoxy, ethoxy, methyl-piperazinyl, methyl, trifluoro-methoxy, chlorine, fluorine, and trifluoromethyl.

Preferred compounds of Formula I are selected from:

4'-Cyano-6-methylbiphenyl-3-carboxylic acid [4- (morpholin-4-sulfonyl) -phenyl] -amide, [6- (2,6-d imeti lm or rf or I n-4- i) -pi ri di n-3-yl] -am id a of 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, (6-azepan-1-yl-pyridin-3-yl) -amide 4'-Methoxy-2-methyl-biphenyl-3-carboxylic acid, (6-azepan1-i lp i rid i n-3-yl) -am 4'-methoxy-2-methyl-b-phenyl acid -315 4'-Methoxy-2-methylbiphenyl-3-carboxylic acid (4-cyclohexyl-phenyl) -amide, [6- (2-met i l-morf ol i n-4-I) -pi rid i n-3-il] -am id a of 4'-methoxy-2-methyl-biphenyl acid -3-carboxylic, 4'-dimethyl-amino-2-methyl-biphenyl-3-carboxylic acid, (4-morpholin-4-phenyl) -amide (4-cyclohexyl-fen¡I) -amide 4'-dimethyl-amino-2-methyl-biphenyl-3-carboxylic acid amide, (6- [1,4] -oxazepan-4-yl-pridin-3-yl) -am¡da of 6-chloro-4'-dimethylamino-biphenyl-3-carboxylic acid, (6-morpholin-4-¡lp¡r¡d¡n-3-¡) -amáda of acid 6- chloro-4'-dimethyl-amino-biphenyl-3-carboxylic acid, (6-azepan-1 - ilpi rid i η-3-il) -am id a of 6-chloro-4'-dimethyl-amino-bfen-3-carboxylic acid, [6- (2-methyl 625 chloro-4'-methoxy-biphenyl-3-carboxylic acid -morpholin-4-yl) -pyridin-3-yl] -am¡da, (6- [1,4] -oxazepan-4-yl-pyridin -312-yl) -6-Chloro-4'-methoxy-biphenyl-3-carboxylic acid, (6-azepan1-yl-pyridin-3-yl) -amide of 6-chloro-4'-methoxy acid 6-Chloro-4'methoxy-biphenyl-3-carboxylic acid -6-phenyl-3-carboxylic acid-6-morpholin-4-yl-pyridin-3-yl) -amide, (6-morphol η-4-i lp ¡ridi η-3-yl) -am id of 4'-methoxy-6-methyl-biphenyl-3-carboxylic acid, (6- [1,4] -oxazepan -4-ilp i rid i η-3-il) -am id a of 4'-methoxy-6-methyl-biphenyl-3-carboxylic acid, [6 (2-methyl-morpholin-4-yl ) 4'-Methoxy-6-methylbiphenyl-3-carboxylic acid, [6- (2-methyl-morpholin-4-yl) -pyridin-3-yl] -pyrin-3-1] -amide | ] -am'da of 4'-dimethyl-amino-6-methyl-biphenyl-3-carboxylic acid, 4'-D-methyl-amino-6-methyl-biphenyl-3-carboxylic acid oxazepan-4-yl-pyridin-3-yl) -am¡da, (6 4'dmethyl-amine-6-methyl-biphenyl-3-carboxylic acid -morpholin-4-yl-pyridin-3-yl) -amide, (6-azepan-1-yl- 4'-Methoxy-6-methyl acid pyridin-3-yl) amide 4'-methoxy-6-methyl-biphenyl-3-carboxylic acid, (6-azepan-1yl-pyridin-3-yl) -amide 6-methyl-4'-methyl-sulfanyl-biphenyl-3-carboxylic acid -biphenyl-3-carboxylic, (6-azepan-1-i-I-pyridi n-3-i-I) -amide, (6-azepa n-1 -i l-pi rid i η-3-il) -am 4'dmethyl-am-n-6-metyl-fenb-3-carboxylic acid 6-methyl- [1,1 '; 4', 1 "] -terfenyl-3-carboxylic acid, (6-azepan-1-pyridin-3-yl) amide, (6-azepan1-i-I - 3'-Chloro-6-methyl-biphenyl-3-carboxylic acid, (6-azepan-1-lp¡rid¡n-3-yl) pyridin-3-iI) -am id a 2 ', 4'-Dichloro-6-methylbiphenyl-3-carboxylic acid, (6-azepan-1-l-pyridin-3-yl) -amide of 2'chloro-6-meti acid l-biphen¡l-3-carboxylic, 3'-Chloro-6-methyl-biphenyl-3-carboxylic acid, (6-azepan-1-yl-pyridin3) (6-azepan-1 -i lp i rid i n-3-i I) -a mid a -3) 3'-4-dichloro-6-methyl-biphenyl-3-carboxylic acid, (625 azepan-1-yl-pyridin-3-yl) -amide of 3'-chloro acid 6,4'-Dimethyl-biphenyl-3 acid-6-methyl-4'-trifluoro13 methyl-biphenyl-3-carboxy-I, (6-azepan-1-yl-pyridin-3-yl) -amide -carboxylic, (6-azepan-1-I-pyridine-3-I) -am'da of 4'-et-l-6-methyl-phenyl-3-carboxylic acid , (6-azepan -1-1 lp ¡ridi n-3-11) 4'-terbutyl-6-methyl-biphenyl-3-carboxylic acid amide, (6-azepan-1il-pridrid-3-yl) -6-methyl-4'-propyl-biphenyl-3-carboxylic acid amide, (6-azepan-1-yl-pyridin-3-yl) -am'da of 4'-isobutiI-6-methyl acid -biphenyl-3-carboxylic acid, (6-azepan-1-1-pi rid and η-3-yl) -am 4'-isopropyl-6-methyl-biphenyl-3-carboxylic acid, (6-azepan-1-yl 6,2 ', 6'-trimethyl-biphenyl-3-carboxylic acid -pyridin-3-yl) -amide, 6,2 ', 3'-trimethyl-biphenyl-3-carboxylic acid (6-azepan-1-pyridin-3-yl) amide, (6-azepan-1-ylpi r id i n-3-i I) -am 6-m eti l-4'-t rif I u oro-m et i I-bife ni I-3-carboxylic acid, (6-azepan-1 -i l-pi rid i n-3-I) -am 6-methyl-3'trifluoro-meti l-biphen and l-3-carboxylic acid, (6-azepan-1 - i l-pi ridi n-3- il) -am 6-methyl-3 ', 5'-bis-trifluoro-methyl-biphenyl-3-carboxylic acid, (6azepan-1-yl-pyridin-3-yl) -amide 3'-Isopropoxy-6-methyl-biphenyl3-carboxylic acid, 3'-Ethoxy-6-methyl-biphenyl-3-carboxylic acid (6-azepan-1-yl-pyridin-3-yl) -amide, (6-azepan-1-yl-pyridin-3-yl) -amide of 2 ', 6'-dimethox¡-6-methyl-biphenyl-3-carboxylic acid, (6-azepan-1-1-pi-rid and η-3-1) amide of 6-methyl-4' acid 6-Methyl-3'-trifluoro-methoxybiphenyl-3-carboxylic acid -trifluoro-methoxy-biphenyl-3-carboxylic acid, (6azepan-1-yl-pyridn-3-1l) -amamda, (4 6-methylbiphenyl-3-carboxylic acid -morpholin-4-yl-phenyl) -amide, 4'-Methox¡6-methyl-b¡phenyl-3-carboxylic acid (4-morpholin-4-yl-phenyl) -amide, (4-morphol-4-yl-phenyl) -amide 4'- (2-Dimethyl-amino-ethoxy) -6-acid 3'methoxy-6-methyl-phenyl-3-carboxylic acid, (4-morphol-4-yl-phenyl) -amide -methyl-biphenyl-3-carboxylic acid, (414 morphoI-4-¡-l-fen¡I) -am'da of 3'-dimethyl-amine-6-methlfen acid -3-carboxylic, 4'-fluoro-6-methyl-phenyl-3-carboxylic acid (4-morphol-4-yl-phenyl) -amamide, 3'-Fluoro6-methyl-b¡fen¡l-3-carboxylic acid (4-morphol-4-l-phenol) -am¡da, (4-morphol-4-yl) -fen¡l) -am¡da of the 2'5 fluoro-6-met¡lb¡fen¡l-3-carboxylic acid, 4-metll-N- (4-morpholin-4-¡-l-fen) -3qulnoxalln-6-yl-benzamlda, (4-morphol-4-ll-phenyl) -methyl acid 6metl-4 '- (4-methyl-p'perazin-1-yl) -b 2'-Cyano-6-methyl-b-phenyl-3-carboxylic acid (4-mo rfol i n-4-i-phenyl) -amide, (4morphol! 3'-Cano-6-metll-b-phenyl-310-carboxylic acid -4-¡-phenyl) -am¡da, 4'c¡ano-6-methyl-biphenyl-3-carboxylic acid (6- [1,4] -oxazepan-4-yl-pyridine-3-yl) -amam; azepan-1 -il-p¡rid¡n-3-¡) -am¡da of 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, [6- (2- 4'-Cyano-6-methyl-blfenyl-3-carboxylic acid methal-morphol-4-yl) -pyridin-3-yl] -amide, (3,4,5,6-tetrah 4'-cyano15 6-methyl-blfenyl-3-carboxylic acid dro-2H- [1,2 '] - bipyrinin-5'-¡1) -amide, (6-morpholin-4-¡ lp¡r¡d¡n-3-¡l) -am¡da of 4'-c¡ano-6-methyl-b¡fen¡l-3-carboxylic acid, [6- (4-methyl-pipe-razin-1-yl) p-rid i n-3-i] -am id a of 4'-cyano-6-methyl-biphenyl-3 acid -carboxylic, (4morphol-4-yl-phenol) -am'da of 4'-clano-6-methyl-phenyl-3-carboxylic acid, (3-fluoro-4-morphol-4- 4'-Cano-620-Methyl-620-phenyl-3-carboxylic acid, l-phenyl-3-chloro-4-morphol-4-l-phenyl) - 4'-Cyano-6-methyl-biphenyl-3-carboxylic acid, (3-bromo-4-morphol-4-laphenyl) -amlda of 4'-cyano-6-met acid L-biphen¡l-3-carboxylic, 4'-Clano-6-metyl-3-carboxylic acid (3-metll4-morphol-4-l-phenol) -am¡da, (4-morpholin-4-yl) 4,25-Cano-6-methyl-3-carboxylic acid -3-trifluoro-methyl-phenyl) -amide, (4-cyclohexyl-phenyl) -amide acid

4'-6-m eti lb 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, biphenyl-4-yl-amide, carboxylic acid, (4'-methoxy) 4'-Cyano-6-methyl-biphenyl-3-carboxylic acid, 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, [4- (4-benzyl-piperazin-1-yl) -phenyl] -amide of 4'- acid cyano-6-methyl-biphenyl-3-carboxylic acid, [45 (piperidin-1-sulfonyl) -phei |] -amide of 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, [4- (pi rro I id i n-1 -su Ifo or I) -fe nil] -am id a of 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, 4'-Cyano-6-methoxy-biphenyl-3-carboxylic acid (6-azepan-1-pyridin-3-yl) -amide, (6-azepan-1-yl-pyridin-3-yl ) 4'-cyano-2-methoxy-biphenyl-3-carboxylic acid amide, (6-azepan-110-yl-pyrd-3-yl) -am'da of 4'-cyano-2- acid 3'-fluoro-4'-methoxy-6-methyl-biphenyl- (6-azepan-1-1 l-pi rid and n-3-i-1) methyl-b-phenyl-3-carboxylic acid 4'-Isopropoxy-6-methyl-biphenyl-3-carboxylic acid (6-azepan-1-yl-pyridin-3-yl) -amide; 4'-Butoxy-6-methyl-biphenyl-3-carboxylic acid (6-azepan-1-pyridin-3-yl) amide, (6-azepan-1-yl15 pi rid i n-3- i I) -am id a of 3'-chloro-4'-methoxy-6-methyl-biphenyl-3-carboxylic acid, (6-azepan-1 -i l-pi rid i η-3-i I) -am id 4'-Methox6,3'-dimethyl-biphenyl-3-carboxylic acid, [4- (piperidin-1-sulfonyl) -phenyl] -amide 4'-cyano-2-methyl-biphenyl acid -3-CarboxH¡co, 4'-Cyano-6-fluoro-biphenyl-3-carboxylic acid [4- (piperidin-1sulfonyl) -phenyl] -amide, 6-Bromo-4'-cyanob ifen and l-3-ca rboxylic acid [4- (piperidin-1-sulfonyl) -feni |] -amide, [6- (4-benzyl- [1,4] - 4'-Cyano-6-methyl-biphenyl-3-carboxylic acid diaze pan-1 -yl) -pyridin-3-yl] amide, [6- (4-thiophene-3-methyl-methyl- [1,4 4'-Cyano-6-methyl-biphenyl-3-carboxylic acid, [6- (2,4-di m eti lm or rfo I i n-] -diazepan-1-yl) -pyridin-3-yl] -amide 4-i I) -pi rid i η-3-i I] 25''-cyano-2-methyl-biphenyl-3-carboxylic acid amide, 4'-Methoxy-2-methyl-phenyl-3-carboxylic acid [6- (2,6-dimethyl16 morpholin-4-¡1) -pyridin-3-yl] -amide, [6- (2,6-di met i l-mo rfo I i η-4-i I) -pi rid i n-3-il] -amide of the acid

2-methyl-4'-triflu gold-methyl l-biphenyl-3-carboxylic acid, [6- (2,6-di meti-mo rfoli n 4-i I) -p¡ rid i n-3-¡ l] -am 2-methyl-4'-trifluoro-methoxy-biphenyl-35 carboxylic acid, [6- (2-methyl-morpholin-4-yl) -pyridin-3-yl] -amide of 4 'acid cyano-2-methyl-b¡fen¡l-3-carboxylic acid, [4- (pipe rid i n-1 - i l-su Ifon i I) -fen i |] 4'-cyano- acid amide 4'-Cyano-6-trifluoro-methyl-phenyl-3-carboxylic acid [4- (piperidin-1-sulfonyl) -phenyl] -amide 2'-fluoro-biphenyl-3-carboxylic acid, [6- (4-pyridin-4-yl-metl- [1,4] -d azepan-1 -i I) -p¡ rid iη-3-iI] 10'-4'- acid amide cyano-6-methyl-biphenyl-3-carboxylic, [6- (4-p¡ rid i n-3¡l-methyl- [1,4] -diazepan-1 -il) -pyrin-3-yl ] -am'da of 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, {6- [4- (2,6-dimethoxy-benzyl) - [1,4] -diazepan1-yl] -pyridin-3 -yl} 4'-Cyano-6-methyl-biphenyl-3-carboxylic acid, {6- [4- (2-ethoxy-benzyl) - [1,4] -diazepan-1- yl] -pyridine -3-yl} 4'-cyano-6-methyl-biphenyl-3-carboxylic acid amide, (6- {4- [2- (4meti l-pipe raz i n-1 -yl) -benzyl] - [1,4] -di azepan-1-yl} -pyridin-3-yl) -amide of the acid 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, {6- [4- (4-methoxy¡-2,3-dimethyl-benzyl) - [1,4] -diazepan-1-i |] -pyridin- 4'cyano-6-methyl-biphenyl-3-carboxylic acid 3-yl} -amide, {6- [4- (2,3-dihydro-benzo- [1,4] 20 di oxy n-6 -¡ lm eti I) - [1,4] -d iaze pa n -1 -il] -pi rid i n-3-il} -am id a of 4'c¡ano-6-methyl-biphenyl- 3-carboxylic, 4'-Cyano-6-methyl-biphenyl-3-carboxylic acid [6- (4-pyridin-2-yl-methyl- [1,4] diazepan-1-yl) -pyridin-3-yl] -amide, [ 4'-Cyano acid 6- (4-benzo- [1,3] -dioxol-4-yl-methyl- [1,4] -d¡azepan-1-iI) pyridin-3-yl] -amide 6-methyl-biphenyl-3-carboxylic acid, {625 [4- (2-trifluoro-methoxy-benzyl) - [1,4] -di azepan-1-yl] -pyrn-3-yl } -am'da of 4'-cyano-6-methyl-b-phenyl-3-carboxylic acid, {6- [4- (2-dimethylamino-benzyl) - [1,4] -diazepan-1-yl] 4'ciano-6-methyl-biphenyl-3-carboxylic acid -pyridin-3-yl} -amide, {6- [4- (2-Chloro-5-trlfluoro-methylbe nci I) - [1,4] -d ¡aze pa n-1 -i I] -pi rid i η-3-il} -am id a of 4'-cyano-65 methyl-biphenyl-3-carboxylic acid, {6- [4- (2,3-difluoro-benzyl) - [1,4] -diazepan1-i |] -pyridin -3-yl} -amide of 4'-cyano-6-methyl-b-phenyl-3-carboxylic acid, {6- [4- (2-chloro-4-fluoro-benzyl) - [1,4] -d¡ 4'-cyano-6-methyl-4-methyl-3-carboxylic acid azepan-1 -iI] -pyridin-3-yl} -am¡da, 4'10 cyano-6-met acid {6- [4- (2,6 difioro-benzyl) - [1,4] -d¡azepan-1-yl] -pyridin-3-yl} -amide 2-Chloro-4'-cyano-b¡phenyl-3-carboxylic acid [4- (p ¡perid ¡n-1-sulfonyl) -phenyl] 2-chloro-3-carboxylic acid, [6 - 4'-Cyano-6-trifluoro-methylbiphenyl-3-carboxylic acid -pyrid-4-¡1) -pyrn-3-i |] -amide 2-Chloro-4'-cyano-b-phenyl acid [6- (2,6-dimethl-morpholin-4-yl) -p¡r¡n-3-l] -am¡da -3-carboxylic, [6- (2,6-D-methyl15 mo rfo I i n-4-l) -pi ri d ¡n-3-l] -am of 4'-cyano-6-ethyl-biphen acid L-3-carboxylic acid, 4'-cyano-6-methane {6- [4- (3-fluoro-benzyl) -p perazi n-1-yl] -pi ridi n-3-yl} -amide 4-phenyl-3-carboxylic acid, {6- [4- (2-trifluoromethox-benzyl) -plperaz-1 -i |] -pyridin-3-α-4-acid acid amide -c¡ano-6met¡lb¡phenyl-3-carboxylic, {6- [4- (3-chloro-benzyl) -p, eg razin-1 -i I] 20 pyrid i n-3-yl} -am outflow of 4'-cyano-6-methyl-phenyl-3-carboxylic acid, {6 [4- (4-¡butbut¡l-benc¡l) -piperaz¡n-1-¡l] -p¡r¡d¡n-3-¡l} -am¡da of 4'ciano acid -6-methyl-biphenyl-3-carboxylic acid, {6- [4- (4-terbuti l-be nci l) -pi perazin-1-l] -pyridin-3-yl} -am'da acid 4'- cyano-6-methyl-biphenyl-3-carboxylic acid, {6- [4- (7-methoxy-benzo- [1,3] -d ¡oxo l-5-yl-methyl) -p¡perazi n-1 25 yl] 4'-cyano-6-methyl-biphenyl-3-carboxylic acid -pyrn-3-yl} -amide, [6- (4-benzyl-piperazin-1-iI) -p¡ r ¡d ¡n-3-¡|] -am of 4'-Cano-6-methyl-biphenyl-3-carboxylic acid, 4'-Cyano-6-methyl-b-phenyl-3-carboxylic acid [6- (4-pyridin-3-yl-methyl-piperazin-1-II) pyridine-3-yl] -amide , {6 [4- (4-d¡fluoro-methox¡-benzyl) -p¡peraz¡n-1 - l] -p¡ridin-3-yl} -am'da acid 4'- cano-6-methyl-phenyl-3-carboxylic acid, {6- [4- (4-cano-benzyl) plperazln-1 - i I] -pi rid ¡η-3-il} -am of the 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, [6- (4-what not I n-5-lm eti l-pipe reason n-1 - 1) 4'-Cyano-6-methyl-b-phenyl-3-carboxylic acid -pyrid-3-yl] -am¡da, [6- (4-pir¡d¡n-4-¡-met¡lp¡peraz¡n-1 -il) -p¡rid¡η-3-il] -am¡da of 4'-cyano acid -6-methyl-biphen¡l-310 carboxylic, [6- (4-pi rid i η-2-i lm eti l-pipe razi n-1 -il) -pir¡d¡n-3-¡l] -amide of 4'-cyano-6-methyl-biphenyl-3-carboxylic acid, {6- [4- (4-midazol-1-ylbe nc I) -pi pe raz ¡N-1 - il] -p ¡rid I η-3-il} -am id a of 4'-c¡ano-6-met¡lb¡fen¡l-3-carboxylic acid, {6- 4'-Cano-6-methal-acid-4-yl-benzyl-p-perazin-1 -i |] -pyridin-3-l | 3-carboxylic, [6- (415 soqu¡nol¡n-5-¡-me¡l-piperaz¡n-1 -il) -p¡rid¡η-3-il] -am¡da of 4'ciano- 6-methyl-b¡fen¡l-3-carboxylic, (R) -2-met¡lN- (6- (2-methylmorphol) -pyrn-3-¡l) -4 '- (trfluoro-methox¡) -b¡fen¡l-3-carboxam¡da, 4'c¡ano-2-met¡lN- (6-sulfonyl-morpholno-pyridin-3-l ) -b¡feriil-3carboxamlda, (S) -4'-cyano-2-methyl¡- (6- (2-methyl-morphol) -p¡r¡d¡n-320¡I) -b ¡¡¡¡¡¡¡L-3-Carboxa mida, (R) -6-Chloro-N- (6- (2-methyl-morphol) -pyridin-3-yl) -4 '- (trfluoro-methoxy) - biphenol-3-carboxamide, 4'-clano-2-methyl-N- (6-sulfulf-morphol-pyrid-3-yl) -b-phen-3-carboxamide, 4'-cyano-N- (6 - (d¡sobutyl-amino) -pyrin-3-yl) -2-methyl-biphenyl-3-carboxamide, 4'-cyano-N (2 - ((2 S, 6R) -2, 6-d ¡meí l-mo rfol¡ no) -piri m id ¡n-5-il) -2-met¡ I-bife or I-325 carboxamide, N- (2 - ((2S, 6R) -2 , 6-dlmet¡l-morfol¡no) -p¡rim¡din-5-¡l) -219 met¡l-4^,- (trifluoro-methyl) -biphenyl-3-carboxamide, N- (2 - ((2S, 6R) -2,6d-methyl-morpholino) -pyrimidin-5-yl) -2-methyl-4^,- (trifluoro-methoxy) -biphenyl-3carboxamide, N- (2- (bis (2-hydroxy-ethyl) -amino) -pyrimin-5-yl) -2-methyl4 '- (trifluoro-methox¡ ) -b¡phenyl-3-carboxam¡da, 2-methyl-N- (6- (tetrahydro5 2H-pyran-4-ylox¡) -pyrn-3-yl) -4 '- (trifluoro- methoxy) -biphenyl-3carboxamide, N- (5-chloro-6 - ((2S, 6R) -2,6-dimethyl-morpholino) -pyridin-3-yl) -2-methyl-4^,- (trifluoro-methox¡) -biphenyl-3-carboxamide, N- (6 - ((2R, 6S) 2,6-dimethyl-tetrahydro-2H-pyran-4-yl) -p¡r¡din- 3-¡) -2-methyl-4 '- (trifluoromethoxy) -biphenyl-3-carboxamide, N- (6- (4-ethyl-piperazin-1-carbon I) 10 p¡rid¡ n-3-yl) -2-methyl-4 '- (trifluoro-methoxy) -biphenyl-3-carboxamide, 2-methyl-N- (6- (2-oxopiperazin-1-yl) -pyridin-3- il) -4^,- (trifluoro-methox¡) biphenyl-3-carboxamide, 2-methyl- (6- (1 - (pyridin-4-i immeti 1) -piperidine-4l) -pyridin-3-yl )-4^,- (trifluoro-methoxy) -b¡fen¡l-3-carboxamide, 2-methyl N (6- (2-oxo-4- (pyrdin-4-ylmethyl) -p¡perazin-1 -i I) -pi rid i n-3-iI) -4 '- (trifI or gold 15 methoxy) -biphenyl-3-carboxamide, 2-methyl-N- (6- (1- (pyridin-4-ylmethyl) piperidin-3- il) -pyridin-3-yl) -4 '- (trifluoro-methoxy) -b¡fen¡l-3-carboxamide, N-íe-O-ethyl-piperidin-S-iO-pyridin-S-iO- Z-methyl ^ '- ^ rifluoro-methoxy) biphenyl-3-carboxamide, and N- (6 - ((2R, 6S) -2,6-dimethyl-morphol) -pridrid-3-yl) - 2-methyl-4 '- (trifluoro-methoxy) -biphenyl-3-carboxamide.

Therefore, it is specifically contemplated that compounds of Formula I that interfere with aspects of Hedgehog, Ptc, or Smoothened signal transduction activity, in the same way will be able to inhibit proliferation (or other biological consequences) in normal cells, and / or in cells that have a Patched function loss phenotype, a Hedgehog function gain phenotype, a Smoothened function gain phenotype, or a function gain phenotype of Gli. Accordingly, it is contemplated that, in certain embodiments, these compounds may be useful for inhibiting Hedgehog activity in normal cells, for example that they do not have a genetic mutation that activates the Hedgehog path. In preferred embodiments, the compounds are capable of inhibiting at least some of the biological activities of Hedgehog proteins, preferably in a specific manner in the target cells.

Therefore, the methods of the present invention include the use of compounds of Formula I that agonize the inhibition of Ptc from Hedgehog signaling, such as by inhibiting the activation of Smoothened components or downstream of the path. signal, in the regulation of repair and / or functional performance of a large number of cells, tissues, and organs, including normal cells, tissues, and organs, as well as those with the Ptc loss of function phenotype, Hedgehog function gain, Smoothened function gain, or Gli function gain. For example, the present method has therapeutic and cosmetic applications from the regulation of neural tissues, the formation and repair of bone and cartilage, the regulation of spermatogenesis, the regulation of smooth muscle, the regulation of the lung, liver and other organs that occur from the primitive intestine, the regulation of hematopoietic function, the regulation of skin and hair growth, etc. Moreover, the present methods can be carried out in cells that are provided in culture (in vitro), or in cells in an entire animal (in vivo).

In another embodiment, the present method may be to treat epithelial cells that have a phenotype of loss of Ptc function, gain of Hedgehog function, gain of Smoothened function, or gain of Gli function. For example, the present method can be used for the treatment or prevention of basal cell carcinoma or other disorders related to the Hedgehog path.

In certain embodiments, a compound of Formula I can inhibit the activation of a Hedgehog path by linking with Smoothened proteins or downstream. In certain embodiments, an object antagonist can inhibit the activation of a Hedgehog path by linking with a Patched.

In another preferred embodiment, the present method can be used as part of a treatment regimen for malignant medulloblastomas and other primary malignant neuroectodermal tumors of the central nervous system.

In another aspect, the present invention provides pharmaceutical preparations comprising, as an active ingredient, a Hedgehog signaling modulator, such as a compound of Formula I, a Ptc agonist, a Smoothened antagonist, or an antagonist of the path protein

Hedgehog downstream, as described herein, formulated in an amount sufficient to inhibit, in vivo, proliferation or other biological consequences of loss of Ptc function, Hedgehog function gain, Smoothened function gain , or the Gli function gain.

The present treatments using a compound of Formula I, Patched agonists, Smoothened antagonists, or downstream Hedgehog protein antagonists may be effective for both human and animal subjects. The animal subjects to which the present invention is applicable extend to both domestic animals and livestock, or to which it is raised as a pet or for commercial purposes. Examples are dogs, cats, cattle, horses, sheep, pigs, and goats.

Pharmacology and Utility

The present invention makes available methods and compounds to inhibit the activation of the Hedgehog signaling path, for example to inhibit aberrant growth states resulting from phenotypes such as loss of Ptc function, Hedgehog function gain, function gain Smoothened, or Gli function gain, which comprise contacting the cell with a compound of Formula I, in an amount sufficient to agonize a normal Ptc activity, antagonize a normal Hedgehog activity, antagonize a Smoothened activity, or antagonize a Gli activity, for example to reverse or control the aberrant growth state.

Hedgehog signaling members mediate many short and long range pattern formation processes during vertebrate development. Pattern formation is the activity by which embryonic cells form ordered spatial arrangements of differentiated tissues. The physical complexity of higher organisms occurs during embryogenesis through the intrinsic lineage of the cell and the extrinsic signaling of the cell. Inductive interactions are essential for the formation of embryonic patterns in the development of vertebrates from the earliest establishment of the body plan, to the formation of patterns of organ systems, and to the generation of various types of cells during differentiation. of the tissues. The effects of developmental cell interactions are varied: the responding cells are diverted from one cell differentiation path to another by inducing the cells that differ from both the uninducted and induced state of the responding cells (inductions). Sometimes, cells induce their neighbors to differentiate themselves (homeogenetic induction); In other cases, a cell inhibits its neighbors from differentiation like itself. Cellular interactions in early development can be sequential, such that an initial induction between two cell types leads to a progressive amplification of diversity. Moreover, inductive interactions occur not only in embryos, but also in adult cells, and can act to establish and maintain morphogenetic patterns, as well as to induce differentiation.

The vertebrate Hedgehog family of genes includes three members that exist in mammals, known as the Hedgehogs Desert (Dhh), Sonic (Shh), and Indian (Ihh), all of which encode secreted proteins. These different Hedgehog proteins consist of a signal peptide, a highly conserved N-terminal region, and a more divergent C-terminal domain. Biochemical studies have shown that Hh precursor protein auto-proteolytic cleavage proceeds through an internal thioester intermediate that subsequently dissociates into a nucleophilic substitution. It is likely that the nucleophile is a small lipophilic molecule that covalently bonds with the C-terminal end of the N-peptide, attaching it to the cell surface. The biological implications are profound. As a result of this binding, a high local concentration of the N-terminal Hedgehog peptide is generated on the surface of the Hedgehog producing cells. It is this N-terminal peptide that is both necessary and sufficient for Hedgehog signaling activities of short and long range.

An inactive Hedgehog signaling path is where the Patched transmembrane protein receptor (Ptc) inhibits the activity of Smoothened (Smo), a seven transmembrane protein. The transcription factor Gli, a downstream component of Hh signaling, is prevented from entering the nucleus through interactions with cytoplasmic proteins, including Fused and Fused Supressor (Sufu). As a consequence, the activation of transcription of Hedgehog target genes is repressed. Path activation is initiated through the binding of any of the three mammalian ligands (Dhh, Shh, or Ihh) with Ptc. The ligand binding results in the reversal of Smo repression, thereby activating a cascade that leads to translocation of the active form of the transcription factor Gli to the nucleus. The nuclear Gil activates the expression of the target gene, including Ptc and Gil themselves.

The highest levels of Hedgehog signaling are sufficient to initiate cancer formation, and are required for tumor survival. These cancers include, but are not limited to, prostate cancer ("Hedgehog signaling in prostate regeneration, neoplasia and metastasis", Karhadkar SS, Bova GS, Abdallah N, Dhara S, Gardner D, Maitra A, Isaacs JT, Berman DM, Beachy PA, Nature. 7 October 2004; 431 (7009): 707-12; “Inhibition of prostate cancer proliferation by interference with SONIC HEDGEHOG-GL11 signaling”, Sánchez P, Hernández AM, Stecca B, Kahler AJ, DeGueme AM, Barrett A, Beyna M, Datta MW, Datta S., Ruiz i Altaba A., Proc. Nati Acad. Sci. USA. August 24, 2004; 101 (34): 12561-6), breast cancer (“Hedgehog signaling pathway is a new therapeutic target for patients with breast cancer”,

Kubo M, Nakamura M, Tasaki A, Yamanaka N, Nakashima H, Nomura

M, Kuroki S, Katano M., Cancer Res. September 1, 2004;

64 (17): 6071-4), medulloblastoma ("Medulloblastoma growth inhibition by hedgehog pathway blockade", Berman DM, Karhadkar SS,

Hallahan AR, Pritchard JI, Eberhart CG, Watkins DN, Chen JK, Cooper Μ. K., Taipale J, Olson JM, Beachy PA, Science. August 30, 2002; 297 (5586): 1559-61), basal cell carcinoma ("Identification of a small molecule inhibitor of the hedgehog signaling pathway: effects on basal cell carcinoma-like lesions", Williams JA, Guicherit OM, Zaharian Β. I. , Xu Y, Chai L, Wichterle H, Kon C, Gatchalian C, Porter JA, Rubin LL, Wang FY, Proc. Nati. Acad. Sci. USA. fifteen April 2003; 100 (8): 4616-21; "Activating Smoothened mutations in sporadic basal-cell carcinoma", Xie J, Murone M, Luoh SM, Ryan A, Gu Q, Zhang C, Bonifas JM, Lam C. W, Hynes M, Goddard A, Rosenthal A, Epstein Ε. H. Jr., from Sauvage FJ, Nature. January 1, 1998; 391 (6662): 90-2), pancreatic cancer (“Hedgehog is an early and late mediator of pancreatic cancer tumorigenesis”, Thayer SP, di Magliano P. P., Heiser PW, Nielsen CM, Roberts DJ, Lauwers GY, Qi Y. P., Gysin S, Fernandez-del Castillo C, Yajnik V, Antoniu B, McMahon M, Warshaw AL, Hebrok M., Nature. October 23, 2003; 425 (6960): 851-6; “Widespread requirement for Hedgehog ligand stimulation in growth of digestive tract tumors”, Berman DM, Karhadkar SS, Maitra A, Montes De Oca R, Gerstenblith MR, Briggs K, Parker AR, Shimada Y, Eshleman JR, Watkins DN, Beachy PA, Nature 2. 3 October 2003; 425 (6960): 846-51), and small cell lung cancer ("Hedgehog signalllng within airway epithelial progenitors and in small-cell lung cancer", Watkins D. N, Berman D.

M., Burkholder SG, Wang B, Beachy PA, Baylln SB, Nature. March 20, 2003; 422 (6929): 313-7).

In accordance with the foregoing, the present invention further provides a method for preventing or treating any of the diseases or disorders described above, in a subject in need of such treatment, whose method comprises administering to this subject a therapeutically effective amount ("Administration and Pharmaceutical Compositions ”, infra) of a compound of Formula I, or a pharmaceutically acceptable salt thereof. For any of the above uses, the required dosage will vary depending on the mode of administration, the particular condition to be treated, and the desired effect.

Administration and Pharmaceutical Compositions:

In general, the compounds of the invention will be administered in therapeutically effective amounts by any of the usual and acceptable ways known in the art, either alone or in combination with one or more therapeutic agents. A therapeutically effective amount can vary widely, depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. In general, it is indicated that systemically satisfactory results are obtained with daily dosages of approximately 0.03 to 2.5 milligrams / kilogram of body weight. A daily dosage indicated in the upper mammal, for example in humans, is in the range of about 0.5 milligrams to about 100 milligrams, conveniently administered, for example, in divided doses up to four times a day, or in a delayed form. Unit dosage forms suitable for oral administration comprise about 1 to 50 milligrams of active ingredient.

The compounds of the invention can be administered as pharmaceutical compositions by any conventional route, in particular enterally, for example orally, for example in the form of tablets or capsules, or parenterally, for example in the form of injectable solutions or suspensions, topically, for example in the form of lotions, gels, ointments or creams, or in a nasal or suppository form. Pharmaceutical compositions comprising a compound of the present invention in free form or in the form of a pharmaceutically acceptable salt, in association with at least one pharmaceutically acceptable carrier or diluent, can be manufactured in a conventional manner by mixing, granulating, or coating methods. . For example, the oral compositions may be gelatin tablets or capsules comprising the active ingredient together with: a) diluents, for example lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine; b) lubricants, for example silica, talc, stearic acid, its magnesium or calcium salt, and / or polyethylene glycol; for tablets also c) binders, for example magnesium aluminum silicate, starch paste, gelatin, tragacanth, methyl cellulose, sodium carboxy methyl cellulose, and / or polyvinyl pyrrolidone; if desired d) disintegrants, for example starches, agar, alginic acid or its sodium salt, or effervescent mixtures; and / or e) adsorbents, colorants, flavorings, and sweeteners. Injectable compositions can be aqueous sotonic solutions or suspensions, and suppositories can be prepared from fatty emulsions or suspensions. The compositions may be sterilized and / or may contain adjuvants, such as preservatives, stabilizers, humectants, or emulsifiers, solution promoters, salts for regulating osmotic pressure, and / or pH regulators. In addition, they may also contain other therapeutically valuable substances. Formulations suitable for transdermal applications include an effective amount of a compound of the present invention with a vehicle. A vehicle may include absorbable pharmacologically acceptable solvents to aid passage through the skin of the host. For example, the transdermal devices are in the form of a patch comprising a backing member, a reservoir containing the compound optionally with vehicles, optionally a speed control barrier to deliver the compound in the host's skin at a controlled speed and previously determined for a prolonged period of time, and elements to secure the device to the skin. Transdermal matrix formulations can also be used. Formulations suitable for topical administration, for example to the skin and eyes, are preferably aqueous solutions, ointments, creams, or gels, well known in this field. These may contain solubilizers, stabilizers, tonicity enhancing agents, pH regulators, and preservatives.

The compounds of the invention can be administered in therapeutically effective amounts in combination with one or more therapeutic agents (pharmaceutical combinations). For example, synergistic effects with immunomodulatory or anti-inflammatory substances or other anti-tumor therapeutic agents may occur. When the compounds of the invention are co-administered with other therapies, the dosages of the co-administered compounds will, of course, vary depending on the type of co-drug used, the specific drug used, the condition being treated, etc. .

The invention also provides pharmaceutical combinations, for example a kit, which comprises: a) a first agent that is a compound of the invention as disclosed herein, in free form or in the form of a pharmaceutically acceptable salt, and b) At least one co-agent. The kit may include instructions for administration.

The terms "co-administration" or "combined administration", or the like, as used herein, are intended to encompass the administration of selected therapeutic agents to a single patient, and are intended to include treatment regimens where the agents are not necessarily administer by the same route of administration or at the same time.

The term "pharmaceutical combination, as used herein, means a product that results from the mixing or combination of more than one active ingredient, and includes both fixed and non-fixed combinations of the Active Ingredients. The term "fixed combination" means that the active ingredients, for example a compound of Formula I and a co-agent, are both administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that the active ingredients, for example a compound of Formula I and a co-agent, are both administered to a patient as separate entities, either concurrently, concurrently, or in sequence, no specific time limits, where this administration provides therapeutically effective levels of the two compounds in the patient's body. The latter also applies to cocktail therapy, for example the administration of three or more active ingredients.

Processes to Prepare the Compounds of the Invention

The present invention also includes processes for the preparation of the compounds of the invention. In the reactions described, it may be necessary to protect the reactive functional groups, for example hydroxyl, amino, imino, thio, or carboxyl groups, where these are desired in the final product, in order to avoid their unwanted participation in the reactions. Conventional protecting groups can be used in accordance with standard practice, for example, see TW Greene and PGM Wuts in "Protective Groups in Organic Chemistry", John Wiley and Sons,

1991.

The compounds of Formula I can be prepared by proceeding as in the following Reaction Scheme I:

Reaction Scheme I

<img file="CU23760B7_D0002.tif" />

where and_b Y₂, Ri, R₂, R₃, R₄, Rs, Re, R7, Rs, and R9 are as defined for Formula I in the Brief Description of the Invention. A compound of Formula I may be prepared by reacting a compound of Formula 2 (or 2 ') with a compound of Formula 3 in the presence of a suitable solvent (eg, dichloromethane, N, N- dimethyl formamide, or the like), in a temperature range of about -20 ° C to about 100 ° C. The reaction may take up to about 20 hours to complete.

Detailed examples of the synthesis of the compounds of Formula I can be found in the Examples below.

Additional Processes for the Preparation of the Compounds of the Invention

A compound of the invention can be prepared as a pharmaceutically acceptable acid addition salt, by reacting the free base form of the compound with a pharmaceutically acceptable inorganic or organic acid. Alternatively, a pharmaceutically acceptable base addition salt of a compound of the invention can be prepared by reacting the free acid form of the compound with a pharmaceutically acceptable inorganic or organic base.

Alternatively, the salt forms of the compounds of the invention can be prepared using the salts of the starting materials or Intermediaries.

The free acid or free base forms of the compounds of the invention can be prepared from the form of base addition salt or corresponding acid addition salt, respectively. For example, a compound of the invention in the form of an acid addition salt can be converted to the corresponding free base by treatment with a suitable base (for example, a solution of ammonium hydroxide, sodium hydroxide, and the like). A compound of the invention in the form of a base addition salt can be converted to the corresponding free acid by treatment with a suitable acid (for example, hydrochloric acid, etc.).

The compounds of the invention in a non-oxidized form can be prepared from the N-oxides of the compounds of the invention, by treatment with a reducing agent (for example, sulfur, sulfur dioxide, trife or l-phosphorus). ina, lithium borohydride, sodium borohydride, phosphorus trichloride, tribromide, or the like) in a suitable inert organic solvent (eg, acetonitrile, ethanol, aqueous dioxane, or the like), from 0 ° C to 80 ° C.

The pro-drug derivatives of the compounds of the invention can be prepared by methods known to those of ordinary skill in the art (for example, for additional details, see Saulnler et al. (1994), Bioorganic and Medicinal Chemistry Letters, Volume 4 , page 1985). For example, the appropriate pro-drugs can be prepared by reacting a compound not derived from the invention with a suitable carbamilating agent (for example, 1,1-acyloxy-alkyl carbonate hydrochloride, para-nitro-phenyl carbonate, or Similar).

Protected derivatives of the compounds of the invention can be made by means known to those of ordinary skill in this field. A detailed description of the techniques applicable to the creation of protective groups and their removal can be found in

TW Greene, “Protecting Groups ¡n Organic Chemistry”, 3^to Edition, John Wiley and Sons, Inc., 1999.

The compounds of the present invention may conveniently be prepared or formed during the process of the invention, as solvates (eg, hydrates). The hydrates of the compounds of the present invention can be conveniently prepared by recrystallization from a mixture of aqueous / organic solvents, using organic solvents such as dioxin, tetrahydrofuran, or methanol.

The compounds of the invention can be prepared as their individual stereoisomers by reacting a racemic mixture of the compound with an optically active resolving agent, to form a pair of diastereoisomeric compounds, separating the diastereomers, and recovering the optically pure enantiomers. Although resolution of the enantiomers can be carried out using divalent diastereomeric derivatives of the compounds of the invention, dissociable complexes (for example, crystalline diastereomeric salts) are preferred. The diastereomers have different physical properties (for example, melting points, boiling points, solubilities, reactivity, etc.), and can be easily separated by taking advantage of these differences. The diastereomers can be separated by chromatography, or preferably by separation / resolution techniques based on differences in solubility. The optically pure enantiomer is then recovered, together with the resolution agent, by any practical means that does not result in racemization. A more detailed description of the techniques applicable to the resolution of stereoisomers of the compounds from their racemic mixtures can be found in Jean Jacques, Andre Collet, Samuel H. Wilen, "Enantiomers, Racemates and Resolutions", John Wiley And Sons,

Inc., 1981.

In summary, the compounds of Formula I can be done through a process that involves:

(a) those of Reaction Scheme I; and (b) optionally converting a compound of the invention into a pharmaceutically acceptable salt;

(c) optionally converting a salt form of a compound of the invention to a non-salt form;

(d) optionally converting an non-oxidized form of a compound of the invention into a pharmaceutically acceptable N-oxide;

(e) optionally converting an N-oxide form of a compound of the invention to its non-oxidized form;

(f) optionally resolving an individual isomer of a compound of the invention from a mixture of isomers;

(g) optionally converting a compound not derived from the invention into a pharmaceutically acceptable pro-drug derivative; and (h) optionally converting a pro-drug derivative of a compound of the invention to its non-derivative form.

As far as production of the starting materials is not particularly described, the compounds are known or can be prepared in a manner analogous to the methods known in the art, or as disclosed in the Examples which are found later here. .

One skilled in the art will appreciate that the above transformations are only representative of the methods for the preparation of the compounds of the present invention, and that other well known methods may similarly be employed.

Examples

The present invention is further exemplified, but not limited, by the following Example illustrating the preparation of the compounds of Formula I according to the Invention.

Example 1 4'-cyano-4-methylbipheni 1-3-carboxylic acid r4- (morpholin-4-sulfonyl) -phenyl1-amide

I.

NC

<img file="CU23760B7_D0003.tif" />

H₂SW₄

MeOH

I.

_xOMe

Pd (OAc) 2, ligand KF, dioxane

B (OH)₂

<img file="CU23760B7_D0004.tif" />

<img file="CU23760B7_D0005.tif" />

Et₃N, CH₂CI₂

Example 1

Zn

Pasol: To a solution of 3-iodo-4-methyl-benzoic acid (10.0 grams, 38.2 mmol) in methanol (70 milliliters), concentrated sulfuric acid (0.5 milliliters) is added. The reaction mixture is heated at 70 ° C for 48 hours, cooled to room temperature, and then concentrated. After that, ethyl acetate (100 milliliters), and an aqueous solution of NaHCO are added₃ (saturated, 100 milliliters) to the residue. The organic layer is separated and washed again with an aqueous NaHCO solution.₃ (saturated, 100 milliliters). The organic layer is separated, dried over Na₂SW₄ anhydrous, and concentrated, to provide the 3-iodo-4-methyl-benzolco 1 methyl ester. This is used without further purification in the next step. ¹H NMR (400 MHz, DMSO-d₆) δ 8.31 (s, 1 H), 7.87 (d, 1 H, J = 8.4 Hz), 7.48 (d, 1 H, J = 8.4 Hz), 3.85 (s, 3 H), 3.35 (s, 3H) ; LC-MS m / z: 277.0 (M + 1).

Step 2: To a round bottom flask containing the methyl ester of 3-iodo-4-methyl-benzoic acid (1.38 grams, 5.00 mmol), 4-cyano-phenyl boronic acid (1.10 grams, 7.48 mmol), acetate palladium (168 milligrams, 0.748 millimoles), 2 (dicyclo-hexyl-phosphine) -b¡phenyl (0.526 grams, 1.50 millimoles), and potassium fluoride (0.870 grams, 15.0 millimoles), is added 1,4 anhydrous dioxane (15 milliliters). The flask is purged with argon and sealed. The mixture is stirred at 130 ° C for 18 hours, cooled to room temperature, and then water (20 milliliters) and ethyl acetate (20 milliliters) are added. The solid is removed by vacuum filtration. The filtrate is extracted with EtOAc (20 milliliters, twice). The organic layers are combined, washed with aqueous HCI (5 percent, 20 milliliters), and NaHCO₃ saturated (20 milliliters). They are dried over MgSO₄, and concentrate. The residue is purified by silica gel column chromatography (EtOAc / hexane, gradient), to give the 4'-cyano-6-methyl-biphenyl-3-carboxylic acid methyl ester 2: LC-MS m / z: 252.1 (M + 1).

Step 3: To a solution of the 4'-cyano-6-methyl-biphenyl-3-carboxylic acid methyl ester 2 (2.56 grams, 10.3 mmol) in 1,4-dioxane-H₂Or (mixture of 1: 1, 20 milliliters), NaOH (1.22 grams, 30.2 mmol) is added. The reaction is stirred at room temperature for 24 hours. To this mixture is added aqueous HCI (1N, 36 milliliters), and then extracted with ethyl acetate (140 milliliters, three times). The organic layers are combined, and dried over Na₂SW₄ anhydrous. The solvent is removed. The solid obtained is washed with a small amount of acetonitrile, and air dried, to give 4'-cyano-6-methyl-biphenyl-3-carboxylic acid 3:¹H NMR (DMSO-d₆) δ 7.94 (d, 2 H, J = 8.0 Hz), 7.84 (dd, 1 H, J₁ = 8.4 Hz, J₂= 1.2 Hz), 7.75 (d, 1 H, J = 1.2 Hz), 7.61 (d, 2 H, J = 8.0 Hz), 7.48 (d, 1 H, J = 8.4 Hz), 2.29 (s, 3 H ); LC-MS m / z 238.1 (M + 1).

Step 4: To a suspension of 4'-cyano-6-methyl-biphenyl3-carboxylic acid 3 (40 milligrams, 0.17 mmol) in anhydrous methylene chloride (5 milliliters), 2 drops of dimethylformamide are added. Then oxalyl chloride (32 milligrams, 22 microliters, 0.25 millimoles) is added. The mixture is stirred at room temperature until it becomes transparent. After that, it is concentrated, re-dissolved in anhydrous methylene chloride (3 milliliters), and added to a solution of 4- (morpholin-4-sulfonyl) -phenylamine (61 milligrams, 0.25 mmol) and triethyl amine (34 milligrams, 47 microliters, 0.33 millimoles) in methylene chloride (2 milliliters). The mixture is stirred for 2 hours, concentrated, and the residue is purified by mass directed preparation HPLC (column C18, eluted with CH₃CN-H₂Or containing 0.05 percent trifluoroacetic acid), to give the [4- (morfolln-4-sulfonyl) -fen¡ |] -amide of 4'-cyano-6-methyl-b-phenyl-3 acid -carboxylic: ¹H NMR (DMSO-d₆) δ 10.64 (s, 1 H), 8.07 (d, 2 H, J = 8.8 Hz), 7.97 (d, 2 H, J = 8.4 Hz), 7.95 (d, 1 H, J = 8.8 Hz), 7.89 (s, 1 H), 7.43 (d, 2 H, J = 8.4 Hz), 7.67 (d, 2 H, J = 8.8 Hz), 7.53 (d, 2 H, J = 8.8 Hz), 3.63 (m, 4 H), 2.84 (m, 4 H) 2.32 (s, 3 H); LC-MS m / z: 462.1 (M + 1).

Example 2 4'-Canoano acid f6- (2,6-dimethyl-morphol-4-yl) -pyridin-3-ill-amda₂n

Cl

<img file="CU23760B7_D0006.tif" />

<img file="CU23760B7_D0007.tif" />

<img file="CU23760B7_D0008.tif" />

Example 2

Step 1; To a solution of 2-chloro-5-n -tro-p¡r¡d¡na 4 (2.38 grams, 15 millimoles), and c / s-2,6-dimethyl-morpholine (1.73 grams, 15 millimoles), K is added₂CO₃ (4.14 grams, 30 mmol). The mixture is heated at 50 ° C overnight. After concentration, the residue is divided between EtOAc and water. The EtOAc layer is dried over Na₂SW₄ anhydrous, and concentrated, to give the crude product 6 as a yellow solid. The crude product is used directly in the next step without further purification. LCMS m / z: 238.1 (M + 1).

Step 2: The above crude material 6 is hydrogenated in the presence of Pd-C (0.2 grams) in MeOH (100 milliliters) under hydrogen for 10 hours. The suspension is filtered through Celite, and the filtrate is concentrated, to give the crude product 7 as a dark brown oil, which is used directly in the next step without further purification. LC-MS m / z: 208.1 (M + 1).

Step 3: To a solution of 3-bromo-4-methyl-benzoic acid (108 milligrams, 0.5 millimoles), 6- (2,6-dim et i lm or rfo I i n-4-¡l) -pi r id i n-3-amine 7 (104 milligrams, 0.5 millimoles), and HATU (190 milligrams, 0.5 millimoles) in dry dimethylformamide (5 milliliters), triethyl amine (139 microliters, 1.0 millimoles) is added drip . The resulting mixture is stirred at room temperature for 2 hours. After concentration, the residue is divided between EtOAc and water. The organic layer is dried and concentrated to give the crude product. The final compound is purified by column chromatography by flash evaporation, using 50 percent EtOAc in hexane as eluent, to give 8 as a white solid. LC-MS m / z: 404.1 (M + 1).

Step 4: A mixture of 4-cyano-phenyl-boronic acid (18 milligrams, 0.12 millimoles), 3-bromo-N- [6- (2,6-d-methyl-morpholin-4-yl) 5 p D¡n-3-¡] -4-methyl-benzam¡da 8 (40 milligrams, 0.1 millimoles),

Pd (PPh₃)₄ (11 milligrams, 0.01 millimoles), and Na₂CO₃ (42 milligrams, 0.4 millimoles) in a combined solvent system of toluene (0.2 milliliters) and water (0.2 milliliters) and ethanol (0.05 milliliters), heated at 140 ° C under microwave irradiation for 30 minutes.

The reaction mixture is diluted with EtOAc and water. The aqueous layer is extracted with EtOAc. The combined organic layer is washed with brine and concentrated, to give the crude product, which is purified by mass directed preparation HPLC (column C18, eluted with CH₃CN-H₂Or containing trifluoroacetic acid at

0.05 percent), to give 4'-cyano-6-methyl-b-phenyl acid [6- (2,6-dimethyl-morphoi-4-iI) -p¡rid¡n-3-iI] -3-carboxylic. LC-MS m / z:

427.2 (M + 1).

By repeating the procedures described in the previous Examples, using the appropriate starting materials, the following compounds of Formula I are obtained, as identified in Table 1.

Table 1

Data

Compound

Physical Structure

Number

MS (m / z)

<img file="CU23760B7_D0009.tif" />

<img file="CU23760B7_D0010.tif" />

<img file="CU23760B7_D0011.tif" />

<img file="CU23760B7_D0012.tif" />

LC-MS m / z

411.2 (M + 1).

LC-MS m / z

416.2 (M + 1).

LC-MS m / z

400.2 (M + 1).

LC-MS m / z

418.2 (M + 1).

LC-MS m / z

413.2 (M + 1).

Compound

Number

Data

Physical Structure

MS (m / z)

<img file="CU23760B7_D0013.tif" />

<img file="CU23760B7_D0014.tif" />

<img file="CU23760B7_D0015.tif" />

<img file="CU23760B7_D0016.tif" />

LC-MS m / z

416.2 (M + 1).

LC-MS m / z

451.2 (M + 1).

LC-MS m / z

437.2 (M + 1).

LC-MS m / z

449.2 (M + 1).

Compound

Number

Structure

<img file="CU23760B7_D0017.tif" />

NO - OR

<img file="CU23760B7_D0018.tif" />

Data

MS physicists (m / z)

LC-MS m / z

438.2 (M + 1).

LC-MS m / z

438.2 (M + 1).

LC-MS m / z

436.2 (M + 1).

LC-MS m / z

424.1 (M + 1).

LC-MS m / z

404.2 (M + 1).

Compound.,. Structure

Number

Data

MS physicists (m / z)

18

<img file="CU23760B7_D0019.tif" />

LC-MS m / z

418.2 (M + 1).

LC-MS m / z

418.2 (M + 1).

LC-MS m / z

431.2 (M + 1).

<img file="CU23760B7_D0020.tif" />

LC-MS m / z

431.2 (M + 1).

<img file="CU23760B7_D0021.tif" />

<img file="CU23760B7_D0022.tif" />

LC-MS m / z

417.2 (M + 1).

Ό

Compound

Number

Structure

<img file="CU23760B7_D0023.tif" />

<img file="CU23760B7_D0024.tif" />

<img file="CU23760B7_D0025.tif" />

MS Physical Data (m / z)

LC-MS m / z

416.2 (M + 1).

LC-MS m / z

430.2 (M + 1).

LC-MS m / z

432.2 (M + 1).

LC-MS m / z

429.2 (M + 1).

LC-MS m / z

462.2 (M + 1).

Compound

Number

Structure

<img file="CU23760B7_D0026.tif" />

<img file="CU23760B7_D0027.tif" />

<img file="CU23760B7_D0028.tif" />

<img file="CU23760B7_D0029.tif" />

Data

MS physicists (m / z)

LC-MS m / z

454.1 (M + 1).

LC-MS m / z

420.2 (M + 1).

LC-MS m / z

420.2 (M + 1).

LC-MS m / z

420.2 (M + 1).

LC-MS m / z

454.2 (M + 1).

LC-MS m / z

488.1 (M + 1).

Compound

Structure

Number

<img file="CU23760B7_D0030.tif" />

Data

MS physicists (m / z)

LC-MS m / z

400.2 (M + 1).

LC-MS m / z

414.2 (M + 1).

<img file="CU23760B7_D0031.tif" />

<img file="CU23760B7_D0032.tif" />

<img file="CU23760B7_D0033.tif" />

LC-MS m / z

442.2 (M + 1).

LC-MS m / z

428.2 (M + 1).

LC-MS m / z

442.2 (M + 1).

Data

Compound

Physical Structure

Number

MS (m / z)

<img file="CU23760B7_D0034.tif" />

<img file="CU23760B7_D0035.tif" />

LC-MS m / z

428.2 (M + 1).

LC-MS m / z

414.2 (M + 1).

<img file="CU23760B7_D0036.tif" />

LC-MS m / z

414.2 (M + 1).

LC-MS m / z

454.2 (M + 1).

LC-MS m / z

454.2 (M + 1).

<img file="CU23760B7_D0037.tif" />

F

LC-MS m / z

522.2 (M + 1).

F

Compound

Structure

Number

Data

MS physicists (m / z)

<img file="CU23760B7_D0038.tif" />

<img file="CU23760B7_D0039.tif" />

<img file="CU23760B7_D0040.tif" />

F

<img file="CU23760B7_D0041.tif" />

N

H

<img file="CU23760B7_D0042.tif" />

LC-MS m / z

444.2 (M + 1).

LC-MS m / z

430.2 (M + 1).

LC-MS m / z

446.2 (M + 1).

LC-MS m / z

470.2 (M + 1).

LC-MS m / z

470.2 (M + 1).

<img file="CU23760B7_D0043.tif" />

373.2

Compound

Number

Structure

<img file="CU23760B7_D0044.tif" />

<img file="CU23760B7_D0045.tif" />

<img file="CU23760B7_D0046.tif" />

Data

MS physicists (m / z) (M + 1).

LC-MS m / z

403.2 (M + 1).

LC-MS m / z

403.2 (M + 1).

LC-MS m / z

460.2 (M + 1).

<img file="CU23760B7_D0047.tif" />

LC-MS m / z

416.2 (M + 1).

LC-MS m / z

391.2 (M + 1).

LC-MS m / z

391.2 (M + 1).

Compound

Structure

Number

<img file="CU23760B7_D0048.tif" />

<img file="CU23760B7_D0049.tif" />

N

<img file="CU23760B7_D0050.tif" />

xx

V^N^

XX

<img file="CU23760B7_D0051.tif" />

OR

MS Physical Data (m / z)

LC-MS m / z

391.2 (M + 1).

LC-MS m / z

425.2 (M + 1).

LC-MS m / z

471.2 (M + 1).

LC-MS m / z

398.2 (M + 1).

LC-MS m / z

398.2 (M + 1).

Compound

Number

<img file="CU23760B7_D0052.tif" />

<img file="CU23760B7_D0053.tif" />

MS Physical Data (m / z)

LC-MS m / z 413.2 (M + 1)

LC-MS m / z

411.2 (M + 1).

LC-MS m / z 413.2 (M + 1)

LC-MS m / z

397.2 (M + 1)

LC-MS m / z 399.2 (M + 1)

LC-MS m / z 412.2 (M + 1).

Compound

Structure

Number

<img file="CU23760B7_D0054.tif" />

<img file="CU23760B7_D0055.tif" />

<img file="CU23760B7_D0056.tif" />

<img file="CU23760B7_D0057.tif" />

<img file="CU23760B7_D0058.tif" />

Data

MS physicists (m / z)

LC-MS m / z

398.2 (M + 1).

LC-MS m / z

416.2 (M + 1).

LC-MS m / z

432.1 (M + 1).

LC-MS m / z

476.1 (M + 1).

LC-MS m / z

412.2 (M + 1).

LC-MS m / z

466.2 (M + 1).

Compound

Structure

Number

<img file="CU23760B7_D0059.tif" />

Data

MS physicists (m / z)

LC-MS m / z

385.2 (M + 1).

LC-MS m / z

389.1 (M + 1).

<img file="CU23760B7_D0060.tif" />

LC-MS m / z

419.2 (M + 1).

<img file="CU23760B7_D0061.tif" />

LC-MS m / z

487.2 (M + 1).

<img file="CU23760B7_D0062.tif" />

<img file="CU23760B7_D0063.tif" />

LC-MS m / z

460.2 (M + 1).

Compound

Structure

Number

<img file="CU23760B7_D0064.tif" />

<img file="CU23760B7_D0065.tif" />

<img file="CU23760B7_D0066.tif" />

Data

MS physicists (m / z)

LC-MS m / z

446.1 (M + 1).

LC-MS m / z

427.2 (M + 1).

LC-MS m / z

427.2 (M + 1)

LC-MS m / z

411.2 (M + 1)

LC-MS m / z

434.2 (M + 1)

Compound

Number

Structure

Data

MS physicists (m / z)

<img file="CU23760B7_D0067.tif" />

LC-MS m / z

444.3 (M + 1)

LC-MS m / z

458.3 (M + 1)

<img file="CU23760B7_D0068.tif" />

LC-MS m / z

450.2 (M + 1)

LC-MS m / z

430.2 (M + 1)

<img file="CU23760B7_D0069.tif" />

LC-MS m / z

460.2 (M + 1)

Compound

Number

Data

Physical Structure

MS (m / z)

<img file="CU23760B7_D0070.tif" />

<img file="CU23760B7_D0071.tif" />

LC-MS m / z

464.1 (M + 1)

LC-MS m / z

524.1 (M + 1)

LC-MS m / z

502.3 (M + 1)

LC-MS m / z

508.2 (M + 1)

<img file="CU23760B7_D0072.tif" />

OR

OR

427.2 (M + 1)

Data

Compound

Physical Structure

Number

MS (m / z)

<img file="CU23760B7_D0073.tif" />

LC-MS m / z

432.2 (M + 1)

<img file="CU23760B7_D0074.tif" />

LC-MS m / z

470.2 (M + 1)

LC-MS m / z

486.2 (M + 1)

<img file="CU23760B7_D0075.tif" />

LC-MS m / z

413.2 (M + 1)

<img file="CU23760B7_D0076.tif" />

LC-MS m / z

464.1 (M + 1)

Data

Compound

Physical Structure

Number

MS (m / z)

100

<img file="CU23760B7_D0077.tif" />

LC-MS m / z

514.1 (M + 1)

LC-MS m / z

503.3 (M + 1)

LC-MS m / z

503.3 (M + 1)

101

<img file="CU23760B7_D0078.tif" />

LC-MS m / z

562.3 (M + 1)

102

<img file="CU23760B7_D0079.tif" />

OR

LC-MS m / z

546.3 (M + 1)

Data

Compound

Physical Structure

Number

MS (m / z)

103

104

105

<img file="CU23760B7_D0080.tif" />

LC-MS m / z

600.3 (M + 1)

LC-MS m / z

560.3 (M + 1)

LC-MS m / z

560.3 (M + 1)

<img file="CU23760B7_D0081.tif" />

LC-MS m / z

503.3 (M + 1)

106

Data

Compound

Physical Structure

Number

MS (m / z)

107

108

109

<img file="CU23760B7_D0082.tif" />

LC-MS m / z

546.2 (M + 1)

LC-MS m / z

586.2 (M + 1)

LC-MS m / z

545.3 (M + 1)

LC-MS m / z

604.2 (M + 1)

F

110

Data

Compound

Physical Structure

Number

MS (m / z)

111

112

<img file="CU23760B7_D0083.tif" />

<img file="CU23760B7_D0084.tif" />

<img file="CU23760B7_D0085.tif" />

LC-MS m / z

538.2 (M + 1)

LC-MS m / z

554.2 (M + 1)

<img file="CU23760B7_D0086.tif" />

LC-MS m / z

538.2 (M + 1)

113

Compound

Structure

Number

Data

MS physicists (m / z)

<img file="CU23760B7_D0087.tif" />

LC-MS m / z

480.1 (M + 1)

LC-MS m / z

481.2 (M + 1)

LC-MS m / z

447.2 (M + 1)

Compound

Number

Structure

Data

MS physicists (m / z)

<img file="CU23760B7_D0088.tif" />

LC-MS m / z

441.2 (M + 1)

LC-MS m / z 506.2 (M + 1)

LC-MS m / z 572.2 (M + 1)

LC-MS m / z 522.2 (M + 1)

OR

Data

Compound

Physical Structure

Number

MS (m / z)

121

<img file="CU23760B7_D0089.tif" />

<img file="CU23760B7_D0090.tif" />

LC-MS m / z

544.3 (M + 1)

122

123

<img file="CU23760B7_D0091.tif" />

LC-MS m / z

544.3 (M + 1)

LC-MS m / z

562.2 (M + 1)

124

<img file="CU23760B7_D0092.tif" />

LC-MS m / z

488.2 (M + 1)

<img file="CU23760B7_D0093.tif" />

OR

Data

Compound

Physical Structure

Number

MS (m / z)

126

<img file="CU23760B7_D0094.tif" />

<img file="CU23760B7_D0095.tif" />

F

LC-MS m / z

554.2 (M + 1)

127

128

<img file="CU23760B7_D0096.tif" />

LC-MS m / z

513.2 (M + 1)

LC-MS m / z

539.3 (M + 1)

129

<img file="CU23760B7_D0097.tif" />

/=^Nx // ^ N

LC-MS m / z

489.2 (M + 1)

<img file="CU23760B7_D0098.tif" />

LC-MS m / z

489.2 (M + 1)

130

Data

Compound

Physical Structure

Number

MS (m / z)

<img file="CU23760B7_D0099.tif" />

<img file="CU23760B7_D0100.tif" />

LC-MS m / z

554.3 (M + 1)

132

133

<img file="CU23760B7_D0101.tif" />

LC-MS m / z

513.2 (M + 1)

LC-MS m / z

539.3 (M + 1)

<img file="CU23760B7_D0102.tif" />

LC-MS m / z

472.1 (M + 1)

134

<img file="CU23760B7_D0103.tif" />

OR

Compound

Structure

Number

Data

MS physicists (m / z)

135

<img file="CU23760B7_D0104.tif" />

<img file="CU23760B7_D0105.tif" />

LC-MS m / z

447.1 (M + 1)

136

<img file="CU23760B7_D0106.tif" />

LC-MS m / z

413.1 (M + 1)

<img file="CU23760B7_D0107.tif" />

LC-MS m / z

492.1 (M + 1) \ _, LC-MS m / z

138 \ = / 431.1 (M + 1)

<img file="CU23760B7_D0108.tif" />

OR

Compound

Structure

Number

<img file="CU23760B7_D0109.tif" />

Data

MS physicists (m / z)

LC-MS m / z

441.1 (M + 1)

LC-MS m / z

428.2 (M + 1)

LC-MS m / z

471.2 (M + 1)

LC-MS m / z

487.2 (M + 1)

OR

Data

Compound

Physical Structure

Number

MS (m / z)

143

144

145

146

<img file="CU23760B7_D0110.tif" />

<img file="CU23760B7_D0111.tif" />

<img file="CU23760B7_D0112.tif" />

F

<img file="CU23760B7_D0113.tif" />

LC-MS m / z

477.2 (M + 1)

LC-MS m / z

513.2 (M + 1)

LC-MS m / z

473.2 (M + 1)

LC-MS m / z

520.2 (M + 1)

OR

Compound

Structure

Number

Data

MS physicists (m / z)

147

F

<img file="CU23760B7_D0114.tif" />

LC-MS m / z

445.2 (M + 1)

148

<img file="CU23760B7_D0115.tif" />

LC-MS m / z

471.2 (M + 1)

149

<img file="CU23760B7_D0116.tif" />

LC-MS m / z

547.2 (M + 1)

150

<img file="CU23760B7_D0117.tif" />

LC-MS m / z

562.2 (M + 1)

N

Data

Compound

Physical Structure

Number

MS (m / z)

151

<img file="CU23760B7_D0118.tif" />

LC-MS m / z

547.2 (M + 1)

152

<img file="CU23760B7_D0119.tif" />

LC-MS m / z

484.2 (M + 1)

153

<img file="CU23760B7_D0120.tif" />

LC-MS m / z

486.2 (M + 1)

The compounds of the present invention are tested to assess their ability to inhibit the Hedgehog signaling path.

Gli-Luc Reporter's Essay for the Hh Path Inhibition

TM3 mouse cells (obtained in the American Type

Culture Collection, ATCC, Manassas, VA) are grown in DMEM / F12 medium (Gibco / lnvitrogen, Carlsbad, CA) supplemented with 5 percent heat-inactivated horse serum and 2.5 percent fetal bovine serum (Gibco / lnvitrogen, Carlsbad, CA), 50 units / milliliter of penicillin, and 50 micrograms / milliliter of streptomycin (Gibco / lnvitrogen, Carlsbad, CA) at 37 ° C, with CO₂ 5 percent in an air atmosphere. TM3 cells are transfected with the reporter plasmid pTA-8xGli-Luc. A stably transfected clone called TMHh-12 was selected. The clone

TMHh-12 showed a good response to the Shh-N stimulus. In order to evaluate the CI₅₀s of the antagonists, 8,000 TMHh-12 cells were applied to each well of 384 well plates with 50 percent DMEM / F12 medium supplemented with 2 percent fetal bovine serum. After 12 hours, the Hh path is activated by the addition of the recombinant mouse Shh protein (expressed in E. coli, 8 micrograms / milliliter), or by the addition of Smo agonists. Test compounds are added to the plates in different concentrations. After 48 hours, the firefly luciferase activities are tested with the Bright-Glo Luciferase Test System^MR (Promise, Madison, Wl). IC₅₀ It is measured when the effect of the compound reduces the luminescence signal by 50 percent. The toxicity of these compounds in TM3 cells is evaluated using the CelITiter Glo assays, or through the TM3-Luc cell line (a TM3 cell stably transfected with a constitutive luciferase expression vector).

The compounds of Formula I preferably have an EC₅or less than 500 nM, more preferably less than 200 nM.

Cytotoxicity Test

A cytotoxicity test is performed to compare the effects of a compound of the invention on medulloblastoma cells (Daoy cells), basal cell carcinoma cells (TE354.T cells), and control cells (human fibroblasts normal), according to the following procedure:

Daoy cells (medulloblastoma cell line) are acquired in the ATCC, and grown in a minimal essential medium (Eagle) with 2 mM L-glutamine, and Earle BSS adjusted to contain 1.5 grams / l of sodium bicarbonate, amino acids 0.1 mM non-essential, and 1.0 mM sodium pyruvate, and 10 percent fetal bovine serum, at 37 ° C, with CO₂ 5 percent in an air atmosphere.

TE354.T cells (from ATCC) are cultured in an Eagle medium modified by Dulbecco, with 4 mM L-glutamine, and 10 percent fetal bovine serum.

Normal human dermal fibroblast cells (Clonetics) are grown in a Fibroblast Growth Medium (Clonetics).

Each of the above cell lines is independently sown in 96-well plates, and grown to a density of 5,000 to 10,000 cells / well. A compound of the invention, in different concentrations, is added to cell cultures. After 2 days, cell viability is assessed with the

Cell Titer-Glo Luminescent Cell Viability Test Kit (Promega), following the manufacturer's protocol. Cell viability is measured directly by luminescent signaling, and EC₅₀s are measured when the signal is inhibited by 50 percent.

The compounds of Formula I preferably have an EC₅₀less than 500 nM, more preferably less than 200 nM.

It is understood that the Examples and the modalities described herein are for illustrative purposes only, and that different modifications or changes will be suggested in light thereof for those skilled in the art, and will be included within the spirit and scope of this application, and within the scope of the appended claims. All publications, patents, and patent applications cited herein are incorporated herein by reference for all purposes.