WO2009009059A1 - Spiro compounds as antagonists of tgf-beta - Google Patents

Abstract

The invention is related to compounds of formula (I) These compounds can be used as antagonists of the TGF-beta family tyle I receptors, Alk5 or Alk4. The compounds of formula (I) can be employed in the prevention and/or treatment of diseases such as fibrosis (e. g. renal fibrosis, pulmonary fibrosis and hepatic fibrosis), progressive cancers or other diseases for which reduction of TGF-beta family signalling activity is desirable.

Description

SPIRO COMPOUNDS AS ANTAGONISTS OF TGF-BETA

CROSS REFERNCE

[01] This application claims priority to U.S. Application No. 60/948,632, filed on July 9, 2007, the content of which is incorporated herein by reference in its entirety.

BACHGROUND OF THE INVENTION

[02] TGF/3 (Transforming Growth Factor /3) is a member of a large family of dimeric polypeptide growth factors that includes, for example, activins, inhibins, bone morphogenetic proteins (BMPs), growth and differentiation factors (GDFs) and mullerian inhibiting substance (MIS). TGF/3 exists in three isoforms (TGF/31, TGF/32, and TGF/33) and is present in most cells, along with its receptors. Each isoform is expressed in both a tissue-specific and developmentally regulated fashion. Each TGF/3 isoform is synthesized as a precursor protein that is cleaved intracellularly into a C-terminal region (latency associated peptide (LAP)) and an N-terminal region known as mature or active TGF/3. LAP is typically non-covalently associated with mature TGF/3 prior to secretion from the cell. The LAP-TGF/3 complex cannot bind to the TGF/3 receptors and is not biologically active. TGF/3 is generally released (and activated) from the complex by a variety of mechanisms including, for example, interaction with thrombospondin-1 or plasmin.

[03] Following activation, TGF/3 binds at high affinity to the type II receptor (TGF/3RII), a constitutively active serine/threonine kinase. The ligand-bound type II receptor phosphorylates the TGF/3 type I receptor (Alk5) in a glycine/serine rich domain, which allows the type I receptor to recruit and phosphorylate downstream signaling molecules, Smad2 or Smad3. See, e.g., Huse, M. et al., MoI. Cell, 8: 671-682 (2001). Phosphorylated Smad2 or Smad3 can then complex with Smad4, and the entire hetero-Smad complex translocates to the nucleus and regulates transcription of various TGF/3-responsive genes. See, e.g., Massague, J. Ann. Rev .Biochem. Med., 67: 773 (1998).

[04] Activins are also members of the TGF/3 superfamily, which are distinct from TGF/3 in that they are homo- or heterodimers of activin /3a or /3b. Activins signal in a manner similar to TGFjS , that is, by binding to a constitutive serine-threonine receptor kinase, activin type II receptor (ActRIIB), and activating a type I serine-threonine receptor, Alk4, to phosphorylate Smad2 or Smad3. The consequent formation of a hetero-Smad complex with Smad4 also results in the activin-induced regulation of gene transcription. [05] Indeed, TGF/3 and related factors such as activin regulate a large array of cellular processes, e.g., cell cycle arrest in epithelial and hematopoietic cells, control of mesenchymal cell proliferation and differentiation, inflammatory cell recruitment, immunosuppression, wound healing, and extracellular matrix production. See, e.g., Massague, J. Ann. Rev .Cell. Biol., 6: 594-641 (1990); Roberts, A. B. and Sporn M. B., Peptide Growth Factors and Their Receptors, 95: 419-472 Berlin: Springer- Verlag (1990); Roberts, A. B. and Sporn M. B. Growth Factors 8:1-9 (1993); and Alexandrow, M. G., Moses, H. L. Cancer Res. 55: 1452- 1457 (1995). Hyperactivity of TGF/3 signaling pathway underlies many human disorders (e.g., excess deposition of extracellular matrix, an abnormally high level of inflammatory responses, fϊbrotic disorders, and progressive cancers). Similarly, activin signaling and overexpression of activin is linked to pathological disorders that involve extracellular matrix accumulation and fibrosis (see, e.g., Matsuse, T. et al., Am. J. Respir. Cell MoI. Biol., 13: 17- 24 (1995); Inoue, S. et al., Biochem. Biophys. Res. Comm. 205: 441-448 (1994); Matsuse, T. et al., Am. J. Pathol, 148: 707-713 (1996); De Bleser et al., Hepatology 26: 905-912 (1997); Pawlowski, J.E., et al., J. Clin. Invest. 100: 639-648 (1997); Sugiyama, M. et al., Gastroenterology 114: 550-558 (1998); Munz, B. βt al, EMBO J. 18: 5205-5215 (1999)), inflammatory responses (see, e.g., Rosendahl, A. et al., Am. J. Repir. Cell MoI. Biol. 25: 60- 68 (2001)), cachexia or wasting (see Matzuk, M. M. et al., Proc. Nat. Acad. Sci. USA 91: 8817-8821 (1994); Coerver, K.A. et al, MoI. Endocrinol. 10: 534-543 (1996); Cipriano, S.C. et al. Endocrinology 141: 2319-27 (2000)), diseases of or pathological responses in the central nervous system (see Logan, A. et al. Eur. J. Neurosci. 11 : 2367-2374 (1999); Logan, A. et al. Exp. Neurol. 159: 504-510 (1999); Masliah, E. et al., Neurochem. Int. 39: 393-400 (2001); De Groot, C. J. A. et al., J. Neuropathol. Exp. Neurol. 58: 174-187 (1999), John, G. R. et al, Nat Med. 8: 1115-21 (2002)) and hypertension (see Dahly, A. J. et al., Am. J. Physiol. Regul. Integr. Comp. Physiol. 283: R757-67 (2002)). Studies have shown that TGF]S and activin can act synergistically to induce extracellular matrix production (see, e.g., Sugiyama, M. et al., Gastroenterology 114: 550-558, (1998)). It is therefore desirable to develop modulators (e.g., antagonists) to members of the TGF/3 family to prevent and/or treat disorders involving this signaling pathway.

SUMMARY OF THE INVENTION

[06] The invention is based on the discovery that compounds of Formula (I) are potent antagonists of the TGF/3 family type I receptors, Alk5 and/or Alk4. Thus, compounds of Formula (I) can be employed in the prevention and/or treatment of diseases such as fibrosis (e.g., renal fibrosis, pulmonary fibrosis, and hepatic fibrosis), progressive cancers, or other diseases for which reduction of TGF/3 family signaling activity is desirable. [07] In one aspect, the present invention provides compounds of Formula (I)

(I) or a pharmaceutically acceptable salt thereof. In Formula (I): each of R¹ and R² independently is an optionally substituted aryl or heteroaryl;

A¹ is NR³ or CR³;

A² is N when A¹ is CR³, or A² is C when A¹ is NR³;

X is O or NR³; each R³ is independently hydrogen, aliphatic, cycloaliphatic, aryl, or heteroaryl; each of R' and R" is independently an aliphatic, halo, cyano, or alkoxy; each of p and q is independently 0, 1 , or 2, provided that the sum of p and q is 2, 3, or 4; r is 1, 2, or 3; and each of m and n is independently 0, 1, or 2. [08] In some embodiments, R¹ is an optionally substituted aryl. [09] In some embodiments, R¹ is phenyl optionally substituted with 1 to 3 substituents each independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, cycloalkylcarbonyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, heterocycloalkylcarbonyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl, heteroaryloxy, heteroarylsulfanyl, and heteroaroyl.

[10] In other embodiments, R¹ is a heteroaryl optionally substituted with 1 to 3 substitutents each independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, cycloalkylcarbonyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, heterocycloalkylcarbonyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl, heteroaryloxy, heteroarylsulfanyl, and heteroaroyl.

[11] In further embodiments, R¹ is benzo[l,3]dioxolyl, benzo[b]thiophenyl, benzo oxadiazolyl, benzothiadiazolyl, benzoimidazolyl, benzooxazolyl, benzothiazolyl, 2-oxo- benzooxazolyl, pyridyl, pyrimidinyl, 2,3-dihydrobenzo[l,4]dioxyl, 2,3-dihydrobenzofuryl, 2,3-dihydrobenzo[b]thiophenyl, 3,4-dihydrobenzo[l,4]oxazinyl, 3-oxo-benzo[l,4]oxazinyl, 1 , 1 -dioxo-2,3-dihydrobenzo[b]thiophenyl, [ 1 ,2,4]triazolo[ 1 ,5-α]pyridyl, [ 1 ,2,4]triazolo[4,3- α]pyridyl, quinolinyl, quinoxalinyl, quinazolinyl, isoquinolinyl, or cinnolinyl, and is optioanlly substituted.

[12] In some embodiments, R² is an optioanlly substituted aryl.

[13] In other embodiments, R² is phenyl optionally substituted with 1 to 3 substitutents each independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, cycloalkylcarbonyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, heterocycloalkylcarbonyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl, heteroaryloxy, heteroarylsulfanyl, and heteroaroyl.

[14] In other embodiments, R² is an heteroaryl optionally substituted with 1 to 3 substitutents each independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, cycloalkylcarbonyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, heterocycloalkylcarbonyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl, heteroaryloxy, heteroarylsulfanyl, and heteroaroyl.

[15] In some embodiments, the heteroaryl in R² is benzo[l,3]dioxolyl, benzo[b]thiophenyl, benzooxadiazolyl, benzothiadiazolyl, benzoimidazolyl, benzooxazolyl, benzothiazolyl, 2- oxo-benzooxazolyl, pyridyl, pyrimidinyl, 2,3-dihydrobenzo[l,4]dioxyl, 2,3- dihydrobenzofuryl, 2,3-dihydrobenzo[b]thiophenyl, 3,4-dihydrobenzo[l,4]oxazinyl, 3-oxo- benzo[ 1 ,4]oxazinyl, 1 , 1 -dioxo-2,3-dihydrobenzo[b]thiophenyl, [ 1 ,2,4]triazolo[ 1 ,5-α]pyridyl,

[l,2,4]triazolo[4,3-α]pyridyl, quinolinyl, quinoxalinyl, quinazolinyl, isoquinolinyl, or cinnolinyl.

[16] In some embodiments, each of p, q, and r is independently 1.

[17] In other embodiments, R³ is H.

[18] Examples of the compound of Formula (I) include, but are not limited to,

8-(5-([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-4-(6-methylpyridin-2-yl)-l H-imidazol-2-yl)- 2-oxaspiro[4.5]decan-l-one;

8-(5-([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-4-(5-fluoro-6-methylpyridin-2-yl)- 1 H- imidazol-2-yl)-2-oxaspiro[4.5]decan-l-one;

8-(3-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)- 1 H-pyrazol- 1 -yl)-2- oxaspiro [4.5] decan- 1 -one ;

8-(4-([l,2,4]triazolo[l,5-a]pyridin-6-yl)-3-(5-fluoro-6-methylpyridin-2-yl)-lH- pyrazol- 1 -yl)-2-oxaspiro[4.5]decan- 1 -one;

8-(4-([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-3-(6-methylpyridin-2-yl)- 1 H-pyrazol- 1 -yl)-2- oxaspiro [4.5] decan- 1 -one ;

8-(4-([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-3-(pyridin-2-yl)- 1 H-pyrazol- 1 -yl)-2- oxaspiro[4.5]decan-l-one; and

2-benzyl-8-(3-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)-lH-pyrazol-l-yl)-2- azaspiro [4.5 ]decan- 1 -one .

[19] In another aspect, the invention pertains to a pharmaceutical composition which includes a compound of Formula (I), in a pharmaceutically effective amount, and a pharmaceutically acceptable carrier.

[20] In another aspect, the invention relates to a method of inhibiting the TGF/3 signaling pathway in a subject in need of the inhibition. This method includes administering to the subject an effective amount of a compound of Formula (I).

[21] In another aspect, the invention relates to a method of inhibiting the TGF/3 type I receptor in a cell, and the method includes contacting the cell with an effective amount of a compound of Formula (I).

[22] In another aspect, the invention relates to a method of reducing the accumulation of excess extracellular matrix induced by TGF/3 in a subject in need of the reduction of the accumulation of excess extracellular matrix. The method includes administering to the subject an effective amount of a compound of Formula (I).

[23] In another aspect, the invention relates to a method of treating or preventing fibrotic condition in a subject in need thereof. The method includes administering to the subject an effective amount of a compound of Formula (I). The fibrotic condition can be, for example, scleroderma, lupus nephritis, connective tissue disease, wound healing, surgical scarring, spinal cord injury, CNS scarring, acute lung injury, pulmonary fibrosis (such as idiopathic pulmonary fibrosis), chronic obstructive pulmonary disease, adult respiratory distress syndrome, drug-induced lung injury, glomerulonephritis, diabetic nephropathy, hypertension- induced nephropathy, alimentary track or gastrointestinal fibrosis, renal fibrosis, hepatic or biliary fibrosis (such as liver cirrhosis, primary biliary cirrhosis, fatty liver disease, primary sclerosing cholangitis), restenosis, cardiac fibrosis, opthalmic scarring, fibrosclerosis, fibrotic cancers, fibroids, fibroma, fibroadenomas, fibrosarcomas, transplant arteriopathy, or keloid. The fibrotic condition can be idiopathic in nature, genetically linked, or induced by radiation. [24] In another aspect, the invention relates to a method of inhibiting growth or metastasis of tumor cells or cancers in a subject in need thereof. The method includes administering to the subject an effective amount of a compound of Formula (I).

[25] In another aspect, the invention relates to a method of treating a disease or disorder mediated by an overexpression of TGF/3 in a subject in need thereof. The method includes administering to a subject in need of such treatment an effective amount of a compound of Formula (I). The disease or disorder can be, for example, demyelination of neurons in multiple sclerosis, Alzheimer's disease, cerebral angiopathy, squamous cell carcinomas, multiple myeloma, melanoma, glioma, glioblastomas, leukemia, sarcomas, leiomyomas, mesothelioma, or carcinomas of the lung, breast, ovary, cervix, liver, biliary tract, gastrointestinal tract, pancreas, prostate, and head and neck.

[26] An N-oxide derivative or a pharmaceutically acceptable salt of each of the compounds of Formula (I) is also within the scope of this invention. For example, a nitrogen ring atom of the imidazole core ring or a nitrogen-containing heterocyclyl substituent can form an oxide in the presence of a suitable oxidizing agent such as /w-chloroperbenzoic acid or H₂O₂. [27] A compound of Formula (I) that is acidic in nature (e.g., having a carboxyl or phenolic hydroxyl group) can form a pharmaceutically acceptable salt such as a sodium, potassium, calcium, or gold salt. Also within the scope of the invention are salts formed with pharmaceutically acceptable amines such as ammonia, alkyl amines, hydroxyalkylamines, and Ν-methylglycamine. A compound of Formula (I) can be treated with an acid to form acid addition salts. Examples of such acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, methanesulfonic acid, phosphoric acid, /7-bromophenyl- sulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, oxalic acid, malonic acid, salicylic acid, maleic acid, fumaric acid, ascorbic acid, maleic acid, acetic acid, and other mineral and organic acids well known to those skilled in the art. The acid addition salts can be prepared by treating a compound of Formula (I) in its free base form with a sufficient amount of an acid (e.g., hydrochloric acid) to produce an acid addition salt (e.g., a hydrochloride salt). The acid addition salt can be converted back to its free base form by treating the salt with a suitable dilute aqueous basic solution (e.g., sodium hydroxide, sodium bicarbonate, potassium carbonate, or ammonia). Compounds of Formula (I) can also be, e.g., in a form of achiral compounds, racemic mixtures, optically active compounds, pure diastereomers, or a mixture of diastereomers.

[28] Compounds of Formula (I) in general exhibit surprisingly high affinity to the TGF/3 family type I receptors, Alk5 or Alk4, e.g., with IC₅₀ and Ki values of less than 10 μM under conditions as described below in Examples 8 and 10, respectively. Some compounds of Formula (I) exhibit IC₅₀ and Kj values of less than 1 μM (such as below 50 nM). [29] Compounds of Formula (I) can also be modified by appending appropriate functionalities to enhance selective biological properties. Such modifications are known in the art and include those that increase biological penetration into a given biological system (e.g., blood, lymphatic system, or central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism, or alter rate of excretion. Examples of these modifications include, but are not limited to, esterification with polyethylene glycols, derivatization with pivolates or fatty acid substituents, conversion to carbamates, hydroxylation of aromatic rings, and heteroatom-substitution in aromatic rings. [30] The present invention also features a pharmaceutical composition including a compound of Formula (I) (or a combination of two or more compounds of Formula (I)) and at least one pharmaceutically acceptable carrier. Also included in the present invention is a medicament composition including any of the compounds of Formula (I), alone or in a combination, together with a suitable excipient.

[31] The invention also features a method of inhibiting the TGFjS family type I receptors, Alk5 or Alk4 (e.g., with an IC₅₀ value of less than 10 μM; such as, less than 1 μM; and for example, less than 5 nM) in a cell, which includes the step of contacting the cell with an effective amount of one or more compounds of Formula (I). Also within the scope of the invention is a method of inhibiting the TGF/3 or activin signaling pathway in a cell or in a subject (e.g., a mammal such as a human), which includes the step of contacting the cell with or administering to the subject an effective amount of one or more of the compounds of Formula (I).

[32] Also within the scope of the present invention is a method of treating a subject or preventing a subject from suffering a condition characterized by or resulted from an elevated level of TGF/3 and/or activin activity. The method includes the step of administering to the subject in need thereof an effective amount of one or more of a compound of Formula (I).

The conditions include an accumulation of excess extracellular matrix; a fibrotic condition (which can be induced by drug or radiation), e.g., scleroderma, lupus nephritis, connective tissue disease, wound healing, surgical scarring, spinal cord injury, CNS scarring, acute lung injury, pulmonary fibrosis (such as idiopathic pulmonary fibrosis and radiation-induced pulmonary fibrosis), chronic obstructive pulmonary disease, adult respiratory distress syndrome, acute lung injury, drug-induced lung injury, glomerulonephritis, diabetic nephropathy, hypertension-induced nephropathy, alimentary track or gastrointestinal fibrosis, renal fibrosis, hepatic or biliary fibrosis, liver cirrhosis, primary biliary cirrhosis, cirrhosis due to fatty liver disease (alcoholic and nonalcoholic steatosis), primary sclerosing cholangitis, restenosis, cardiac fibrosis, opthalmic scarring, fibrosclerosis, fibrotic cancers, fibroids, fibroma, fibroadenomas, fibrosarcomas, transplant arteriopathy, and keloid); TGF/3- induced growth or metastasis of tumor/cancer cells; and carcinomas (e.g, squamous cell carcinomas, multiple myeloma, melanoma, glioma, glioblastomas, leukemia, sarcomas, leiomyomas, mesothelioma, and carcinomas of the lung, breast, ovary, cervix, liver, biliary tract, gastrointestinal tract, pancreas, prostate, and head and neck); and other conditions such as cachexia, hypertension, ankylosing spondylitis, demyelination in multiple sclerosis, cerebral angiopathy and Alzheimer's disease.

[33] Examples of fibrotic conditions include, but are not limited to, mesothelioma, acute respiratory distress syndrome (ARDS), atherosclerosis, scleroderma, keloids, glomerulonephritis, diabetic nephropathy, lupus nephritis, hypertension-induced nephropathy, idiopathic pulmonary fibrosis, cholangitis, restenosis, ocular scarring, corneal scarring, hepatic fibrosis, biliary fibrosis, liver cirrhosis, cirrhosis due to fatty liver disease (alcoholic and nonalcoholic steatosis), pulmonary fibrosis, renal fibrosis, sarcoidosis, acute lung injury, drug-induced lung injury, spinal cord injury, CNS scarring, systemic lupus erythematosus, Wegener's granulomatosis, cardiac fibrosis, post-infarction cardiac fibrosis, post-surgical fibrosis, connective tissue disease, radiation-induced fibrosis, chemotherapy-induced fibrosis, transplant arteriopathy, fibrosclerosis, fibrotic cancers, fibroids, fibroma, fibroadenomas, and fibrosarcomas.

[34] Also within the scope of the invention is a method of treating or preventing a TGF/3 related disease in a subject in need thereof, which includes administering to the subject an effective amount of a compound of Formula (I). Exampels of such diease include, but are not limited to, acute lung injury, adult respiratory distress syndrome, alimentary track or gastrointestinal fibrosis, atherosclerosis, biliary fibrosis, cardiac fibrosis, chemotherapy- induced fibrosis, cholangitis, chronic obstructive pulmonary disease, cirrhosis due to fatty liver disease, CNS scarring, connective tissue disease, corneal scarring, diabetic nephropathy, drug-induced lung injury, fatty liver disease, fibroadenoma, fibroid, fibroma, fibrosarcoma, fibrosclerosis, fibrotic cancer, glomerulonephritis, hepatic or biliary fibrosis, hypertension- induced nephropathy, idiopathic pulmonary fibrosis, keloids, liver cirrhosis, lupus nephritis, mesothelioma, opthalmic scarring, post-infarction cardiac fibrosis, primary biliary cirrhosis, post-surgical fibrosis, primary sclerosing cholangitis, pulmonary fibrosis, radiation-induced fibrosis, radiation-induced pulmonary fibrosis, renal fibrosis, restenosis, sarcoidosis, scleroderma, spinal cord injury, surgical scarring, transplant arteriopathy, wound healing, scleroderma, spinal cord injury, systemic lupus erythematosus, and Wegener's granulomatosis.

[35] In some embodiments, the compounds of this invention may be administered locally. [36] In some other embodiments, the compounds of this invention can be administered by the way of an implantable device such as, for example, a delivery pump or a stent. [37] For the purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. Additionally, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "Advanced Organic Chemistry", 5th Ed. (Ed.: M.B. Smith and J. March), John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference. [38] As described herein, compounds of the invention may optionally be substituted with one or more substituents, such as are illustrated generally above, or as exemplified by particular classes, subclasses, and species of the invention.

[39] As used herein, the term "aliphatic" encompasses the terms alkyl, alkenyl, and alkynyl, each of which being optionally substituted as set forth below. [40] As used herein, an "alkyl" group refers to a linear (i.e., noncyclic) saturated aliphatic hydrocarbon group containing 1 to 12 (e.g., 1 to 10, 1 to 8, 1 to 6, or 1 to 4) carbon atoms. An alkyl group can be straight or branched. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec -butyl, tert-butyl, n-pentyl, n- heptyl, or 2-ethylhexyl. An alkyl group can be substituted (i.e., optionally substituted) with one or more substituents such as halo, phospho, cycloaliphatic (e.g., cycloalkyl or cycloalkenyl), heterocycloaliphatic (e.g., heterocycloalkyl or heterocycloalkenyl), aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl (e.g., (aliphatic)carbonyl, (cycloaliphatic)carbonyl, or (heterocycloaliphatic)carbonyl), nitro, cyano, amido (e.g., (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino alkylaminocarbonyl, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, arylaminocarbonyl, or heteroarylaminocarbonyl), amino (e.g., aliphaticamino, cycloaliphaticamino, or heterocycloaliphaticamino), sulfonyl (e.g., aliphatic-SO₂-), sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, carboxy, carbamoyl, cycloaliphaticoxy, heterocycloaliphaticoxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, or hydroxy. Without limitation, some examples of substituted alkyls include carboxyalkyl (such as HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl), cyanoalkyl, hydroxyalkyl, alkoxyalkyl, acylalkyl, aralkyl, (alkoxyaryl)alkyl, (sulfonylamino)alkyl (such as (alkyl-SO₂-amino)alkyl), aminoalkyl, amidoalkyl, (cycloaliphatic)alkyl, or haloalkyl.

[41] As used herein, an "alkenyl" group refers to a linear (i.e., noncyclic) aliphatic carbon group that contains 2 to 12 (e.g., 2 to 8, 2 to 6, or 2 to 4) carbon atoms and at least one double bond. Like an alkyl group, an alkenyl group can be straight or branched. Examples of an alkenyl group include, but are not limited to, allyl, isoprenyl, 2-butenyl, and 2-hexenyl. An alkenyl group can be optionally substituted with one or more substituents such as halo, phospho, cycloaliphatic (e.g., cycloalkyl or cycloalkenyl), heterocycloaliphatic (e.g., heterocycloalkyl or heterocycloalkenyl), aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl (e.g., (aliphatic)carbonyl, (cycloaliphatic)carbonyl, or (heterocycloaliphatic)carbonyl), nitro, cyano, amido (e.g., (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino alkylaminocarbonyl, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, arylaminocarbonyl, or heteroarylaminocarbonyl), amino (e.g., aliphaticamino, cycloaliphaticamino, heterocycloaliphaticamino, or aliphaticsulfonylamino), sulfonyl (e.g., alkyl-SO₂-, cycloaliphatic-SO₂-, or aryl-SO₂-), sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, carboxy, carbamoyl, cycloaliphaticoxy, heterocycloaliphaticoxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkoxy, alkoxycarbonyl, alkylcarbonyloxy, or hydroxy. Without limitation, some examples of substituted alkenyls include cyanoalkenyl, alkoxyalkenyl, acylalkenyl, hydroxyalkenyl, aralkenyl, (alkoxyaryl)alkenyl, (sulfonylamino)alkenyl (such as (alkyl-SO₂-amino)alkenyl), aminoalkenyl, amidoalkenyl, (cycloaliphatic)alkenyl, and haloalkenyl.

[42] As used herein, an "alkynyl" group refers to an aliphatic carbon group that contains 2 to 12 (e.g., 2 to 8, 2 to 6, or 2 to 4) carbon atoms and has at least one triple bond. An alkynyl group can be straight or branched. Examples of an alkynyl group include, but are not limited to, propargyl and butynyl. An alkynyl group can be optionally substituted with one or more substituents such as aroyl, heteroaroyl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, nitro, carboxy, cyano, halo, hydroxy, sulfo, mercapto, sulfanyl (e.g., aliphaticsulfanyl or cycloaliphaticsulfanyl), sulfinyl (e.g., aliphaticsulfinyl or cycloaliphaticsulfinyl), sulfonyl (e.g., aliphatic-SO₂-, aliphaticamino-SO₂-, or cycloaliphatic- SO₂-), amido (e.g., aminocarbonyl, alkylaminocarbonyl, alkylcarbonylamino, cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl, cycloalkylcarbonylamino, arylaminocarbonyl, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (cycloalkylalkyl)carbonylamino, heteroaralkylcarbonylamino, heteroarylcarbonylamino or heteroarylaminocarbonyl), urea, thiourea, sulfamoyl, sulfamide, alkoxycarbonyl, alkylcarbonyloxy, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, acyl (e.g., (cycloaliphatic)carbonyl or (heterocycloaliphatic)carbonyl), amino (e.g., aliphaticamino), sulfoxy, oxo, carboxy, carbamoyl, (cycloaliphatic)oxy, (heterocycloaliphatic)oxy, and (heteroaryl)alkoxy. [43] As used herein, an "amido" encompasses both "aminocarbonyl" and "carbonylamino". These terms when used alone or in connection with another group refers to an amido group such as -N(R^X)-C(O)-R^Y or -C(O)-N(R^X)₂, when used terminally, and -C(O)-N(R^X)- or -N(R^X)-C(O)- when used internally, wherein R^x and R^γ are defined below. Examples of amido groups include, but are not limited to, alkylamido (such as alkylcarbonylamino or alkylaminocarbonyl), (heterocycloaliphatic)amido, (heteroaralkyl)amido, (heteroaryl)amido, (heterocycloalkyl)alkylamido, arylamido, aralkylamido, (cycloalkyl)alkylamido, and cycloalkylamido.

[44] As used herein, an "amino" group refers to -NR^XR^Y wherein each of R^x and R^γ is independently hydrogen, aliphatic, cycloaliphatic, (cycloaliphatic)aliphatic, aryl, araliphatic, heterocycloaliphatic, (heterocycloaliphatic)aliphatic, heteroaryl, carboxy, sulfanyl, sulfinyl, sulfonyl, (aliphatic)carbonyl, (cycloaliphatic)carbonyl, ((cycloaliphatic)aliphatic)carbonyl, arylcarbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl,

((heterocycloaliphatic)aliphatic)carbonyl, (heteroaryl)carbonyl, or (heteroaraliphatic)carbonyl, each of which being defined herein and being optionally substituted. Examples of amino groups include, but are not limited to, alkylamino, dialkylamino, and arylamino. When the term "amino" is not the terminal group (e.g., alkylcarbonylamino), it is represented by -NR^X-. R^x has the same meaning as defined herein.

[45] As used herein, an "aryl" group used alone or as part of a larger moiety such as in "aralkyl", "aralkoxy", or "aryloxyalkyl" refers to monocyclic (e.g., phenyl); bicyclic (e.g., indenyl, naphthalenyl, tetrahydronaphthyl, tetrahydroindenyl); and tricyclic (e.g., fluorenyl tetrahydrofluorenyl, or tetrahydroanthracenyl, anthracenyl) ring systems in which the monocyclic ring system is aromatic or at least one of the rings in a bicyclic or tricyclic ring system is aromatic. The bicyclic and tricyclic groups include benzofused systems having 2 to3 carbocyclic rings. For example, a benzofused group includes phenyl fused with two or more C_4-8 carbocyclic moieties. An aryl is optionally substituted with one or more substituents including aliphatic (e.g., alkyl, alkenyl, or alkynyl); cycloaliphatic; (cycloaliphatic)aliphatic; heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; oxo (on a non-aromatic carbocyclic ring of a benzofused bicyclic or tricyclic aryl); nitro; carboxy; amido; acyl [e.g., (aliphatic)carbonyl, (cycloaliphatic)carbonyl, ((cycloaliphatic)aliphatic)carbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl,

((heterocycloaliphatic)aliphatic)carbonyl, or (heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphatic-SO₂- or amino-SO₂-]; sulfinyl (e.g., aliphatic-S(O)- or cycloaliphatic-S(O)-); sulfanyl (e.g., aliphatic-S-); cyano; halo; hydroxy; mercapto; sulfoxy; urea; thiourea; sulfamoyl; sulfamide; or carbamoyl. Alternatively, an aryl can be unsubstituted. [46] Non-limiting examples of substituted aryls include haloaryl (e.g., mono-, di-(such as /7,rø-dihaloaryl), and tri-(such as halo) aryl); (carboxy)aryl (e.g., (alkoxycarbonyl)aryl, ((aralkyl)carbonyloxy)aryl, and (alkoxycarbonyl)aryl); (amido)aryl (e.g., (aminocarbonyl)aryl, (((alkylamino)alkyl)aminocarbonyl)aryl, (alkylcarbonyl)aminoaryl, (arylaminocarbonyl)aryl, and (((heteroaryl)amino)carbonyl)aryl); aminoaryl (e.g., ((alkylsulfonyl)amino)aryl or ((dialkyl)amino)aryl); (cyanoalkyl)aryl; (alkoxy)aryl; (sulfamoyl)aryl (e.g., (aminosulfonyl)aryl); (alkylsulfonyl)aryl; (cyano)aryl; (hydroxyalkyl)aryl; ((alkoxy)alkyl)aryl; (hydroxy)aryl, ((carboxy)alkyl)aryl; (((dialkyl)amino)alkyl)aryl; (nitroalkyl)aryl; (((alkylsulfonyl)amino)alkyl)aryl; ((heterocycloaliphatic)carbonyl)aryl; ((alkylsulfonyl)alkyl)aryl; (cyanoalkyl)aryl; (hydroxyalkyl)aryl; (alkylcarbonyl)aryl; alkylaryl; (trihaloalkyl)aryl; p-amino-m- alkoxycarbonylaryl; p-amino-w-cyanoaryl; p-halo-m-aminoaryl; or (/w-(heterocycloaliphatic)- o-(alkyl))aryl.

[47] As used herein, an "araliphatic" such as an "aralkyl" group refers to an aliphatic group (e.g., a C_M alkyl group) that is substituted with an aryl group. "Aliphatic," "alkyl," and "aryl" are as defined herein. An example of an araliphatic such as an aralkyl group is benzyl.

[48] As used herein, an "aralkyl" group refers to an alkyl group (e.g., a Ci_-4 alkyl group) that is substituted with an aryl group. Both "alkyl" and "aryl" are as defined herien. An example of an aralkyl group is benzyl. An aralkyl is optionally substituted with one or more substituents such as aliphatic (e.g., alkyl, alkenyl, or alkynyl, including carboxyalkyl, hydroxyalkyl, or haloalkyl such as trifluoromethyl); cycloaliphatic (e.g., cycloalkyl or cycloalkenyl); (cycloalkyl)alkyl; heterocycloalkyl; (heterocycloalkyl)alkyl; aryl; heteroaryl; alkoxy; cycloalkyloxy; heterocycloalkyloxy; aryloxy; heteroaryloxy; aralkyloxy; heteroaralkyloxy; aroyl; heteroaroyl; nitro; carboxy; alkoxycarbonyl; alkylcarbonyloxy; amido (e.g., aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, or heteroaralkylcarbonylamino); cyano; halo; hydroxy; acyl; mercapto; alkylsulfanyl; sulfoxy; urea; thiourea; sulfamoyl; sulfamide; oxo; or carbamoyl. [49] As used herein, a "bicyclic ring system" includes 8- to 12- (e.g., 9-, 10-, or H-) membered structures that form two rings, wherein the two rings have at least one atom in common (e.g., 1 atom or 2 atoms in common). Bicyclic ring systems include bicycloaliphatics (e.g., bicycloalkyl or bicycloalkenyl), spiroaliphatics, spiroheteroaliphatics, bicycloheteroaliphatics, bicyclic aryls, and bicyclic heteroaryls.

[50] As used herein, a "cycloaliphatic" group encompasses a "cycloalkyl" group and a "cycloalkenyl" group, each of which being optionally substituted as set forth below. [51] As used herein, a "cycloalkyl" group refers to a saturated carbocyclic fused or bridged ring system of 3-15 (e.g., 5-12) carbon atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, norbornyl, cubyl, octahydro-indenyl, decahydro-naphthyl, spiro[5.5]undecanyl, spiro[2.5]octanyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl, bicyclo[3.3.2.]decyl, bicyclo[2.2.2]octyl, adamantyl, or ((aminocarbonyl)cycloalkyl)cycloalkyl. [52] A "cycloalkenyl" group, as used herein, refers to a non-aromatic carbocyclic mono- or bicyclic ring of 3 to 12 (e.g., 4 to 8) carbon atoms having one or more double bonds. Examples of cycloalkenyl groups include, but are not limited to, cyclopentenyl, 1,4- cyclohexa-di-enyl, cycloheptenyl, cyclooctenyl, hexahydro-indenyl, octahydro-naphthyl, cyclohexenyl, cyclopentenyl, spiro[5.5]]undec-3-enyl, spiro[2.5]oct-5-enyl, bicyclo[2.2.2]octenyl, and bicyclo[3.3.1]nonenyl.

[53] A cycloalkyl or cycloalkenyl group can be optionally substituted with one or more substituents such as phosphoryl; aliphatic (e.g., alkyl, alkenyl, or alkynyl); cycloaliphatic; (cycloaliphatic) aliphatic; heterocycloaliphatic; (heterocycloaliphatic) aliphatic; aryl; heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; amido (e.g., (aliphatic)carbonylamino, (cycloaliphatic)carbonylamino, ((cycloaliphatic)aliphatic)carbonylamino, (aryl)carbonylamino, (araliphatic)carbonylamino,

(heterocycloaliphatic)carbonylamino, ((heterocycloaliphatic)aliphatic)carbonylamino, (heteroaryl)carbonylamino, or (heteroaraliphatic)carbonylamino); nitro; carboxy (e.g., HOOC-, alkoxycarbonyl, or alkylcarbonyloxy); acyl (e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic) aliphatic)carbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl, ((heterocycloaliphatic)aliphatic)carbonyl, or (heteroaraliphatic)carbonyl); cyano; halo; hydroxy; mercapto; sulfonyl (e.g., alkyl-SO₂- and aryl-SO₂-); sulfinyl (e.g., alkyl-S(O)-); sulfanyl (e.g., alkyl-S-); sulfoxy; urea; thiourea; sulfamoyl; sulfamide; oxo; or carbamoyl. [54] As used herein, the term "heterocycloaliphatic" encompasses a heterocycloalkyl group and a heterocycloalkenyl group, each of which being optionally substituted as set forth below.

[55] As used herein, a "heterocycloalkyl" group refers to a 3- to 12-membered mono- or bicylic (fused or bridged) (e.g., 5- to 12-membered mono- or bicyclic) saturated ring structure, in which one or more of the ring atoms is a heteroatom (e.g., N, O, S, or combinations thereof). Examples of a heterocycloalkyl group include piperidyl, piperazyl, tetrahydropyranyl, tetrahydrofuryl, 1 ,4-dioxolanyl, 1 ,4-dithianyl, 1,3-dioxolanyl, oxazolidyl, isoxazolidyl, morpholinyl, thiomorpholyl, octahydrobenzofuryl, octahydrochromenyl, octahydrothiochromenyl, octahydroindolyl, octahydropyrindinyl, decahydroquinolinyl, octahydrobenzo[ό]thiopheneyl, 2-oxa-bicyclo[2.2.2]octyl, l-aza-bicyclo[2.2.2]octyl, 3-aza- bicyclo[3.2.1]octyl, 3-oxaspiro[5.5]]undec-3-enyl, 6-oxaspiro[2.5]oct-5-enyl,and 2,6-dioxa- tricyclo[3.3.1.0^3>7]nonyl.

[56] As used herein, a "heterocycloalkenyl" group refers to a mono- or bicylic (e.g., 5- to 10-membered mono- or bicyclic) non-aromatic ring structure having one or more double bonds, and wherein one or more of the ring atoms is a heteroatom (e.g., N, O, or S). [57] Monocyclic and bicycloheteroaliphatics are numbered according to standard chemical nomenclature. A heterocycloalkyl or heterocycloalkenyl group can be optionally substituted with one or more substituents such as phosphor; aliphatic (e.g., alkyl, alkenyl, or alkynyl); cycloaliphatic; (cycloaliphatic)aliphatic; heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; amido [e.g., (aliphatic)carbonylamino, (cycloaliphatic)carbonylamino, ((cycloaliphatic) aliphatic)carbonylamino, (aryl)carbonylamino, (araliphatic)carbonylamino, (heterocycloaliphatic)carbonylamino, ((heterocycloaliphatic) aliphatic)carbonylamino, (heteroaryl)carbonylamino, or (heteroaraliphatic)carbonylamino]; nitro; carboxy [e.g., HOOC-, alkoxycarbonyl, or alkylcarbonyloxy]; acyl [e.g., (cycloaliphatic)carbonyl, ((cycloaliphatic) aliphatic)carbonyl, (araliphatic)carbonyl, (heterocycloaliphatic)carbonyl,

((heterocycloaliphatic)aliphatic)carbonyl, or (heteroaraliphatic)carbonyl]; nitro; cyano; halo; hydroxy; mercapto; sulfonyl (e.g., alkylsulfonyl or arylsulfonyl); sulfinyl (e.g., alkylsulfinyl); sulfanyl (e.g., alkylsulfanyl); sulfoxy; urea; thiourea; sulfamoyl; sulfamide; oxo; or carbamoyl.

[58J A "heteroaryl" group, as used herein, refers to a monocyclic, bicyclic, or tricyclic ring system having 4 to 15 ring atoms wherein one or more of the ring atoms is a heteroatom (e.g., N, O, S, or combinations thereof) and in which the monocyclic ring system is aromatic or at least one of the rings in the bicyclic or tricyclic ring systems is aromatic. A heteroaryl group includes a benzofused ring system having 2 to 3 rings. For example, a benzofused group includes benzo fused with one or two 4 to 8 membered heterocycloaliphatic moieties (e.g., indolizyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[6]furyl, benzo[ό]thiophenyl, quinolinyl, or isoquinolinyl). Some examples of heteroaryl are azetidinyl, pyridyl, IH- indazolyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuryl, isoquinolinyl, benzthiazolyl, xanthene, thioxanthene, phenothiazine, dihydroindole, 1,2,3,4- tetrahydroisoquinoline, isoindoline, benzo[l,3]dioxole, benzo[b]furyl, benzo[b]thiophenyl, indazolyl, benzimidazolyl, benzthiazolyl, puryl, cinnolyl, quinolyl, quinazolyl,cinnolyl, phthalazyl, quinazolyl, quinoxalyl, isoquinolyl, 4H-quinolizyl, benzo-l,2,5-thiadiazolyl, or 1,8-naphthyridyl.

[59] Without limitation, monocyclic heteroaryls include furyl, thiophenyl, 2H-pyrrolyl, pyrrolyl, oxazolyl, thazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4-H-pranyl, pyridyl, pyridazyl, pyrimidyl, pyrazolyl, pyrazyl, or 1,3,5-triazyl. Monocyclic heteroaryls are numbered according to standard chemical nomenclature. [60] Without limitation, bicyclic heteroaryls include indolizyl, indolyl, isoindolyl, 3H- indolyl, indolinyl, benzo[6]furyl, benzo[6]thiophenyl, quinolinyl, isoquinolinyl, indolizyl, isoindolyl, indolyl, benzo[6]furyl, bexo[ό]thiophenyl, indazolyl, benzimidazyl, benzthiazolyl, purinyl, 4H-quinolizyl, quinolyl, isoquinolyl, cinnolyl, phthalazyl, quinazolyl, quinoxalyl, 1,8-naphthyridyl, or pteridyl. Bicyclic heteroaryls are numbered according to standard chemical nomenclature.

[61] A heteroaryl is optionally substituted with one or more substituents such as aliphatic [e.g., alkyl, alkenyl, or alkynyl]; cycloaliphatic; (cycloaliphatic)aliphatic; heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; oxo (on a non-aromatic carbocyclic or heterocyclic ring of a bicyclic or tricyclic heteroaryl); carboxy; amido; acyl [e.g., aliphaticcarbonyl; (cycloaliphatic)carbonyl; ((cycloaliphatic)aliphatic)carbonyl;

(araliphatic)carbonyl ; (heterocycloaliphatic)carbonyl ;

((heterocycloaliphatic)aliphatic)carbonyl; or (heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphaticsulfonyl or aminosulfonyl]; sulfinyl [e.g., aliphaticsulfinyl]; sulfanyl [e.g., aliphaticsulfanyl]; nitro; cyano; halo; hydroxy; mercapto; sulfoxy; urea; thiourea; sulfamoyl; sulfamide; or carbamoyl. Alternatively, a heteroaryl can be unsubstituted. [62] Non-limiting examples of substituted heteroaryls include (halo)heteroaryl (e.g., mono- and di-(halo)heteroaryl); (carboxy)heteroaryl (e.g., (alkoxycarbonyl)heteroaryl); cyanoheteroaryl; aminoheteroaryl (e.g., ((alkylsulfonyl)amino)heteroaryl and ((dialkyl)amino)heteroaryl); (amido)heteroaryl [e.g., aminocarbonylheteroaryl, ((alkylcarbonyl)amino)heteroaryl, ((((alkyl)amino)alkyl)aminocarbonyl)heteroaryl, (((heteroaryl)amino)carbonyl)heteroaryl, ((heterocycloaliphatic)carbonyl)heteroaryl, and ((alkylcarbonyl)amino)heteroaryl] ; (cyanoalkyl)heteroaryl; (alkoxy)heteroaryl; (sulfamoyl)heteroaryl (e.g., (aminosulfonyl)heteroaryl); (sulfonyl)heteroaryl (e.g., (alkylsulfonyl)heteroaryl); (hydroxyalkyl)heteroaryl; (alkoxyalkyl)heteroaryl; (hydroxy)heteroaryl; ((carboxy)alkyl)heteroaryl; (((dialkyl)amino)alkyl]heteroaryl; (heterocycloaliphatic)heteroaryl; (cycloaliphatic)heteroaryl; (nitroalkyl)heteroaryl; (((alkylsulfonyl)amino)alkyl)heteroaryl; ((alkylsulfonyl)alkyl)heteroaryl; (cyanoalkyl)heteroaryl; (acyl)heteroaryl [e.g., (alkylcarbonyl)heteroaryl]; (alkyl)heteroaryl, and (haloalkyl)heteroaryl [e.g., trihaloalkylheteroaryl].

[63] As used herein, a "heteroaraliphatic" (such as a heteroaralkyl group) refers to an aliphatic group (e.g., a Ci_-4 alkyl group) that is substituted with a heteroaryl group. "Aliphatic," "alkyl," and "heteroaryl" have been defined above.

[64] As used herein, a "heteroaralkyl" group refers to an alkyl group (e.g., a Ci_-4 alkyl group) that is substituted with a heteroaryl group. Both "alkyl" and "heteroaryl" are as defined herien. A heteroaralkyl is optionally substituted with one or more substituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoromethyl), alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino, cyano, halo, hydroxy, acyl, mercapto, alkylsulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, or carbamoyl. [65] As used herein, an "acyl" group refers to a formyl group or R^X-C(O)- (such as alkyl-C(O)-, also referred to as "alkylcarbonyl") wherein R^x and "alkyl" are as defined herein.

Acetyl and pivaloyl are examples of acyl groups.

[66] As used herein, an "aroyl" or "heteroaroyl" refers to an aryl-C(O)- or a heteroaryl-

C(O)-. The aryl and heteroaryl portion of the aroyl or heteroaroyl is optionally substituted as previously defined.

[67] As used herein, an "alkoxy" group refers to an alkyl-O- group where "alkyl" are as defined herein.

[68] As used herein, a "carbamoyl" group refers to a group having the structure -O-CO-

NR^XR^Y or -NR^X-CO-O-R^Z wherein R^x and R^γ are as defined herein, and R^z can be aliphatic, aryl, araliphatic, heterocycloaliphatic, heteroaryl, or heteroaraliphatic.

[69] As used herein, a "carboxy" group refers to -COOH, -C00R^x, -OC(O)H, or

-OC(O)R^X when used as a terminal group; or -OC(O)- or -C(O)O- when used as an internal group.

[70] As used herein, a "haloaliphatic" group refers to an aliphatic group substituted with 1 to 3 halogen atoms. For instance, the term haloalkyl includes the group -CF₃.

[71] As used herein, a "mercapto" group refers to -SH.

[72] As used herein, a "sulfo" group refers to -SO₃H or -SO₃R^X when used terminally or

-S(O)₃- when used internally.

[73] As used herein, a "sulfamide" group refers to the structure -NR^X-S(O)₂-NR^YR^Z when used terminally or -NR^X-S(O)₂-NR^Y- when used internally, wherein R^x, R^γ, and R^z are as defined above.

[74] As used herein, a "sulfonamide" group refers to the structure -S(O)₂-NR^XR^Y or

-NR^x-S(0)₂-R^z when used terminally; or -S(O)₂-NR^X- or -NR^X -S(O)₂- when used internally, wherein R^x, R^γ, and R^z are as defined herein.

[75] As used herein a "sulfanyl" group refers to -S-R^x when used terminally or -S- when used internally, wherein R is as defined herein. Examples of sulfanyls include aliphatic-S-, cycloaliphatic-S-, aryl-S-, or the like.

[76] As used herein a "sulfinyl" group refers to -S(O)-R^X when used terminally or -S(O)- when used internally, wherein R^x is as defined above. Exemplary sulfinyl groups include, but are not limited to, aliphatic-S(O)-, aryl-S(O)-, (cycloaliphatic(aliphatic))-S(O)-, cycloalkyl-S(O)-, heterocycloaliphatic-S(O)-, heteroaryl-S(O)-, and the like.

[77] As used herein, a "sulfonyl" group refers to-S(O)₂-R^x when used terminally or

-S(O)₂- when used internally, wherein R^x is as defined herein. Exemplary sulfonyl groups include, but are not limited to, aliphatic-S(0)2-, aryl-S(O)₂-, (cycloaliphatic(aliphatic))-S(O)₂-, cycloaliphatic-S(O)₂-, heterocycloaliphatic-S(O)₂-, heteroaryl-S(O)₂-,

(cycloaliphatic(amido(aliphatic)))-S(O)₂-, and the like.

[78] As used herein, a "sulfoxy" group refers to -O-SO-R^X or -SO-O-R* when used terminally, or -0-S(O)- or -S(O)-O- when used internally, where R^x as as defined herein.

[79] As used herein, a "halogen" or "halo" group refers to fluorine, chlorine, bromine, or iodine.

[80] As used herein, an "alkoxycarbonyl" group, which is encompassed by the term carboxy, used alone or in connection with another group, refers to a group such as alkyl-O-

C(O)- wherein alkyl is as defined herein.

[81] As used herein, an "alkoxyalkyl" group refers to an alkyl group such as alkyl-O-alkyl-, wherein alkyl is as defined herein.

[82] As used herein, a "carbonyl" group refer to -C(O)-.

[83] As used herein, an "oxo" group refers to =0.

[84] As used herein, the term "phosphor" refers to phosphinates and phosphonates.

Examples of phosphinates and phosphonates include, but are not limited to, -P(O)(R^P)₂, wherein R^p is aliphatic, alkoxy, aryloxy, heteroaryloxy, (cycloaliphatic)oxy,

(heterocycloaliphatic)oxy aryl, heteroaryl, cycloaliphatic or amino.

[85] As used herein, an "aminoalkyl" group refers to the structure (R^x)₂N-alkyl- wherein

R^x and alkyl are as defined herein.

[86] As used herein, a "cyanoalkyl" group refers to the structure (NC)-alkyl-.

[87] As used herein, a "urea" group refers to the structure -NR^X-CO-NR^YR^Z and a

"thiourea" group refers to the structure -NR^X-CS-NR^YR^Z when used terminally and -NR^X-

C0-NR^γ- or -NR^X-CS-NR^Y- when used internally, wherein R^x, R^γ, and R^z have been defined above.

[88] As used herein, a "guanidine" group refers to the structure -N=C(N(R^XR^Y))N(R^XR^Y) or -NR^X-C(=NR^X)NR^XR^Y wherein R^x and R^γ are as defined herien.

[89] As used herein, the term "amidino" group refers to the structure -C=(NR^X)N(R^XR^Y) wherein R^x and R^γ have been defined above.

[90] In general, the term "vicinal" refers to the placement of substituents on a group that includes two or more carbon atoms, wherein the substituents are attached to adjacent carbon atoms.

[91] In general, the term "geminal" refers to the placement of substituents on a group that includes two or more carbon atoms, wherein the substituents are attached to the same carbon atom.

[92] As used herein, the terms "terminally" and "internally" refer to the location of a group within a substituent. A group is terminal when the group is present at the end of the substituent not further bonded to the rest of the chemical structure. Carboxyalkyl, i.e., R^xO(O)C-alkyl, is an example of a carboxy group used terminally. A group is internal when the group is present in the middle of a substituent of the chemical structure. Alkylcarboxy (e.g., alkyl-C(O)O- or alkyl-OC(O)-) and alkylcarboxyaryl (e.g., alkyl-C(O)O-aryl- or alkyl- O(CO)-aryl-) are examples of carboxy groups used internally.

[93] As used herein, a "cyclic group" or "cyclic moiety" includes mono-, bi-, and tri-cyclic ring systems including cycloaliphatic, heterocycloaliphatic, aryl, or heteroaryl, each of which is as defined herein.

[94] As used herein, a "bridged cyclic ring system" refers to a cyclic heterocycloalipahtic ring system or cyclic cycloaliphatic ring system in which at least two rings have two atoms in common. Examples of bridged cyclic ring systems include, but are not limited to, adamantanyl, norbornanyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl, bicyclo[3.2.3]nonyl, 2-oxabicyclo[2.2.2]octyl, l-azabicyclo[2.2.2]octyl, 3- azabicyclo[3.2.1]octyl, and 2,6-dioxa-tricyclo[3.3.1.0^3>7]nonyl. A bridged cyclic ring system can be optionally substituted with one or more substituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoromethyl), alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino, cyano, halo, hydroxy, acyl, mercapto, alkylsulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, or carbamoyl. [95] As used herein, an "aliphatic chain" refers to a branched or straight aliphatic group (e.g., alkyl groups, alkenyl groups, or alkynyl groups). A straight aliphatic chain has the structure -(CH₂)_V-, where v is 1-6. A branched aliphatic chain is a straight aliphatic chain that is substituted with one or more aliphatic groups. A branched aliphatic chain has the structure -(CHQ)v- where Q is hydrogen or an aliphatic group; however, Q shall be an aliphatic group in at least one instance. The term aliphatic chain includes alkyl chains, alkenyl chains, and alkynyl chains, where alkyl, alkenyl, and alkynyl are defined above. [96] As used herein, the term "optionally substituted" is used interchangeably with the term "substituted or unsubstituted." As described herein, compounds of the invention can optionally be substituted with one or more substituents, such as are illustrated generally above, or as exemplified by particular classes, subclasses, and species of the invention. As described herein, the variables A, B, R₁, R₂, R₃, Y and Y', and other variables contained in formulae described herein encompass specific groups, such as alkyl and aryl. Unless otherwise noted, each of the specific groups for the variables A, B, Ri, R₂, R₃, Y and Y', and other variables contained therein can be optionally substituted with one or more substituents described herein. Each substituent of a specific group is further optionally substituted with one to three of halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, cycloaliphatic, heterocycloaliphatic, heteroaryl, haloalkyl, and alkyl. For instance, an alkyl group can be substituted with alkylsulfanyl and the alkylsulfanyl can be optionally substituted with one to three of halo, cyano, oxo, alkoxy, hydroxy, amino, nitro, aryl, haloalkyl, and alkyl. As an additional example, the cycloalkyl portion of a (cycloalkyl)carbonylamino can be optionally substituted with one to three of halo, cyano, alkoxy, hydroxy, nitro, haloalkyl, and alkyl. When two alkoxy groups are bound to the same atom or adjacent atoms, the two alkoxy groups can form a ring together with the atom(s) to which they are bound. [97] In general, the term "substituted", whether preceded by the term "optionally" or not, refers to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. Specific substituents are described above in the definitions and below in the description of compounds and examples thereof. Unless otherwise indicated, an optionally substituted group can have a substituent at each substitutable position of the group, and when more than one position in any given structure can be substituted with more than one substituent selected from a specified group, the substituent can be either the same or different at every position. A ring substituent, such as a heterocycloalkyl, can be bound to another ring, such as a cycloalkyl, to form a spiro-bicyclic ring system, e.g., both rings share one common atom. As one of ordinary skill in the art will recognize, combinations of substituents envisioned by this invention are those combinations that result in the formation of stable or chemically feasible compounds.

[98] The term "stable or chemically feasible," as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and preferably their recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is one that is not substantially altered when kept at a temperature of 40 ⁰C or less, in the absence of moisture or other chemically reactive conditions, for at least a week.

[99] An "antagonist," as used herein, refers to a molecule that binds to the receptor without activating the receptor. It competes with the endogenous ligand(s) or substrate(s) for binding site(s) on the receptor and, thus inhibits the ability of the receptor to transduce an intracellular signal in response to endogenous ligand binding.

[100] As compounds of Formula (I) are antagonists of TGF/3 receptor type I (Alk5) and/or activin receptor type I (Alk4), these compounds are useful in inhibiting the consequences of TGF/3 and/or activin signal transduction such as the production of extracellular matrix (e.g., collagen and fibronectin), the differentiation of stromal cells to myofibroblasts, and the stimulation of and migration of inflammatory cells. Thus, compounds of Formula (I) inhibit pathological inflammatory and fibrotic responses and possess the therapeutic utility of treating and/or preventing disorders or diseases for which reduction of TGF/3 and/or activin activity is desirable (e.g., various types of fibrosis or progressive cancers). In addition, the compounds of Formula (I) are useful for studying and researching the role of TGF/3 receptor type I (Alk5) and/or activin receptor type I (Alk4), such as their role in cellular processes, for example, signal transduction, production of extracellular matrix, the differentiation of stromal cells to myofibroblasts, and the stimulation of and migration of inflammatory cells. [101] As used herein, "an effective amount" (or "a pharmaceutically effective amount") is defined as the amount required to confer a therapeutic effect on the treated patient, and is typically determined based on age, surface area, weight, and condition of the patient. The interrelationship of dosages for animals and humans (based on milligrams per meter squared of body surface) is described by Freireich et al., Cancer Chemother. Rep., 50: 219 (1966). Body surface area may be approximately determined from height and weight of the patient. See, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, New York, 537 (1970). As used herein, "patient" refers to a mammal, including a human.

[102] Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C -enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools or probes in biological assays, or as therapeutic agents. [103] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. DETAILED DESCRIPTION OF THE INVENTION

[104] In general, the invention features compounds of Formula (I), which exhibit surprisingly high affinitiy for the TGF/3 family type I receptors, Alk5 and/or Alk4. Synthesis of the Compounds of Formula (T)

[105] Compounds of the invention may be prepared using known methodology. In one method, the compounds wherein X is O, Ai is NH and A₂ is C may be prepared as illustrated below in Scheme 1.

Scheme 1

[106] Referring to Scheme 1, reaction of the aldehyde 1 with ethylene glycol in the presence of a catalytic amound of/>-toluenesulfonic acid provides the acetal 2. Alkylation of 2 with a bromochloro alkane, e.g. l-bromo-2-chloroethane, provides the alkylated acetal 3. Treatment of 3 with silver nitrate in water at elevated temperature results in acetal hydrolysis and ring closure to provide the spirolactone 5. Reaction of 5 with a dione 6 and ammonium acetate provides imidazole compounds of the invention (i.e., Formula (I)).

[107] Alternatively, compounds of Formula (I) wherein X is O, Ai is C ,and A₂ is N, may be prepared as illustrated below in Scheme 2. Scheme 2

Pyridine

[108] Referring to Scheme 2, protection of the hydroxyester 7 is achieved by reaction with ϊ-bytyldimethylsilyl chloride in the presence of imidazole to provide 8. Alkylation of 8 with a bromochloro alkane, e.g. l-bromo-2-chloroethane, provides the alkylated ester 9. Treatment of compound 9 with silver nitrate in water at elevated temperature results in deprotection and ring closure to provide the spirolactone 10. Reaction of 10 with/?- toluenesulfonyl chloride in the presence of pyridine provides the tosylate 11. Reaction of 11 with the imidazole 12 provides imidazole compounds of the invention (i.e., Formula (I)). [109] Compounds of the Formula (I) wherein X is N-R₃ may be prepared from the corresponding lactones illustrated in Schemes 1 and 2 by reaction of the lactone with an amine, N-R₃, in the presence of aluminum trichloride.

Uses of Compounds of Formula (T)

[110] As discussed above, hyperactivity of the TGF/3 family signaling pathways can result in excess deposition of extracellular matrix and increased inflammatory responses, which can then lead to fibrosis in tissues and organs (e.g., lung, kidney, and liver) and ultimately result in organ failure. See, e.g., Border, W.A. and Ruoslahti E. J. Clin. Invest. 90: 1-7 (1992) and Border, W.A. and Noble, N.A. N. Engl. J. Med. 331 : 1286-1292 (1994). Studies have been shown that the expression of TGF/3 and/or activin mRNA and the level of TGFjS and/or activin are increased in patients suffering from various fibrotic disorders, e.g., fϊbrotic kidney diseases, alcohol-induced and autoimmune hepatic fibrosis, myelofibrosis, bleomycin- induced pulmonary fibrosis, and idiopathic pulmonary fibrosis. Elevated TGFβ and/or activin is has also been demonstrated in cachexia, demyelination of neurons in multiple sclerosis, Alzheimer's disease, cerebral angiopathy and hypertension.

[Ill] Compounds of Formula (I), which are antagonists of the TGF/3 family type I receptors Alk5 and/or Alk4, and inhibit TGFjS and/or activin signaling pathway, are therefore useful for treating and/or preventing fibrotic disorders or diseases mediated by an increased level of TGF/3 and/or activin activity. As used herein, a compound inhibits the TGFjS family signaling pathway when it binds (e.g., with an IC₅₀ value of less than 10 μM; such as, less than 1 μM; and for example, less than 5 nM) to a receptor of the pathway (e.g., Alk5 and/or AIk 4), thereby competing with the endogenous ligand(s) or substrate(s) for binding site(s) on the receptor and reducing the ability of the receptor to transduce an intracellular signal in response to the endogenous ligand or substrate binding. The aforementioned disorders or diseases include any condition (a) marked by the presence of an abnormally high level of TGF/3 and/or activin; and/or (b) an excess accumulation of extracellular matrix; and/or (c) an increased number and synthetic activity of myofibroblasts. These disorders or diseases include, but are not limited to, fibrotic conditions such as scleroderma, glomerulonephritis, diabetic nephropathy, lupus nephritis, hypertension-induced nephropathy, ocular or corneal scarring, alimentary track or gastrointestinal fibrosis, renal fibrosis, hepatic or biliary fibrosis, acute lung injury, pulmonary fibrosis (such as idiopathic pulmonary fibrosis and radiation- induced pulmonary fibrosis), post-infarction cardiac fibrosis, fibrosclerosis, fibrotic cancers, fibroids, fibroma, fibroadenomas, and fibrosarcomas. Other fibrotic conditions for which preventive treatment with compounds of Formula (I) can have therapeutic utility include radiation-induced fibrosis, chemotherapy-induced fibrosis, and surgically-induced scarring including surgical adhesions, laminectomy, and coronary restenosis.

[112] Increased TGFjS activity is also found to manifest in patients with progressive cancers. Studies have shown that in many cancers, the tumor cells, stromal cells, and/or other cells within a tumor generally overexpress TGF/3. This leads to stimulation of angiogenesis and cell motility, suppression of the immune system, and/or increased interaction of tumor cells with the extracellular matrix. See, e.g., Hojo, M. et al., Nature 397: 530-534 (1999) and Lammerts E. et al., Int. J. Cancer 102: 453-462 (2002). As a result, the tumors grow more readily, become more invasive and metastasize to distant organs. See, e.g., Maehara, Y. et al., J. CHn. Oncol. 17: 607-614 (1999) and Picon, A. et al., Cancer Epidemiol. Biomarkers Prev. 7: 497-504 (1998). Thus, compounds of Formula (I), which are antagonists of the TGF/3 type I receptor and inhibit TGF/3 signaling pathways, are also useful for treating and/or preventing various cancers which overexpress TGF/3 or benefit from TGF/3's above-mentioned pro- tumor activities. Such cancers include carcinomas of the lung, breast, liver, biliary tract, gastrointestinal tract, head and neck, pancreas, prostate, cervix as well as multiple myeloma, melanoma, glioma, and glioblastomas.

[113] Importantly, it should be pointed out that because of the chronic, and in some cases localized, nature of disorders or diseases mediated by overexpression of TGF/3 and/or activin (e.g., fibrosis or cancers), small molecule treatments (such as treatment disclosed in the present invention) are favored for long-term treatment.

[114] Not only are compounds of Formula (I) useful in treating disorders or diseases mediated by high levels of TGF/3 and/or activin activity, these compounds can also be used to prevent the same disorders or diseases. It is known that polymorphisms leading to increased TGF/3 and/or activin production have been associated with fibrosis and hypertension. Indeed, high serum TGF/3 levels are correlated with the development of fibrosis in patients with breast cancer who have received radiation therapy, chronic graft-versus-host-disease, idiopathic interstitial pneumonitis, veno-occlusive disease in transplant recipients, and peritoneal fibrosis in patients undergoing continuous ambulatory peritoneal dialysis. Thus, the levels of TGF/3 and/or activin in serum and of TGF/3 and/or activin mRNA in tissue can be measured and used as diagnostic or prognostic markers for disorders or diseases mediated by overexpression of TGF/3 and/or activin, and polymorphisms in the gene for TGFjS that determine the production of TGF/3 and/or activin can also be used in predicting susceptibility to disorders or diseases. See, e.g., Blobe, G.C. et al., N. Engl. J. Med. 342(18): 1350-1358 (2000); Matsuse, T. et al., Am. J. Respir. Cell MoI. Biol. 13: 17-24 (1995); Inoue, S. et al., Biochem. Biophys. Res. Comm. 205: 441-448 (1994); Matsuse, T. et al, Am. J. Pathol. 148: 707-713 (1996); De Bleser et al., Hepatology 26: 905-912 (1997); Pawlowski, J.E., et al., J. Clin. Invest. 100: 639-648 (1997); and Sugiyama, M. et al., Gastroenterology 114: 550-558 (1998).

Administration of Compounds of Formula (I)

[115] As defined above, an effective amount is the amount required to confer a therapeutic effect on the treated patient. For a compound of Formula (I), an effective amount can range, for example, from about 1 mg/kg to about 150 mg/kg (e.g., from about 1 mg/kg to about 100 mg/kg). Effective doses will also vary, as recognized by those skilled in the art, dependant on route of administration, excipient usage, and the possibility of co-usage with other therapeutic treatments including use of other therapeutic agents and/or radiation therapy. [116] Compounds of Formula (I) can be administered in any manner suitable for the administration of pharmaceutical compounds, including, but not limited to, pills, tablets, capsules, aerosols, suppositories, liquid formulations for ingestion or injection or for use as eye or ear drops, dietary supplements, and topical preparations. The pharmaceutically acceptable compositions include aqueous solutions of the active agent, in an isotonic saline, 5% glucose or other well-known pharmaceutically acceptable excipient. Solubilizing agents such as cyclodextrins, or other solubilizing agents well-known to those familiar with the art, can be utilized as pharmaceutical excipients for delivery of the therapeutic compounds. As to route of administration, the compositions can be administered orally, intranasally, transdermally, intradermally, vaginally, intraaurally, intraocularly, buccally, rectally, transmucosally, or via inhalation, implantation (e.g., surgically), or intravenous administration. The compositions can be administered to an animal (e.g., a mammal such as a human, non-human primate, horse, dog, cow, pig, sheep, goat, cat, mouse, rat, guinea pig, rabbit, hamster, gerbil, or ferret, or a bird, or a reptile, such as a lizard). [117] Optionally, compounds of Formula (I) can be administered in conjunction with one or more other agents that inhibit the TGF/3 signaling pathway or treat the corresponding pathological disorders (e.g., fibrosis or progressive cancers) by way of a different mechanism of action. Examples of these agents include angiotensin converting enzyme inhibitors, nonsteroid and steroid anti-inflammatory agents, immunotherapeutics, chemotherapeutics, as well as agents that antagonize ligand binding or activation of the TGFjS receptors, e.g., anti-

TGFjS, anti-TGFjS receptor antibodies, or antagonists of the TGF/3 type II receptors.

Compounds of Formula (I) can also be administered in conjunction with other treatments, e.g., radiation.

[118] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

Example 1 : 8-(5-([l,2,4]triazolo[l,5-a]pyridin-6-yl)-4-(6-methylpyridin-2-yl)-lH- imidazol-2-yl)-2-oxaspiro[4.5]decan-l-one

Step 1: Dioxolan-2-yl-cyclohexanecarboxylic acid methyl ester.

[119] A solution of 4-(methoxycarbonyl)-cyclohexanecarboxaldehyde (10.0 g, 0.0588 mol), 1 ,2-ethanediol (16.4 mL, 0.294 mol), p-toluenesulfonic acid monohydrate (0.7 g, 0.004 mol) in benzene (200 mL, 2 mol; ) was refluxed under a Dean-Stark trap overnight. The mixture was cooled to room temperature, diluted with ether and washed with water followed by aqueous NaHCO₃. The organic phase was dried over MgSO₄ and concentrated to give a mix of isomers (1:3) (10.2 g, 80%). MS: ES+/214.63 (M+l) The major isomer: ¹H-NMR, CDCl₃, 300MHz, d 4.55 (d, IH), 3.85-3.80 (m, 4H), 3.63 (s,

3H), 2.20 (m, IH), 2.05-1.85 (m, 4H), 1.50-1.15 (m, 4H)

Step 2: methyl l-(2-chloroethyl)-4-(l,3-dioxolan-2-yl)cyclohexanecarboxylate.

[120] To a solution of 2.0M LDA in heptane (17 mL) and tetrahydrofuran (40 mL, 0.5 mol) was added 4-[l,3]dioxolan-2-yl-cyclohexanecarboxylic acid methyl ester (5.0 g, 0.023 mol) at -78 ⁰C. The mixture was stirred at -78 °C for 30 minutes. l-Bromo-2-chloro-ethane, (5.02 g, 0.0350 mol) was then added to the solution at -78 ⁰C and the mixture stirred for 1 hour, then warmed to room temperature and held overnight. The mixture was concentrated and the residue was partitioned between water and CH₂Cl₂. The aqueous layer was extracted with CH₂Cl₂, the organic layers were combined, dried, and concentrated to give the desired product as a light orange colored liquid (6.45 g, 100%).

¹H-NMR, CDCl₃, 300MHz, d 4.50 (d, IH), 3.85-3.70 (m, 4H), 3.63 (s, 3H), 3.36 (t, 2H), 2.21 (m, IH), 1.88 (t, 2H), 1.65 (m, 2H), 1.45 (m, 3H), 1.07 (m, 2H)

Step 3: l-oxo-2-oxaspiro[4.51decane-8-carbaldehyde.

[121] A mixture of silver nitrate (1.23 g, 0.00723 mol) in water (15 mL, 0.83 mol) and 1 ,4- dioxane (20 mL, 0.2 mol) was heated to reflux. Methyl l-(2-chloroethyl)-4-(l,3-dioxolan-2- yl)cyclohexanecarboxylate (1.00 g, 0.00361 mol) in 1,4-dioxane (5 mL, 0.06 mol) was added. The mixture was refluxed for 30 minutes then cooled to room temperature. The mixture was concentrated and the residue partitioned between CH₂Cl₂ and water. The organic layer was dried and concentrated to give a mixture of 8-(l,3-dioxolan-2-yl)-2-oxaspiro[4.5]decan-l-one and l-oxo-2-oxaspiro[4.5]decane-8-carbaldehyde (¹H-NMR, CDCl₃, 300MHz, d 9.61 (s, - CHO) which was used in the next step without further purification.

Step 4: 8-(5-([l,2,4]triazolo[l,5-alpyridin-6-yl)-4-(6-methylpyridin-2-yl)-lH-imidazol-2- yl)-2-oxaspiro [4.5] decan-1-one.

[122] To a suspension of 1 -([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-2-(6-methylpyridin-2- yl)ethane-l,2-dione (450 mg, 0.0017 mol), l-oxo-2-oxaspiro[4.5]decane-8-carbaldehyde (204 mg, 0.00112 mol) , ammonium acetate (650 mg, 0.0084 mol; ) and 2-methoxy-2- methylpropane (10 mL, 0.1 mol) was added acetic acid (0.6 mL, 0.01 mol). The mixture was heated to reflux overnight. LC/MS indicated completion of the reaction and the formation of the desired product. The mixture was concentrated and the residue was dissolved in CH₂Cl₂ and washed with water. The organic phase was concentrated and the residue purified by preparative HPLC to give the desired product as a yellow solid (326 mg). 1H-NMR, MeOD, 300MHz, d 9.20 (s, IH), 8.42 (s, IH), 7.82-7.61 (m, 3H), 7.26 (m, 2H), 4.25 (t, 2H), 3.12 (m, IH), 2.5 (s, 3H), 2.25 (t, 2H), 2.1-2.0 (m, 2H), 1.85-1.65 (m, 4H) MS: ES+/429.06 (M+l).

Example 2: 8-(5-([l,2,4]triazolo[l,5-a]pyridin-6-yl)-4-(5-fluoro-6-methyIpyridin-2-yl)- lH-imidazol-2-yl)-2-oxaspiro [4.5] decan-1-one

[123] To a suspension of l-(5-Fluoro-6-methyl-pyridin-2-yl)-2-[l,2,4]triazolo[l,5- a]pyridin-6-yl-ethane-l,2-dione (477 mg, 0.00168 mol), l-oxo-2-oxaspiro[4.5]decane-8- carbaldehyde (200 mg, 0.001 mol), ammonium acetate (650 mg, 0.0084 mol) and 2-methoxy- 2-methylpropane (10 mL, 0.08 mol) was added acetic acid (0.6 mL, 0.01 mol). The mixture was heated to reflux overnight. LC/MS indicated completion of the reaction and the formation of the desired product. The mixture was concentrated and the residue was dissolved in CH₂Cl₂ and washed with water. The organic phase was concentrated and the residue purified by preparative HPLC to give the desired product as a yellow solid (263mg).

¹H-NMR, MeOD, 300MHz, d 9.20 (s, IH), 8.42 (s, IH), 7.85-7.70 (m, 2H), 7.40 (t, IH), 7.31

(m, IH), 4.25 (t, 2H), 3.15 (m, IH), 2.46 (d, 3H), 2.25 (t, 2H), 2.15-2.0 (m, 2H), 1.85-1.65 (m,

4H).

MS: ES+/446.94 (M+ 1).

Example 3 : 8-(3-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)-lH-pyrazol-l-yl)-2- oxaspiro [4.5] decan-1-one

Step 1: ethyl 4-(tert-butyldimethyIsilyloxy)cyclohexanecarboxylate.

[124] tert-Butyldimethylsilyl chloride (3.61 g, 0.0240 mol) was added to a solution of 4- hydroxy-cyclohexanecarboxylic acid ethyl ester (3.44 g, 0.0200 mol; ), and lH-imidazole (3.40 g, 0.0499 mol; ) in N,N-Dimethylformamide (5.0 mL, 0.064 mol). The mixture was stirred at the room temperature for 3 hours. The mixture was partitioned between ethyl acetate and water. The ethyl acetate phase was washed with brine, dried over sodium sulfate, filtered, and then concentrated to give oily crude product. Column chromatography eluting with 0-20% ethyl acetate/hexanes gave 5.5 g (96%) of the title compound as colorless oil. ¹H-NMR (300 MHz, CD₃Cl) δ 4.12 (m, 2H), 3.72 (m, IH), 2.29 (m, IH), 1.93 (m, 2H), 1.65 (m, 2H), 1.56 (m, 2H), 1.48 (m, 2H), .125 (m, 3H), 0.88 (d, 6H), 0.03 (d, 6H). Step 2: ethyl 4-(tert-butyldimethylsilyloxy)-l-(2-chloroethyl)cyclohexanecarboxylate.

[125] Lithium diisopropylamide (23.0 mmol, 0.0230 mol) (2M solution in heptane/THF/ethyl benzene) was added to a solution of 4-(terf-butyldimethylsilanyloxy)- cyclohexanecarboxylic acid ethyl ester (5.50 g, 0.0192 mol) in tetrahydrofuran (31.1 mL, 0.384 mol). The mixture was stirred at -78 °C for 30 minutes. l-Bromo-2-chloroethane, (4.13 g, 0.0288 mol) was added into the solution at -78 ⁰C and the mixture then stirred for 1 hour. The mixture was warmed to the room temperature over 2 hours. The mixture was concentrated and residue was partitioned between ethyl acetate and water. The ethyl acetate phase was washed with brine, dried over sodium sulfate, filtered, and concentrated. Column chromatography eluting with 0 to 20% ethyl acetate gave 2.0 g (30%) of the title compound as slightly yellow oil.

¹H-NMR (300 MHz, CD₃Cl) δ 4.15 (m, 2H), 3.56 (m, IH), 3.46 (m, 2H), 2.22 (m, 2H), 1.96 (m, 2H), 1.74 (m, 2H), 1.26 (m, 7H), 0.88 (d, 9H), 0.03 (d, 6H).

Step 2: 8-hydroxy-2-oxaspiro[4.5]decan-l-one.

[126] Silver nitrate (1.95 g, 0.0115 mol) was added to 1 ,4-dioxane (17.9 mL, 0.229 mol) and water (10.32 mL, 0.5731 mol). The mixture was heated to reflux. Ethyl 4-(tert- butyldimethylsilyloxy)-l-(2-chloroethyl)cyclohexanecarboxylate (2.00 g, 0.00573 mol) in 1,4-Dioxane (4.47 mL, 0.0573 mol) was added to the reaction mixture. The reaction mixture was then refluxed for 30 min., cooled to room temperature, filtered through celite. and concentrated. The residue was partitioned between ethyl acetate and water. The ethyl acetate phase was washed with brine, dried over sodium sulfate, filtered, and concentrated to give 0.75 g (77%) of a crude product as yellow oil.

¹H-NMR (300 MHz, Methanol-^) δ 3.82 (m, 2H), 3.62 (m, IH), 1.98 (m, 2H), 1.82 (m, 2H), 1.64 (m, 2H), 1.39 (m, 2H), 1.19 (m, 2H). Step 3: l-oxo-2-oxaspiro[4.5]decan-8-yl 4-methylbenzenesulfonate.

[127] /?-Toluenesulfonyl chloride (1.26 g, 0.00661 mol) was added to a solution of 8- hydroxy-2-oxaspiro[4.5]decan-l-one (0.750 g, 0.00441 mol) in pyridine (7.13 mL, 0.0881 mol). The mixture was stirred at 0 ⁰C for 3 hours, and then partitioned between ethyl acetate and water. The ethyl acetate phase was washed with brine, dried over sodium sulfate, filtered, and concentrated to give yellow oil. The oil was puried by silica gel column eluting with 5- 100% ethyl acetate/hexane to give 0.5 g (35%) of the title compound as oil. 1H-NMR (300 MHz, CD₃Cl) δ 7.45 (d, IH, J = 8.1 Hz), 7.03 (d, IH, J = 8.4 Hz), 4.36 (m, IH), 3.94 (m, 2H), 2.12 (s, 3H), 1.79 (m, 2H), 1.67 (m, 4H), 1.32 (m, 2H), 1.12 (m, 2H).

5te2_4_i 8-(3-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yI)-lH-pyrazol-l-yl)-2- oxaspiro [4.5] decan-1-one.

[128] Toluene-4-sulfonic acid l-oxo-2-oxaspiro[4.5]dec-8-yl ester (250.0 mg, 0.0007707 mol) was added to a solution of 6-[5-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoxaline (221 mg, 0.000771 mol) and cesium carbonate (0.502 g, 0.00154 mol) in N,N- dimethylformamide (5.0 mL, 0.064 mol). The mixture was heated at 100 ⁰C for 18 hours, cooled, and then partitioned between ethyl acetate and water. The ethyl acetate phase was washed with brine, dried over sodium sulfate, filtered, and then concentrated to give yellow oil. The yellow oil was purified by HPLC eluting with acetonitrile/water (with 0.1% TFA) to give 30 mg of the title compound as a yellow solid. ¹H-NMR (300 MHz, Methanol-cL,) δ 8.89 (s, 2H), 8.38 (s, IH), 8.30 (t, IH, J = 8.1 Hz), 8.12 (m, IH), 8.06 (d, IH, J = 1.8 Hz), 7.87 (m, 2H), 7.66 (d, IH, J = 8.1 Hz), 4.53 (m, IH), 4.39 (m, 2H), 2.88 (s, 3H), 2.34 (m, 4H), 2.10 (m, 2H), 1.93 (m, 4H). MS (ESP⁺) m/z 440.45.

Example 4 : 8-(4-([l ,2,4] triazolo [1 ,5-a] pyridin-6-yl)-3-(5-fluoro-6-methylpyridin-2-yl)- 1 H-pyrazol-l-yI)-2-oxaspiro [4.5] decan-1-one

[129] Toluene-4-sulfonic acid l-oxo-2-oxa-spiro[4.5]dec-8-yl ester (165 mg, 0.000510 mol) was added to a solution of 6-[3-(5-fluoro-6-methyl-pyridin-2-yl)-lH-pyrazol-4-yl]- [l,2,4]triazolo[l,5-a]pyridine (150.0 mg, 0.0005097 mol) and cesium carbonate (0.415 g, 0.00127 mol) in N,N-dimethylfbrmamide (5.0 mL, 0.064 mol). The mixture was heated at 100 ⁰C for 18 hours, cooled, filtered and concentrated. The residue was partitioned between ethyl acetate and water. The ethyl acetate phase was washed with brine, dried over sodium sulfate, filtered, and concentrated to give brown oil. The oil was purified on HPLC eluting with acetonitrile/water (with 0.1% TFA) to 35 mg of the title compound as a yellow solid. ¹H-NMR (300 MHz, Methanol-^) δ 9.19 (s, IH), 8.75 (s, IH), 8.15 (s, IH), 8.00 (m, IH), 7.86 (d, IH, J = 9.3 Hz), 7.74 (m, IH), 7.06 (m, IH), 4.37 (m, 3H), 2.39 (m, 5H), 2.25 (m, 2H), 2.05 (m, 2H), 1.91 (m, 4H). MS (ESP⁺) m/z 447.40.

Example 5: 8-(4-([l,2,4]triazolo[l,5-a]pyridin-6-yl)-3-(6-methylpyridin-2-yl)-lH- pyrazol-l-yl)-2-oxaspiro[4.5]decan-l-one

[130] Toluene-4-sulfonic acid l-oxo-2-oxa-spiro[4.5]dec-8-yl ester (250.0 mg, 0.0007707 mol) was added to a solution of 6-[3-(6-Methyl-pyridin-2-yl)-lH-pyrazol-4-yl]- [l,2,4]triazolo[l,5-a]pyridine (213 mg, 0.000771 mol) and cesium carbonate (0.502 g,

0.00154 mol) in N,N-dimethylfbrrnamide (5.0 mL, 0.064 mol). The mixture was heated at

100 ⁰C for 18 hours, cooled, filtered, and then concentrated to give yellow oil. The oil was purified on HPLC eluting with acetonitrile/water (with 0.1% TFA) to give 25 mg of the title compound as yellow oil.

¹H-NMR (300 MHz, Methanol-cU) δ: 8.9 (d, IH, J = 1.5 Hz), 8.55 (s, IH), 8.28 (m, 2H), 7.76

(m, 4H), 4.53 (m, IH), 4.37 (m, 2H), 2.88 (s, 3H), 2.41 (m, 2H), 2.02 (m, 8H).

MS (ESP⁺) m/z 429.43.

Example 6: 8-(4-([l,2,4]triazolo[l,5-a]pyridin-6-yl)-3-(pyridin-2-yl)-lH-pyrazol-l-yl)-2- oxaspiro [4.5] decan-1-one

[131] Toluene-4-sulfonic acid l-oxo-2-oxa-spiro[4.5]dec-8-yl ester (5.00E2 mg, 0.00154 mol) was added to a solution of 6-(3-(pyridin-2-yl)-lH-pyrazol-4-yl)-[l,2,4]triazolo[l,5- ajpyridine (404 mg, 0.00154 mol) and cesium carbonate (1.00 g, 0.00308 mol) in N,N- dimethylformamide (10.0 mL, 0.129 mol). The mixture was heated at 100 ⁰C for 18 hours cooled, filtered and concentrated to yellow oil. The oil was purified on HPLC eluting with acetonitrile/water (with 0.1% TFA) to give 100 mg of the title compound as yellow oil. 1H-NMR (300 MHz, Methanol^): δ 9.10 (s, IH), 8.84 (s, IH), 8.70 (m, IH), 8.45 (m, IH), 8.31 (m, IH), 7.89 (m, 3H), 4.52 (m, IH), 4.36 (m, 2H), 2.19 (m, 10H). MS (ESP⁺) m/z 415.18.

Example 7: 2-benzyl-8-(3-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)-lH-pyrazol-l-yl)-2- azaspiro [4.5] decan-1-one

[132] Aluminum trichloride (2 mg, 0.00001 mol) was added to a solution of 8-[3-(6-methyl- pyridin-2-yl)-4-quinoxalin-6-yl-pyrazol-l-yl]-2-oxa-spiro[4.5]decan-l-one (50.0 mg, 0.000114 mol) and benzylamine (3.0 mL, 0.027 mol). The mixture was heated at 110 °C for

3 hours, cooled, and then concentrated to give a yellow residue. The yellow residue was purified on HPLC eluting with acetonitrile/water (with 0.1% TFA) to give 6 mg of the title compound as a yellow solid.

¹H-NMR (300 MHz, Methanol^) δ: 8.89 (m, 2H), 8.11 (m, 5H), 7.33 (m, 7H), 4.49 (m, 3H),

4.10 (s, 2H), 2.64 (s, 3H), 2.09 (m, 10H).

MS (ESP⁺) m/z 529.30.

Example 8: Cell-Free Assay for Evaluating Inhibition of Autophosphorylation of TGF/S Type I Receptor

[133] The serine-threonine kinase activity of TGF/3 type I receptor was measured as the autophosphorylation activity of the cytoplasmic domain of the receptor containing an N- terminal poly histidine, TEV cleavage site-tag, e.g., His-TGFjSRI. The His-tagged receptor cytoplasmic kinase domains were purified from infected insect cell cultures using the Gibco- BRL FastBac HTb baculovirus expression system.

[1341 To a 96-well Nickel FlashPlate (NEN Life Science, Perkin Elmer) was added 20 μL of 1.25 μCi ³³P-ATP/25 μM ATP in assay buffer (50 mM Hepes, 60 mM NaCl, 1 mM MgCl₂, 2 mM DTT, 5 mM MnCl₂, 2% glycerol, and 0.015% Brij^® 35). 10 μL of each test compound of Formula (I) prepared in 5% DMSO solution were added to the FlashPlate. The assay was then initiated with the addition of 20 μL of assay buffer containing 12.5 pmol of His-TGFjSRI to each well. Plates were incubated for 30 minutes at room temperature and the reactions were then terminated by a single rinse with TBS. Radiation from each well of the plates was read on a TopCount (Packard). Total binding (no inhibition) was defined as counts measured in the presence of DMSO solution containing no test compound and non-specific binding was defined as counts measured in the presence of EDTA or no-kinase control. [135] Alternatively, the reaction performed using the above reagents and incubation conditions but in a microcentrifuge tube was analyzed by separation on a 4-20% SDS-PAGE gel and the incorporation of radiolabel into the 40 kDa His-TGFβRI SDS-PAGE band was quantitated on a Storm Phosphoimager (Molecular Dynamics).

[136] Compounds of Formula (I) typically exhibited IC₅₀ values of less than 10 μM; some exhibited IC₅₀ values of less than 1 μM; and some even exhibited IC₅₀ values of less than 50 nM. Example 9: Cell-Free Assay for Evaluating Inhibition of Activin Type I Receptor Kinase Activity

[137] Inhibition of the Activin type I receptor (AIk 4) kinase autophosphorylation activity by test compounds of Formula (I) can be determined in a similar manner to that described above in Example 8 except that a similarly His-tagged form of Alk4 (His-Alk 4) is used in place of the His-TGF/3RI.

Example 10: TGF/S Type I Receptor Ligand Displacement FlashPlate Assay

[138] 50 nM of tritiated 4-(3-pyridin-2-yl-lH-pyrazol-4-yl)-quinoline (custom-ordered from PerkinElmer Life Science, Inc., Boston, MA) in assay buffer (50 mM Hepes, 60 mM NaCl₂, 1 mM MgCl₂, 5 mM MnCl₂, 2 mM 1,4-dithiothreitol (DTT), 2% Brij^® 35; pH 7.5) was premixed with a test compound of Formula (I) in 1% DMSO solution in a v-bottom plate. Control wells containing either DMSO without any test compound or control compound in DMSO were used. To initiate the assay, His-TGF/3 Type I receptor in the same assay buffer (Hepes, NaCl₂, MgCl₂, MnCl₂, DTT, and 30% Brij^® added fresh) was added to a nickel coated FlashPlate (PE, NEN catalog number: SMP 107), while the control wells contained only buffer (i.e., no His-TGF/3 Type I receptor). The premixed solution of tritiated 4-(3- pyridin-2-yl-lH-pyrazol-4-yl)-quinoline and test compound of Formula (I) was then added to the wells. The wells were aspirated after an hour at room temperature and radioactivity in wells (emitted from the tritiated compound) was measured using TopCount (PerkinElmer Lifesciences, Inc., Boston MA).

[139] Compounds of Formula (I) typically exhibited Kj values of less than 10 μM; some exhibited Kj values of less than 1 μM; and some even exhibited Ki values of less than 50 nM.

Example 11: Assay for Evaluating Cellular Inhibition of TGFjS Signaling and Cytotoxicity

[140] Biological activity of the compounds of Formula (I) was determined by measuring their ability to inhibit TGF/3-induced PAI-Luciferase reporter activity in HepG2 cells. [141] HepG2 cells were stably transfected with the PAI-luciferase reporter grown in DMEM medium containing 10% FBS, penicillin (100 U/mL), streptomycin (100 μg/mL), L- glutamine (2 mM), sodium pyruvate (1 mM), and non-essential amino acids (Ix). The transfected cells were then plated at a concentration of 2.5 x 10⁴ cells/well in 96 well plates and starved for 3-6 hours in media with 0.5% FBS at 37 ⁰C in a 5% CO₂ incubator. The cells were then stimulated with 2.5 ng/mL TGF/3 ligand in the starvation media containing 1% DMSO either in the presence or absence of a test compound of Formula (I) and incubated as described above for 24 hours. The media was washed out the following day and the luciferase reporter activity was detected using the LucLite Luciferase Reporter Gene Assay kit (Packard, Cat. No. 6016911) as recommended. The plates were read on a Wallac Microbeta plate reader, the reading of which was used to determine the IC₅₀ values of compounds of Formula (I) for inhibiting TGF/3-induced PAI-Luciferase reporter activity in HepG2 cells. Compounds of Formula (I) typically exhibited IC₅₀ values of less 10 μM. [142] Cytotoxicity was determined using the same cell culture conditions as described above. Specifically, cell viability was determined after overnight incubation with the CytoLite cell viability kit (Packard, Cat. No. 6016901). Compounds of Formula (I) typically exhibited LD₂₅ values greater than 10 μM.

Example 12: Assay for Evaluating Inhibition of TGFβ Type I Receptor Kinase Activity in Cells

[143] The cellular inhibition of activin signaling activity by the test compounds of Formula (I) is determined in a similar manner as described above in Example 11 except that 100 ng/mL of activin is added to serum starved cells in place of the 2.5 ng/mL TGF/3.

Example 13: Assay for TGF/5-Induced Collagen Expression

Preparation of Immortalized Collagen Promotor-Green Fluorescent Protein Cells [144] Fibroblasts are derived from the skin of adult transgenic mice expressing Green Fluorescent Protein (GFP) under the control of the collagen IAl promoter (see Krempen, K. et al., Gene Exp. 8: 151-163 (1999)). Cells are immortalized with a temperature sensitive large T antigen that is in an active stage at 33 ⁰C. Cells are expanded at 33 ⁰C and then transferred to 37 ⁰C at which temperature the large T antigen becomes inactive (see Xu, S. et al., Exp. Cell Res. 220: 407-414 (1995)). Over the course of about 4 days and one split, the cells cease proliferating. Cells are then frozen in aliquots sufficient for a single 96 well plate.

Assay of TGFβ-induced Collagen-GFP Expression

[145] Cells are thawed, plated in complete DMEM (contains non-essential amino acids, ImM sodium pyruvate and 2mM L-glutamine) with 10 % fetal calf serum, and then incubated for overnight at 37 ⁰C, 5% CO₂. The cells are trypsinized in the following day and transferred into 96 well format with 30,000 cells per well in 50 μL complete DMEM containing 2 % fetal calf serum, but without phenol red. The cells are incubated at 37 ⁰C for 3 to 4 hours to allow them to adhere to the plate. Solutions containing a test compound of Formula (I) are then added to wells with no TGF/3 (in triplicates), as well as wells with 1 ng/mL TGF/3 (in triplicates). DMSO is also added to all of the wells at a final concentration of 0.1%. GFP fluorescence emission at 530 nm following excitation at 485 run is measured at 48 hours after the addition of solutions containing a test compound on a CytoFluor microplate reader (PerSeptive Biosystems). The data are then expressed as the ratio of TGF/3-induced to non-induced for each test sample.

OTHER EMBODIMENTS

[146] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

WHAT IS CLAIMED IS:

(I) or a pharmaceutically acceptable salt thereof, wherein: each of R¹ and R² independently is an optionally substituted aryl or heteroaryl; A¹ is NR³ or CR³;

A² is N when A¹ is CR³, or A² is C when A¹ is NR³; X is O or NR³; each R³ is independently hydrogen, aliphatic, cycloaliphatic, aryl, or heteroaryl; each of R' and R" is independently an aliphatic, halo, cyano, or alkoxy; each of p and q is independently 0, 1, or 2, provided that the sum of p and q is 2, 3, or 4; r is 1, 2, or 3; and each of m and n is independently 0, 1, or 2.

2. The compound of claim 1, wherein R¹ is an optionally substituted aryl.

3. The compound of claim 2, wherein R¹ is optionally substituted phenyl.

4. The compound of claim 3, wherein R¹ is phenyl substituted with 1 to 3 substituents each independently selected from the group consisting of aliphatic, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, amido, alkylamido, arylamido, heteroarylamido, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, cycloalkylcarbonyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, heterocycloalkylcarbonyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl, heteroaryloxy, heteroarylsulfanyl, and heteroaroyl.

5. The compound of claim 1 , wherein R is an optionally substituted heteroaryl.

6. The compound of claim 5, wherein R¹ is an optionally substituted bicyclic heteroaryl.

7. The compound of claim 6, wherein R¹ is benzo[l,3]dioxolyl, benzo[b]thiophenyl, benzooxadiazolyl, benzothiadiazolyl, benzoimidazolyl, benzooxazolyl, benzothiazolyl, 2- oxo-benzooxazolyl, 2,3-dihydrobenzo[l,4]dioxyl, 2,3-dihydrobenzofuryl, 2,3- dihydrobenzo[b]thiophenyl, 3,4-dihydrobenzo[l,4]oxazinyl, 3-oxo benzo[l,4]oxazinyl,

1 ,1 -dioxo-2,3 dihydrobenzo[b]thiophenyl, [1 ,2,4]triazolo[l ,5-α]pyridyl, [l,2,4]triazolo[4,3-α]pyridyl, quinolinyl, quinoxalinyl, quinazolinyl, isoquinolinyl, or cinnolinyl, and all of which being optionally substituted.

8. The compound of claim 7, wherein R¹ is optionally substituted [l,2,4]triazolo[l,5- α]pyridin-6-yl.

9. The compound of claim 7, wherein R¹ is optionally substituted quinoxalin-6-yl.

10. The compound of claim 5, wherein R¹ is an optionally substituted monocyclic heteroaryl.

11. The compound of claim 10, wherein R¹ is optionally substituted pyridyl or pyrimidinyl,

12. The compound of any of claims 5 to 11, wherein R¹ is substituted with 1 to 3 substituents each independently selected from the group consisting of aliphatic, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfϊnyl, alkylsulfonyl, amido, alkylamido, arylamido, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, cycloalkylcarbonyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, heterocycloalkylcarbonyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl, heteroaryloxy, heteroarylsulfanyl, and heteroaroyl.

13. The compound of claim 1, wherein R² is an optionally substituted aryl.

14. The compound of claim 13, wherein R² is optionally substituted phenyl.

15. The compound of claim 14, wherein R² is phenyl is substituted with 1 to 3 substituents each independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, amido, alkylamido, arylamido, heteroarylamido, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, cycloalkylcarbonyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, heterocycloalkylcarbonyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl, heteroaryloxy, heteroarylsulfanyl, and heteroaroyl.

16. The compound of claim 1, wherein R² is an optionally substituted heteroaryl.

17. The compound of claim 16, wherein R² is an optionally substituted bicyclic heteroaryl.

18. The compound of claim 17, wherein R² is benzo[l,3]dioxolyl, benzo[b]thiophenyl, benzooxadiazolyl, benzothiadiazolyl, benzoimidazolyl, benzooxazolyl, benzothiazolyl, 2- oxo-benzooxazolyl, 2,3-dihydrobenzo[l,4]dioxyl, 2,3-dihydrobenzofuryl, 2,3- dihydrobenzo[b]thiophenyl, 3,4-dihydrobenzo[l,4]oxazinyl, 3-oxo benzo[l,4]oxazinyl,

1 , 1 -dioxo-2,3-dihydrobenzo[b]thiophenyl, [ 1 ,2,4]triazolo[ 1 ,5-α]pyridyl, [l,2,4]triazolo[4,3-α]pyridyl, quinolinyl, quinoxalinyl, quinazolinyl, isoquinolinyl, or cinnolinyl, and is optionally substituted.

19. The compound of claim 16, wherein R² is an optionally substituted monocyclic heteroaryl.

20. The compound of claim 19, wherein R² is optionally substituted pyridyl or optionally substituted pyrimidinyl.

21. The compound of any of claims 16 to 20, wherein R² is substituted with 1 to 3 substituents each independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, amido, alkylamido, arylamido, heteroarylamido, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, cycloalkylcarbonyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, heterocycloalkylcarbonyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl, heteroaryloxy, heteroarylsulfanyl, and heteroaroyl.

22. The compound of claim 20, wherein R² is 6-methylpyridine-2-yl.

23. The compound of claim 20, wherein R² is 5-fluoro-6-methylpyridine-2-yl.

24. The compound of any of claims 1 to 23, wherein each of p, q, and r is 1.

25. The compound of any of claims 1 to 23, wherein R³ is H.

26. The compound of any of claims 1 to 25, wherein m is 0 and n is 0.

27. The compound of any of claims 1 to 25, wherein each of R' and R" is independently an alkyl, halo, cyano, or alkoxy.

28. The compound of claim 1, wherein the compound is

8-(5-([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-4-(6-methylpyridin-2-yl)- 1 H-imidazol-2-yl)- 2-oxaspiro[4.5]decan-l-one;

8-(5-([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-4-(5-fluoro-6-methylpyridin-2-yl)- 1 H- imidazol-2-yl)-2-oxaspiro[4.5]decan-l-one;

8-(3-(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)-lH-pyrazol-l-yl)-2- oxaspiro[4.5]decan-l-one;

8-(4-([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-3-(5-fluoro-6-methylpyridin-2-yl)- 1 H- pyrazol-l-yl)-2-oxaspiro[4.5]decan-l-one;

8-(4-([ 1 ,2,4]triazolo[ 1 ,5-a]pyridin-6-yl)-3-(6-methylpyridin-2-yl)-l H-pyrazol- 1 -yl)-2- oxaspiro[4.5]decan-l -one;

8-(4-([l,2,4]triazolo[l,5-a]pyridin-6-yl)-3-(pyridin-2-yl)-lH-pyrazol-l-yl)-2- oxaspiro[4.5]decan-l-one; or

2-benzyl-8-(3 -(6-methylpyridin-2-yl)-4-(quinoxalin-6-yl)- 1 H-pyrazol- 1 -yl)-2- azaspiro[4.5]decan-l-one.

29. A pharmaceutical composition comprising a compound of any of claims 1 to 28, and a pharmaceutically acceptable carrier.

30. A method of inhibiting the TGF/3 signaling pathway in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any of claims 1 to 28.

31. A method of inhibiting the TGF/3 type I receptor in a cell, comprising contacting the cell with an effective amount of a compound of any of claims 1 to 28.

32. A method of reducing the accumulation of excess extracellular matrix induced by TGF/3 in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any of claims 1 to 28.

33. A method of treating or preventing a fibrotic condition in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any of claims 1 to 28.

34. The method of claim 33, wherein the fibrotic condition is selected from the group consisting of scleroderma, lupus nephritis, connective tissue disease, wound healing, surgical scarring, spinal cord injury, CNS scarring, acute lung injury, idiopathic pulmonary fibrosis, radiation-induced pulmonary fibrosis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, acute lung injury, drug-induced lung injury, glomerulonephritis, diabetic nephropathy, hypertension-induced nephropathy, alimentary track or gastrointestinal fibrosis, renal fibrosis, hepatic or biliary fibrosis, liver cirrhosis, primary biliary cirrhosis, fatty liver disease, primary sclerosing cholangitis, restenosis, radiation-induced fibrosis, chemotherapy-induced fibrosis, cardiac fibrosis, opthalmic scarring, fibrosclerosis, a fibrotic cancer, a fibroid, fibroma, a fibroadenoma, a fibrosarcoma, transplant arteriopathy, mesothelioma, and keloids.

35. A method of inhibiting growth or metastasis of tumor cells or cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any of claims 1 to 28.

36. A method of treating a disease or disorder mediated by an overexpression of TGF/3 in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any of claims 1 to 28.

37. The method of claim 36, wherein the disease or disorder is selected from the group consisting of demyelination of neurons in multiple sclerosis, Alzheimer's disease, cerebral angiopathy, squamous cell carcinomas, multiple myeloma, melanoma, glioma, glioblastomas, leukemia, sarcomas, leiomyomas, and mesothelioma.

38. The method of claim 36, wherein the disease or disorder is selected from the group consisting of acute lung injury, adult respiratory distress syndrome, alimentary track or gastrointestinal fibrosis, atherosclerosis, biliary fibrosis, cardiac fibrosis, chemotherapy- induced fibrosis, cholangitis, chronic obstructive pulmonary disease, cirrhosis due to fatty liver disease, CNS scarring, connective tissue disease, corneal scarring, diabetic nephropathy, drug-induced lung injury, fatty liver disease, fibroadenoma, fibroid, fibroma, fibrosarcoma, fibrosclerosis, fibrotic cancer, glomerulonephritis, hepatic or biliary fibrosis, hypertension-induced nephropathy, idiopathic pulmonary fibrosis, keloids, liver cirrhosis, lupus nephritis, mesothelioma, opthalmic scarring, post-infarction cardiac fibrosis, primary biliary cirrhosis, post-surgical fibrosis, primary sclerosing cholangitis, pulmonary fibrosis, radiation-induced fibrosis, radiation-induced pulmonary fibrosis, renal fibrosis, restenosis, sarcoidosis, scleroderma, spinal cord injury, surgical scarring, transplant arteriopathy, wound healing, scleroderma, spinal cord injury, systemic lupus erythematosus, and Wegener's granulomatosis.

39. A method of treating carcinomas mediated by an overexpression of TGF/3 in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any of claims 1 to 28.

40. The method of claim 39, wherein said carcinomas is selected from the group consisting of carcinomas of the lung, breast, ovary, liver, biliary tract, gastrointestinal tract, head, neck, pancreas, prostate, cervix, multiple myeloma, melanoma, glioma, and glioblastomas.

41. A method of treating or preventing restinosis, vascular disease, or hypertension in a subject in need thereof, comprising administering to the subject a compound of any of claims 1 to 28.

42. The method of claim 41, wherein the restinosis is coronary restenosis, peripheral restenosis, or carotid restenosis.

43. The method of claim 41, wherein the vascular disease is intimal thickening, vascular remodeling, or organ transplant-related vascular disease.

44. The method of claim 41, wherein the hypertension is primary or secondary hypertension, systolic hypertension, pulmonary hypertension, or hypertension-induced vascular remodeling.

45. The method of claim 41, wherein the compound is administered locally.

46. The method of claim 41, wherein the compound is administered via an implantable device.

47. The method of claim 46, wherein the device is a delivery pump.

48. The method of claim 46, wherein the device is a stent.