Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
  • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
  • C07D471/04—Ortho-condensed systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
  • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
  • C07D487/04—Ortho-condensed systems

Definitions

• the invention relates to modulators of Transforming Growth Factor ⁇ (TGF ⁇ ), compositions thereof and methods of use.
• TGF ⁇ Transforming Growth Factor ⁇
• the invention also relates to the treatment of TGF ⁇ -related diseases.
• TGF ⁇ Transforming Growth Factor ⁇
• BMPs bone morphogenetic proteins
• GDFs growth and differentiation factors
• MIS mullerian inhibiting substance
• Each TGF ⁇ 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 ⁇ .
• LAP latency associated peptide
• LAP-TGF ⁇ complex cannot bind to the TGF ⁇ receptors and is not biologically active.
• TGF ⁇ is generally released (and activated) from the complex by a variety of mechanisms including, for example- interaction with thrombospondin-1 or plasmin.
• TGF ⁇ binds at high affinity to the type II receptor (TGF ⁇ RII), a constitutively active serine/threonine kinase.
• TGF ⁇ RII type II receptor
• the ligand-bound type II receptor phosphorylates the TGF ⁇ 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.
• TGF ⁇ RII type II receptor
• Alk5 TGF ⁇ type I receptor
• 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 ⁇ -responsive genes. See, e.g., Massague, J., Ann. Rev. Biochem. Med., 67: 773 (1998).
• Activins are also members of the TGF ⁇ superfamily, which are distinct from TGF ⁇ in that they are homo- or heterodimers of activin ⁇ a or ⁇ b. Activins signal in a manner similar to TGF ⁇ , 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.
• ActRIIB activin type II receptor
• Alk4 type I serine-threonine receptor
• TGF ⁇ 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.
• 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.
• 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.
• TGF ⁇ signaling pathway underlies many human disorders (e.g., excess deposition of extracellular matrix, an abnormally high level of inflammatory responses, fibrotic disorders, and progressive cancers).
• 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.
• the invention is based on the discovery that compounds of formula (I) are potent antagonists of the TGF ⁇ family type I receptors, Alk5 and/or Alk4.
• 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 ⁇ family signaling activity is desirable.
• diseases such as fibrosis (e.g., renal fibrosis, pulmonary fibrosis, and hepatic fibrosis), progressive cancers, or other diseases for which reduction of TGF ⁇ family signaling activity is desirable.
• fibrosis e.g., renal fibrosis, pulmonary fibrosis, and hepatic fibrosis
• progressive cancers e.g., hepatic fibrosis
• the invention features compounds of formula (I) or a pharmaceutically acceptable salt or mixtures thereof,
• a pharmaceutical composition of this invention includes a compound of formula (I) and a pharmaceutically acceptable carrier.
• the invention also relates to a method of inhibiting the TGF ⁇ signaling pathway in a subject, which includes administering to said subject in need thereof an effective amount of a compound of formula (I).
• the invention in another aspect, relates to a method of inhibiting the TGF ⁇ type I receptor in a cell, which includes contacting said cell with an effective amount of a compound of formula (I).
• the invention relates to a method of reducing the accumulation of excess extracellular matrix induced by TGF ⁇ in a subject in need thereof, which includes administering to said subject an effective amount of a compound of formula (I).
• the invention relates to a method of treating or preventing flbrotic condition in a subject, which includes administering to said subject in need thereof an effective amount of a compound of formula (I).
• the fibrotic condition can be, e.g., mesothelioma, acute respiratory distress syndrome (ARDS), atherosclerosis, scleroderma, keloids, glomerulonephritis, diabetic nephropathy, lupus nephritis, hypertension-induced nephropathy, cholangitis, restenosis (e.g., coronary restenosis, peripheral restinosis, or carotid restenosis), ocular scarring, corneal scarring, hepatic fibrosis, biliary fibrosis, liver cirrhosis, cirrhosis due to fatty liver disease (alcoholic and nonalcoholic steatosis), primary sclerosing cholangitis, pulmonary fibrosis (such
• the fibrotic condition can be idiopathic in nature, genetically linked, or induced by radiation.
• the invention relates to a method of using the compounds of the invention for treating and preventing a vascular disease such as intimal thickening, vascular remodeling, or organ transplant-related vascular disease.
• the invention relates to a method of inhibiting growth or metastasis of tumor cells and/or cancers in a subject, which includes administering to said subject in need thereof an effective amount of a compound of formula (I).
• the invention relates to a method of treating a disease or disorder mediated by an overexpression of TGF ⁇ . 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.
• 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 H 2 O 2 .
• 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.
• salts formed with pharmaceutically acceptable amines such as ammonia, alkyl amines, hydroxyalkylamines, and N- methylglycamine.
• a compound of formula (I) can be treated with an acid to form acid addition salts.
• acids examples include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, methanesulfonic acid, phosphoric acid, /j-bromophenyl- sulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, oxalic acid, malonic acid, salicylic acid, malic 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).
• an acid e.g., hydrochloric acid
• 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.
• Compounds of formula (I) exhibit high affinity to the TGF ⁇ family type I receptors, Alk5 and/or Alk4, e.g., with IC5 0 and Ki values of less than 10 ⁇ M under conditions described below.
• Some compounds of formula (1) exhibit IC 50 and K; values of less than 1 ⁇ M (such as below 200 nM).
• compounds of formula (I) can be employed in the prevention and treatment of diseases mediated by TGF ⁇ such as fibrosis (e.g., renal fibrosis, pulmonary fibrosis, or hepatic fibrosis), progressive cancers, or other diseases for which reduction of TGF ⁇ family signaling activity is desirable.
• diseases mediated by TGF ⁇ such as fibrosis (e.g., renal fibrosis, pulmonary fibrosis, or hepatic fibrosis), progressive cancers, or other diseases for which reduction of TGF ⁇ family signaling activity is desirable.
• implantable devices each comprising a compound of formula (I) described above.
• implantable devices can be, e.g., a delivery pump or a stent, and can also be used for treating a disease or disorder mediated by an overexpression of TGF ⁇ .
• modulating means increasing or decreasing, e.g. activity, by a measurable amount.
• Compounds that modulate TGF ⁇ activity by increasing the activity of the TGF ⁇ receptors are called agonists.
• Compounds that modulate TGF ⁇ activity by decreasing the activity of the TGF ⁇ receptors are called antagonists.
• An agonist interacts with a TGF ⁇ receptor to increase the ability of the receptor to transduce an intracellular signal in response to endogenous ligand binding.
• An antagonist interacts with a TGF ⁇ receptor and competes with the endogenous ligand(s) or substrate(s) for binding site(s) on the receptor to decrease the ability of the receptor to transduce an intracellular signal in response to endogenous ligand binding.
• aliphatic encompasses the terms alkyl, alkenyl, and alkynyl, each of which is optionally substituted as set forth below.
• an "alkyl” group refers to a saturated aliphatic hydrocarbon group containing 1-8 (e.g., 1-6 or 1-4) carbon atoms.
• An alkyl group can be straight or branched. Examples of such alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, «-pentyl, n-heptyl, or 2-ethylhexyl.
• An alkyl group can be substituted (i.e., optionally substituted) with one or more substituents such as halo, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, cycloaliphaticcarbonyl, (heterocycloaliphatic)carbonyl, nitro, cyano, amino, amido, acyl, sulfonyl, sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, thioxo, carboxy, carbamoyl, cycloaliphaticoxy, heterocycloaliphaticoxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, or hydroxy.
• substituents such as halo, cycloaliphatic, heterocycloaliphatic, ary
• substituted alkyls include carboxyalkyl (such as HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl), cyanoalkyl, hydroxyalkyl, alkoxyalkyl, acylalkyl, hydroxyalkyl, aralkyl, (alkoxyaryl)alkyl, (sulfonylamino)alkyl (such as (alkylsulfonylamino)alkyl), aminoalkyl, amidoalkyl, (cycloaliphatic)alkyl, cyanoalkyl, or haloalkyl.
• carboxyalkyl such as HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl
• cyanoalkyl such as HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl
• cyanoalkyl such as HO
• an "alkenyl” group refers to an aliphatic carbon group that contains 2-8 (e.g., 2-6 or 2-4) 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.
• alkenyl group can be optionally substituted with one or more substituents such as halo, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, nitro, cyano, amino, amido, acyl (e.g., cycloaliphaticcarbonyl and (heterocycloaliphatic)carbonyl), sulfonyl, sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, thioxo, oxo, carboxy, carbamoyl, cycloaliphaticoxy, heterocycloaliphaticoxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, or hydroxy [0026] As used herein, an "alkynyl" group refers to an aliphatic carbon group that contains 2-8
• 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 halo, cycloaliphatic, heterocycloaliphatic, aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, (cycloaliphatic)carbonyl, (heterocycloaliphatic)carbonyl, nitro, cyano, amino, amido, acyl, sulfonyl, sulf ⁇ nyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, thioxo, carboxy, carbamoyl, (cycloaliphatic)oxy, (heterocycloaliphatic)oxy, aryloxy, heteroaryloxy,
• 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 ) 2 -C(O)- or R Y C(O)-N(R X )- when used terminally; and - C(O)-N(R X )- or -N(R X )-C(O)- when used internally, wherein R x and R ⁇ are defined below.
• amido groups include alkylamido (such as alkylcarbonylamino and alkylcarbonylamino), (heterocycloaliphatic)amido, (heteroaralkyl)amido, (heteroaryl)amido, (heterocycloalkyl)alkylamido, arylamido, aralkylamido, (cycloalkyl)alkylamido, and cycloalkylamido
• an "amino" group refers to -NR X R Y wherein each of R x and R ⁇ is independently hydrogen, alkyl, cycloaliphatic, aryl, heterocycloaliphatic, or heteroaryl, each of which being defined herein and being optionally substituted.
• Examples of amino groups include alkylamino, dialkylamino, and arylamino.
• 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, or tetrahydroindenyl); and tricyclic (e.g., fluorenyl tetrahydrofluorenyl, tetrahydroanthracenyl, or anthracenyl).
• the bicyclic and tricyclic groups include benzofused 2-3 membered carbocyclic rings.
• 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
• sulfonyl e.g., aliphaticsulfonyl and aminosulfonyl
• sulfinyl e.g., ali
• an aryl can be unsubstituted.
• substituted aryls include haloaryl (e.g., mono-, di- (such asp.m-dihaloaryl), or (trihalo)aryl); (carboxy)aryl (e.g., (alkoxycarbonyl)aryl, ((aryalkyl)carbonyloxy)aryl, or (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 and ((dialkyl)amino)aryl);
• an "araliphatic” such as an “aralkyl” group refers to an aliphatic group (e.g., a Ci 4 alkyl group) that is substituted with an aryl group.
• "Aliphatic,” “alkyl,” and “aryl” are defined herein.
• An example of an araliphatic such as an aralkyl group is benzyl.
• an "aralkyl” group refers to an alkyl group (e.g., a C 1 . 4 alkyl group) that is substituted with an aryl group. Both “alkyl” and “aryl” have been defined above.
• An example of an aralkyl group is benzyl.
• An aralkyl 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, cycloalkylcarbonyl amino, (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalky
• a "bicyclic ring system” includes 8-12 (e.g., 9, 10, or 11) membered structures that form two rings, wherein the two rings have at least one atom in common (e.g., 2 atoms in common).
• Bicyclic ring systems include bicycloaliphatics (e.g., bicycloalkyl or bicycloalkenyl), bicycloheteroaliphatics, bicyclic aryls, and bicyclic heteroaryls.
• a "cycloaliphatic” group encompasses a "cycloalkyl” group and a “cycloalkenyl” group, each of which being optionally substituted as set forth below.
• a "cycloalkyl” group refers to a saturated carbocyclic mono- or bicyclic (fused or bridged) ring of 3-10 (e.g., 5-10) carbon atoms.
• Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, norbomyl, cubyl, octahydro-indenyl, decahydro-naphthyl, 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, azacycloalkyl, or ((aminocarbonyl)cycloalkyl)cycloalkyl.
• a "cycloalkenyl” group refers to a non-aromatic carbocyclic ring of 3-10 (e.g., 4-8) carbon atoms having one or more double bonds.
• Examples of cycloalkenyl groups include cyclopentenyl, 1,4- cyclohexa-di-enyl, cycloheptenyl, cyclooctenyl, hexahydro-indenyl, octahydro-naphthyl, cyclohexenyl, cyclopentenyl, bicyclo[2.2.2]octenyl, and bicyclo[3.3.1]nonenyl.
• a cycloalkyl or cycloalkenyl group can be optionally substituted with one or more substituents such as aliphatic (e.g., alkyl, alkenyl, and 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)carbonyl amino, (heterocycloaliphatic)carbony
• a "cyclic moiety" includes cycloaliphatic, heterocyclo aliphatic, aryl, or heteroaryl, each of which has been defined previously.
• heterocycloaliphatic encompasses a heterocycloalkyl group and a heterocycloalkenyl group, each of which being optionally substituted as set forth below.
• heterocycloalkyl refers to a 3-10 membered mono- or bicylic (fused or bridged) (e.g., 5- to 10-membered mono- or bi cyclic) saturated ring structure, in which one or more of the ring atoms is a heteroatom (e.g., N, O, S, or combinations thereof).
• heterocycloalkyl group examples 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[£]thiopheneyl, 2-oxa-bicyclo[2.2.2]octyl, l-aza-bicyclo[2.2.2]octyl, 3-aza- bicyclo[3.2.1]octyl, anad 2,6-d
• a monocyclic heterocycloalkyl group can be fused with a phenyl moiety such as tetrahydroisoquinoline.
• 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).
• 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 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,
• a “heteroaryl” group refers to a monocyclic, bicyclic, or tricyclic ring structure 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 wherein one or more rings of the bicyclic or tricyclic ring structure is aromatic.
• a heteroaryl group includes a benzofused ring system having 2. to 3 rings.
• 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[ ⁇ ]thiophenyl, quinolinyl, or isoquinolinyl).
• heterocycloaliphatic moieties e.g., indolizyl, indolyl, isoindolyl, 3H- indolyl, indolinyl, benzo[6]furyl, benzo[ ⁇ ]thiophenyl, quinolinyl, or isoquinolinyl.
• heteroaryl examples include azetidinyl, pyridyl, 1 H-indazolyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuryl, isoquinolinyl, benzthiazolyl, xanthene, thioxanthene, phenothiazine, dihydroindole, benzo[l,3]dioxole, benzo[b]furyl, benzo[b]thiophenyl, indazolyl, benzimidazolyl, benzthiazolyl, puryl, cinnolyl, quinolyl, quinazolyljCinnolyl, phthalazyl, quinazolyl, quinoxalyl, isoquinolyl, 4H-quinolizyI, benzo- 1,2,5-thiadiazolyl, or
• monocyclic heteroaryls include furyl, thiophenyl, 2H-pyrrolyl, pyrrolyl, oxazolyl, thazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,3,4-thiadiazolyl, 2H- ⁇ yranyl, 4-H-pranyl, pyridyl, pyridazyl, pyrimidyl, pyrazolyl, pyrazyl, or 1,3,5-triazyl.
• Monocyclic heteroaryls are numbered according to standard chemical nomenclature.
• bicyclic heteroaryls include indolizyl, indolyl, isoindolyl, 3H- indolyl, indolinyl, benzo[&]furyl, benzo[6] thiophenyl, quinolinyl, isoquinolinyl, indolizyl, isoindolyl, indolyl, benzo[ ⁇ ]furyl, bexo[&]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.
• 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, (arali ⁇ hatic)oxy, (heteroaraliphatic)oxy, aroyl, heteroaroyl, amino, oxo (on a non-aromatic carbocyclic or heterocyclic ring of a bicyclic or tricyclic heteroaryl), nitro, carboxy, amido, acyl (e.g., aliphaticcarbonyl, (cycloaliphatic)carbonyl, ((cycloaliphatic)aliphatic)carbonyl, (araliphathat
• 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, ((heteroaryl)amino)carbonyl)heteroaryl, ((heter
• heteroaralkyl refers to an aliphatic group (e.g., a C ⁇ alkyl group) that is substituted with a heteroaryl group.
• aliphatic group e.g., a C ⁇ alkyl group
• heteroaryl e.g., a C ⁇ alkyl group
• heteroaryl refers to an alkyl group (e.g., a Q -4 alkyl group) that is substituted with a heteroaryl group. Both “alkyl” and “heteroaryl” have been defined above.
• a heteroaralkyl is optionally substituted with one or more substituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifiuoromethyl), 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, (heterocycl
• a "cyclic moiety" includes cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, or heteroaryl, each of which has been defined previously.
• an "acyl” group refers to a formyl group or R W -C(O)- (such as alkyl-
• alkylcarbonyl C(O)-, also referred to as "alkylcarbonyl" where alkyl has been defined previously and R w is aliphatic, cylcoaliphatic, heterocycloaliphatic, each of which can be optionally substituted with aliphatic (e.g., alkyl, alkenyl, or alkynyl), cycloaliphatic, (cycloaliphatic)aliphatic,; heterocycloaliphatic, (heterocycloaliphatic)aliphatic, aryl, heteroaryl, alkoxy,
• alkoxy refers to an alkyl-O- group where “alkyl” has been defined previously.
• an “aroyl” (or “arylcarbonyl”) group referes to an Ar-CO- group, wherein Ar is an aryl group as previously defined.
• heteroaroyl or “heteroarylcarbonyl” group refers to a HetAr-
• a "carbamoyl” group refers to a group having the structure -O-CO-
• R x and R ⁇ have been defined above and R z can be aliphatic, aryl, araliphatic, heterocycloaliphatic, heteroaryl, or heteroaraliphatic.
• a "carboxy” group refers to -COOH, -COOR X , -OC(O)H, -0C(O)R x when used as a terminal group; or -OC(O)- or -C(O)O- when used as an internal group.
• haloaliphatic refers to an aliphatic group substituted with
• haloalkyl includes the group -CF 3 .
• mercapto refers to -SH.
• a "sulfo" group refers to -SO 3 H or -S ⁇ 3 R x when used terminally, or
• a "sulfamide” group refers to the structure -NR X -S(O) 2 -NR Y R Z when used terminally and -NR X - S(O) 2 -NR ⁇ - when used internally, wherein R x , R ⁇ , and R z have been defined above.
• a "sulfamoyl” group refers to the structure -S(O) 2 -NR X R Y or -NR X -
• sulfanyl refers to -S-R x and encompasses mercapto (-SH) when used terminally, and -SR X - when used internally, wherein R x has been defined above.
• sulfanyls examples include alkylsulfanyl.
• sulfinyl refers to -S(O)-R X when used terminally and -
• a "sulfonyl” group refers to-S(0)2-R x when used terminally and -
• a "sulfoxy" group refers to -O-SO-R X or -SO-O-R X , when used terminally and -O-S(O)- or -S(O)-O- when used internally, where R x has been defined above.
• halogen or halo group refers to fluorine, chlorine, bromine or iodine.
• alkoxycarbonyl 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)-.
• alkoxyalkyl refers to an alkyl group such as alkyl-O-alkyl-, wherein alkyl has been defined above.
• aminoalkyl refers to the structure (R x ) 2 N-alkyl-.
• cyanoalkyl refers to the structure (NC)-alkyl-
• urea refers to the structure -NR X -CO-NR Y R Z and a
• thiourea group refers to the structure -NR X -CS-NR Y R Z when used terminally and -NR X -
• bridged bicyclic ring system refers to a bicyclic heterocyclicalipahtic ring system or bicyclic cycloaliphatic ring system in which the rings are bridged.
• bridged bicyclic 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-oxa-bicyclo[2.2.2]octyl, l-aza-bicyclo[2.2.2]octyl, 3-aza- bicyclo[3.2.1]octyl, and 2,6-dioxa-tricyclo[3.3.1.03,7]nonyl.
• a bridged bicyclic 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, (cycloalkylalky ⁇ carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (hetero
• terminal refers to the location of a group within a substiruent.
• 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 x O(O)C-alkyK is an example of a carboxy group used terminally.
• a group is internal when the group is present in the middle of a substituent to at the end of the substituent bound to the rest of the chemical structure.
• Alkylcarboxy e.g., alkyl-C(O)O- or alkyl-OC(O)-
• alkylcarboxyaryl e.g., alkyl-C(O)O-aryl- or alkyl-O(CO)-aryl-
• Each substituent of a specific group is further optionally substituted with one to three of halo, cyano, oxoalkoxy, hydroxyl, amino, nitro, aryl, haloalkyl, and alkyl.
• an alkyl group can be substituted with alkylsulfanyl and the alkylsulfanyl can be optionally substituted with one to three of halo, cyano, oxoalkoxy, hydroxyl, amino, nitro, aryl, haloalkyl, and alkyl.
• the cycloalkyl portion of a (cycloalkyl)carbonylamino can be optionally substituted with one to three of halo, cyano, alkoxy, hydroxyl, nitro, haloalkyl, and alkyl.
• the two alkxoy groups can form a ring together with the atom(s) to which they are bound.
• substituted 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.
• 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.
• substituents envisioned by this invention are those combinations that result in the formation of stable or chemically feasible compounds.
• stable or chemically feasible 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.
• an 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 can be approximately determined from height and weight of the patient. See, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, New York, 537 (1970).
• a "patient” or “subject” refers to a mammal, including a human.
• the term "electron withdrawing group” refers to a substituent that draws electrons to itself more than a hydrogen atom would if it occupied the same position in a molecule. See, e.g., March, J., "Advanced Organic Chemistry," 4 th Ed., pp. 18-19 and 36, Wiley-Interscience, New York (1992). Examples of electron withdrawing groups include but are not limited to -C(O)-, -S(O)-, or -S(O) 2 -.
• 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. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention.
• structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
• compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a 13 C- or 14 C-enriched carbon are within the scope of this invention.
• Such compounds are useful, for example, as analytical tools or probes in biological assays.
• Ri is aryl or heteroaryl each optionally substituted with 1 to 3 R a ;
• R 2 is aryl or heteroaryl, each optionally substituted with 1 to 3 R b , provided that at least one of Ri and R2 is heteroaryl optionally substituted with 1 to 3 Rb ;
• Ring A is a saturated or partially unsaturated 5 to 8 membered cycloaliphatic, a 5 to 8 membered heterocycloaliphatic containing 1 to 3 heteroatoms, phenyl, or a 5 to 6 membered heteroaryl containing 1 to 3 heteroatoms;
• Ring B is a partially unsaturated non-aromatic 5 to S membered heterocycloaliphatic containing one to three heteroatoms; each of Ra and R b is independently aliphatic, alkoxy, acyl, halo, hydroxy, amino, amido (e.g., aminocarbonyl or alkylcarbonylamino), nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy (e.g., alkoxycarbonyl, or alkylcarbonyloxy), sulfo, sulfanyl (e.g., mercaptoalkylsulfanyl, cycloalkylsulfanyl, heterocycloalkylsulfanyl, arylsulfanyl, or heteroarylsulfanyl), sulf ⁇ nyl, sulfonyl, urea, thiourea, sulfamoyl (e.g
• Xi is C and X 2 is N or Xi is N and X 2 is C; i is 0 to 4; j is 0 to 4; and each m is independently 0 to 2.
• the ring juncture between rings A and B is in the trans configuration.
• Xi is C and X 2 is N to provide imidazole compounds of the invention.
• Ri is an optionally substituted aryl, such as a mono- or bi- carbocyclic aromatic group.
• Ri can be an optionally substituted mono-carbocyclic aromatic ("monocyclic aryl") group, e.g., an optionally substituted phenyl.
• Rj can be an optionally substituted bi-carbocyclic aromatic (“bicyclic aryl") group, e.g., naphthyl, indenyl, or azulenyl.
• Ri can be an optionally substituted benzofused bicyclic aryl moiety, e.g., tetrahydronaphthalyl.
• Ri is an optionally substituted heteroaryl, such as a mono- or bi-heterocyclic aromatic group.
• Ri can be an optionally substituted mono-heterocyclic aromatic ("monocyclic heteroaryl") group, e.g., furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazaloyl, isoxazolyl, isothiazolyl, triazolyl, pyridinyl, pyridazinyl, pyrimidinyl, and pyrazinyl, each of which are optionally substituted.
• monocyclic heteroaryl e.g., furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazaloyl, isoxazolyl, isothiazolyl, triazolyl, pyridinyl, pyridazinyl, pyrimidinyl, and
• Rj can be an optionally substituted 5-membered mono-heterocyclic aromatic group, e.g., furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazaloyl, isoxazolyl, isothiazolyl, and triazolyl, each of which is optionally substituted.
• Ri can be an optionally substituted 6-membered mono-heterocyclic aromatic group, e.g., pyridinyl, pyridazinyl, pyrimidinyl, and pyrazinyl, each of which is optionally substituted.
• Ri is an optionally substituted bicyclic heteroaryl, e.g., indolizinyl, indolyl, isoindolyl, benzofuranyl, benzothiopenyl, lH-indazolyl, benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and pteridinyl, each of which is optionally substituted.
• Ri can be an optionally substituted 9-membered bi-heterocyclic aromatic group, e.g., indolizinyl, indolyl, isoindolyl, benzofuranyl, benzothiopenyl, lH-indazolyl, benzimidazolyl, benzthiazolyl, and purinyl, each of which is optionally substituted.
• Ri can be an optionally substituted 10-membered bi-heterocyclic aromatic group, e.g., 4H- quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and pteridinyl, each of which are optionally substituted.
• Ri can be an optionally substituted benzofused bicyclic heteroaryl moiety covered under the term heteroaryl, e.g., indolinyl and tetrahydoquinolinyl.
• Ri is an optionally substituted pyridinyl or pyrimidinyl.
• Ri is an optionally substituted pyridin-2-yl (e.g., 6-aliphatic pyridin-2-yl).
• R 2 is an optionally substituted aryl, such as a mono- or bi- carbocyclic aromatic group.
• R 2 can be an optionally substituted mono-carbocyclic aromatic ("monocyclic aryl") group, e.g., an optionally substituted phenyl.
• R 2 can be an optionally substituted bi-carbocyclic aromatic ("bicyclic aryl") group, e.g., naphthyl, indenyl, or azulenyl.
• R 2 can be an optionally substituted benzofused bicyclic aryl moiety, e.g., tetrahydronaphthalyl.
• R 2 is an optionally substituted heteroaryl, such as a mono- or bi-heterocyclic aromatic group.
• R 2 can be an optionally substituted mono-heterocyclic aromatic ("monocyclic heteroaryl") group, e.g., furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazaloyl, isoxazolyl, isothiazolyl, triazolyl, pyridinyl, pyridazinyl, pyrimidinyl, and pyrazinyl, each of which are optionally substituted.
• monocyclic heteroaryl mono-heterocyclic aromatic
• R 2 can be an optionally substituted 5-membered mono-heterocyclic aromatic group, e.g., furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazaloyl, isoxazolyl, isothiazolyl, and triazolyl, each of which is optionally substituted.
• R 2 can be an optionally substituted 6-membered mono-heterocyclic aromatic group, e.g., pyridinyl, pyridazinyl, pyrimidinyl, and pyrazinyl, each of which is optionally substituted.
• R 2 is an optionally substituted bicyclic heteroaryl, e.g., indolizinyl, indolyl. isoindolyl, benzofaranyl, benzothiopenyl, lH-indazolyl, benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and pteridinyl, each of which is optionally substituted.
• R 2 can be an optionally substituted 9-membered bi-heterocyclic aromatic group, e.g., indolizinyl, indolyl, isoindolyl, benzofuranyl, benzothiopenyl, lH-indazolyl, benzimidazolyl, benzthiazolyl, and purinyl, each of which is optionally substituted.
• indolizinyl indolyl
• isoindolyl benzofuranyl
• benzothiopenyl lH-indazolyl
• benzimidazolyl benzimidazolyl
• benzthiazolyl and purinyl, each of which is optionally substituted.
• R 2 can be an optionally substituted 10-membered bi-heterocyclic aromatic group, e.g., 4H- quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and pteridinyl, each of which are optionally substituted.
• R 2 can be an optionally substituted benzofused bicyclic herteroaryl moiety covered under the term heteroaryl, e.g., indolinyl and tetrahydoquinolinyl.
• R 2 is an optionally substituted pyridinyl or pyrimidinyl.
• R 2 is an optionally substituted pyridin-2-yl (e.g., 6-aliphatic pyridin-2-yl).
• Examples of bicyclic heteroaryl Ri or R 2 substituents include, but are not limited to
• Ring A is a 5- or 6-membered saturated, or partially unsaturated cycloaliphatic or heterocycloaliphatic ring wherein Ring B, Ri, R 2 , R3 and R4 are as previously described.
• Ring A is a 6-membered aryl or heteroaryl ring wherein
• Ring B, Ri, R 2 , R 3 and R 4 are as previously described.
• Ring A contains one degree of unsaturation. [0099] In some embodiments, Ring A contains two degrees of unsaturation.
• Ring A is a 5-membered cycloaliphatic or heterocycloaliphatic ring.
• Ring A is a 6-membered cycloaliphatic or heterocycloaliphatic ring.
• Ring A is a 7-membered cycloaliphatic or heterocycloaliphatic ring.
• Ring B is a 5-, 6- or 7-membered partially unsaturated cycloaliphatic or heterocycloaliphatic ring wherein ring A, Ri, R 2 , R 3 and R 4 are as previously described.
• Ring B contains one degree of unsaturation.
• Ring B contains two degrees of unsaturation provided that
• Ring B is non-aromatic.
• Ring B is a 5-membered cycloaliphatic or heterocycloaliphatic ring.
• Ring B is a 6-membered cycloaliphatic or heterocycloaliphatic ring.
• Ring B is a 7-membered cycloaliphatic or heterocycloaliphatic ring.
• Ring A can be a 6-membered cycloaliphatic and Ring B can be a 6-membered partially unsaturated heterocycloaliphatic.
• the intermediate imidazole 1-3 is prepared by reaction of a diketone 1-1 with a cyclic aldehyde 1-2 (step I-A), wherein Q represents a radical which can be directly cyclizied (step I-B) or first derivatized (not shown) and subsequently cyclizied (step I-B) to the tricyclic compound 1-4.
• the radicals R 3 and R 4 can be modified by known methods (step I-C) to give compounds 1-5 wherein R' 3 and R' 4 represents modifications to R 3 and R4.
• Q radicals include, but are not limited to, olefins, protected hydroxyalkyl, alkylsulfonate esters and alkylhalides.
• Step A a diketone 1 is reacted with an allyl-aldehyde 2 in the presence of an ammonium salt and an organic acid in a suitable solvent to provide the imidazole 3.
• the diketones 1 are commercially available or may be prepared according to known procedures. See, e.g., U.S. Pat. No. 6,465,493; U.S. Publication 2004/0110797.
• Suitable ammonium salts include, but are not limited to, ammonium acetate and ammonium chloride.
• suitable solvents include, but are not limited to, dimethoxyethane, methyl-f-butyl ether, dioxane, dimethylformamide, methanol, ethanol, and acetic acid.
• Ring B is a 6-membered cycloaliphatic
• the allyl compound 3 is reacted with a borane hydride followed by oxidation with an oxidizing agent under known reaction conditions (see, e.g., H.C. Brown, Hydroboration, W.A.Benjamin, New York (1962)) to provide the alcohol 4.
• Subsequent cyclization of the alcohol 4 to the 6- membered Ring B compound 7 (step E) may be achieved by contacting an alcohol 4 with iodine in the presence of a tertiary phosphine such as triphenylphosphine and an organic base.
• a tertiary phosphine such as triphenylphosphine and an organic base.
• cyclization can be achieved by contacting an alcohol 4 with a azodicarboxylate such as diethylazodicarboxylate and diisopropylazodicarboylate, in the presence of a tertiary phosphine such as triphenylphosphine.
• a azodicarboxylate such as diethylazodicarboxylate and diisopropylazodicarboylate
• a tertiary phosphine such as triphenylphosphine.
• Suitable borane hydrides include, but are not limited to, diborane and 9-borobicyclo[3.3.1]nonane (9-BBN).
• Suitable oxidizing agents include, but are not limited to, hydrogen peroxide and m-chloroperbenzoic acid.
• suitable phosphines include, but are not limited to, tri-aryl phosphines, e.g., triphenylphosphine.
• the olefin 3 can be converted to the alcohol 5 (step C) by known oxidation-reduction methods.
• olefin 3 can be converted to the alcohol 5 via ozonolysis followed by reduction of the resultant aldehyde.
• olefin 3 can be converted to the alcohol 5 by contacting olefin 3 with osmium tetroxide-periodate followed by a reduction. Cyclization of alcohol 5 to the 5-membered ring compound 6 is achieved under conditions as described above for step E.
• scheme III illustrates preparation of compounds of the invention wherein X 1 is C and X 2 is N and Ring B is a 7-membered cycloaliphatic.
• the di-allylimidazole of formula 8 is subjected to a metathesis reaction using a ruthenium catalyst (Grubb's reaction, see, e.g., Grubbs, et al., J. Org. Chern., 62: 7310 (1997); Grubbs et al., J. Amer. Chem Soc, 125, 11360 (2003); Martin et al., Chem. Rev., 104: 2199 (2004); McReynolds et al., Chem. Rev., 2004, 104, 2239; McDonald et al., J. Am. Chem. Soc, 126, 2495 (2004); J. Am. Chem. Soc, 122.
• Grubb's reaction see, e.g., Grubbs, et al., J. Org. Chern., 62: 7310 (1997); Grubbs et al., J. Amer. Chem Soc, 125, 11360 (2003); Martin e
• ruthenium catalysts include, but are not limited to, benzylidene-bis(tricyclohexylphosphine)dichlororuthenium (Grubbs 1 st catalyst), 1,3-bis- (2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(phenylmethylene) (tricyclohexylphosphine)ruthenium and 1 ,3-(bis(mesityl)-2-imidazolidinylidene)dichloro-(o- isopropoxyphenylmethylene)ruthenium.
• Suitable solvents for this reaction include, e.g., methylenechloride, ethylenedichloride and tetrahydrofuran.
• compounds of structure 9 may be reduced under known olefin hydrogenation conditions to produce saturated analogs of compound 9.
• reaction of the diketone 1 with the cyclicimino compound of formula 10 wherein n is 1 to 3 in the presence of an ammonium salt under conditions similar to step A provides compounds in which ring A is aromatic.
• the imine compounds of formula 10 are commercially available or may be prepared by known methods (see, e.g., Nicolaou, K.C. et al., Angewandt Chemie, International Edition, 42 (34): 4077 (2003); and Larsen, R.D., et al., J. Org. Chem., 56: 6034-6038 (1991).
• compounds of formula (I) are produced by modifying the substituents on rings A and/or B.
• R 3 is a protected hydroxyl (-OPg)
• compound analogs are generated by removing the hydroxyl protecting group, Pg, to provide the corresponding alcohol.
• the alcohol can be converted by known methods to alcohol derivatives such esters, thioesters, carbamates, halides, nitriles, alkylethers, arylethers, and the like.
• the alcohol may also be converted to the corresponding amine, ketone or olefin.
• R 3 is a ketone. Reduction of the ketone with, for example, sodium borohydride leads to the corresponding alcohol.
• the stereochemistry of the derived alcohol may be inverted by known methods, e.g., the Mitsunobu Reaction (O. Mitsunobu, Synthesis, 1981, pp. 1-28).
• the amines and ketones maybe further derivatized to provide substituted amines or homologated lactams.
• R 3 may be -CHiOPg. Removal of the protecting group provides the alcohol which can be converted to alcohol derivatives described above. Additionally, the alcohol can be oxidized to provide compounds containing an aldehyde or a carboxylic acid functionality. The aldehyde and carboxylic acid functionality can be further modified by known methods.
• the cyclic aldehydes 2 of scheme II may be produced by known methods.
• a cyclohexane carboxaldehyde of formula 2 wherein two of R 3 together form a cyclic ketal can be prepared as outlined in scheme V and as further illustrated, e.g., in Preparation 1.
• a cyclopentane carboxaldehyde of formula 2 can be prepared as outlined in scheme VI and as further illustrated in the Examples.
• Piperidine aldehyde 12 can be made by oxidation of aldehyde 13, which in turn, is formed by reaction of the hydroxymethyl piperidine 13a with chloroacetyl chloride. See, e.g., Example
• Embodiments wherein Xi is N and X 2 is C, Ring B is heterocycloaliphatic and Ring A is cycloaliphatic can, in general, be prepared as described above but substituting a cycloaliphatic hydrazide which may be obtained as shown in Scheme IX.
• TGF ⁇ 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.
• tissues and organs e.g., lung, kidney, and liver
• TGF ⁇ and/or activin mRNA and the level of TGF ⁇ and/or activin are increased in patients suffering from various fibrotic disorders, e.g., fibxotic kidney diseases, alcohol- induced and autoimmune hepatic fibrosis, myelofibrosis, and bleomycin-induced pulmonary fibrosis.
• various fibrotic disorders e.g., fibxotic kidney diseases, alcohol- induced and autoimmune hepatic fibrosis, myelofibrosis, and bleomycin-induced pulmonary fibrosis.
• Compounds of formula (I), which are antagonists of the TGF ⁇ family type I receptors Alk5 and/or Alk4, and inhibit TGF ⁇ and/or activin signaling pathway, are therefore useful for treating and/or preventing fibrotic disorders or diseases mediated by an increased level of TGF ⁇ and/or activin activity.
• a compound inhibits the TGF ⁇ family signaling pathway when it binds (e.g., with an IC 50 value of less than 10 ⁇ M; such as, less than 1 ⁇ M; and for example, less than 5 nM) to a receptor of the pathway (e.g., Alk5 and/or Alk4), 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.
• a receptor of the pathway e.g., Alk5 and/or Alk4
• the aforementioned disorders or diseases include any condition (a) marked by the presence of an abnormally high level of TGF ⁇ and/or activin; and/or (b) an excess accumulation of extracellular matrix; and/or (c) an increased number and synthetic activity of myofibroblasts.
• fibrotic conditions such as mesothelioma, acute respiratory distress syndrome (ARDS), atherosclerosis, scleroderma, keloids, glomerulonephritis, diabetic nephropathy, lupus nephritis, hypertension-induced nephropathy, cholangitis, restinosis, ocular or corneal scarring, hepatic or biliary fibrosis, liver cirrhosis, cirrhosis due to fatty liver disease (alcoholic and nonalcoholic steatosis), renal fibrosis, sarcoidosis, acute lung injury, drug-induced lung injury, spinal cord injury, CNS scarring, systemic lupus erythematosus, Wegener's granulomatosis, pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis), cardiac fibrosis, post-infarction cardiac fibrosis, post-surgical
• ARDS acute respiratory distress syndrome
• fibrotic conditions for which preventive treatment with compounds of formula (I) can have therapeutic utility include radiation therapy-induced fibrosis, chemotherapy-induced fibrosis, and surgically induced scarring including surgical adhesions, transplant arteriopathy, laminectomy, and coronary restenosis.
• TGF ⁇ activity is also found to manifest in patients with progressive cancers. Studies have shown that in late stages of various cancers, both the tumor cells and the stromal cells within the tumors generally overexpress TGF ⁇ . This leads to stimulation of angiogenesis and cell motility, suppression of the immune system, and increased interaction of tumor cells with the extracellular matrix. See, e.g., Hojo, M. et al., Nature 397: 530-534 (1999). As a result, the tumor cells become more invasive and metastasize to distant organs. See, e.g., Maehara, Y. et al., J. Clin. Oncol, 17: 607-614 (1999) and Picon, A.
• compounds of formula (I), which are antagonists of the TGF ⁇ type I receptor and inhibit TGF ⁇ signaling pathways, are also useful for treating and/or preventing various late stage cancers (including carcinomas) which overexpress TGF ⁇ .
• late stage 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.
• TGF ⁇ and/or activin e.g., fibrosis or cancers
• small molecule treatments such as treatment disclosed in the present invention
• TGF ⁇ and/or activin are favored for long-term treatment.
• compounds of formula (I) useful in treating disorders or diseases mediated by high levels of TGF ⁇ 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 ⁇ and/or activin production have been associated with fibrosis and hypertension.
• TGF ⁇ 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.
• the levels of TGF ⁇ and/or activin in serum and of TGF ⁇ 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 ⁇ and/or activin, and polymorphisms in the gene for TGF ⁇ that determine the production of TGF ⁇ 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.
• the inhibitors described herein are effective at treating, preventing, or reducing intimal thickening, vascular remodeling, restenosis (e.g., coronary, peripheral, or carotid restenosis), vascular diseases (e.g., intimal thickening, vascular remodeling, organ transplant-related, cardiac, and renal), and hypertension (e.g., primary and secondary, systolic, pulmonary, and hypertension-induced vascular remodeling resulting in target organ damage).
• restenosis e.g., coronary, peripheral, or carotid restenosis
• vascular diseases e.g., intimal thickening, vascular remodeling, organ transplant-related, cardiac, and renal
• hypertension e.g., primary and secondary, systolic, pulmonary, and hypertension-induced vascular remodeling resulting in target organ damage.
• TGF ⁇ RI TGF ⁇ type I receptor
• Alk4 activin type I receptor
• TGF ⁇ RI kinase activity is required for TGF ⁇ signaling as is Alk4 for activin signaling.
• Kinases have proven to be useful targets for development of small molecule drugs. There is a good structural understanding of the TGF ⁇ RI kinase domain allowing the use of structure-based drug discovery and design to aid in the development of inhibitors.
• TGF ⁇ or activin-mediated pathological changes in vascular flow and tone are often the cause of morbidity and mortality in a number of diseases (Gibbons G.H. and Dzau VJ. N Eng. J. Med 330:1431-1438 (1994)).
• the initial response of the vasculature to injury is an infiltration of adventitial inflammatory cells and induction of activated myofibroblasts or smooth muscle cells (referred to as myofibroblasts from hereon).
• myofibroblasts smooth muscle cells
• TGF ⁇ is initially produced by infiltrating inflammatory cells and activates myofibroblasts or smooth muscle cells. These activated myofibroblasts can also secrete TGF ⁇ as well as respond to it.
• TGF ⁇ vascular remodeling processes, intimal thickening and vascular contraction, restrict blood flow to the tissues supported by the effected vasculature and result in tissue damage.
• Activin is also produced in response to injury and shows very similar actions in inducing activated myofibroblasts or activated smooth muscle cells intimal thickening and vascular remodeling. See, for example, scientific articles by Pawlowski et al. J. Clin. Invest.
• the treatment of arterial stenotic disease by surgical grafts also can elicit restenosis in the grafted vessel.
• vein grafts undergo intimal thickening and vascular remodeling through a similar mechanism involving TGF ⁇ -induced intimal thickening and vascular remodeling.
• the injury is either due to the overdistention of the thin-walled vein graft placed into an arterial vascular context or due to anastamotic or ischemic injury during the transplantation of the graft.
• the loss of patency in arteriovenous or synthetic bridge graft fistulas is another vascular remodeling response involving increased TGF ⁇ production.
• Elevated TGF ⁇ is implicated in chronic allograft vasculopathy both in animals and humans.
• Vascular injury, intimal thickening and vascular remodeling is a characteristic pathology in chronic allograft failure.
• the fibrotic response in chronic allograft failure initiates in the vasculature of the donor organ.
• Chronic allograft vasculopathy in allografted hearts often manifests within 5 years of transplantation and is the main cause of death in long term survivors of cardiac transplant.
• Both early detection of cardiac allograft vasculopathy measured as intimal thickening by intravascular ultrasound as well as the elevation of plasma TGF ⁇ has been suggested as a prognostic marker for late cardiac allograft failure (Mehra, M.R.
• Elevation of TGF ⁇ can be induced by ischemic, immune and inflammatory responses to the allograft organ.
• Animal models of acute and chronic renal allograft rejection identify the elevation of TGF ⁇ as a significant contributor to graft failure and rejection (Nagano, H. et al., Transplantation, 63: 1101 (1997); Paul, L.C. et al, Am. J. Kidney Dis., 28: 441 (1996); Shihab, F.S. et al., Kidney Int., 50: 1904 (1996)).
• Rodent models of chronic allograft nephropathy (CAN) show elevation of TGF ⁇ mRNA and immunostaining.
• TGF ⁇ immunostaining is strongly positive in interstitial inflammatory andfibrotic cells, but also in blood vessels and glomeruli.
• the loss of renal function 1 year post renal allograft correlates with TGF ⁇ staining in the grafted kidney (Cuhaci, B. et al., Transplantation, 68: 785 (1999)).
• Graft biopsies show also that renal dysfunction correlates with chronic vascular remodeling, i.e., vasculopathy, and the degree of TGF ⁇ expression correlates significantly with chronic vasculopathy (Viticiany, O. et al., Physiol Res,, 52: 353 (2003)).
• TGF ⁇ is implicated in chronic allograft rejection in both renal and lung transplants due to the clear TGF ⁇ -related fibrotic pathology of this condition as well as the ability of immune suppressants, especially cyclosporin A, to induce TGF ⁇ (Jain, S. et al., supra).
• TGF ⁇ blockade improved renal function while decreasing collagen deposition, renal TGF ⁇ expression as well as vascular afferent arteriole remodeling in a cyclosporine A-induced renal failure model using an anti-TGF ⁇ monoclonal antibody (Islam, M. et al., Kidney Int. , 59: 498 (2001); Khanna, A.K. et al., Transplantation, 67: 882 (1997)).
• These data are strongly indicative of a causal role for TGF ⁇ in the development and progression of chonic allograft vasculopathy and chronic allograft failure.
• Hypertension is a major cause of morbidity and mortality in the U.S. population affecting approximately 1 in 3 individuals.
• the effect of hypertension on target organs include increased incidence of cardiac failure, myocardial infarction, stroke, renal failure, aneurysm and microvascular hemorrhage.
• Hypertension-induced damage to the vasculature results in vascular remodeling and intimal thickening which are a major causative factor in many of these morbidities (Weber, W.T., Curr. Opin. Cardiol, 15: 264-72 (2000)).
• TGF ⁇ is elevated upon induction of hypertension and anti-TGF ⁇ monoclonal antibody blockade of this pathway decreases blood pressure and renal pathology in hypertensive rats (Xu, C. et al., J. Vase. Surg., 33: 570 (2001); Dahly, AJ. et al., Am. J. Physiol. Regul. Integr. Comp. Physiol, 283: R757 (2002)).
• plasma TGF ⁇ is elevated in hypertensive individuals compared to normotensive controls and plasma TGF ⁇ is also higher in hypertensive individuals with manifest target organ disease compared to hypertensive individuals without apparent target organ damage (Derhaschnig, U. et al., Am. J. Hypertens., 15: 207 (2002); Suthanthiran, M., Proc. Natl. Acad. ScL USA, 97: 3479 (2000)).
• TGF ⁇ TGFy-producing genotypes of TGF ⁇ are a risk factor for development of hypertension.
• the inhibition of the TGF ⁇ pathway may provide an effective therapeutic approach for hypertension or hypertension-induced organ damage.
• the vascular injury response in the pulmonary vasculature results in pulmonary hypertension which can lead to overload of the right heart and cardiac failure (Runo, J.R. et al., Lancet, 361(9368): 1533-44 (2003); Sitbon, O.
• TGF ⁇ RI inhibitors Prevention of pulmonary vascular remodeling by TGF ⁇ RI inhibitors can be of practical utility in diseases such as primary or secondary pulmonary hypertension (Sitbon O et al 2002 Prog Cardiovasc Dis. 45: 115-28; Humbert M et al. J Am Coll Cardiol. 200443:13S-24S). Inhibition of the progression of vascular remodeling over time will prevent the progression of pulmonary pathology in these life threatening diseases.
• Pulmonary hypertension occurs often as a manifestation of scleroderma and is one of the primary causes of morbidity and mortality in scleroderma patients (Denton, CP. et al., Rheum Dis Clin North Am., 29:335-49 (2003)). Pulmonary hypertension is also a sequalae of mixed connective tissue disease, chronic obstructive pulmonary disease (COPD) and lupus erythematosis (Fagan, K.A. et al., Prog. Cardiovasc. Dis., 45: 225-34 (2002); Presberg, K.W. et al., Curr. Opin. PuIm. Med., 9:131-8 (2003)).
• COPD chronic obstructive pulmonary disease
• blockade of TGF ⁇ is of particular utility in diabetic patients at risk for hypertension-related organ failure, diabetic nephropathy, restenosis or vein graft stenosis in coronary or peripheral arteries, and chronic failure of allograft organ transplants (Endemann, D.H. et al., Hypertension, 43 (2): 399-404 (2004); Ziyadeh, F., J. Am. Soc. Nephrol, 15 (Suppl. 1): S55-7 (2004); Jerums, G. et al., Arch. Biochem. Biophys., 419:55-62 (2003)).
• TGF ⁇ RI and Alk4 antagonists are effective at treating, preventing, or reducing intimal thickening, vascular remodeling, restenosis (e.g., coronary, peripheral, or carotid restenosis), vascular diseases (e.g., organ transplant-related, cardiac, and renal), and hypertension (e.g., systolic, pulmonary, and hypertension-induced vascular remodeling resulting in target organ damage).
• vascular remodeling e.g., coronary, peripheral, or carotid restenosis
• vascular diseases e.g., organ transplant-related, cardiac, and renal
• hypertension e.g., systolic, pulmonary, and hypertension-induced vascular remodeling resulting in target organ damage.
• Changes in vascular remodeling and intimal thickening may be qualified by measuring the intimal versus medial vascular thickness.
• an effective amount is the amount required to confer a therapeutic effect on the treated patient.
• 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.
• Compounds of formula (1) 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.
• Solubilizi ⁇ g 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.
• 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).
• 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
• the compounds of formula (I) can be administered by any method that permits the delivery of the compound to combat vascular injuries.
• the compounds of formula (I) can be delivered by any method described above.
• the compounds of formula (I) can be administered by implantation (e.g., surgically) via an implantable device.
• implantable devices include, but are not limited to, stents, delivery pumps, vascular filters, and implantable control release compositions. Any implantable device can be used to deliver the compound provided that: (1) the device, compound and any pharmaceutical composition including the compound are biocompatible, and (2) that the device can deliver or release an effective amount of the compound to confer a therapeutic effect on the treated patient.
• stents Delivery of therapeutic agents via stents, delivery pumps (e.g., mini-osmotic pumps), and other implantable devices is known in the art. See, e.g., Hofma et al., Current Interventional Cardiology Reports, 3: 28-36 (2001), the entire contents of which, including references cited therein, are incorporated herein. Other descriptions of implantable devices, such as stents, can be found in U.S. Pat. Nos.
• a delivery device such as stent, includes a compound of formula (I).
• the compound may be incorporated into or onto the stent using methodologies known in the art.
• a stent can include interlocked meshed cables. Each cable can include metal wires for structural support and polyermic wires for delivering the therapeutic agent.
• the polymeric wire can be dosed by immersing the polymer in a solution of the therapeutic agent.
• the therapeutic agent can be embedded in the polymeric wire during the formation of the wire from polymeric precursor solutions.
• stents or implatable devices can be coated with polymeric coatings that include the therapeutic agent. The polymeric coating can be designed to control the release rate of the therapeutic agent.
• Controlled release of therapeutic agents can utilize various technologies.
• Devices having a monolithic layer or coating incorporating a heterogeneous solution and/or dispersion of an active agent in a polymeric substance, where the diffusion of the agent is rate limiting, as the agent diffuses through the polymer to the polymer-fluid interface and is released into the surrounding fluid.
• a soluble substance is also dissolved or dispersed in the polymeric material, such that additional pores or channels are left after the material dissolves.
• a matrix device is generally diffusion limited as well, but with the channels or other internal geometry of the device also playing a role in releasing the agent to the fluid.
• the channels can be pre-existing channels or channels left behind by released agent or other soluble substances.
• Erodible or degradable devices typically have the active agent physically immobilized in the polymer.
• the active agent can be dissolved and/or dispersed throughout the polymeric material.
• the polymeric material is often hydrolytically degraded over time through hydrolysis of labile bonds, allowing the polymer to erode into the fluid, releasing the active agent into the fluid.
• Hydrophilic polymers have a generally faster rate of erosion relative to hydrophobic polymers. Hydrophobic polymers are believed to have almost purely surface diffusion of active agent, having erosion from the surface inwards. Hydrophilic polymers are believed to allow water to penetrate the surface of the polymer, allowing hydrolysis of labile bonds beneath the surface, which can lead to homogeneous or bulk erosion of polymer.
• the implantable device coating can include a blend of polymers each having a different release rate of the therapeutic agent.
• the coating can include a polylactic acid/polyethylene oxide (PLA-PEO) copolymer and a polylactic acid/polycaprolactone (PLA-PCL) copolymer.
• the polylactic acid/polyethylene oxide (PLA- PEO) copolymer can exhibit a higher release rate of therapeutic agent relative to the polylactic acid/polycaprolactone (PLA-PCL) copolymer.
• the relative amounts and dosage rates of therapeutic agent delivered over time can be controlled by controlling the relative amounts of the faster releasing polymers relative to the slower releasing polymers.
• the stent can be coated by spraying the stent with a solution or dispersion of polymer, active agent, and solvent.
• the solvent can be evaporated, leaving a coating of polymer and active agent.
• the active agent can be dissolved and/or dispersed in the polymer.
• the copolymers can be extruded over the stent body.
• compounds of formula (I) can be administered in conjunction with one or more other agents that inhibit the TGF ⁇ signaling pathway or treat the corresponding pathological disorders (e.g., fibrosis or progressive cancers) by way of a different mechanism of action.
• agents include angiotensin converting enzyme inhibitors, nonsteroid, steroid anti-inflammatory agents, and chemotherapeutics or ⁇ radiation, as well as agents that antagonize l ⁇ gand binding or activation of the TGF ⁇ receptors, e.g., anti-TGF ⁇ , anti-TGF ⁇ receptor antibodies, or antagonists of the TGF ⁇ type II receptors.
• compounds of formula (I) can be administered in conjunction with one or more other agents that inhibit the TGF ⁇ signaling pathway or treat the corresponding pathological disorders (e.g., fibrosis or progressive cancers) by way of a different mechanism of action.
• agents include angiotensin converting enzyme inhibitors, nonsteroid and steroid anti-inflammatory agents, as well as agents that antagonize ligand binding or activation of the TGF ⁇ receptors, e.g., ant ⁇ -TGF ⁇ , anti-TGF ⁇ receptor antibodies, or antagonists of the TGF ⁇ type II receptors.
• Step a [3-(tert-Butyl-dimethyI-siIanyIoxy)-buta-l,3-dienyll-dimethyI-amine: [00165J To a 200 mL flask was added 4-dimethylaminobut-3,4-en-2-one (Aldrich Chemical Co., Milwaukee, WI; 3.0 g; 20.5 mmol) and THF (100 mL). The flask was cooled to -78 0 C and a 0.5 M solution of KHMDS (56 mL; 28.0 mmol) in toluene was added dropwise via addition funnel. The reaction was stirred while warming to 0 0 C over 1 hour.
• 4-dimethylaminobut-3,4-en-2-one Aldrich Chemical Co., Milwaukee, WI; 3.0 g; 20.5 mmol
• THF 100 mL
• the flask was cooled to -78 0 C and a 0.5 M solution of KHM
• reaction mixture was cooled to -78 0 C and a solution of TBSCl (4.39 g; 29.2 mmol) in THF (25 mL) was added via addition funnel. The reaction was allowed to stir while warming to 20 0 C. The reaction was diluted with ether (500 mL) and then filtered through celite and concentrated in vacuo. The resulting oil (5.92 g; 98% yield) was pure and needed no further purification.
• Step b 4-(tert-Butyl-dimethyl-sUanyloxy)-2-dimethylamino-cyclohex-3- enecarboxylic acid methyl ester.
• step Ia A 1.0 L round-bottom flask was charged with the diene of step Ia (56.6 g; 249 mmol) toluene (500 mL) and methyl acrylate (29.3 mL; 323 mmol). The reaction was heated to 50 0 C and stirred overnight. The mixture was concentrated in vacuo to yield product (68.2 g; 87% yield) as a mixture of stereoisomer.
• Step c [4-(tert-Butyl-dimethyl-silanyloxy)-2-dimethylamino-cyclohex-3-eiiyl]- methanol.
• Step d 4-Hydroxymethyl-cyclohex-2-enone.
• step Ic To a 1-L round-bottom flask was added the alcohol of step Ic (22.6 g; 79.0 mmol), THF (300 mL) and a 35% solution of HF in water (8.08 mL; 182 mmol). The reaction was allowed to stir at 20 0 C for 12 hours and then concentrated in vacuo. The residue was diluted with EtOAc (250 mL), washed with 10% HCl (250 mL ) and brine (250 mL). The organic layer was then treated with solid NaHCO 3 for 1 hour, dried (MgSO 4 ), filtered and concentrated in vacuo. The crude oil (5.3 g; 53% yield) was used without further purification.
• Step e 4-(tert-Butyl-dimethyl-s£Ianyloxymethyl)-cyclohex-2-enone.
• a 200 mL round-bottom flask was charged with the alcohol of step Id (3.80 g; 30.1 mmol), DMF (100 mL), imidazole (2.46 g; 36.1 mmol) and TBSCl (5.0 g; 33.1 mmol).
• the reaction was allowed to stir for 72 hours and then diluted with ether (200 mL).
• the mixture was washed with 5% HCl (200 mL), saturated NaCl (200 mL), dried (MgSO 4 ), filtered and concentrated in vacuo.
• Step f 4-(tert-Butyl-dimethyl-siIanyIoxymethyl)-3-vinyl-cyclohexanone.
• copper(I) bromide- dimethylsulfide complex (1.80 g; 8.74 mmol)
• THF (30 mL)
• TMEDA 2.50 mL; 16.6 mmol
• the mixture was stirred until all the CuBr had dissolved and then cooled to -78 0 C.
• To the mixture was added a 1.0 M solution of vinylmagnessium bromide in THF (16.6 mL; 16.6 mmol).
• Step g (7-Vinyl-l,4-dioxa-spiro[4.5]dec-8-yl)-methanol.
• Step h 7-Vinyl-l,4-dioxa-spiro[4.5]decane-8-carbaldehyde.
• Example 9 The compound of Example 9 (0.18 g, 0.00041 mol) was dissoved in CH 2 Cl 2 at RT and 1.0 M of boron tribromide in hexane (1.0 mL) was added to the solution. The mixture was stirred for 2 hours. The reaction was quenched with NaHCO 3 aq. and was extracted with CH 2 CI 2 . The organic layers were combined, dried over Na 2 SO 4 , filtered and rotovaped to give the title compound (0.14g, 83%).
• Example 16 1 H-
• TGF ⁇ inhibitory activity of compounds of formula (I) can be assessed by methods described in the following examples.
• the serine-threonine kinase activity of TGF ⁇ 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-TGF ⁇ RI.
• the His-tagged receptor cytoplasmic kinase domains were purified from infected insect cell cultures using the Gibco- BRL FastBac HTb baculovirus expression system.
• Example 18 Cell-Free Assay for Evaluating Inhibition of Activin Type I Receptor
• Example 20 Assay for Evaluating Cellular Inhibition of TGF ⁇ Signaling
• Biological activity of the compounds of formula (I) was determined by measuring their ability to inhibit TGF ⁇ -induced PAI-Luciferase reporter activity in HepG2 cells.
• HepG2 cells were stably transfected with the PAI-luciferase reporter grown in
• DMEM medium containing 10% FBS, penicillin (100 U/mL), streptomycin (100 ⁇ g/mL), L- glutamine (2 mM), sodium pyruvate (1 mM), and non-essential amino acids (once).
• 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 0 C in a 5% CO 2 incubator.
• the cells were then stimulated with 2.5 ng/mL TGF ⁇ 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.
• test compounds of formula (I) 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 20 except that 100 ng/mL of activin is added to serum starved cells in place of the 2.5 ng/mL TGF ⁇ .
• Example 22 Assay for TGF ⁇ -induced Collagen Expression
• 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)).
• GFP Green Fluorescent Protein
• Cells are immortalized with a temperature sensitive large T antigen that is in an active stage at 33 0 C.
• Cells are expanded at 33 0 C and then transferred to 37 0 C 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.
• 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 0 C, 5% CO 2 .
• the cells are trypsinized in the following day and transferred into 96-well format with 30,000 cells per well in 50 ⁇ L complete DMEM containing fetal calf serum, but without phenol red.
• the cells are incubated at 37 0 C for 3 to 4 hours to allow them to adhere to the plate.
• Stenotic fibrotic response Balloon Catheter Injury of the Rat Carotid Artery
• the ability of compounds of formula (I) to prevent the stenotic fibrotic response is tested by administration of the test compounds to rats that have undergone balloon catheter injury of the carotid artery.
• the test compounds are administerd intervenously, subcutaneously, p.o., orally, or parentally.
• Sprague Dawley rats 400 g, 3 to 4 months old are anesthetized by inter paratenal i.p. injection with 2.2 mg/kg xylazine (AnaSed, Lloyd laboratories) and 50 mg/kg ketamine (Ketalar, Parke-Davis).
• the left carotid artery and the aorta are denuded with a 2F balloon catheter according to the procedure described in Clowes et al., Lab Invest., 49: 327-333 (1983).
• the animals were sacrificed under anesthesia 14 days post-balloon injury.
• Perfusion fixation was carried out under physiological pressure with phosphate buffered (0.1 mol/L, pH 7.4) 4% paraformadehyde.
• the injured carotid artery was excised, post-fixed and embedded for histological and morphometic analysis.
• Sections (5 ⁇ m) were cut from the proximal, middle and distal segments of the denuded vessel and analyzed using image analysis software.
• the circumference of the lumen and the lengths of the internal elastic lamina (IEL) and the external elastic lamina (EEL) were determined by tracing along the luminal surface the perimeter of the neointima (IEL) and the perimeter of the tunica media (EEL) respectively.
• the lumen (area within the lumen), medial (area between the IEL and EEL) and intimal (area between the lumen and the IEL) areas were also determined using morphometric analysis.
• Statistical analysis used ANOVA to determine statistically significant differences between the means of treatment groups (p ⁇ 0.05). Multiple comparisons between groups were then performed using the Scheffe test. The Student t test was used to compare the means between 2 groups, and differences were considered significant if P ⁇ 0.05. All data are shown as mean + SEM.

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