Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
  • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
  • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
  • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
  • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • 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
• • 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
  • A61P9/12—Antihypertensives

Definitions

• 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).
• BMPs bone morphogenetic proteins
• GDFs growth and differentiation factors
• MIS mullerian inhibiting substance
• TGF/3 exists in three isoforms (TGF 1 Sl, TGF
• 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 TGFjS.
• LAP latency associated peptide
• TGFjS N-terminal region
• LAP is typically non-covalently associated with mature TGFjS prior to secretion from the cell.
• the LAP-TGFjS 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.
• TGF 1 S binds at high affinity to the type II receptor (TGFjSRII), a constitutively active serine/threonine kinase.
• TGFjSRII type II receptor
• the ligand-bound type II receptor phosphorylates the TGF/3 type I receptor (AIk 5) in a glycine/serine rich domain, which allows the type I receptor to recruit and phosphorylate downstream signaling molecules, Smad2 or Smad3.
• TGFjSRII type II receptor
• AIk5 TGF/3 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/3-responsive genes. See, e.g., Massague, J. Ann. Rev. Biochem. Med. 67: 773 (1998).
• 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 TGF/3, that is, by binding to a constitutive serine-threonine receptor kinase, activin type II receptor (ActRIIB), and activating a type I serine-threonine receptor, AIk 4, 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
• AIk 4 type I serine-threonine receptor
• TGF 1 S 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.
• TGF 1 S 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). See, e.g. Blobe G.C., Schiemann W.P., Lodish H.F. N. Eng. J. Med. 342: 1350-8 (2000).
• 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.
• TGF/3 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 ' ⁇ family to prevent and/or treat disorders involving this signaling pathway.
• modulators e.g., antagonists
• the compounds of formulae I, II, III, IV, V, and VI, described herein unexpectedly exhibit systemic bioavailability and are useful in combating restenosis (e.g., coronary restenosis, peripheral restenosis, and carotid restenosis), vascular diseases (e.g., intimal thickening, vascular remodeling, or organ transplant-related vascular disease), and hypertension (e.g., primary or secondary, systolic hypertension, pulmonary hypertension or hypertension-induced vascular remodeling) when administered to a subject (e.g., a patient).
• restenosis e.g., coronary restenosis, peripheral restenosis, and carotid restenosis
• vascular diseases e.g., intimal thickening, vascular remodeling, or organ transplant-related vascular disease
• hypertension e.g., primary or secondary, systolic hypertension, pulmonary hypertension or hypertension-induced vascular remodeling
• the invention features an implantable device, such as a delivery pump or stent, including an inhibitor of TGF ⁇ type I receptor or Alk4.
• the inhibitor can be a compound of formula I, II, III, IV, V, or VI.
• 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 an alkyl group include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-heptyl, and 2-ethylhexyl.
• An alkyl group can be optionally substituted with one or more substituents such as alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, amino, nitro, carboxy, cyano, halo, hydroxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, cycloalkyl- alkylcarbonylamino, arylcarbonylamino, aralkylcarbonylamino, heterocycloalkyl- carbonylamino, heterocycloalkyl-alkylcarbonylamino, heteroarylcarbonylammo, heteroaralkylcarbonylamino, urea, thiourea, sulfamoyl,
• 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.
• An alkenyl group can be optionally substituted with one or more substituents such as alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, amino, nitro, carboxy, cyano, halo, hydroxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, cycloalkyl- alkylcarbonylamino, arylcarbonylamino, aralkylcarbonylamino, heterocycloalkyl- carbonylamino, heterocycloalkyl-alkylcarbonylamino, heteroarylcarbonylammo, het ' er ⁇ aralkylcarbonylamino, urea, thiourea, s
• an "alkynyl” group refers to an aliphatic carbon group that contains 2- 8 (e.g., 2-6 or 2-4) 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, proparg;yl and butynyl.
• An alkynyl group can be optionally substituted with one or more substituents such as alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, amino, nitro, carboxy, cyano, halo, hydroxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, cycloalkyl-alkylcarbonylamino, arylcarbonylamino, aralkylcarbonylamino, heterocycloalkyl- carbonylamino, heterocycloalkyl-alkylcarbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino, urea, thiourea, sulfamoyl
• an “amino” group refers to -NR X R Y wherein each of R x and R ⁇ is independently hydrogen, hydroxyl, alkyl, alkoxy, cycloalkyl, (cycloalkyl)alkyl, aryl, aralkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, heteroaryl, or heteroaralkyl.
• R x is independently hydrogen, hydroxyl, alkyl, alkoxy, cycloalkyl, (cycloalkyl)alkyl, aryl, aralkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, heteroaryl, or heteroaralkyl.
• an "aryl” group refers to phenyl, naphthyl, or a benzofused group having 2 to 3 rings.
• a benzofused group includes phenyl fused with one or two C 4-8 carbocyclic moieties, e.g., 1,2,3,4-tetrahydronaphthyl, indanyl, or fluorenyl.
• An aryl 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, amino, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyL, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkyl)alkylcarbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloal
• 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 above. An example of an aralkyl group is benzyl.
• a "cycloalkyl” group refers to an aliphatic carbocyclic ring of 3-10 (e.g., 4-8) carbon atoms.
• Examples of cycloalkyl groups include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, norbornyl, cubyl, octahydro-indenyl, decahydro- naphthyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl, and bicyclo[3.2.3]nonyl.
• cycloalkenyl refers to a non-aromatic carbocyclic ring of 3-10 (e.g., 4-8) carbon atoms having one or more double bond.
• cycloalkenyl groups include cyclopentenyl, 1,4-cyclohexa-di-enyl, cycloheptenyl, cyclooctenyl, hexahydro-indenyl, octahydro-naphthyl, 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 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, amino, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkyl)alkylcarbonylamino, arylcarbonylamino, aralkyl
• heterocycloalkyl refers to a 3- to 10-membered (e.g., 4- to 8-membered) saturated ring structure, in which one or more of the ring atoms is a heteroatom, e.g., N, O, or S.
• heterocycloalkyl group examples include piperidinyl, piperazinyl, tetrahydropyranyl, tetrahydrofuryl, dioxolanyl, oxazolidinyl, isooxazolidinyl, morpholinyl, octahydro-benzofuryl, octahydro-chromenyl, octahydro-thiochromenyl, octahydro-indolyl, octahydro-pyrindinyl, decahydro-quinolinyl, octahydro-benzo[b]thiophenyl, 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[3.3.1.0 3>7 ]nonyl.
• heterocycloalkenyl group refers to a 3- to 10- membered (e.g., 4- to 8-membered) 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.
• a heterocycloalkyl or heterocycloalkenyl group 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, cycloallcyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, amino, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkyl)alkylcarbonylamin.o, arylcarbonylamino, aral
• a "heteroaryl” group refers to a monocyclic, bicyclic, or tricyclic ring structure having 5 to 15 ring atoms wherein one or more of the ring atoms is a heteroatom, e.g., N, O, or S, and wherein one ore more rings of the bicyclic or tricyclic ring structure is aromatic.
• heteroaryl examples include pyridyl, furyl., pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, indolyl, tetrazolyl, benzofuryl, benzthiazolyl, xanthene, thioxanthene, phenothiazine, dihydroindole, and benzo[l,3]dioxole.
• a tieteroaryl is optionally substituted with one or more substituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoromethyl), alkenyl, alkynyl, cyclo alkyl, (cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl, heteroaryl;, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, amino, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalk ⁇ alkylcarbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycl
• heteroaryl group refers to an alkyl group (e.g., a C 1-4 alkyl group) th.at is substituted with a heteroaryl group. Both “alkyl” and “heteroaryl” are as defined above.
• cyclic moiety includes cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocyclo alkenyl, aryl, or heteroaryl, each of which has Been defined previously.
• a “carbamoyl” group refers to a group having the structure
• R x and R ⁇ have been defined above and R z is alkyl, cycloalkyl, (cycloalkyl)alkyl, aryl, aralkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, heteroaryl, or heteroaralkyl.
• a "carboxy” group and a “sulfo” group refer to -COOH and -SO 3 H, respectively.
• alkoxy refers to an alkyl-O- group wherein “alkyl” has been defined previously.
• a "sulfoxy" group refers to -O-SO-R X or -SO-O-R* 1 , wherein R x has been defined above.
• halogen or halo group refers to fluorine, chlorine, bromine or iodine.
• a "sulfamoyl” group refers to the structure -SO 2 -NR X R Y or -NR X - SO 2 -R 2 wherein R x , R ⁇ , and R z have been defined above.
• sulfamide refers to the structure -NR X -S(O) 2 -NR Y R Z wherein R x , R ⁇ , and R z have been defined above.
• urea refers to the structure -NR X -CO-NR ⁇ R Z and a "thiourea” group refers to the structure -NR X -CS-NR Y R Z .
• R x , R ⁇ , and R z have been defined above.
• an effective amount is defined as the amount which is 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).
• a. "patient” refers to a mammal, including a human.
• An antagonist is 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.
• Figure 1 is an illustration of a delivery device.
• the inhibitors described herein are effective at treating, preventing, or reducing intimal thickening, vascular remodeling, restenosis (e.g., coronary, peripheral, carotid restenosis), vascular diseases, (e.g., 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, carotid restenosis
• vascular diseases e.g., 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 ⁇ and activin pathway plays a critical role in the progression of fibrotic diseases.
• TGF ⁇ RT 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.
• T 1 GrFPi or activin-mediated pathological changes in vascular flow and tone are often the cause of morbidity and mortality in a number of diseases (see, e.g., Gibbons G.H. and Dzau V.J. NEng. 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).
• 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.
• Stent placement physically prevents remodeling, but hyperplasia and extracellular matrix deposition by activated myofibroblasts proliferating at the luminal side of the stent results in mtimal thickening within the stented vessel resulting in the eventual impairment of blood flow.
• the treatment of arterial stenotic diseases 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.
• 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.
• 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 (see, e.g., Nagano, H. et al., 1997, Transplantation, 63: 1101; Paul, LC. et al., 1996, Am. J. Kidney Dis., 28: 441; and Shihab, FS. et al., 1996, Kidney M., 50: 1904).
• Rodent models of chronic allograft nephropathy 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. See, e.g., Cuhaci, B. et al., 1999, Transplantation, 68: 785).
• 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 (see, e.g., Viticiany, O. et al., 2003, Physiol Res. 52: 353).
• 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, esp cyclosporin A, to induce TGF ⁇ (Jain, S. et al., 2000 Transplantation, 69: 1759).
• 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., 2001 Kidney Int., 59: 498; Khanna, A.K. et al., 1997 Transplantation, 67: 882).
• 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 ofthese morbidities (Weber, W.T. 2000 Curr. Opin. Cardiol. 15:264-72).
• Animal experiments suggest that TGF ⁇ is elevated upon induction of hypertension and anti-XGF ⁇ monoclonal antibody blockade of this pathway decreases blood pressure and renal pathology in hypertensive rats (Xu, C.
• 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., 2002 Am. Jf. Hypertens., 15:207; Suthanthiran, M. 2000 PrOc. Natl. Acad. ScL U.S.A., 97:3479).
• TGF ⁇ -producing genotypes of TGF ⁇ are a risk factor for development of hypertension (Lijnen, PJ. 2003 Am. J. Hypertens., 16:604; Suthanthiran, M. 2000 Proc. Natl. Acad. ScL U.S.A., 97:3479).
• the inhibition of the TGF ⁇ pathway may provide a effective therapeutic approach for hypertension or hypertension-induced organ damage.
• Pulmonary hypertension is also a sequalae of mixied connective tissue disease, chronic obstructive pulmonary disease (COPD) and lupus erythematosis (Fagan, K. A., Badesch, D.B., 2002, Prog. Cardiovasc. Dis., 45:225-34; and Presberg, K.W., Dincer, H.E., 2003, Curr. Opin. PuIm. Med., 9:131-8).
• COPD chronic obstructive pulmonary disease
• diabetic patients have significantly higher rates of restenosis, vein graft stenosis, peripheral artery disease, chronic allograft nephropathy and chronic allograft vasculopathy (Reginelli, J.P., Bhatt, D. L., 2002, J. Invasive Cardiol., 14 Suppl E:2E-10E; Eisen. H., Ross, H., 2004, J. Heart Lung Transplant., 23:S207- 13; Valentine, H., 2004, J. Heart Lung Transplant., 23:S187-93).
• 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., 2004, Hypertension, 43(2):399- 404; Ziyadeh, FJ., Am. Soc. Nephrol, 2004 15 Suppl l:S55-7; and Jerums, G. et al., 2003, Arch. Bi ⁇ chem. Biophys., 419:55-62).
• TGF ⁇ RI and Alk4 antagonists are effective at treating, preventing, or reducing intimal thickening, vascular remodeling, restenosis (e.g., coronary, peripheral, and 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, and 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.
• TGFjS and activin inhibitory activity of compounds can be assessed by methods described below.
• the antagonists have the structure shown in formula I:
• R 1"1 can be aryl, heteroaryl, aralkyl, or heteroaralkyl.
• Each R 1"8 can be alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, alkylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, svilfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, lieterocycloal
• Xi -J can be cycloalkyl or heterocycloalkyl.
• Yn can be a bond, -C(O)-, -C(O)-O-, -O-C(O)-, -S(O) P -O-, -O-S(O) P -, -C(O)-N(R 1"13 )-, -N(R 1 ⁇ )-C(O)-, -O- C(O)-N(R 1"13 )-, -N(R ⁇ )-C(O)-O-, -0-S(O) P -N(R 1" ")-, -N(R 1 ⁇ )-S(O) P -O-, -N(R 1 ⁇ )-C(O)-N(R 1" c )-, -N(R 1 ⁇ )-S(O) P -N(R 1"0 )-, -C(O)-N(
• R 1"2 can be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, cycloalkenyl, (cycloalkenyl)alkyl, aryl, aralkyl, arylalkenyl, heterocycloalkyl, (heterocycloalkyl)alkyl, heterocycloalkenyl, (heterocycloalkcenyl)alkyl, heteroaryl, heteroaralkyl, or (heteroaryl)alkenyl.
• Each of A 1"1 and A 1"2 can be O, S, N, or NR 1"13 , provided that at least one of A 1"1 and A 1"2 can be N.
• m can be O, 1, 2, or 3.
• the 2- pyridyl ring can be unsubstituted or substituted with 1 to 3 R 1" * 1 groups. Note that when m > 2, two adjacent R ⁇ a groups can optionally together to form a 4- to 8-membered optionally substituted cyclic moiety.
• the 2-pyridyl ring can fuse with a cyclic moiety to form a moiety, e.g., 7H-[2]pyrindiiiyl, 6,7-dihydro-5H-[l]pyrindinyl, 5,6,7,8-tetrahydro- quinolinyl, 5,7-dihydro-furo[3,4-b]pyridinyl, or 3,4-dihydro-lH-thiopyrano[4,3-c]pyridinyl, that can be optionally substituted with one or more substituents such as alkyl (including substituted alkyl such as carboxyalkyl, hydroxyalkyl, and haloalkyl (e.g., trifluoromethyl)), alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, aryl, heteroaryl, aryloxy, heteroaryloxy, aroyl, heteroaroyl, amino,
• Methanesulfonic acid 4-[4-toenzo[l,3]dioxol-5-yl-5-(6-methyl-pyridin-2-yl)-lH- imidazol-2-yl]-bicyclo[2.2.2]oct- 1-ylmethyl ester;
• Butane- 1 -sulfonic acid ⁇ 4-[4-benzo[ l,3]dioxol-5-yl-5-(6-methyl-pyridin-2-yl)-lH- imidazol-2-yl]-cyclohexylmethyl ⁇ -amide;
• the antagonists have the structure shown below in formula II:
• each OfXn -1 , Xn -2 , Xn -3 , and Xn -4 can be independently CR ⁇ "x or N, provided that only two of X ⁇ -i, Xn -2 , Xn -3 , and Xn -4 can be N simultaneously.
• Each OfYn -1 and Y 11-2 can be independently CR ⁇ "y or N, provided that at least one OfYn -1 and Yn -2 must be N.
• the ring having Y n-1 and Yn -2 ring atoms can be a pyrimidinyl or pyridyl.
• Each R 11'1 can be independently alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, alkylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, heterocycloalkyl, heterocycloalkyloxy, heterocycloalkylsulfanyl, aryl, aryloxy, arylsulfanyl, aroyl, heteroaryl,
• Each R 11"2 can be independently alkyl, alkenyl, alkynyl, acyl, halo, hydroxy, -NH 2 , -NH(alkyl), -N(alkyl) 2 , -NH(cycloalkyl), -
• R ⁇ "x and R ⁇ "y can be independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, cycloalkylcarbonyl, (cycloalkyl)alkylcarbonyl, aroyl, aralkylcarbonyl, heterocycloalkylcarbonyl, (heterocycloalkyl)acyl, heteroaroyl, (heteroaryl)acyl, aminocarbonyl, alkylcarbonylamino, (amino)aminocarbonyl, alkylsulfonylaminocarbonyl, alkylsulfonylamino, cycloalkylcarbonylamino, cycloal
• two adjacent R 11"1 groups can optionally together to form a 4- to 8-membered optionally substituted cyclic moiety. That is, tlie 2- pyridyl ring can fuse with a 4- to 8-membered cyclic moiety to form a moiety such as 7H- [l]py ⁇ indinyl, 6,7-dihydro-5H-[l]pyrindinyl, 5,6,7,8-tetrahydro-quinolinyl, 5,7-dihydro- furo[3,4-b]pyridinyl, or 3,4-dihydro-lH-thiopyrano[4,3-c]pyridinyl.
• the fused ring moiety can be optionally substituted with one or more substituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoromethyl; see the defmitoix of "alkyl” below), alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, aryl, heteroaryl, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, aroyl, heteroaroyl, amino, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, eye Io alkylcarbonylamino, cycloalkyl-alkylcarbonylamino, arylcarbonylamino, aralkylcarbonylamino, heterocycloalkyl-carbonylamino, heterocycloalkyl- alkylcarbonylamino,
• n _ ⁇ 2 two adjacent R 11"2 groups can optionally join together to form a 4- to 8-membered optionally substituted cyclic moiety, thereby forming a ring fused with the pyridyl or pyrimidinyl group.
• the 4- to 8-membered cyclic moiety formed by two adjacent R 11"2 groups can be optionally substituted with substituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoromethyl; see the defmiton of "alkyl” below), alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, aryl, heteroaryl, aryloxy, heteroaryloxy, aralkyloxy, heteroarylalkoxy, aroyl, heteroaroyl, amino, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylarnino, cycloalkyl-alkylcarbonylamino, arylcarbonylamino, aralkylcarbonylamino, heterocycloalkyl- carbonylamino, heterocycloal
• the antagonists have the structure shown in formula III:
• each OfXm -1 , Xm -2 , X ⁇ i- 3 , and Xm -4 can be independently CR or N, provided that only two of X ⁇ i -i, Xm-2, X 111 - 3 , and Xm -4 can be N simultaneously.
• ⁇ and Ym -2 can be independently CR. m ⁇ y or N, provided that at least one of Y ⁇ n and Y m-2 must be N.
• Each of R 111"1 can be independently alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, cyano, guanadino, amidino, carboxiy, sulfo, mercapto, alkylsulfanyl, alkylsulfmyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, alkylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl, heterocyclo alkyl, heterocycl
• Each of R 111"2 can be independently alkyl, alkenyl, alkynyl, acyl, halo, hydroxy, -NH 2 , -NH(alkyl), -N(alkyl) 2 , -NH(cycloalkyl), -N(alkyl)(cyclocalkyl), - NH(heterocycloalkyl), -NH(heteroaryl), -NH-alkyl-hetero cycloalkyl, -NH-alkyl- heteroaryl, -NH(aralkyl), cycloalkyl, (cycloalkyl)alkyl, aryl, aralkyl, aroyl, heterocycloalkyl, (heterocycloalkyl)alkyl, heteroaryl, heteroaralkyl, heteroaroyl, nitro, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkoxy,
• R m"x and R rII"y can be independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino ., nitro, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, cycloalkylcarbonyl, (cycloalkyl)alkylcarbonyl, aroyl, aralkylcarbonyl, heterocyeloalkylcarbonyl, (heterocycloalkyl)acyl, heteroaroyl, (heteroaryl)acyl, aminocarbonyl, alkylcarbonylamino, (amino)aminocarbonyl, alkylsulfonylaminocarbonyl, alkyl sulfonylamino, cycloalkylcarbon
• the antagonists have the structure shown in formula IV:
• Each R IV"a can be independently alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfmyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkylsulfanyl
• R ⁇ '1 can be a bond, alkylene, alkenylene, alkynylene, or -(CH 2 ) rl -O-(CH 2 ) r2 -, wherein each of rl and r2 is independently 2 or 3.
• R ⁇ "2 can be cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, or a bond.
• R 1 ⁇ 3 can be -C(O)-, -C(O)O-, -OC(O)-, -C(O)-N(R 1 ⁇ )-, -N(R 1 ⁇ )-C(O)-, -0-C(O)- N(R 1 ⁇ )-, -N(R 1 ⁇ )-C(O)-O-, -0-S(O) P -N(R ⁇ 13 )-, -N(R 1 ⁇ )- S(O) P -O-, -N(R 1 ⁇ )-C(O)- N(R 1 ⁇ 0 )-, -N(R IV - b )-S(0) p -N(R IV - b )-, -C(O)-N(R 1 ⁇ )-S(OV, -S(O) P -N(R ⁇ )-C(O)-, -S(0) p - N(R ⁇ )-, -C
• K W' ⁇ can be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, cycloalkenyl, (cycloalkenyl)alkyl, heterocycloalkenyl, (heterocycloalkenyl)alkyl, aryl, aralkyl, heteroaryl, or heteroaralkyl.
• R ⁇ '5 can be hydrogen, unsubstituted alkyl, halo-substituted alkyl, alkoxy, alkylsulfinyl, amino, alkenyl, alkynyl, cycloalkyl, cycloalko:xy, cycloalkylsulf ⁇ nyl, heterocycloalkyl, heterocycloalkoxy, heterocycloalkylsulf ⁇ n ⁇ l, aryl, aryloxy, arylsulfinyl, heteroaryl, heteroaryloxy, or heteroarylsulfinyl.
• Ring A can be an aromatic ring containing 0-4 hetero ring atoms
• ring B can be a 5- to 7- membered aromatic or nonaromatic ring containing 0-4 hetero ring atoms, provided that at least one of ring A and ring B contains one or more hetero rmg atoms.
• Ring A' can be an aromatic ring containing 0-4 hetero ring atoms
• ring B' can be a 5- to 7-membered saturated or unsaturated ring containing 0-4 hetero ring atoms, provided that at least one of ring A' and ring B' contains one or more hetero ring atoms.
• Each X 1 can be independently N or C.
• R w'h and R IV" ' can be independently alkyl, alkenyl, alkynyL, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonyl-amino, alkylsulfonylamino, arylsulfonylamirLO, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamo>4, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycloalkyl
• R 17"6 is 2-naphthyridinyl, 4-quinolinyl, imidazo[l,2- a] ⁇ yridyl, or benzimidazolyl
• -R ⁇ "1 -R IV"2 -R IV"3 -R IV'4 is not H, unsubstituted alkyl, -CH 2 -C(O)-N(H)-alkyl, -CH 2 -C(O)-N(alkyl) 2 , or benzyl.
• WO 04/072033 which is incorpoarated in its entrity by reference, describes synthetic methods for producing inhibitors of formula IV.
• compounds of formula IV include, but are not limited to,
• the antagonists have the structure shown in formula V.
• R v ⁇ ! can be heteroaryl
• Each R v"a can be alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cya.no, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl, sulfamide, carbamoyl, cycloalkyl, cycloalkyloxy, cycl
• Xv can be cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or a bond.
• Yv can be a bond, -CCO)-, -C(O)-O-, -0-C(O)-, -S(O) P -O-, -O-S(O) P -, -C(O)-N(IT 13 )-, -N(R v"b )-C(O)-, -O-C(O)-N(R v - b )-, -N(R v'b )-C(O)-O-, -C(O)-N(R v"b )-O-, -O-N(R v - b )-C(O)-, -O-S(O)p-N(R v"b )-, -N(R v"b )-S(O)p-N(R v"b )-, -N(R v"b )-S(O)p-N(R v"b )-
• R v ⁇ h and R v"c independently, can be hydrogen, hydroxy, alkyl, alkoxy, amino, aryl, aralkyl, rieterocyclo alkyl, heteroaryl, or heteroaralkyl.
• p can be 1 or 2
• q can be 1-4.
• R v"2 can be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, cycloalkenyl, (cycloalkenyl)alkyl, aryl, aralkyl, arylalkenyl, heterocycloalkyl, (heterocycloalkyl)alkyl, heterocycloalkenyl, (Tieterocycloalkenyl)alkyl, heteroaryl, heteroaralkyl, or (heteroaryl)alkenyl.
• Each of A v"! and A v"2 can be N or NR v"b . It is to be understood that when A v ⁇ ' is NR v"b , A v ⁇ 2 is N, and vice versa.
• the variable, m can be 0, 1, 2, or 3.
• the pyrimidinyl ring can be unsubstituted or substituted with 1-3 R v"a groixps. Note that when m ⁇ 2, two adjacent R v"a groups can. optionally together to form a 4- to 8-membered optionally substituted cyclic moiety.
• the pyrimidinyl ring can fuse with a cyclic moiety to form a moiety, that can be optionally substituted with one or more sub> stituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoronx ethyl), alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, aryl, heteroaryl, aryloxy, heteroaryloxy, aroyl, heteroaroyl, amino, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cyano, halo, hydroxy, acyl, mercapto, alkylthio, sulfoxy, sulfamoyl, oxo, or carbamoyl.
• alkyl including carboxyalkyl, hydroxyalkyl, and haloal
• R v ⁇ 2 is hydrogen or alkyl
• m is 1, 2, or 3
• at least one R v"a is substituted at the 2-p;yrimidinyl position (i.e., the position of the pyrimidinyl ring that is between the two nitrogen ring atoms).
• the metrxoxy-methyl- amide can be prepared by reacting a corresponding acid chloride (i.e., R ⁇ -CO-G) with N, O-dimethylhydroxylamine hydrochloride.
• R ⁇ -CO-G N, O-dimethylhydroxylamine hydrochloride.
• the R v"1 -(6-methylpyrimidinyl)-ke “ tone (III) can then be treated with sodium nitrite in acetic acid to afford an ⁇ -keto-oxime (IV), "which can undergo a further reaction with an appropriate substituted (and optionally protected) aldehyde (VI) in the presence of ammonium acetate to yield a compound of formula V.
• the above-described compounds of formula V can be prepared according to Scheme V-Ib below. Specifically, l,l-dimethoxy-propan-2-one can first react with dimethoxymethyl-dimethyl-amine at an elevated temperature to produce the intermediate 4-dimethylamino-l,l-dimethoxy-but-3-en-2-one, which can then react an R v"a - substituted amidine to form an R v"a -substituted pyrimidine-2-carbaldehyde (Ha).
• This carbaldehyde (Ha) can then react with aniline and diphenyl phosphite to form a resulting N,P- acetal, which can further couple with an R ⁇ -substituted aldehyde to produced an (R v" '- methyl)-pyrimidinyl-ketone (Ilia). See, e.g., Journet et al., Tetrahedron Lett. 39:1717-1720 (1998).
• the above-described compounds of formula V can be prepared according to Scheme V-Ic below. Specifically, an (Ilia) (described above) can be oxidized to form a pyrimidinyl-diketone (FVb), which can undergo reaction with an appropriate substituted (and optionally protected) aldehyde (VI) to yield a compound of formula V (V) as described above.
• an (Ilia) (described above) can be oxidized to form a pyrimidinyl-diketone (FVb), which can undergo reaction with an appropriate substituted (and optionally protected) aldehyde (VI) to yield a compound of formula V (V) as described above.
• a compound of formula V can be prepared by reacting intermediate (IV) or (IVa) with an aldehyde (VII) to yield a further intermediate (VIII), which can then react with compound (IX) to yield a compound of formula V.
• VIII aldehyde
• moieties Y' and Y" are precursors of moiety Yv. See Scheme V-2 below, hi addition, desired substitutions at R v"a can be obtained by selecting, for example, the appropriate compound (Ha) intermediate.
• moiety Xv in compound (VII) is a nitrogen-containing heterocycloalkyl (e.g., piperidine).
• the nitrogen ring atom can be protected by a nitrogen protecting group (e.g., Cbz, Boc, or FMOC) before coupling to compound (IV) or (IVa) and deprotected afterwards (see first step of Scheme 3) to yield compound (Villa).
• This compound can further react with various compounds (IX) to produce a compound of formula V. See second steps of Scheme V-3 below. It should be noted that compound (VIII) or compound (Villa) can be a compound of formula V as well.
• Examples of compounds of formula V include, but are not limited to,
• the antagonists have the structure shown in formula VI:
• each R VI"a can be alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, -NH 2 , -NH(unsubstituted alkyl), -N(unsubstituted alkyl) 2 , nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfinyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarboixyloxy, urea, thiourea,
• R VM can be a bond, alkylene, alkenylene, alkynylene, or -(CH 2 )ri-O-(CH 2 ) r2 -, wherein- each of rl and r2, independently, is 2 or 3.
• R y ⁇ can be cycloalkylene, heterocycloalkylene, cycloalkenylene, heterocycloalkenylene, arylene, heteroaryleae, or a bond.
• R VM can be -C(O)-, -C(O)-O-, -O-C(O)-, -S(O) P -O-, -O-S(O) P -, -C(O)-N(R VI'b )-, -N(R VI"b )-C(O)-, -O-C(O)-N(R VI"b )-, -N(R VI'b )-C(O)-O-, -C(0)-N(R VI"b )-0-, -O-N(R VI" b )-C(O)-, -O-S(O)p-N(R VI"b )-, -N(R VW > S(O) P -O-, -S(O) p -N(R VI - b )-O-, -O-N(R VW >S(O) P -, -N(R VI - -O
• R Vhb and R VI"C independently, can be hydrogen, hydroxy, alkyl, aryl, aralkyl, heterocycloalkyl, heteroaryl, or heteroaralkyl.
• p can be 1 or 2 and q can be 1-4.
• R VM can be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, cycloalkenyl, (cycloalkenyl)alkyl, heterocyclo alkenyl, (heterocycloalkenyl)alkyl, aryl, aralkyl, heteroaryl, or heteroaralkyl.
• R VI"5 can be hydrogen, unsubstituted alkyl, halo-substituted alkyl, alkoxy, alkylsulflnyl, amino, alkenyl, alkynyl, cycloalkoxy, cycloalkylsulfmyl, heterocycloalkoxy, heterocycloalkylsulfinyl, aryloxy, arylsulfinyl, heteroaryloxy, or heteroarylsulfmyl.
• R VI"6 can be a 5- to 6-membered monocyclic heterocyclyl or a 8- to 11-membered bicyclic heteroaryl, and optionally substituted with alkyl, alkenyl, alkynyl, alkoxy, acyl, halo, hydroxy, amino, nitro, oxo, thioxo, cyano, guanadino, amidino, carboxy, sulfo, mercapto, alkylsulfanyl, alkylsulfmyl, alkylsulfonyl, aminocarbonyl, alkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, alkoxycarbonyl, alkylcarbonyloxy, urea, thiourea, sulfamoyl,
• m can be 0-3, and when m ⁇ 2, two adjacent R a groups can optionally together to form a 4- to 8-membered optionally substituted cyclic moiety.
• the intermediate (III) can be further substituted at the 4-position of the pyrazole core ring with a good leaving group such as halo (e.g., iodo or bromo) t>y reacting with a halogenation reagent (e.g., bromination reagent such as Br 2 or iodination reagent such as N-iodosuccinimide) to form a 2-(4-halo-pyrazol-3-yl)-pyrimidine (IV).
• halo e.g., iodo or bromo
• a good leaving group such as halo (e.g., iodo or bromo) t>y reacting with a halogenation reagent (e.g., bromination reagent such as Br 2 or iodination reagent such as N-iodosuccinimide) to form a 2-(4-halo-pyrazol-3-yl)-pyrim
• the iodo substituent can be converted into a boronic acid substituent (see compound (V) below), which can react with a R VI"6 -halide (VI) C ⁇ g-, an aryl halide or a heteroaryl halide) via Suzuki coupling reaction to form a compound of formula VI.
• R VI"6 -halide (VI) C ⁇ g-, an aryl halide or a heteroaryl halide) via Suzuki coupling reaction to form a compound of formula VI.
• Other substitution reactions can also be employed to produce a wide range of compounds of formula VI (see, e.g., via a reaction between the protected iodinated compound. (Wa) and phthalic anhydride to form a di-keto intermediate (VII), which can undergo a cyclization reaction with an R g -substituted hydrazine to form a compound (VI); for reference, see J. Med.
• a compound of formula VI can be prepared according to Scheme VI-3 shown below.
• a dirnethoxymethyl-substituted pyrimidine of formula (IEa) can be prepared by reacting dimethylformamide dimethylacetal with l,l-dimethoxy-propan-2-one to form 4-dimethylamino-l,l-dirnethoxy-but-3-en-2-one as an intermediate, which can further react with an R VI"a -substituted acetamidine (i.e., R VI"a -C(NH)-NH 2 ) to produce a compound of formula (Ha).
• R VI"a -substituted acetamidine i.e., R VI"a -C(NH)-NH 2
• the compound of formula (Ha) can then be deprotected in an acidic medium (e.g., aqueous HBr) and react with aniline and diphenylphosphite to form a compound of formula (lib), which can then react with an R VI"6 -substituted aldehyde to form a compound of formula (lie). Further reaction of a compound of formula (lie) with N,N-dimethylformamide dimettiylacetal (DMFDMA), followed by hydrazine hydrate, yields a compound of formula VI.
• DMFDMA N,N-dimethylformamide dimettiylacetal
• a compound of formula VI can also be prepared via a phenylacetyl pyrimidine compound (IX) as shown in Scheme VI-5 below. Specifically, a pyrimidine- carboxyaldehyde compound (VIII) is converted to the N 5 P acetal intermediate with aniline and diphenylphosphite. This acetal intermediate is then coupled to an aldehyde substituted with R VI"6 in basic condition (e.g., Cs 2 CO 3 ) to afford an enamine intermediate, which is hydrolyzed to a ketone intermediate (DC).
• DC ketone intermediate
• Cyclizing the ketone intermediate (IX) with N 5 N- dimethylformamide dimethyl acetal and hydrazine affords the pyrazole ring of the desired compound of formula VI.
• the pyrazole ring of a compound of formula VI can also be formed by cyclizing the ketone intermediate (IX) with an R VI"5 -substituted carboxylic acid hydrazide (X).
• R VI"5 -substituted carboxylic acid hydrazide (X) For reference, see, e.g., Chemistry of Heterocyclic compounds 35(11): 1319- 1324 (2000).
• a compound of formula VI wherein the 1 -position of the pyrazole core ring is unsubstituted can undergo a conjugate addition reaction as shown in reaction (B) below.
• the electrophile or acceptor in the addition reaction generally contains a double bond connecting to an electron-withdrawing group or a double bond conjugating to groups such as carbonyl, cyano, or r ⁇ itro.
• the -R VI"1 -R VI"2 -R VI"3 -R VI"4 giOup can be further transformed into other functionalities as shown in Scheme VI-8 below.
• a compound of formula VI wherein the -R VM - R VI ⁇ 2 -R VI"3 -R VI"4 group is cyanoalkyl can be reduced to aminoallcyl, which can be further converted to other functionalities such as heteroaralkyl, heterocycloalkylalkyl, and carboxylic acid.
• Substituents at the pyrimidinyl ring can also be converted into other functionalities.
• R VI"a is bromo
• a compound of formula VI wherein R VI"a is bromo can be converted into other functionalities such as alkyl, alkenyl, cycloalkyl and the like, by known methods.
• R VI"6 substituents of the R VI"6 moiety can be further converted into other functionalities as well.
• A-S will be obvious to a skilled person in the art, some starting materials and intermediates may need to be protected before undergoing synthetic steps as described above.
• suitable protecting groups see, e.g., T. W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, Inc., New York (1981).
• Examples of compounds of formula VI include, but are not limited to,
• an effective amount is the amount which is required to confer a therapeutic effect on the treated subject, e.g. a patient.
• an effective amount can range 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, dependent 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.
• the compounds of formulae I, II, III, IV, V, and VI can be administered by any method that permits the delivery of the compounds to combat vascular injuries.
• the compounds of formulae I, II, III, IV, V, and VI can be admininistered via pills, tablets, capsules, aerosols, suppositories, liquid formulations for ingestion or injection or for use as eye or ear drops, dietary supplements, and topical preparations.
• a pharmaceutically acceptable composition includes the aqueous solution of the compound of formula I, II, III, IV, V, or VI, in an isotonic saline, 5% glucose or another 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.
• the compositions can be administered orally, intranasally, transdermally, intradermally, vaginally, intraaurally, intraocularly, buccally, rectally, transmucosally, or via inhalation, or intravenous administration.
• the compositions may be delivered intravenously via a ballon catheter.
• 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, ferret, lizard, reptile, or bird).
• 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, ferret, lizard, reptile, or bird.
• the compounds of formulae I, U, III, IV, V, and VI also can be delivered by implantation (e.g., surgically) with 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 within, a short or over an extended period of time.
• stents Delivery of therapeutic agents via stents, delivery pumps (e.g., mini-osmotic pumps), and other implantable devices is known in the art. See for example, "Recent Developments in Coated Stents” by Hofrna et al., published in Current Interventional Cardiology Reports 2001, 3:28-36, the entire contents of which, including references cited therein, are incorporated herein. Other descriptions of implantable devices, such as stents, can " be found, e.g., in U.S. Patent Nos.
• a delivery device such as stent 10, includes a compound 20 of formula I, II, III, IV, V, or VI.
• the compound, as a therapeutic agent 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 (e.g., a polyester), where the diffusion of the agent is irate ⁇ m ⁇ t ⁇ ngj as ' the agent diffuses through the polymer to the polymer- fluid interface and is released into the surrounding fluid, hi some devices, a soluble substance is also dissolved or dispersed in, or chemically bonded to, 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 or 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 and 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 changing the relative amounts of the faster releasing polymers relative to the slower releasing polymers.
• the proportion of faster releasing polymer can be increased relative to the slower releasing polymer. If most of the dosage is desired to be released over a. long time period, most of the polymer can be the slower releasing polymer.
• 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. In some embodiments., the co ⁇ polymers 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/3 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 ligand binding or activation of the TGFjS receptors, e.g., anti-TGF/3, anti-TGF/3 receptor antibodies, or antagonists of the TGF/? type II receptors.
• Example 1 Balloon Catheter Injury of the Rat Carotid Artery
• Sprague Dawley rats 400g, 3 to 4 months old were anesthetized by i.p. injection with 10 mg/kg xylazine (XyIa- Ject, Phoenix Pharmaceuticals) and 80 mg/kg ketamine (Ketaset, Fort Dodge).
• the left carotid artery and the aorta were denuded with a 2F balloon catheter (Edwards Life Sciences) 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.
• Exsanguination and then perfusion fixation was carried out under physiological pressure with 0.9% sodium chloride, injection USP (310 mOsmol/L, pH 5.6 (4.5-7.0)) and (10% neutral buffered formalin).
• 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 Dunnet's Multiple Comparisons 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.
• the serine-threonine kinase activity of TGF/? type I receptor was measured as th.e autophosphorylation activity of the cytoplasr ⁇ ic domain of the receptor containing an IST- terminal poly histidine, TEV cleavage site-tag, e.g., His-TGF/3RI.
• the His-tagged receptor cytoplasmic kinase domains were purified from infected insect cell cultures using the Gibco- BRL FastBac HTb baculovirus expression system.
• Compounds of formulae I, II, III, IV, V, and VI typically exhibited low IC 50 values of less than 10 ⁇ M; some exhibited IC 50 values of less than 1 ⁇ M; and some even exhibited IC 5O values of less than 50 nM.
• Example 3 Cell-Free Assay for Evaluating Inhibition of Activin Type I Receptor Kinase Activity
• Inhibition of the Activin type I receptor (AIk 4) kinase autophosphorylation activity by compounds of formulae I, II, III, IV, V, and VI, can be determined in a similar manner to that described above in Example 2 except that a similarly His-tagged form of AIk 4 (His- AIk 4) is used in place of the HiS-TGF 1 SRI.
• AIk 4 Activin type I receptor
• Control wells containing either DMSO without any test compound or controL compound in DMSO were used.
• His-TGF ⁇ Type I receptor in the same assay buffer Hepes, NaCl 2 , MgCl 2 , MnCl 2 , DTT, and 30% Brij ® added fresh
• PE nickel coated FlashPlate
• 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 the test compound 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).
• Biological activity of the compounds of formulae I, II, III, IV, V, and VI 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 (Ix).
• the transfected cells were then plated at a concentration of 2.5 x 10 4 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/3 ligand in the starvation media containing 1 % DMSO either in the presence or absence of a test compound of formula I, II, III, IV, V, or VI 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 IC 50 values of the test compounds for inhibiting TGF ⁇ -induced PAI-Luciferase reporter activity in HepG2 cells.
• Compounds of formulae I, II, III, IV, V, and VI typically exhibited IC 50 values of less 10 uM.
• 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, II, III, IV, V, and VI typically exhibited LD 25 values greater than 10 /xM.
• 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, and then expanded at 33°C before being transferred to 37°C at which temperature the large T antigen becomes inactive (see, e.g., 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. The 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°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 ⁇ complete DMEM containing 2 % fetal calf serum, but without phenol red.
• the cells are incubated at 37°C for 3 to 4 h-ours to allow them to adhere to the plate.
• Solutions containing a test compound of formula I, II, III, IV, V, or VI are then added to wells with no TGFjS (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 ran following excitation at 485 urn is measured 48 hours after the addition of solutions containing a test compound on a CytoFluor microplate reader (PerSeptive Biosystems). The data are expressed as the ratio of TGF ⁇ -induced to non-induced for each test sample.

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