Fibroblast Activation Protein Inhibitor Compounds and Methods

Abstract

Amino terminus-blocked peptide boronate compounds of Formulas I and II are useful for inhibiting Fibroblast Activation Protein (FAP) and other proteases, and for treating disorders mediated by FAP. Methods of using the amino terminus blocked peptide boronate compounds, and stereoisomers, tautomers, solvates and pharmaceutically acceptable salts thereof, for in vitro, in situ, and in vivo diagnosis, prevention or treatment of such disorders in mammalian cells, or associated pathological conditions are disclosed.

1 . A compound selected from Formula I: 
       
         
           
           
               
               
           
         
         and stereoisomers, tautomers, solvates and pharmaceutically acceptable salts thereof, wherein: 
         X is C(═O), C(═NR), NRC(═O), NRC(═NR), OC(═O), OC(═NR), P(O)(OR), S(O), and S(O) 2 ; 
         R 1  is selected from H, C 1 -C 8  alkyl, C 2 -C 8  alkenyl, C 2 -C 8  alkynyl, C 2 -C 20  heterocycle, C 3 -C 12  carbocycle, and C 6 -C 20  aryl; 
         R 2  is selected from H, C 1 -C 8  alkyl, C 2 -C 8  alkenyl, C 2 -C 8  alkynyl, C 2 -C 20  heterocycle, C 3 -C 12  carbocycle, and C 6 -C 20  aryl; 
         or R 1  and R 2  form a C 2 -C 20  heterocycle; 
         R 3  is an optionally protected amino acid side chain; 
         R 4  and R 5  are independently selected from H, C 1 -C 8  alkyl, C 2 -C 8  alkenyl, C 2 -C 8  alkynyl, C 2 -C 20  heterocycle, C 3 -C 12  carbocycle, C 6 -C 20  aryl, a prodrug, and a protecting group; or R 4  and R 5  together form a C 6 -C 20  aryl, a C 3 -C 12  carbocycle, a prodrug, or a protecting group; 
         R 6 , R 7 , R 8 , R 9 , R 10 , R 11  and R 12  are independently selected from F, Cl, Br, I, OH, OR, R, —C(═Y)R, —C(═Y)OR, —C(═Y)N(R) 2 , —N(R) 2 , —N + (R) 3 , —N(R)C(═Y)R, —N(R)C(═Y)OR, —N(R)C(═Y)N(R) 2 , —SR, —OC(═Y)R, —OC(═Y)OR, —OC(═Y)(N(R) 2 ), —OS(O) 2 (OR), —OP(═Y)(OR) 2 , —OP(OR) 2 , —P(═Y)(OR) 2 , —P(═Y)(OR)NR 2 , —S(O)R, —S(O) 2 R, —S(O) 2 NR, —S(O)(OR), —S(O) 2 (OR), —SC(═Y)R, —SC(═Y)OR, and —SC(═Y)NR 2 ; 
         each alkyl, alkenyl, alkynyl, aryl, carbocycle, and heterocycle is optionally and independently substituted with one or more substituents selected from F, Cl, Br, I, OH, OR, R, ═O, ═S, ═NR, ═N + (O)(R), ═N(OR), ═N + (O)(OR), ═N—N(R) 2 , —C(═Y)R, —C(═Y)OR, —C(═Y)N(R) 2 , —N(R) 2 , —N + (R) 3 , —N(R)C(═Y)R, —N(R)C(═Y)OR, —N(R)C(═Y)N(R) 2 , —SR, —OC(═Y)R, —OC(═Y)OR, —OC(═Y)(N(R) 2 ), —O S(O) 2 (OR), —OP(═Y)(OR) 2 , —OP(OR) 2 , —P(═Y)(OR) 2 , —P(═Y)(OR)NR 2 , —S(O)R, —S(O) 2 R, —S(O) 2 NR, —S(O)(OR), —S(O) 2 (OR), —SC(═Y)R, —SC(═Y)OR, and —SC(═Y)NR 2 ; 
         R is H, C 1 -C 8  alkyl, C 2 -C 8  alkenyl, C 2 -C 8  alkynyl, C 6 -C 20  aryl, C 1 -C 20  heterocyclyl, or a protecting group; and 
         Y is independently O, S, NR, N + (O)(R), N(OR), N + (O)(OR), or N—N(R) 2 ; 
         with the provisos that when R 1  is methyl and X is C(═O), then R 3  is not lysine or acetyl-lysine; and that when R is tert-butyl and X is OC(═O), then R 3  is not methyl. 
       
     
     
         2 . The compound of  claim 1  wherein R 1  is selected from methyl, isobutyl, tert-butyl, phenyl, and 2,4,6-trimethylphenyl; and X is C(═O). 
     
     
         3 . The compound of  claim 1  wherein R 1  is C 1 -C 8  alkyl or C 6 -C 20  aryl; and X is S(O) 2 . 
     
     
         4 . The compound of  claim 1  wherein R 2  is H. 
     
     
         5 . The compound of  claim 1  wherein R 2  is methyl. 
     
     
         6 . The compound of  claim 1  wherein R 3  is selected from hydrogen, methyl, isopropyl, n-propyl, isobutyl, sec-butyl, n-butyl, benzyl, p-hydroxybenzyl, —CH 2 OH, —CH(OH)CH 3 , —CH 2 CH 2 SCH 3 , —CH 2 CONH 2 , —CH 2 COOH, —CH 2 CH 2 CONH 2 , —CH 2 CH 2 COOH, —(CH 2 ) 3 NHC(═NH)NH 2 , —(CH 2 ) 3 NH 2 , —(CH 2 ) 3 NHCOCH 3 , —(CH 2 ) 3 NHCHO, —(CH 2 ) 4 NHC(═NH)NH 2 , —(CH 2 ) 4 NH 2 , —(CH 2 ) 4 NHCOCH 3 , —(CH 2 ) 4 NHCHO, —(CH 2 ) 3 NHCONH 2 , —(CH 2 ) 4 NHCONH 2 , —CH 2 CH 2 CH(OH)CH 2 NH 2 , 2-pyridylmethyl-, 3-pyridylmethyl-, 4-pyridylmethyl-, phenyl, cyclohexyl, and the structures: 
       
         
           
           
               
               
           
         
       
     
     
         7 . The compound of  claim 1  wherein R 3  is hydrogen. 
     
     
         8 . The compound of  claim 1  wherein R 4  and R 5  are each hydrogen. 
     
     
         9 . The compound of  claim 1  wherein R 4  and R 5  together form a cyclic boronate ester. 
     
     
         10 . The compound of  claim 9  wherein the cyclic boronate ester is selected from pinanediol, pinacol, and catechol. 
     
     
         11 . The compound of  claim 1  wherein the carbon atom bearing R 3  is in the R configuration. 
     
     
         12 . The compound of  claim 1  wherein the carbon atom bearing R 3  is in the S configuration. 
     
     
         13 . The compound of  claim 1  wherein the carbon atom bearing the boron atom is in the R configuration. 
     
     
         14 . The compound of  claim 1  wherein the carbon atom bearing the boron atom is in the S configuration. 
     
     
         15 . The compound of  claim 1  wherein R is a protecting group selected from a trialkylsilyl, a dialkylphenylsilyl, benzoate, benzyl, benzyloxymethyl, methyl, methoxymethyl, a triarylmethyl, phthalimido and tetrahydropyranyl. 
     
     
         16 . The compound of  claim 1  wherein R 1  is phenyl optionally substituted with one or more substituents selected from F, Cl, Br, I, OH, OR, R, —C(═Y)R, —C(═Y)OR, —C(═Y)NR 2 , —NR 2 , —N + (R) 3 , —N(R)C(═Y)R, —N(R)C(═Y)OR, —N(R)C(═Y)NR 2 , —SR, —OC(═Y)R, —OC(═Y)OR, —OC(═Y)NR 2 , —OS(O) 2 (OR), —OP(═Y)(OR) 2 , —OP(OR) 2 , —P(═Y)(OR) 2 , P(═Y)(OR)NR 2 , —S(O)R, —S(O) 2 R, —S(O) 2 NR, —S(O)(OR), —S(O) 2 (OR), —SC(═Y)R, —SC(═Y)OR, and —SC(═Y)NR 2 . 
     
     
         17 . The compound of  claim 1  wherein R 1  is heterocyclyl selected from 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-imidazolyl, 4-imidazolyl, 3-pyrazolyl, 4-pyrazolyl, 2-pyrrolyl, 3-pyrrolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl, 2-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, and substituted forms thereof. 
     
     
         18 . The compound of  claim 1  wherein X is C(═O) and R 12  is H, and the compound is selected from Formula Ia: 
       
         
           
           
               
               
           
         
       
     
     
         19 . The compound of  claim 18  wherein R 6 , R 7 , R 8 , R 9 , R 10  and R 11  are each H and the compound is selected from Formula Ib: 
       
         
           
           
               
               
           
         
       
     
     
         20 . The compound of  claim 19  selected from the structures: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         21 . The compound of  claim 18  selected from the structures: 
       
         
           
           
               
               
           
         
       
     
     
         22 . The compound of  claim 19  selected from the structures: 
       
         
           
           
               
               
           
         
       
     
     
         23 . The compound of  claim 19  having the structure: 
       
         
           
           
               
               
           
         
       
     
     
         24 . The compound of  claim 1  wherein X is S(O) 2  and R 12  is H, and the compound is selected from Formula Ic: 
       
         
           
           
               
               
           
         
       
     
     
         25 . The compound of  claim 24  selected from the structures: 
       
         
           
           
               
               
           
         
       
     
     
         26 . A compound selected from Formula II: 
       
         
           
           
               
               
           
         
         and stereoisomers, tautomers, solvates and pharmaceutically acceptable salts thereof, wherein: 
         Z is 
       
       
         
           
           
               
               
           
         
         R 1  is selected from H, C 1 -C 8  alkyl, C 2 -C 8  alkenyl, C 2 -C 8  alkynyl, C 2 -C 20  heterocycle, C 3 -C 12  carbocycle, and C 6 -C 20  aryl; 
         each R 2  is independently selected from H, C 1 -C 8  alkyl, C 2 -C 8  alkenyl, C 2 -C 8  alkynyl, C 2 -C 20  heterocycle, C 3 -C 12  carbocycle, and C 6 -C 20  aryl; 
         or R 1  and R 2  form a C 2 -C 20  heterocycle; 
         each R 3  is independently an optionally protected amino acid side chain; 
         R 4  and R 5  are independently selected from H, C 1 -C 8  alkyl, C 2 -C 8  alkenyl, C 2 -C 8  alkynyl, C 2 -C 20  heterocycle, C 3 -C 12  carbocycle, C 6 -C 20  aryl, a prodrug, and a protecting group; or R 4  and R 5  together form a C 6 -C 20  aryl, a C 3 -C 12  carbocycle, a prodrug, or a protecting group; 
         R 6 , R 7 , R 8 , R 9 , R 10 , R 11  and R 12  are independently selected from F, Cl, Br, I, OH, OR, R, —C(═Y)R, —C(═Y)OR, —C(═Y)N(R) 2 , —N(R) 2 , —N + (R) 3 , —N(R)C(═Y)R, —N(R)C(═Y)OR, —N(R)C(═Y)N(R) 2 , —SR, —OC(═Y)R, —OC(═Y)OR, —OC(═Y)(N(R) 2 ), —OS(O) 2 (OR), —OP(═Y)(OR) 2 , —OP(OR) 2 , —P(═Y)(OR) 2 , —P(═Y)(OR)NR 2 , —S(O)R, —S(O) 2 R, —S(O) 2 NR, —S(O)(OR), —S(O) 2 (OR), —SC(═Y)R, —SC(═Y)OR, and —SC(═Y)NR 2 ; 
         R 13  is independently selected from C 1 -C 8  alkyl, C 1 -C 8  alkenyl, C 1 -C 8  alkynyl, C 6 -C 20  aryl, C 1 -C 20  heterocyclyl, or a protecting group; 
         each alkyl, alkenyl, alkynyl, aryl, carbocycle, and heterocycle is optionally and independently substituted with one or more substituents selected from F, Cl, Br, I, OH, OR, R, ═O, ═S, ═NR, ═N + (O)(R), ═N(OR), ═N + (O)(OR), ═N—N(R) 2 , —C(═Y)R, —C(═Y)OR, —C(═Y)N(R) 2 , —N(R) 2 , —N + (R) 3 , —N(R)C(═Y)R, —N(R)C(═Y)OR, —N(R)C(═Y)N(R) 2 , —SR, —OC(═Y)R, —OC(═Y)OR, —OC(═Y)(N(R) 2 ), —O S(O) 2 (OR), —OP(═Y)(OR) 2 , —OP(OR) 2 , —P(═Y)(OR) 2 , —P(═Y)(OR)NR 2 , —S(O)R, —S(O) 2 R, —S(O) 2 NR, —S(O)(OR), —S(O) 2 (OR), —SC(═Y)R, —SC(═Y)OR, and —SC(═Y)NR 2 ; 
         R is H, C 1 -C 8  alkyl, C 2 -C 8  alkenyl, C 2 -C 8  alkynyl, C 6 -C 20  aryl, C 1 -C 20  heterocyclyl, or a protecting group; 
         Y is independently O, S, NR, N + (O)(R), N(OR), N + (O)(OR), or N—N(R) 2 ; and 
         n is 1, 2, 3, 4, 5, 6, 7, or 8. 
       
     
     
         27 . The compound of  claim 26  selected from Formula IIa: 
       
         
           
           
               
               
           
         
       
     
     
         28 . The compound of  claim 27  wherein R 1  is C 1 -C 8  alkyl or C 6 -C 20  aryl. 
     
     
         29 . The compound of  claim 27  wherein R 2  is H. 
     
     
         30 . The compound of  claim 27  wherein at least one of R 2  is methyl. 
     
     
         31 . The compound of  claim 27  wherein R 4  and R 5  are each H. 
     
     
         32 . The compound of  claim 31  selected from the structures: 
       
         
           
           
               
               
           
         
       
     
     
         33 . The compound of  claim 26  selected from Formula IIb: 
       
         
           
           
               
               
           
         
       
     
     
         34 . The compound of  claim 33  wherein R 13  is C 1 -C 8  alkyl or C 6 -C 20  aryl. 
     
     
         35 . The compound of  claim 33  wherein each R 2  is H. 
     
     
         36 . The compound of  claim 33  wherein R 4  and R 5  are each H. 
     
     
         37 . A pharmaceutical composition comprising a compound according to  claim 1  or  claim 26 , and one or more pharmaceutically acceptable carriers or excipients. 
     
     
         38 . The composition according to  claim 37 , additionally comprising an additional therapeutic agent selected from an anti-proliferative agent, an anti-inflammatory agent, an immunomodulatory agent, a neurotrophic factor, an agent for treating cardiovascular disease, an agent for treating liver disease, an anti-viral agent, an agent for treating blood disorders, an agent for treating diabetes, or an agent for treating immunodeficiency disorders. 
     
     
         39 . A composition comprising the compound according to  claim 1  or  claim 26  in an amount to detectably inhibit FAP activity, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. 
     
     
         40 . A composition comprising the compound according to  claim 1  or  claim 26  in an amount to detectably inhibit POP activity, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. 
     
     
         41 . A method of treating or lessening the severity of a disease or condition selected from the group consisting of cancer, stroke, diabetes, hepatomegaly, cardiovascular disease, Alzheimer's disease, cystic fibrosis, viral disease, autoimmune diseases, atherosclerosis, restenosis, psoriasis, allergic disorders, inflammation, neurological disorders, a hormone-related disease, conditions associated with organ transplantation, immunodeficiency disorders, destructive bone disorders, proliferative disorders, infectious diseases, conditions associated with cell death, thrombin-induced platelet aggregation, chronic myelogenous leukemia (CML), liver disease, pathologic immune conditions involving T cell activation, and CNS disorders in a patient, comprising the step of administering to said patient a composition according to  claim 37 . 
     
     
         42 . A method of treating or preventing cancer in a mammal in need of such treatment which is comprised of administering to said mammal a therapeutically effective amount of a composition according to  claim 37 . 
     
     
         43 . The method of  claim 41  wherein the cancer is selected from breast, ovary, cervix, prostate, testis, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, non-small cell lung carcinoma (NSCLC), small cell carcinoma, lung adenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidney carcinoma, myeloid disorders, lymphoid disorders, hairy cells, buccal cavity and pharynx (oral), lip, tongue, mouth, pharynx, small intestine, colon-rectum, large intestine, rectum, brain and central nervous system, Hodgkin's and leukemia. 
     
     
         44 . Use of a compound according to  claim 1  or  claim 26  in the preparation of a pharmaceutical composition for the treatment of cancer. 
     
     
         45 . A method of treatment of cancer, comprising administering a pharmaceutical composition according to  claim 37  to a patient. 
     
     
         46 . Use of a compound according to  claim 1  or  claim 26  for the manufacture of a medicament for the treatment of cancer. 
     
     
         47 . A kit for treating a hyperproliferative disorder, comprising:
 a) a container holding a compound of  claim 1  or  claim 26 ; and   b) a label or package insert with instructions for use.   
     
     
         48 . The kit of  claim 47  further comprising a second pharmaceutical composition, wherein the second pharmaceutical composition comprises a second compound having anti-hyperproliferative activity.