Process for making modulators of cystic fibrosis transmembrane conductance regulator

Abstract

The invention provides a process for the preparation of a compound of Formula 1, comprising coupling a carboxylic acid of Formula 2 with an aniline of Formula 3 in the presence of a coupling agent.

1 . A process for the preparation of a compound of Formula 1, 
       
         
           
           
               
               
           
         
       
       comprising coupling a carboxylic acid of Formula 2 
       
         
           
           
               
               
           
         
       
       with an aniline of Formula 3 
       
         
           
           
               
               
           
         
       
       in the presence of a coupling agent selected from the group consisting of 2-chloro-1,3-dimethyl-2-imidazolium tetrafluoroborate, HBTU, HCTU, 2-chloro-4,6-dimethoxy-1,3,5-triazine, HATU, HOBT/EDC, and T3P®; wherein
 each R 2  and R 4  is independently selected from hydrogen, CN, CF 3 , halo, C 1-6  straight or branched alkyl, 3-12 membered cycloaliphatic, phenyl, C 5-10  heteroaryl or C 3-7  heterocyclic, wherein said heteroaryl or heterocyclic has up to 3 heteroatoms selected from O, S, or N, and each C 1-6  straight or branched alkyl, 3-12 membered cycloaliphatic, phenyl, C 5-10  heteroaryl or C 3-7  heterocyclic is independently and optionally substituted with up to three substituents selected from —OR′, —CF 3 , —OCF 3 , SR′, S(O)R′, SO 2 R′, —SCF 3 , halo, CN, —COOR′, —COR—, —O(CH 2 ) 2 N(R′)(R′), —O(CH 2 )N(R′)(R′), —CON(R′)(R′), —(CH 2 ) 2 OR′, —(CH 2 )OR′, CH 2 CN, optionally substituted phenyl or phenoxy, —N(R′)(R′), —NR′C(O)OR′, —NR′C(O)R′, —(CH 2 ) 2 N(R′)(R′), or —(CH 2 )N(R′)(R′); 
 each R 5  is independently selected from hydrogen, —OH, NH 2 , CN, CHF 2 , NHR′, N(R′) 2 , —NHC(O)R′, NHC(O)OR′, NHSO 2 R′, —OR′, OC(O)OR′, OC(O)NHR′, OC(O)NR′ 2 , CH 2 OH, CH 2 N(R′) 2 , C(O)OR′, SO 2 NHR′, SO 2 N(R′) 2 , or CH 2 NHC(O)OR′, or 
 R 4  and R 5  are taken together form a 5-7 membered ring containing 0-3 three heteroatoms selected from N, O, or S, wherein said ring is optionally substituted with up to three R 3  substituents; 
 each X is independently a bond or is an optionally substituted C 1-6  alkylidene chain wherein up to two methylene units of X are optionally and independently replaced by —CO—, —CS—, —COCO—, —CONR′—, —CONR′NR′—, —CO 2 —, —COO—, —NR′CO 2 —, —O—, —NR′CONR′—, —OCONR′—, —NR′NR′, —NR′NR′CO—, —NR′CO—, —S—, —SO, —SO 2 —, —NR′—, —SO 2 NR′—, NR′SO 2 —, or —NR′SO 2 NR′—; 
 each R X  is independently R′, halo, NO 2 , CN, CF 3 , or OCF 3 ; 
 y is an integer from 0-4; 
 each R′ is independently selected from hydrogen or an optionally substituted group selected from a C 1-8  aliphatic group, a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or two occurrences of R′ are taken together with the atom(s) to which they are bound to form an optionally substituted 3-12 membered saturated, partially unsaturated, or fully unsaturated monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from N, O, or S; and 
 each R 3  is independently —C 1-3  alkyl, C 1-3  perhaloalkyl, —O(C 1-3  alkyl), —CF 3 , —OCF 3 , —SCF 3 , —F, —Cl, —Br, or —COOR′, —COR′, —O(CH 2 ) 2 N(R′)(R′), —O(CH 2 )N(R′)(R′), —CON(R′)(R′), —(CH 2 ) 2 OR′, —(CH 2 )OR′, optionally substituted monocyclic or bicyclic aromatic ring, optionally substituted arylsulfone, optionally substituted 5-membered heteroaryl ring, —N(R′)(R′), —(CH 2 ) 2 N(R′)(R′), or —(CH 2 )N(R′)(R′). 
 
     
     
         2 . The process of  claim 1 , wherein R 5  is independently —OC(O)OR′, —OC(O)NHR′, or
 —OC(O)N(R) 2 , wherein R′ is not hydrogen. 
 
     
     
         3 . The process of  claim 2 , further comprising cleaving the —OC(O)OR′, —OC(O)NHR′, or —OC(O)N(R′) 2  to form —OH. 
     
     
         4 . The process of  claim 3 , wherein the cleavage is performed by treating a compound of Formula 1 with an alcoholic solvent in the presence of NaOH, KOH or sodium methoxide. 
     
     
         5 . The process of  claim 4 , wherein the alcoholic solvent is methanol. 
     
     
         6 . The process of  claim 1 , wherein at least one of R 4  or R 2  is independently a C 1-6  straight or branched alkyl which is substituted with —COOR′ or —CON(R′) 2 , wherein R′ is not hydrogen. 
     
     
         7 . The process of  claim 6 , further comprising hydrolyzing each —COOR′, or —CON(R′) 2  to form —COOH. 
     
     
         8 . The process of  claim 7 , wherein the hydrolysis is performed by treating a compound of Formula 1 with an alcoholic solvent in the presence of NaOH, KOH or sodium methoxide. 
     
     
         9 . The process of  claim 8 , wherein the alcoholic solvent is methanol. 
     
     
         10 . The process of  claim 6 , wherein R 5  is independently —OC(O)OR′, —OC(O)NHR′, or —OC(O)N(R′) 2 , wherein R′ is not hydrogen. 
     
     
         11 . The process of  claim 10 , further comprising cleaving the —OC(O)OR′, —OC(O)NHR′, or —OC(O)N(R′) 2  to form —OH. 
     
     
         12 . The process of  claim 11 , wherein the cleavage is performed by treating a compound of Formula 1 with an alcoholic solvent in the presence of NaOH, KOH or sodium methoxide. 
     
     
         13 . The process of  claim 12 , wherein the alcoholic solvent is methanol. 
     
     
         14 . The process of  claim 1 , wherein the coupling is performed in the presence of a base. 
     
     
         15 . The process of  claim 14 , wherein the base is K 2 CO 3 , Et 3 N, NMM, pyridine or DIEA. 
     
     
         16 . The process of  claim 1 , wherein the coupling is performed in the presence of a solvent. 
     
     
         17 . The process of  claim 16 , wherein the solvent is acetonitrile. 
     
     
         18 . The process of  claim 16 , wherein the solvent is DMF. 
     
     
         19 . The process of  claim 16 , wherein the solvent is 2-methyltetrahydrofuran. 
     
     
         20 . The process of  claim 1 , wherein the coupling is performed at a reaction temperature which is maintained between about 10° C. and 78° C. 
     
     
         21 . The process of  claim 20 , wherein the coupling is performed at a reaction temperature which is maintained between about 20° C. and 30° C. 
     
     
         22 . The process of  claim 20 , wherein the coupling is performed at a reaction temperature which is maintained between about 40° C. and 50° C. 
     
     
         23 . The process of  claim 20 , wherein the coupling is performed at a reaction temperature which is maintained between about 42° C. and 53° C. 
     
     
         24 . The process of  claim 1 , wherein the coupling reaction is stirred for at least 2 hours. 
     
     
         25 . The process of  claim 24 , wherein the coupling reaction is stirred for at least 70 hours. 
     
     
         26 . The process of  claim 24 , wherein the coupling reaction is stirred for at least 3 days. 
     
     
         27 . The process of  claim 1 , wherein y is 0. 
     
     
         28 . The process of  claim 1 , wherein R 2  is tert-butyl. 
     
     
         29 . The process according to  claim 1 , further comprising the step of contacting a compound of Formula 4 
       
         
           
           
               
               
           
         
       
       with an aqueous acid to produce a compound of Formula 2. 
     
     
         30 . The process of  claim 1 , wherein the aniline of Formula 3 is a compound of Formula 40 
       
         
           
           
               
               
           
         
       
     
     
         31 . The process of  claim 30  further comprising the step of contacting a compound of Formula 41 
       
         
           
           
               
               
           
         
       
       with methyl trimethylsilyl dimethylketene acetal (MTDA) 
       
         
           
           
               
               
           
         
       
       to produce a compound of Formula 42 
       
         
           
           
               
               
           
         
       
     
     
         32 . The process of  claim 31 , further comprising the step of reducing a compound of Formula 42 to produce a compound of Formula 40. 
     
     
         33 . The process of  claim 1 , wherein the aniline of Formula 3 is an aniline of Formula 43 
       
         
           
           
               
               
           
         
       
     
     
         34 . The process of  claim 33 , comprising the step of contacting a compound of Formula 44 
       
         
           
           
               
               
           
         
       
       with methyl trimethylsilyl dimethylketene acetal (MTDA) 
       
         
           
           
               
               
           
         
       
       to produce a compound of Formula 45 
       
         
           
           
               
               
           
         
       
     
     
         35 . The process of  claim 34  further comprising the step of reducing a compound of Formula 45 to produce an aniline of Formula 43. 
     
     
         36 . A process for the preparation of a compound of Formula 2 
       
         
           
           
               
               
           
         
       
       comprising contacting a compound of Formula 4 
       
         
           
           
               
               
           
         
       
       with an aqueous acid, wherein
 each X is independently a bond or is an optionally substituted C 1-6  alkylidene chain wherein up to two methylene units of X are optionally and independently replaced by —CO—, —CS—, —COCO—, —CONR′—, —CONR′NR′—, —CO 2 —, —OCO—, —NR′CO 2 —, —O—, —NR′CONR′—, —NR′NR′, —NR′NR′CO—, —NR′CO—, —S—, —SO, —SO 2 —, —NR′—, —SO 2 NR′—, NR′SO 2 —, or —NR′SO 2 NR′—; 
 each R X  is independently R′, halo, NO 2 , CN, CF 3 , or OCF 3 ; 
 y is an integer from 0-4; and 
 each R′ is independently selected from hydrogen or an optionally substituted group selected from a C 1-8  aliphatic group, a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or two occurrences of R′ are taken together with the atom(s) to which they are bound to form an optionally substituted 3-12 membered saturated, partially unsaturated, or fully unsaturated monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from N, O, or S. 
 
     
     
         37 . A process for the preparation of a compound of Formula 40 
       
         
           
           
               
               
           
         
       
       comprising the step of contacting a compound of Formula 41 
       
         
           
           
               
               
           
         
       
       with methyl trimethylsilyl dimethylketene acetal (MTDA) 
       
         
           
           
               
               
           
         
       
       to produce a compound of Formula 42 
       
         
           
           
               
               
           
         
       
       wherein
 each R 2  is independently selected from hydrogen, CN, CF 3 , halo, C 1-6  straight or branched alkyl, 3-12 membered cycloaliphatic, phenyl, C 5-10  heteroaryl or C 3-7  heterocyclic, wherein said heteroaryl or heterocyclic has up to 3 heteroatoms selected from O, S, or N, and each C 1-6  straight or branched alkyl, 3-12 membered cycloaliphatic, phenyl, C 5-10  heteroaryl or C 3-7  heterocyclic is independently and optionally substituted with up to three substituents selected from —OR′, —CF 3 , —OCF 3 , SR′, S(O)R′, SO 2 R′, —SCF 3 , halo, CN, —COOR′, —COR—, —O(CH 2 ) 2 N(R′)(R′), —O(CH 2 )N(R′)(R′), —CON(R′)(R′), —(CH 2 ) 2 OR′, —(CH 2 )OR′, CH 2 CN, optionally substituted phenyl or phenoxy, —N(R′)(R′), —NR′C(O)OR′, —NR′C(O)R′, —(CH 2 ) 2 N(R′)(R′), or —(CH 2 )N(R′)(R′); 
 each R 5  is independently selected from hydrogen, —OH, NH 2 , CN, CHF 2 , NHR′, N(R′) 2 , —NHC(O)R′, NHC(O)OR′, NHSO 2 R′, —OR′, OC(O)OR′, OC(O)NHR′, OC(O)NR′ 2 , CH 2 OH, CH 2 N(R′) 2 , C(O)OR′, SO 2 NHR′, SO 2 N(R′) 2 , or CH 2 NHC(O)OR′; and 
 each R′ is independently selected from hydrogen or an optionally substituted group selected from a C 1-8  aliphatic group, a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or two occurrences of R′ are taken together with the atom(s) to which they are bound to form an optionally substituted 3-12 membered saturated, partially unsaturated, or fully unsaturated monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from N, O, or S. 
 
     
     
         38 . The process of  claim 37  further comprising the step of reducing a compound of Formula 42 to produce a compound of Formula 40. 
     
     
         39 . A process for the preparation of an aniline of Formula 43 
       
         
           
           
               
               
           
         
       
       comprising the step of contacting a compound having the Formula 44 
       
         
           
           
               
               
           
         
       
       with methyl trimethylsilyl dimethylketene acetal (MTDA) 
       
         
           
           
               
               
           
         
       
       to produce a compound of Formula 45 
       
         
           
           
               
               
           
         
       
       wherein
 each R 2  is independently selected from hydrogen, CN, CF 3 , halo, C 1-6  straight or branched alkyl, 3-12 membered cycloaliphatic, phenyl, C 5-10  heteroaryl or C 3-7  heterocyclic, wherein said heteroaryl or heterocyclic has up to 3 heteroatoms selected from O, S, or N, and each C 1-6  straight or branched alkyl, 3-12 membered cycloaliphatic, phenyl, C 5-10  heteroaryl or C 3-7  heterocyclic is independently and optionally substituted with up to three substituents selected from —OR′, —CF 3 , —OCF 3 , SR′, S(O)R′, SO 2 R′, —SCF 3 , halo, CN, —COOR′, —COR—, —O(CH 2 ) 2 N(R′)(R′), —O(CH 2 )N(R′)(R′), —CON(R′)(R′), —(CH 2 ) 2 OR′, —(CH 2 )OR′, CH 2 CN, optionally substituted phenyl or phenoxy, —N(R′)(R′), —NR′C(O)OR′, —NR′C(O)R′, —(CH 2 ) 2 N(R′)(R′), or —(CH 2 )N(R′)(R′); and 
 each R′ is independently selected from hydrogen or an optionally substituted group selected from a C 1-8  aliphatic group, a 3-8-membered saturated, partially unsaturated, or fully unsaturated monocyclic ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an 8-12 membered saturated, partially unsaturated, or fully unsaturated bicyclic ring system having 0-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or two occurrences of R′ are taken together with the atom(s) to which they are bound to form an optionally substituted 3-12 membered saturated, partially unsaturated, or fully unsaturated monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from N, O, or S. 
 
     
     
         40 . The process of  claim 39  further comprising the step of reducing a compound of Formula 45 to produce an aniline of Formula 43. 
     
     
         41 . A process for the preparation of a compound 34 
       
         
           
           
               
               
           
         
       
       comprising:
 (a) reacting compound 26 
 
       
         
           
           
               
               
           
         
       
       with compound 32 
       
         
           
           
               
               
           
         
       
       in the presence of T3P® and pyridine using 2-methyl tetrahydrofuran as the solvent, wherein the reaction temperature is maintained between about 42° C. and 53° C., and wherein the reaction is allowed proceed for at least 2 hours, to produce compound 33 
       
         
           
           
               
               
           
         
       
       and
 (b) treating compound 33 with NaOMe/MeOH in 2-methyl tetrahydrofuran. 
 
     
     
         42 . The process of  claim 41 , wherein the reaction is allowed to proceed for at least 6 hours. 
     
     
         43 . The process of  claim 41 , further comprising forming a slurry of compound 34 in a mixture of acetonitrile and water. 
     
     
         44 . The process of  claim 43 , wherein the ratio of acetonitrile to water is about 9:1. 
     
     
         45 . The process of  claim 43 , wherein the slurry is heated to a temperature between about 73° C. and 83° C. 
     
     
         46 . The process of  claim 43 , wherein compound 34 is in the slurry for at least about 3 hours. 
     
     
         47 . The process of  claim 43 , further comprising forming a slurry of compound 34 in Isopropyl acetate. 
     
     
         48 . The process of  claim 46 , wherein the slurry is heated to reflux temperature. 
     
     
         49 . The process of  claim 41 , further comprising dissolving compound 34 in a biphasic solution of 2-methyltetrahydrofuran and 0.1N HCl; stirring said biphasic solution; separating the organic phase from said biphasic solution; filtering and removing solid matter from said organic phase; reducing the volume of said organic phase by approximately 50% using distillation; performing thrice the procedure of: adding MeOAc, EtOAc, IPAc, t-BuOAc, tetrahydrofuran (THF), Et 2 O or methyl-t-butyl ether (MTBE) to the organic phase until the volume of said organic phase increases by 100% and reducing the volume of the organic phase by 50% using distillation; adding MeOAc, EtOAc, IPAc, t-BuOAc, tetrahydrofuran (THF), Et 2 O or methyl-t-butyl ether (MTBE) to the organic phase until the volume of said organic phase increases by 100%; heating the organic phase to reflux temperature, and maintaining said reflux temperature for a time at least about 5 hours; and cooling the organic phase to a temperature between about
 −5° C. and 5° C. over a time period of about 4.5 hours to 5.5 hours.   
     
     
         50 . The process of  claim 41 , further comprising quenching the reaction mixture with 1.2 N HCl; thereby creating a biphasic mixture; agitating said biphasic mixture; separating the organic phase from said biphasic mixture; adding 0.1N HCl to the organic layer thereby creating a biphasic mixture; agitating said biphasic mixture; separating the organic phase; filtering and removing solid matter from said organic phase; reducing the volume of the organic phase by approximately 50% using distillation; performing thrice the steps of: adding acetonitrile to the organic phase until the volume of said organic phase increases by 100% and reducing the volume of the organic phase by approximately 50%; increasing the volume of the organic phase by approximately 100% by adding acetonitrile and then adding water, to form a slurry wherein the final solvent ratio is 9:1 acetonitrile/water; heating said slurry to a temperature between about 73° C. and 83° C.; stirring said slurry for at least 5 hours; and cooling said slurry to a temperature between about 20° C. and 25° C.; filtering and removing solid matter from said slurry; washing the solid matter with acetonitrile having a temperature of between about 20° C. and 25° C. four times; and drying the solid material under vacuum at a temperature of from about 45° C. to about 55° C. 
     
     
         51 . A compound produced by the process of  claim 1 . 
     
     
         52 . A pharmaceutical composition comprising a compound produced by the process of  claim 1 . 
     
     
         53 . A method of modulating CFTR activity in a biological sample comprising the step of contacting said biological sample with a compound produced by the process of  claim 1 . 
     
     
         54 . A method of treating or lessening the severity of a disease in a patient comprising administering to said patient an effective amount of a compound produced by the process of  claim 1 , wherein said disease is selected from cystic fibrosis, asthma, smoke induced COPD, chronic bronchitis, rhinosinusitis, constipation, pancreatitis, pancreatic insufficiency, male infertility caused by congenital bilateral absence of the vas deferens (CBAVD), mild pulmonary disease, idiopathic pancreatitis, allergic bronchopulmonary aspergillosis (ABPA), liver disease, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, such as protein C deficiency, Type 1 hereditary angioedema, lipid processing deficiencies, such as familial hypercholesterolemia, Type 1 chylomicronemia, abetalipoproteinemia, lysosomal storage diseases, such as I-cell disease/pseudo-Hurler, mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II, polyendocrinopathy/hyperinsulinemia, Diabetes mellitus, Laron dwarfism, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, glycanosis CDG type 1, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), neurohypophyseal DI, nephrogenic DI, Charcot-Marie Tooth syndrome, Pelizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, Pick's disease, several polyglutamine neurological disorders such as Huntington, spinocerebellar ataxia type I, spinal and bulbar muscular atrophy, dentatorubral pallidoluysian atrophy, and myotonic dystrophy, as well as spongiform encephalopathies, such as hereditary Creutzfeldt-Jakob disease (due to prion protein processing defect), Fabry disease, F Gerstmann-Sträussler-Scheinker syndrome, COPD, dry-eye disease, or Sjogren's disease. 
     
     
         55 . The method of  claim 54 , wherein said disease is cystic fibrosis. 
     
     
         56 . A kit for use in measuring the activity of CFTR or a fragment thereof in a biological sample in vitro or in vivo, comprising:
 i. a composition comprising a compound produced by the process of  claim 1 ; and   ii. instructions for:
 a. contacting the composition with the biological sample; and 
 b. measuring the activity of said CFTR or a fragment thereof. 
   
     
     
         57 . The kit of  claim 56 , further comprising instructions for:
 i. contacting an additional compound with the biological sample;   ii. measuring the activity of said CFTR or a fragment thereof in the presence of said additional compound; and   iii. comparing the activity of the CFTR or a fragment thereof in the presence of the additional compound with the activity of the CFTR or a fragment thereof in the presence of a composition comprising a compound of Formula 1.   
     
     
         58 . The kit according to  claim 57 , wherein the step of comparing the activity of said CFTR or a fragment thereof provides a measure of the density of said CFTR or a fragment thereof.