Benzothia(di)azepine compounds and their use as bile acid modulators

Abstract

The present invention relates to certain 1,5-benzothiazepine and 1,2,5-benzothiadiazepine derivatives, as defined herein. These compounds are bile acid modulators that have apical sodium-dependent bile acid transporter (ASBT) and / or hepatic bile acid transport (LBAT) inhibitory activity. The present invention also relates to pharmaceutical compositions containing these compounds and the use of these compounds in the treatment of cardiovascular diseases, fatty acid metabolism and glucose utilization disorders, gastrointestinal diseases, and liver diseases.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to Indian Patent Application No. 202011033169, filed on August 3, 2020, the disclosure of which is incorporated herein by reference in its entirety.

[0002] The present invention relates to certain 1,5-benzothiazepine and 1,2,5-benzothiadiazepine derivatives, as defined herein. These compounds are bile acid modulators that have apical sodium-dependent bile acid transporter (ASBT) and / or hepatic bile acid transport (LBAT) inhibitory activity. The present invention also relates to pharmaceutical compositions containing these compounds and the use of these compounds in the treatment of cardiovascular diseases, fatty acid metabolism and glucose utilization disorders, gastrointestinal diseases, and liver diseases. [Background technology]

[0003] Bile acids are physiological surfactants that play an important role in the intestinal absorption and transport of lipids, nutrients, and vitamins. They are also signaling molecules that activate nuclear receptors and cell signaling pathways that regulate lipid, glucose, and energy metabolism. Bile acids are steroid acids synthesized from cholesterol in the liver and stored in the gallbladder as mixed micelles. During digestion, the duodenum triggers the release of hormones that cause gallbladder contraction, releasing bile acids into the small intestine, where they enable the absorption of fat-soluble vitamins and cholesterol. Upon reaching the ileum, bile acids are reabsorbed from the intestine and secreted into the portal vein and returned to the liver via the portal circulation. In this way, over 90% of bile acids are recycled and returned to the liver. These bile acids are then transported across the sinusoidal membrane of hepatocytes and resecreted across the canalicular membrane into bile. During this first pass, 75–90% of bile acids are taken up by hepatocytes, completing a single enterohepatic circulation. Some bile acids that are not removed by the liver enter the systemic circulation, where they are filtered by the renal glomerulus, efficiently recovered by the proximal tubule, and returned to the systemic circulation. Interestingly, the majority of bile acids secreted across the canalicular membrane into bile originate from the recycling pool, with less than 10% resulting from de novo hepatic synthesis. The small fraction of bile acids that are not reabsorbed in the ileum reach the colon. Within the intestinal lumen, primary bile acids are converted to secondary bile acids by the action of intestinal bacteria, primarily through single or double dehydroxylation of the steroid nucleus. Bile acids that are not absorbed in the intestine are then excreted in the feces.

[0004] Overall, an efficient transport system helps maintain a constant bile acid pool, thereby ensuring a sufficiently high level of conjugated bile acids in the intestine to promote lipid absorption and reduce bacterial load in the small intestine. This system also minimizes the loss of bile acids in feces and urine, and protects the intestine and hepatobiliary tract by eliminating potentially cytotoxic surfactants (as reviewed by Kosters and Karpen (Xenobiotica 2008, Vol. 38, pp. 1043-1071); Chiang (J. Lipid Res. 2009, Vol. 50, pp. 1955-1966); and Dawson (Handb. Exp. Pharmacol. 2011, Vol. 201, pp. 169-203)).

[0005] Regulation of the bile acid pool size by converting cholesterol to bile acids in the liver has been found to play an important role in cholesterol homeostasis, and this corresponds to the main pathway for cholesterol excretion from the body. The liver plays an essential role in removing endogenous and xenobiotic compounds from the body. Normal hepatic bile secretion and enterohepatic circulation are required to excrete endogenous compounds such as cholesterol and bilirubin and their metabolites from the body, thereby maintaining lipid and bile acid homeostasis. (Kosters and Karpen, Xenobiotica 2008, Vol. 38, pp. 1043-1071)

[0006] Ileal reabsorption of bile acids can be inhibited by apical sodium-dependent bile acid transporter (ASBT) inhibitor compounds. Inhibition of bile acid reabsorption has been reported to be useful in the treatment of several diseases, including dyslipidemia, diabetes, obesity, constipation, cholestatic liver disease, nonalcoholic steatohepatitis, and other liver diseases. Several ASBT inhibitor compounds have been disclosed over the past several decades. For example, WO 93 / 16055, WO 94 / 18183, WO 94 / 18184, WO 96 / 05188, WO 96 / 08484, WO 96 / 16051, WO 97 / 33882, WO 98 / 03818, WO 98 / 07449, WO 98 / 40375, WO 99 / 35135, WO 99 / 64409, WO 99 / 64410, WO 00 / 47568, WO 00 / 61568, WO 00 / 38725, WO 00 / 38726, WO 00 / 38727, WO 00 / 38728, WO 00 / 38729, WO 01 / 66533, WO 01 / 68096, WO 02 / 32428, WO 02 / 50051, WO 03 / 020710, WO 03 / 022286, WO 03 / 022825, WO 03 / 022830, WO 03 / 061663, WO 03 / 091232, WO 03 / 106482, WO 2004 / 006899, WO 2004 / 076430, WO 2007 / 009655, WO 2007 / 009656, WO 2011 / 137135, WO 2019 / 234077, WO 2020 / 161216, WO 2020 / 161217, WO 2021 / 110883, WO 2021 / 110884, WO 2021 / 110885, WO 2021 / 110886, WO 2021 / 110887, DE 19825804, EP 864582, EP 489423, EP 549967, EP 573848, EP 624593, EP 624594, EP 624595, EP 624596, EP 0864582, EP 1173205, EP 1535913 and EP 3210977. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] WO 93 / 16055 [Patent Document 2] WO 94 / 18183 [Patent Document 3] WO 94 / 18184 [Patent Document 4] WO 96 / 05188 [Patent Document 5] WO 96 / 08484 [Patent Document 6] WO 96 / 16051 [Patent Document 7] WO 97 / 33882 [Patent Document 8] WO 98 / 03818 [Patent Document 9] WO 98 / 07449 [Patent Document 10] WO 98 / 40375 [Patent Document 11] WO 99 / 35135 [Patent Document 12] WO 99 / 64409 [Patent Document 13] WO 99 / 64410 [Patent Document 14] WO 00 / 47568 [Patent Document 15] WO 00 / 61568 [Patent Document 16] WO 00 / 38725 [Patent Document 17] WO 00 / 38726 [Patent Document 18] WO 00 / 38727 [Patent Document 19] WO 00 / 38728 [Patent Document 20] WO 00 / 38729 [Patent Document 21] WO 01 / 66533 [Patent Document 22] WO 01 / 68096 [Patent Document 23] WO 02 / 32428 [Patent Document 24] WO 02 / 50051 [Patent Document 25] WO 03 / 020710 [Patent Document 26] WO 03 / 022286 [Patent Document 27] WO 03 / 022825 [Patent Document 28] WO 03 / 022830 [Patent Document 29] WO 03 / 061663 [Patent Document 30] WO 03 / 091232 [Patent Document 31] WO 03 / 106482 [Patent Document 32] WO 2004 / 006899 [Patent Document 33] WO 2004 / 076430 [Patent Document 34] WO 2007 / 009655 [Patent Document 35] WO 2007 / 009656 [Patent Document 36] WO 2011 / 137135 [Patent Document 37] WO 2019 / 234077 [Patent Document 38] WO 2020 / 161216 [Patent Document 39] WO 2020 / 161217 [Patent Document 40] WO 2021 / 110883 [Patent Document 41] WO 2021 / 110884 [Patent Document 42] WO 2021 / 110885 [Patent Document 43] WO 2021 / 110886 [Patent Document 44] WO 2021 / 110887 [Patent Document 45] DE 19825804 [Patent Document 46] EP 864582 [Patent Document 47] EP 489423 [Patent Document 48] EP 549967 [Patent Document 49] EP 573848 [Patent Document 50] EP 624593 [Patent Document 51] EP 624594 [Patent Document 52] EP 624595 [Patent Document 53] EP 624596 [Patent Document 54] EP 0864582 [Patent Document 55] EP 1173205 [Patent Document 56] EP 1535913 [Patent Document 57] EP 3210977 [Patent Document 58] U.S. Patent Application Publication No. 2018 / 0140219 [Patent Document 59] U.S. Patent Application Publication No. 2016 / 146715 [Patent Document 60] U.S. Patent Application Publication No. 2005 / 0215882 [Patent Document 61] U.S. Patent No. 9,872,844 [Patent Document 62] WO 2017 / 138877 [Patent Document 63] WO 2017 / 138878 [Patent Document 64] WO 2019 / 032026 [Patent Document 65] WO 2019 / 032027 [Non-patent literature]

[0008] [Non-Patent Document 1] Kosters and Karpen (Xenobiotica 2008, Vol. 38, pp. 1043-1071) [Non-patent document 2] Chiang (J. Lipid Res. 2009, Vol. 50, pp. 1955-1966) [Non-patent document 3] Dawson (Handb. Exp. Pharmacol. 2011, Vol. 201, pp. 169-203) [Non-patent document 4] Dong et al., Mol. Pharm. 2013, Vol. 10, pp. 1008-1019 [Non-Patent Document 5] Vaz et al., Hepatology 2015, Vol. 61, pp. 260-267 [Non-patent document 6] Karpen and Dawson, Hepatology 2015, Vol. 61, pp. 24-27 [Non-Patent Document 7] Liu et al., Scientific Reports 2017, 7: 9214, pp. 1-7 [Non-patent document 8] Danese et al., PLoS One. 2017, Vol. 12(6): e0179200 [Non-Patent Document 9] Kooistra et al., "KLIFS: A structural kinase-ligand interaction database," Nucleic Acids Res. 2016, Vol. 44, No. D1, pp. D365-D371 [Non-Patent Document 10] Gunaydin, M. et al., Hepat Med. 2018, Vol. 10, pp. 95-104 [Non-Patent Document 11] Ferslew et al., Dig Dis Sci. 2015, Vol. 60, pp. 3318-3328 [Non-Patent Document 12] Chalasani et al., Hepatology 2018, Vol. 67(1), pp. 328-357 [Non-Patent Document 13] Kleiner et al., Hepatology. 2005, 41(6):1313-1321 [Non-Patent Document 14] Di Lascio et al., Ultrasound Med Biol. 2018, 44(8), 1585-1596; [Non-Patent Document 15] Lv et al., J Clin Transl Hepatol. 2018, Vol. 6(2), pp. 217-221; [Non-Patent Document 16] Reeder et al., J Magn Reson Imaging. 2011, Vol. 34(4), spcone; [Non-Patent Document 17] de Ledinghen V et al., J Gastroenterol Hepatol. 2016, 31(4), 848-855 [Non-Patent Document 18] Brunt et al., Am J Gastroenterol 1999, Vol. 94, pp. 2467-2474 [Non-Patent Document 19] Angulo et al., Hepatology 2007, 45(4), 846-54 [Non-Patent Document 20] Ishak et al., J. Hepatol. 1995, Vol. 22, pp. 696-699 [Non-Patent Document 21] McPherson et al., Gut 2010, 59(9), 1265-9 [Non-Patent Document 22] Adams et al., Clin. Chem. 2005, 51(10), 1867-1873 [Non-Patent Document 23] Lichtinghagen R et al. J Hepatol. 2013 Aug;59(2):236-42 [Non-Patent Document 24] Neuman et al., Can. J. Gastroenterol. Hepatol. 2014, 28(11), 607-618 [Non-Patent Document 25] Perez MJ, Briz O. World J. Gastroenterol. 2009, No. 15(14), pp. 1677-1689 [Non-Patent Document 26] Sorrentino P et al., Dig. Dis. Sci. 2005, 50(6), 1130-1135 [Non-Patent Document 27] Satapathy SK and Sanyal AJ. Semin. Liver Dis. 2015, 35(3), 221-235 [Non-patent document 28] Greene's Protective Groups in Organic Synthesis, 4th ed., by P. G. M. Wutz and T. W. Greene, John Wiley & Sons, Hoboken, 2006 Summary of the Invention

[0009] Despite several previously reported ASBT inhibitor compounds, there is a need for additional bile acid modulating compounds with optimized profiles of potency, selectivity, and bioavailability.

[0010] It has been discovered that certain benzothiazepine and benzothiadiazepine derivatives are potent inhibitors of the apical sodium-dependent bile acid transporter (ASBT) and / or hepatic bile acid transporter (LBAT) and may be useful in treating diseases in which inhibition of bile acid cycling is desirable. DETAILED DESCRIPTION OF THE INVENTION

[0011] Thus, in a first aspect, the present invention provides a compound of formula (I)

[0012] [ka]

[0013] (In the formula, M, R 1 , R 2 , R 3 , R 4 , R 5A and R 5B (See Table 1 below) or a pharmaceutically acceptable salt thereof:

[0014] [Table 1A]

[0015] [Table 1B]

[0016] [Table 1C]

[0017] [Table 1D]

[0018] [Table 1E]

[0019] [Table 1F]

[0020] [Table 1G]

[0021] [Table 1H]

[0022] [Table 1I]

[0023] [Table 1J]

[0024] In certain embodiments, the compound of formula (I) is (S)-3-(((S)-3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; (S)-3-(((R)-3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; (R)-3-(((S)-3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; (R)-3-(((R)-3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; 1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (S)-1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (R)-1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; (S)-3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; (R)-3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; 3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid; (S)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid; (R)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid; 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid; 3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid; (S)-3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid; (R)-3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid; 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoic acid; 3-((3,3-dibutyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoic acid; 3-((3,3-diethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; and 2-hydroxy-3-((7-(methylthio)-1,1-dioxide-5-phenyl-3,3-dipropyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)propanoic acid; or a pharmaceutically acceptable salt thereof.

[0025] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that are suitable for human pharmaceutical use and are generally safe, non-toxic, and not biologically or otherwise undesirable.

[0026] As used herein, the term "about" refers herein to a value or parameter, including (and describing) embodiments directed to the value or parameter itself. For example, a statement referring to "about 20" includes the statement "20." Numerical ranges include the numbers defining the range. Generally speaking, the term "about" refers to the value recited by the variable, to any value within experimental error of the value recited by the variable (e.g., within a 95% confidence interval of the mean), or within 10 percent of the value recited by the variable, whichever is greater.

[0027] The 1,5-benzothiazepine and 1,2,5-benzothiadiazepine compounds of Formula (I), or pharmaceutically acceptable salts thereof, are inhibitors of the apical sodium-dependent bile acid transporter, the hepatic bile acid transporter, or both the apical sodium-dependent bile acid transporter and the hepatic bile acid transporter (ASBT inhibitors, LBAT inhibitors, and dual ASBT / LBAT inhibitors, respectively). As such, they are useful for treating or preventing conditions, disorders, and diseases in which inhibition of bile acid circulation is desirable, such as cardiovascular diseases, fatty acid metabolism and glucose utilization disorders, gastrointestinal diseases, and liver diseases.

[0028] Cardiovascular diseases and disorders of fatty acid metabolism and glucose utilization include, but are not limited to, hypercholesterolemia; disorders of fatty acid metabolism; type 1 and type 2 diabetes mellitus; complications of diabetes, such as cataracts, micro- and macrovascular disease, retinopathy, neuropathy, nephropathy, and delayed wound healing, tissue ischemia, diabetic foot, arteriosclerosis, myocardial infarction, acute coronary syndrome, unstable angina, stable angina, stroke, peripheral arterial occlusive disease, cardiomyopathy, heart failure, heart rhythm disorders, and vascular restenosis; diabetes-related diseases, such as insulin resistance (impaired glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, obesity, dyslipidemia, hyperlipidemia, e.g., hypertriglyceridemia, metabolic syndrome (syndrome X), atherosclerosis, and hypertension; and increased high-density lipoprotein levels.

[0029] Gastrointestinal diseases and disorders include constipation (including chronic constipation, functional constipation, chronic idiopathic constipation (CIC), intermittent / sporadic constipation, constipation secondary to diabetes mellitus, constipation secondary to stroke, constipation secondary to chronic kidney disease, constipation secondary to multiple sclerosis, constipation secondary to Parkinson's disease, constipation secondary to systemic sclerosis, drug-induced constipation, irritable bowel syndrome with constipation (IBS-C), mixed irritable bowel syndrome (IBS-M), pediatric functional constipation, and opioid-induced constipation); Crohn's disease; primary bile acid malabsorption; irritable bowel syndrome (IBS); inflammatory bowel disease (IBD); ileal inflammation; and reflux disease and its complications, such as Barrett's esophagus, bile reflux esophagitis, and bile reflux gastritis.

[0030] Liver disease, as defined herein, is any disease of the liver and its associated organs, such as the pancreas, portal vein, liver parenchyma, intrahepatic biliary system, extrahepatic biliary system, and gallbladder. In some cases, the liver disease is a bile acid-dependent liver disease. Liver diseases and disorders include inherited metabolic disorders of the liver; congenital anomalies of bile acid synthesis; congenital abnormalities of bile duct course; biliary atresia; biliary atresia after Kasai operation; biliary atresia after liver transplantation; neonatal hepatitis; neonatal cholestasis; inherited forms of cholestasis; cerebrotendinous xanthomatosis; secondary defects of BA synthesis; Zellweger syndrome; cystic fibrosis-related liver disease; alpha-1 antitrypsin deficiency; Alagille syndrome (ALGS); Byler syndrome; primary defects of bile acid (BA) synthesis; progressive familial intrahepatic cholestasis (PFIC), e.g., PFIC-1, PFIC-2, PFIC-3, and non-specific PFIC, PFIC after bile diversion, and PFIC after liver transplantation; benign recurrent intrahepatic cholestasis (BRIC), e.g., BRIC1 , BRIC2, and non-specific BRIC, BRIC after bile diversion, and BRIC after liver transplant; autoimmune hepatitis; primary biliary cirrhosis (PBC); liver fibrosis; nonalcoholic fatty liver disease (NAFLD); nonalcoholic steatohepatitis (NASH); portal hypertension; cholestasis; cholestasis of Down syndrome; drug-induced cholestasis; intrahepatic cholestasis of pregnancy (jaundice in pregnancy); intrahepatic cholestasis; extrahepatic cholestasis; parenteral nutrition-associated cholestasis (PNAC); low phospholipid-associated cholestasis; lymphedema cholestasis syndrome 1 (LCS1); primary sclerosing cholangitis (PSC); immunoglobulin G4-associated cholangitis; primary biliary cholangitis; cholelithiasis (gallstones); biliary lithiasis (biliary lithiasis); choledocholithiasis; gallstone pancreatitis; Caroli disease; malignant tumors of the bile duct; malignant tumors causing obstruction of the biliary tree; bile duct stenosis; AIDS cholangiopathy; ischemic cholangiopathy; pruritus due to cholestasis or jaundice; pancreatitis; chronic autoimmune liver disease leading to progressive cholestasis; hepatic steatosis; alcoholic hepatitis; acute fatty liver; fatty liver of pregnancy; drug-induced hepatitis; iron overload; congenital bile acid metabolism disorder type 1 (BAS disorder type 1); drug-induced liver injury (DILI); hepatic fibrosis; congenital hepatic fibrosis; cirrhosis; Langerhans cell histiocytosis (LCH); neonatal ichthyotic sclerosing cholangitis (NISCH); erythropoietic protoporphyria (EPP); idiopathic adult bile duct paucity (IAD); idiopathic neonatal hepatitis (INH);These include, but are not limited to, nonsyndromic intrahepatic cholangiopancreas (NS PILBD); autosomal recessive inherited intrahepatic cholestasis (North American Indian childhood cirrhosis) (NAIC); hepatic sarcoidosis; amyloidosis; necrotizing enterocolitis; serum bile acid-induced toxicity, such as cardiac rhythm disorders (e.g., atrial fibrillation) in the setting of abnormal serum bile acid profiles, cardiomyopathy associated with cirrhosis ("cholecardia"), and skeletal muscle wasting associated with cholestatic liver disease; polycystic hepatic disease; viral hepatitis (including hepatitis A, hepatitis B, hepatitis C, hepatitis D, and hepatitis E); hepatocellular carcinoma (hepatocellular carcinoma); cholangiocarcinoma; bile acid-related gastrointestinal cancers; and cholestasis caused by tumors and neoplasms of the liver, biliary tract, and pancreas. The compounds of formula (I) or pharmaceutically acceptable salts thereof are also useful for augmenting corticosteroid therapy in liver disease;

[0031] Other diseases that can be treated or prevented by the compound of formula (I) or a pharmaceutically acceptable salt thereof include hyperabsorption syndromes (including abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicron retention disease (CRD), and sitosterolemia); hypervitaminosis and osteopetrosis; hypertension; glomerular hyperfiltration; polycystic kidney disease (PKD), such as autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD); and pruritus in renal failure. The compounds are also useful for protecting against kidney damage associated with liver or metabolic disease.

[0032] The transport of bile acids in the human body is mediated by members of the SLC10 family of solute transporter proteins, particularly Na β-glucanase (Na β-glucanase) expressed on the sinusoidal plasma membrane of hepatocytes. +It is regulated by the apical sodium-dependent bile acid transporter (ASBT, also called ileal bile acid transporter (IBAT), ISBT, ABAT, or NTCP2; gene symbol SLC10A2), which is expressed in the apical membrane of ileal enterocytes, proximal tubule cells, bile duct epithelium, large duct cells, and gallbladder epithelial cells. Within the liver, bile acids are efficiently extracted from portal blood by the hepatic bile acid transporter (LBAT) and resecreted across the canalicular membrane by the bile salt export pump (BSEP; gene symbol ABCB11). Reabsorption of bile acids in the ileum is handled by the apical sodium-dependent bile acid transporter (ASBT), commonly referred to as the ileal bile acid transporter (IBAT). Both LBAT and ASBT transport more than two sodium salts per solute molecule. + It functions as an electrogenic sodium-solute cotransporter, moving ions.

[0033] Biosubstances, including xenobiotics and bile acids, are taken up by the liver from the portal blood and secreted into bile by distinct transport proteins with individualized substrate specificities. Glycine- and taurine-conjugated bile acids exist in anionic form and cannot cross membranes by diffusion, thus relying entirely on membrane transport proteins for transport into and out of hepatocytes (Kosters and Karpen, Xenobiotica 2008, Vol. 38, pp. 1043-1071). ASBT and LBAT prefer glycine- and taurine-conjugated bile salts over their unconjugated counterparts and show higher affinity for dihydroxy bile salts than trihydroxy bile salts. Although non-bile acid substrates of ASBT have yet to be identified, LBAT has also been found to transport various steroid sulfates, hormones, and xenobiotics.

[0034] LBAT has not been as thoroughly characterized as ASBT in terms of drug inhibition requirements. Dong et al. identified FDA-approved drugs that inhibit human LBAT and compared the inhibition requirements of LBAT and ASBT. A series of LBAT inhibition studies were conducted using FDA-approved drugs in conjunction with the development of an iterative computational model. Screening studies identified 27 drugs as novel LBAT inhibitors, including irbesartan (Ki = 11.9 μM) and ezetimibe (Ki = 25.0 μM). Common pharmacophore features indicated that two hydrophobic groups and one hydrogen-bonding receptor are important for LBAT inhibition. Of the 72 drugs screened in vitro, a total of 31 drugs inhibited LBAT, while 51 drugs (i.e., more than half) inhibited ASBT. Thus, despite the inhibitor overlap, ASBT is unexpectedly more tolerant to drug inhibition than LBAT, which may be related to LBAT having fewer pharmacophore features (Dong et al., Mol. Pharm. 2013, Vol. 10, pp. 1008-1019).

[0035] Vaz et al. describe the identification of LBAT deficiency as a new congenital anomaly with a relatively mild clinical phenotype. Identification of LBAT deficiency confirms this transporter as the primary import system for conjugated bile salts into the liver, but also demonstrates that in its absence, auxiliary transporters can sustain enterohepatic circulation (Vaz et al., Hepatology 2015, Vol. 61, pp. 260-267). These findings support the hypothesis that LBAT inhibition is a safe mechanism of action, since hepatocytes still have the potential to uptake the necessary amounts of bile acids.

[0036] Liu et al. described the identification of a new type of hypercholanic acidemia associated with homozygosity for the p.Ser267Phe mutation in SLC10A1 (LBAT). The allele frequency of this mutation in the gene SLC10A1 varies by ethnic group, with the highest incidence occurring in southern China (8% and 12% in the Chinese Han and Dai populations, respectively) and Vietnam (11%). This "hidden" hypercholanic acidemia was thought to affect 0.64% of the southern Chinese Han population, 1.44% of the Chinese Dai population, and 1.21% of the Vietnamese population. Increased levels of conjugated and unconjugated serum BAs were also observed in homozygous individuals. Liu et al. suggested that this finding was most likely due to reduced BA transport from the portal circulation to hepatocytes. This supports the hypothesis that the physiological function of the enterohepatic circulation is not only to recycle bile acids but also to remove bile acids from the circulation to achieve homeostasis (Karpen and Dawson, Hepatology 2015, Vol. 61, pp. 24-27). Alternatively, in homozygous carriers, the liver may synthesize increased levels of bile acids to compensate for the reduced enterohepatic recirculation. Because LBAT also transports unconjugated bile acids, the increase in unconjugated bile acids in this study was not surprising (Liu et al., Scientific Reports 2017, 7: 9214, pp. 1-7).

[0037] LBAT has been found to be downregulated in several forms of cholestatic liver injury and cholestasis, while ASBT has been found to be downregulated in various gastrointestinal disorders such as Crohn's disease, primary bile acid malabsorption, inflammatory bowel disease, and ileal inflammation, but upregulated in cholestasis. LBAT also functions as a cellular receptor for viral entry of hepatitis B virus (HBV) and hepatitis D virus (HDV), which in turn is a major cause of liver disease and hepatocellular carcinoma.

[0038] ASBT inhibition has been investigated for reducing plasma cholesterol levels, improving insulin resistance, and reducing the burden of bile acids on the liver in cholestatic liver disease.In addition, ASBT inhibition has been found to restore insulin levels and normal blood glucose levels, thereby establishing ASBT inhibition as a promising treatment for type 2 diabetes mellitus.ASBT inhibitors are also used to treat functional constipation.

[0039] Because ASBT is primarily expressed in the ileum (often referred to as IBAT in the ileum), ASBT inhibitors do not need to be systemically absorbed. However, ASBT is also expressed in proximal tubule cells in the kidney. Therefore, systemically absorbable ASBT inhibitors may also inhibit renal bile acid reuptake. This would increase urinary bile acid levels and enhance bile acid removal from the body via urine. Therefore, systemically absorbable ASBT inhibitors that act not only in the ileum but also in the kidney are predicted to result in greater reductions in bile acid levels than non-systemically absorbed ASBT inhibitors that act only in the ileum.

[0040] Compounds with high ASBT inhibitory potency are particularly suitable for the treatment of liver diseases that cause cholestasis, such as progressive familial intrahepatic cholestasis (PFIC), Alagille syndrome, biliary atresia, and non-alcoholic steatohepatitis (NASH).

[0041] Biliary atresia is a rare pediatric liver disease involving partial or total obstruction (or even absence) of the large bile duct. This obstruction or absence causes cholestasis, which leads to the accumulation of bile acids that damage the liver. In some embodiments, the accumulation of bile acids occurs in the extrahepatic bile duct system. In some embodiments, the accumulation of bile acids occurs in the intrahepatic bile duct system. The current standard of care is the Kasai procedure, a surgical procedure in which the obstructed bile duct is removed and a portion of the small intestine is connected directly to the liver. Currently, there is no approved drug therapy for this disorder.

[0042] Provided herein are methods for treating biliary atresia in a subject in need thereof, the methods comprising administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is administered a compound of Formula (I) or a pharmaceutically acceptable salt thereof after undergoing Kasai's procedure. In some embodiments, the subject is administered a compound of Formula (I) or a pharmaceutically acceptable salt thereof before undergoing Kasai's procedure. In some embodiments, treating biliary atresia reduces serum bile acid levels in the subject. In some embodiments, serum bile acid levels are determined, for example, by an ELISA enzyme assay or a total bile acid measurement assay such as that described in Danese et al., PLoS One. 2017, Vol. 12(6): e0179200, which is incorporated herein by reference in its entirety. In some embodiments, serum bile acid levels may be reduced by, for example, 10% to 40%, 20% to 50%, 30% to 60%, 40% to 70%, 50% to 80%, or greater than 90% of the serum bile acid levels prior to administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, treating biliary atresia includes treating pruritus.

[0043] PFIC is a rare genetic disorder that is estimated to affect one in 50,000 to 100,000 live births worldwide and causes progressive, life-threatening liver disease.

[0044] One symptom of PFIC is pruritus, which often significantly reduces quality of life. In some cases, PFIC leads to cirrhosis and liver failure. Current therapies include partial external biliary drainage (PEFD) and liver transplantation, but these options carry substantial risks of postoperative complications as well as potential psychological and social problems.

[0045] Three alternative gene defects have been identified that correlate with three distinct PFIC subtypes, known as types 1, 2, and 3: PFIC type 1, sometimes called "Byler's disease," is caused by impaired bile secretion due to mutations in the ATP8B1 gene, which encodes a protein that helps maintain the proper balance of fats known as phospholipids in cell membranes within the bile ducts. An imbalance of these phospholipids is associated with cholestasis and elevated bile acids in the liver. Subjects with PFIC type 1 typically develop cholestasis within the first month of life and, without surgical treatment, progress to cirrhosis and end-stage liver disease before the first decade of life. PFIC type 2, sometimes called "Byler syndrome," is caused by impaired bile salt secretion due to mutations in the ABCB11 gene, which encodes a protein known as the bile salt export pump, which moves bile acids out of the liver. Individuals with PFIC type 2 often develop liver failure within the first few years of life and are at increased risk of developing a type of liver cancer known as hepatocellular carcinoma. PFIC type 3 typically presents with progressive cholestasis in the first few years of childhood and is caused by mutations in the ABCB4 gene, which encodes a transporter that moves phospholipids across cell membranes.

[0046] Additionally, mutations in the TJP2, NR1H4, or Myo5b genes have been proposed as a cause of PFIC. Additionally, some subjects with PFIC do not have mutations in any of the ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b genes. In these cases, the cause of the condition is unknown.

[0047] Exemplary mutations of the ATP8B1 gene or resulting protein are numbered based on the human wild-type ATP8B1 protein (e.g., SEQ ID NO: 1) or gene (e.g., SEQ ID NO: 2) and are listed in Tables 2 and 3. Exemplary mutations of the ABCB11 gene or resulting protein are numbered based on the human wild-type ABCB11 protein (e.g., SEQ ID NO: 3) or gene (e.g., SEQ ID NO: 4) and are listed in Tables 4 and 5.

[0048] As will be appreciated by one of ordinary skill in the art, amino acid positions in a reference protein sequence that correspond to particular amino acid positions in SEQ ID NO: 1 or 3 can be determined by aligning the reference protein sequence with SEQ ID NO: 1 or 3 (e.g., using a software program such as ClustalW2). Changes to these residues (referred to herein as "mutations") can include single or multiple amino acid substitutions, insertions, and deletions within or adjacent to the sequence. As will be appreciated by one of ordinary skill in the art, nucleotide positions in a reference gene sequence that correspond to particular nucleotide positions in SEQ ID NO: 2 or 4 can be determined by aligning the reference gene sequence with SEQ ID NO: 2 or 4 (e.g., using a software program such as ClustalW2). Changes to these residues (referred to herein as "mutations") can include single or multiple nucleotide substitutions, insertions, and deletions within or adjacent to the sequence. See also Kooistra et al., "KLIFS: A structural kinase-ligand interaction database," Nucleic Acids Res. 2016, Vol. 44, No. D1, pp. D365-D371, which is incorporated herein by reference in its entirety.

[0049] Canonical protein sequence of ATP8B1 (SEQ ID NO:1) - Uniprot ID O43520

[0050] [ka]

[0051] Canonical DNA sequence of ATP8B1 (SEQ ID NO: 2)

[0052] [ka]

[0053] [ka]

[0054] Table 2A

[0055] Table 2B

[0056] Table 2C

[0057] Table 2D

[0058] Table 2E

[0059] Table 2F

[0060] Table 2G

[0061]

Table 2H

[0062]

Table 2I

[0063] Table 3A

[0064] Table 3B

[0065] Table 3C

[0066] Table 3D

[0067] References for Table 2 and Table 3 1 Folmer et al., Hepatology. 2009, vol. 50(5), p. 1597-1605. 2 Hsu et al., Hepatol Res. 2009, vol. 39(6), p. 625-631. 3 Alvarez et al., Hum Mol Genet. 2004, vol. 13(20), p. 2451-2460. 4 Davit-Spraul et al., Hepatology 2010, vol. 51(5), p. 1645-1655. 5 Vitale et al., J Gastroenterol. 2018, vol. 53(8), p. 945-958. 6 Klomp et al., Hepatology 2004, vol. 40(1), p. 27-38. 7 Zarenezhad et al., Hepatitis Monthly: 2017, vol. 17(2); e43500. 8 Dixon et al., Scientific Reports 2017, vol. 7, 11823. 9 Painter et al., Eur J Hum Genet. 2005, vol. 13(4), p. 435-439. 10 Deng et al., World J Gastroenterol. 2012, vol. 18(44), p. 6504-6509. 11 Giovannoni et al., PLoS One. 2015, vol. 10(12): e0145021. 12 Li et al., Hepatology International 2017, vol. 11, No. 1, Supp. Supplement 1, pp. S180. Abstract Number: OP284. 13 Togawa et al., Journal of Pediatric Gastroenterology and Nutrition 2018, vol. 67, Supp. Supplement 1, pp. S363. Abstract Number: 615. 14 Miloh et al., Gastroenterology 2006, vol. 130, No. 4, Suppl. 2, pp. A759-A760. Meeting Info.: Digestive Disease Week Meeting / 107th Annual Meeting of the American-Gastroenterological-Association. Los Angeles, CA, USA. May 19. 15Droege et al., Journal of Gastroenterology 2015, vol. 53, no. 12. Abstract Number: A3-27. Meeting info: 32nd Annual Meeting of the German Working Community for the Study of the Liver. Dusseldorf, Germany. 22 Jan 2016-23 Jan 2016 16 Mizuochi et al., Clin Chim Acta. 2012, Vol. 413(15-16), p. 1301-1304. 17 Liu et al., Hepatology International 2009, vol. 3, no. 1, p. 184-185. Abstract Number: PE405. Meeting Info: 19th Conference of the Asian Pacific Association for the Study of the Liver. Hong Kong, China. 13 Feb 2009-16 Feb 2009 18 McKay et al., Version 2. F1000Res. 2013; 2: 32. DOI: 10.12688 / f1000research.2-32.v2 19 Hasegawa et al., Orphanet J Rare Dis. 2014, Vol. 9:89. 20 Stone et al., J Biol Chem. 2012, Vol. 287(49), p. 41139-51. 21 Kang et al., J Pathol Transl Med. 2019 May 16. doi: 10.4132 / jptm.2019.05.03. [Epub ahead of print] 22Sharma et al., BMC Gastroenterol. 2018, vol. 18(1), p. 107. 23 Uegaki et al., Intern Med. 2008, vol. 47(7), p. 599-602. 24 Goldschmidt et al., Hepatol Res. 2016, vol. 46(4), p. 306-311. 25 Liu et al., J Pediatr Gastroenterol Nutr. 2010, vol. 50(2), p. 179-183. 26 Jung et al., J Pediatr Gastroenterol Nutr. 2007, vol. 44(4), p. 453-458. 27 Bounford. University of Birmingham. Dissertation Abstracts International, (2016) Vol. 75, No. 1C. Order No.: AAI10588329. ProQuest Dissertations & Theses. 28 Stolz et al., Aliment Pharmacol Ther. 2019, vol. 49(9), p. 1195-1204. 29 Ivashkin et al., Hepatology International 2016, vol. 10, No. 1, Supp. SUPPL. 1, pp. S461. Abstract Number: LBO-38. Meeting Info: 25th Annual Conference of the Asian Pacific Association for the Study of the Liver, APASL 2016. Tokyo, Japan. 20 Feb 2016-24 Feb 2016 30 Blackmore et al., J Clin Exp Hepatol. 2013, vol. 3(2), p. 159-161. 31 Matte et al., J Pediatr Gastroenterol Nutr. 2010, vol. 51(4), p. 488-493. 32 Squires et al., J Pediatr Gastroenterol Nutr. 2017, vol. 64(3), p. 425-430. 33 Hayshi et al., EBioMedicine. 2018, vol. 27, p. 187-199. 34 Nagasaka et al., J Pediatr Gastroenterol Nutr. 2007, vol. 45(1), p. 96-105. 35 Wang et al., PLoS One. 2016; vol. 11(4): e0153114. 36 Narchi et al., Saudi J Gastroenterol. 2017, vol. 23(5), p. 303-305. 37 Alashkar et al., Blood 2015, vol. 126, No. 23. Meeting Info.: 57th Annual Meeting of the American-Society-of-Hematology. Orlando, FL, USA. December 05 -08, 2015. Amer Soc Hematol. 38Ferreira et al., Pediatric Transplantation 2013, vol. 17, Supp. SUPPL. 1, pp. 99. Abstract Number: 239. Meeting Info: IPTA 7th Congress on Pediatric Transplantation. Warsaw, Poland. 13 Jul 2013-16 Jul 2013. 39 Pauli-Magnus et al., J Hepatol. 2005, vol. 43(2), p. 342-357. 40 Jericho et al., Journal of Pediatric Gastroenterology and Nutrition 2015, vol. 60(3), p. 368-374. 41 van der Word et al., PLoS One. 2013, vol. 8(11): e80553. 42 Copeland et al., J Gastroenterol Hepatol. 2013, vol. 28(3), p. 560-564. 43 Droege et al., J Hepatol. 2017, vol. 67(6), p. 1253-1264. 44 Chen et al., Journal of Pediatrics 2002, vol. 140(1), p. 119-124. 45 Jirsa et al., Hepatol Res. 2004, vol. 30(1), p. 1-3. 46 van der Word et al., Hepatology 2015, vol. 61(4), p. 1382-1391.

[0068] In some embodiments, the mutation in ATP8B1 is selected from L127P, G308V, T456M, D554N, F529del, I661T, E665X, R930X, R952X, R1014X, and G1040R.

[0069] Canonical protein sequence of ABCB11 (SEQ ID NO:3) - Uniprot ID O95342

[0070] [ka]

[0071] Canonical DNA sequence of ABCB11 (SEQ ID NO: 4)

[0072] [ka]

[0073] [ka]

[0074] [Table 4A]

[0075] [Table 4B]

[0076] [Table 4C]

[0077] [Table 4D]

[0078] [Table 4E]

[0079] [Table 4F]

[0080] Table 4G

[0081]

Table 4H

[0082]

Table 4I

[0083] Table 4J

[0084] Table 4K

[0085] Table 4L

[0086] Table 4M

[0087] Table 4N

[0088] Table 5A

[0089] Table 5B

[0090] Table 5C

[0091] Table 5D

[0092] Table 5E

[0093] Table 5F

[0094] Table 5G

[0095]

Table 5H

[0096] References for Table 4 and Table 5 1 Noe et al., J Hepatol. 2005, vol. 43(3), p. 536-543. 2 Lam et al., Am J Physiol Cell Physiol. 2007, vol. 293(5), p. C1709-16. 3 Stindt et al., Liver Int. 2013, vol. 33(10), p. 1527-1735. 4Gao et al., Shandong Yiyao 2012, vol. 52(10), p. 14-16. 5 Strautnieks et al., Gastroenterology. 2008, vol. 134(4), p. 1203-1214. 6 Kagawa et al., Am J Physiol Gastrointest Liver Physiol. 2008, vol. 294(1), p. G58-67. 7 Byrne et al., Hepatology. 2009, vol. 49(2), p. 553-567. 8 Chen et al., J Pediatr. 2008, vol. 153(6), p. 825-832. 9 Davit-Spraul et al., Hepatology 2010, vol. 51(5), p. 1645-1655. 10 Droege et al., Sci Rep. 2016, vol. 6: 24827. 11 Lang et al., Pharmacogenet Genomics. 2007, vol. 17(1), p. 47-60. 12 Ellinger et al., World J Gastroenterol. 2017, vol. 23(29), p. :5295-5303. 13 Vitale et al., J Gastroenterol. 2018, vol. 53(8), p. 945-958. 14 Knisely et al., Hepatology. 2006, vol. 44(2), p. 478-86. 15Ellis et al., Hepatology. 2018, vol. 67(4), p. 1531-1545. 16 Lam et al., J Hepatol. 2006, vol. 44(1), p. 240-242. 17 Varma et al., Hepatology 2015, vol. 62(1), p. 198-206. 18 Treepongkaruna et al., World J Gastroenterol. 2009, vol. 15(34), p. 4339-4342. 19 Zarenezhad et al., Hepatitis Monthly: 2017, vol. 17(2); e43500. 20 Hayashi et al., Hepatol Res. 2016, vol. 46(2), p. 192-200. 21 Guorui et al., Linchuang Erke Zazhi 2013, vol. 31(10), 905-909. 22 van Mil et al., Gastroenterology. 2004, vol. 127(2), p. 379-384. 23 Anzivino et al., Dig Liver Dis. 2013, vol. 45(3), p. 226-232. 24 Park et al., World J Gastroenterol. 2016, vol. 22(20), p. 4901-4907. 25 Imagawa et al., J Hum Genet. 2018, vol. 63(5), p. 569-577. 26Giovannoni et al., PLoS One. 2015, vol. 10(12): e0145021. 27 Hu et al., Mol Med Rep. 2014, vol. 10(3), p. 1264-1274. 28 Lang et al,. Drug Metab Dispos. 2006, vol. 34(9), p. 1582-1599. 29 Masahata et al., Transplant Proc. 2016, vol. 48(9), p. 3156-3162. 30 Holz et al., Hepatol Commun. 2018, vol. 2(2), p. 152-154. 31 Li et al., Hepatology International 2017, vol. 11, No. 1, Supp. Supplement 1, pp. S180. Abstract Number: OP284. 32 Francalanci et al., Laboratory Investigation 2011, vol. 91, Supp. SUPPL. 1, pp. 360A. Abstract Number: 1526. 33 Francalanci et al., Digestive and Liver Disease 2010, vol. 42, Supp. SUPPL. 1, pp. S16. Abstract Number: T.N.5. 34 Shah et al., J Pediatr Genet. 2017, vol. 6(2), p. 126-127. 35 Gao et al., Hepatitis Monthly 2017, vol. 17(10), e55087 / 1-e55087 / 6. 36 Evason et al., Am J Surg Pathol. 2011, vol. 35(5), p. 687-696. 37 Davit-Spraul et al., Mol Genet Metab. 2014, vol. 113(3), p. 225-229. 38 Maggiore et al., J Hepatol. 2010, vol. 53(5), p. 981-6. 39 McKay et al., Version 2. F1000Res. 2013; 2: 32. DOI: 10.12688 / f1000research.2-32.v2 40 Liu et al., Pediatr Int. 2013, vol. 55(2), p. 138-144. 41 Waisbourd-Zinman et al., Ann Hepatol. 2017, vol. 16(3), p. 465-468. 42 Griffin, et al., Canadian Journal of Gastroenterology and Hepatology 2016, vol. 2016. Abstract Number: A200. Meeting Info: 2016 Canadian Digestive Diseases Week, CDDW 2016. Montreal, QC, United States. 26 Feb 2016-29 Feb 2016 43 Qiu et al., Hepatology 2017, vol. 65(5), p. 1655-1669. 44 Imagawa et al., Sci Rep. 2017, 7:41806. 45Kang et al., J Pathol Transl Med. 2019 May 16. doi: 10.4132 / jptm.2019.05.03. [Epub ahead of print] 46 Takahashi et al., Eur J Gastroenterol Hepatol. 2007, vol. 19(11), p. 942-6. 47 Shimizu et al., Am J Transplant. 2011, vol. 11(2), p. 394-398. 48 Krawczyk et al., Ann Hepatol. 2012, vol. 11(5), p. 710-744. 49 Sharma et al., BMC Gastroenterol. 2018, vol. 18(1), p. 107. 50 Sattler et al., Journal of Hepatology 2017, vol. 66, No. 1, Suppl. S, pp. S177. Meeting Info.: International Liver Congress / 52nd Annual Meeting of the European-Association-for-the-Study-of-the-Liver. Amsterdam, NETHERLANDS. April 19 -23, 2017. European Assoc Study Liver. 51 Jung et al., J Pediatr Gastroenterol Nutr. 2007, vol. 44(4), p. 453-458. 52Sciveres. Digestive and Liver Disease 2010, vol. 42, Supp. SUPPL. 5, pp. S329. Abstract Number: CO18. Meeting Info: 17th National Congress SIGENP. Pescara, Italy. 07 Oct 2010-09 Oct 2010 53 Sohn et al., Pediatr Gastroenterol Hepatol Nutr. 2019, vol. 22(2), p. 201-206. 54 Ho et al., Pharmacogenet Genomics. 2010, vol. 20(1), p. 45-57. 55 Wang et al., Hepatol Res. 2018, vol. 48(7), p. 574-584. 56 Shaprio et al., J Hum Genet. 2010, vol. 55(5), p. 308-313. 57 Bounford. University of Birmingham. Dissertation Abstracts International, (2016) Vol. 75, No. 1C. Order No.: AAI10588329. ProQuest Dissertations & Theses. 58 Stolz et al., Aliment Pharmacol Ther. 2019, vol. 49(9), p. 1195-1204. 59 Jankowska et al., J Pediatr Gastroenterol Nutr. 2014, vol. 58(1), p. 92-95. 60Kim. Journal of Pediatric Gastroenterology and Nutrition 2016, vol. 62, Supp. SUPPL. 1, pp. 620. Abstract Number: H-P-045. Meeting Info: 49th Annual Meeting of the European Society for Paediatric Gastroenterology, Hepatology and Nutrition, ESPGHAN 2016. Athens, Greece. 25 May 2016-28 May 2016. 61 Pauli-Magnus et al., Hepatology 2003, vol. 38, No. 4 Suppl. 1, pp. 518A. print. Meeting Info.: 54th Annual Meeting of the American Association for the Study of Liver Diseases. Boston, MA, USA. October 24-28, 2003. American Association for the Study of Liver Diseases. 62 Li et al., Hepatology International 2017, vol. 11, No. 1, Supp. Supplement 1, pp. S362. Abstract Number: PP0347. Meeting Info: 26th Annual Conference of the Asian Pacific Association for the Study of the Liver, APASL 2017. Shanghai, China. 15 Feb 2017-19 Feb 2017. 63Rumbo et al., Transplantation 2018, vol. 102, No. 7, Supp. Supplement 1, pp. S848. Abstract Number: P.752. Meeting Info: 27th International Congress of The Transplantation Society, TTS 2018. Madrid, Spain. 30 Jun 2018-05 Jul 2018. 64 Lee et al., Pediatr Gastroenterol Hepatol Nutr. 2017, vol. 20(2), p. 114-123. 65 Sherrif et al., Liver international: official journal of the International Association for the Study of the Liver 2013, vol. 33, No. 8, pp. 1266-1270. 66 Blackmore et al., J Clin Exp Hepatol. 2013, vol. 3(2), p. 159-161. 67 Matte et al., J Pediatr Gastroenterol Nutr. 2010, vol. 51(4), p. 488-493. 68 Lin et al., Zhongguo Dang Dai Er Ke Za Zhi. 2018, vol. 20(9), p. 758-764. 69Harmanci et al., Experimental and Clinical Transplantation 2015, vol. 13, Supp. SUPPL. 2, pp. 76. Abstract Number: P62. Meeting Info: 1st Congress of the Turkic World Transplantation Society. Astana, Kazakhstan. 20 May 2015-22 May 2015. 70 Herbst et al., Mol Cell Probes. 2015, vol. 29(5), p. 291-298. 71 Moghadamrad et al., Hepatology. 2013, vol. 57(6), p. 2539-2541. 72 Holz et al., Journal of Gastroenterology 2016, vol. 54, no. 8. Abstract number: KV275. Meeting info: Visceral Medicine 2016, 71st Annual Meeting of the German Society for Gastroenterology, Digestive and Metabolic Diseases with Endoscopy Section - 10th Autumn Meeting of the German Society for General and Visceral Surgery. Hamburg, Germany. September 21–24, 2016. 73 Wang et al., PLoS One. 2016; vol. 11(4): e0153114. 74Hao et al., International Journal of Clinical and Experimental Pathology 2017, vol. 10(3), p. 3480-3487. 75 Arnell et al., J Pediatr Gastroenterol Nutr. 2010, vol. 51(4), p. 494-499. 76 Sharma et al., Indian Journal of Gastroenterology 2017, vol. 36, No. 1, Supp. Supplement 1, pp. A99. Abstract Number: M-20. Meeting Info: 58th Annual Conference of the Indian Society of Gastroenterology, ISGCON 2017. Bhubaneswar, India. 14 Dec 2017-17 Dec 2017. 77 Beausejour et al., Can J Gastroenterol. 2011, vol. 25(6), p. 311-314. 78 Imagawa et al., Journal of Pediatric Gastroenterology and Nutrition 2016, vol. 63, Supp. Supplement 2, pp. S51. Abstract Number: 166. Meeting Info: World Congress of Pediatric Gastroenterology, Hepatology and Nutrition 2016. Montreal, QC, Canada. 05 Oct 2016-08 Oct 2016. 79Peng et al., Zhonghua er ke za zhi (Chinese journal of pediatrics) 2018, vol. 56, No. 6, pp. 440-444. 80 Tibesar et al., Case Rep Pediatr. 2014, vol. 2014: 185923. 81 Ng et al., Journal of Pediatric Gastroenterology and Nutrition 2018, vol. 66, Supp. Supplement 2, pp. 860. Abstract Number: H-P-127. Meeting Info: 51st Annual Meeting European Society for Paediatric Gastroenterology, Hepatology and Nutrition, ESPGHAN 2018. Geneva, Switzerland. 09 May 2018-12 May 2018. 82 Wong et al., Clin Chem. 2008, vol. 54(7), p. 1141-1148. 83 Pauli-Magnus et al., J Hepatol. 2005, vol. 43(2), p. 342-357. 84 Jericho et al., Journal of Pediatric Gastroenterology and Nutrition. 60, vol. 3, p. 368-374. 85Scheimann et al., Gastroenterology 2007, vol. 132, No. 4, Suppl. 2, pp. A452. Meeting Info.: Digestive Disease Week Meeting / 108th Annual Meeting of the American-Gastroenterological-Association. Washington, DC, USA. May 19 -24, 2007. Amer Gastroenterol Assoc; Amer Assoc Study Liver Dis; Amer Soc Gastrointestinal Endoscopy; Soc Surg Alimentary Tract. 86 Jaquotot-Haerranz et al., Rev Esp Enferm Dig. 2013, vol. 105(1), p. 52-54. 87 Khosla et al., American Journal of Gastroenterology 2015, vol. 110, No. Suppl. 1, pp. S397. Meeting Info.: 80th Annual Scientific Meeting of the American-College-of-Gastroenterology. Honolulu, HI, USA. October 16 -21, 2015. 88 Droege et al., J Hepatol. 2017, vol. 67(6), p. 1253-1264. 89 Liu et al., Liver International 2010, vol. 30(6), p. 809-815. 90 Chen et al., Journal of Pediatrics 2002, vol. 140(1), p. 119-124. 91US Patent No. 9,295,677

[0097] In some embodiments, the mutation in ABCB11 is selected from A167T, G238V, V284L, E297G, R470Q, R470X, D482G, R487H, A570T, N591S, A865V, G982R, R1153C, and R1268Q.

[0098] Methods for treating PFIC (e.g., PFIC-1 and PFIC-2) in a subject are provided, the methods including: performing an assay on a sample obtained from the subject to determine whether the subject has a PFIC-associated mutation (e.g., an ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b mutation); and administering (e.g., specifically or selectively administering) a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to the subject determined to have a PFIC-associated mutation. In some embodiments, the mutation is an ATP8B1 or ABCB11 mutation, such as a mutation as set forth in any one of Tables 1-4. In some embodiments, the ATP8B1 mutation is selected from L127P, G308V, T456M, D554N, F529del, I661T, E665X, R930X, R952X, R1014X, and G1040R. In some embodiments, the mutation in ABCB11 is selected from A167T, G238V, V284L, E297G, R470Q, R470X, D482G, R487H, A570T, N591S, A865V, G982R, R1153C, and R1268Q.

[0099] Also provided are methods for treating PFIC (e.g., PFIC-1 and PFIC-2) in a subject in need thereof, the methods comprising: (a) detecting a PFIC-associated mutation (e.g., an ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b mutation) in the subject; and (b) administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the method for treating PFIC may comprise administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject having a PFIC-associated mutation (e.g., an ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b mutation). In some embodiments, the mutation is an ATP8B1 or ABCB11 mutation, e.g., a mutation as set forth in any one of Tables 1-4. In some embodiments, the mutation in ATP8B1 is selected from L127P, G308V, T456M, D554N, F529del, I661T, E665X, R930X, R952X, R1014X, and G1040R. In some embodiments, the mutation in ABCB11 is selected from A167T, G238V, V284L, E297G, R470Q, R470X, D482G, R487H, A570T, N591S, A865V, G982R, R1153C, and R1268Q.

[0100] In some embodiments, a subject is determined to have a mutation associated with PFIC in the subject or in a biopsy specimen from the subject through the use of a test recognized by those skilled in the art, including next generation sequencing (NGS). In some embodiments, a subject is determined to have a mutation associated with PFIC using a regulatory agency-approved, e.g., FDA-approved, test or assay for identifying a mutation associated with PFIC in a subject or in a biopsy specimen from the subject, or by performing any of the non-limiting examples of assays described herein. Further methods for diagnosing PFIC are described in Gunaydin, M. et al., Hepat Med. 2018, Vol. 10, pp. 95-104 (incorporated herein by reference in its entirety).

[0101] In some embodiments, treatment of a PFIC (e.g., PFIC-1 or PFIC-2) reduces serum bile acid levels in a subject. In some embodiments, serum bile acid levels are determined, for example, by an ELISA enzyme assay or by an assay for measuring total bile acids such as that described in Danese et al., PLoS One. 2017, Vol. 12(6): e0179200, which is incorporated herein by reference in its entirety. In some embodiments, serum bile acid levels may be reduced by, for example, 10% to 40%, 20% to 50%, 30% to 60%, 40% to 70%, 50% to 80%, or greater than 90% of serum bile acid levels prior to administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, treatment of PFIC includes treatment of pruritus.

[0102] Because LBAT is expressed in hepatocytes, LBAT and dual ASBT / LBAT inhibitor substances should have at least some degree of bioavailability and free fraction in the blood. Because LBAT inhibitor compounds only need to survive from the intestine to the liver, relatively low systemic exposure of such compounds is expected to be sufficient, thereby minimizing the potential risk of any side effects elsewhere in the body. Inhibition of LBAT and ASBT is expected to have at least an additive effect on reducing bile acid concentrations in the liver. It is also expected that dual ASBT / LBAT inhibitors may be able to reduce bile acid levels without inducing diarrhea, as is sometimes observed with ASBT inhibitors.

[0103] Compounds with high LBAT inhibitory potency and sufficient bioavailability are expected to be particularly suitable for the treatment of hepatitis. Compounds with dual ASBT / LBAT inhibitory potency and sufficient bioavailability are expected to be particularly suitable for the treatment of non-alcoholic steatohepatitis (NASH).

[0104] NASH (nonalcoholic steatohepatitis, or NASH) is a common and serious chronic liver disease similar to alcoholic liver disease, but occurs in people who rarely or never drink alcohol. In patients with NASH, the accumulation of fat in the liver, known as nonalcoholic fatty liver disease (NAFLD) or steatosis, and other factors such as high LDL cholesterol and insulin resistance, can trigger chronic inflammation in the liver, leading to progressive scarring of tissue known as fibrosis, cirrhosis, and ultimately liver failure and death. Patients with NASH have been found to have significantly higher total serum bile acid concentrations than healthy controls at fasting and all postprandial time points (a 2.2- to 2.4-fold increase in NASH at fasting and a 1.7- to 2.2-fold increase in NASH at all postprandial time points). These are driven by increases in taurine- and glycine-conjugated primary and secondary bile acids. Patients with NASH exhibited significant variations in their fasting and postprandial bile acid profiles. These results indicate that patients with NASH are more exposed to bile acids, including more hydrophobic and cytotoxic secondary species, during fasting and after meals. Increased exposure to bile acids may contribute to liver injury and the pathogenesis of NAFLD and NASH (Ferslew et al., Dig Dis Sci. 2015, Vol. 60, pp. 3318-3328). Therefore, ASBT and / or LBAT inhibition may be beneficial for the treatment of NASH.

[0105] NAFLD is characterized by hepatic steatosis in the absence of secondary causes, including excessive alcohol consumption, other known liver diseases, or long-term use of steatogenic medications (Chalasani et al., Hepatology 2018, Vol. 67(1), pp. 328-357). NAFLD can be classified as nonalcoholic fatty liver disease (NAFL) and nonalcoholic steatohepatitis (NASH). According to Chalasani et al., NAFL is defined as the presence of ≥5% hepatic steatosis without evidence of hepatocellular injury in the form of hepatocyte ballooning. NASH is defined as the presence of ≥5% hepatic steatosis and inflammation with hepatocellular injury (e.g., ballooning), with or without any liver fibrosis. NASH is also generally associated with liver inflammation and fibrosis, which can progress to cirrhosis, end-stage liver disease, and hepatocellular carcinoma. Liver fibrosis is not always present in NASH, but when present, the severity of fibrosis may be related to long-term outcome.

[0106] There are many techniques used to evaluate and assess the severity of disease, including whether a subject has NAFLD and, if so, whether the NAFLD is NAFL or NASH. In some embodiments, the severity of NAFLD can be evaluated using the NAS. In some embodiments, treatment of NAFLD can be evaluated using the NAS. In some embodiments, the NAS can be determined as described in Kleiner et al., Hepatology. 2005, 41(6):1313-1321, which is incorporated herein by reference in its entirety. See, for example, Table 6 for a simplified NAS scheme adapted from Kleiner.

[0107] [Table 6]

[0108] In some embodiments, the NAS is determined noninvasively, for example, as described in U.S. Patent Application Publication No. 2018 / 0140219, which is incorporated herein by reference in its entirety. In some embodiments, the NAS is determined for a sample from a subject prior to administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the NAS is determined during or after a period of administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a lower NAS score during or after a period of administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof compared to before administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof indicates treatment of NAFLD (e.g., NASH). For example, a decrease in NAS of 1, 2, 3, 4, 5, 6, or 7 indicates treatment of NAFLD (e.g., NASH). In some embodiments, the NAS after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is 7 or less. In some embodiments, the NAS during administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the NAS during administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 7 or less. In some embodiments, the NAS during administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the NAS after administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 7 or less. In some embodiments, the NAS after administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is 5 or less, 4 or less, 3 or less, or 2 or less.

[0109] Further methods for evaluating or assessing NASH in a subject include determining one or more biomarkers (for example, serum markers and panels) that indicate one or more of hepatic steatosis (for example, fat accumulation in the liver); liver inflammation; liver damage, liver inflammation, liver fibrosis, and / or liver cirrhosis.Further examples of physiological indicators of NASH can include liver morphology, liver stiffness, and size or mass of the liver of the subject.In some embodiments, NASH in a subject is confirmed by detecting biomarkers that indicate hepatic fat accumulation and liver damage.For example, elevated serum ferritin and low titer of serum autoantibody can be common characteristics of NASH.

[0110] In some embodiments, methods for assessing NASH include magnetic resonance imaging (either by spectroscopy or proton density fat fraction measurement (MRI-PDFF)) to quantify steatosis, transient elastography (FIBROSCAN®) to diagnose significant liver fibrosis and / or cirrhosis, hepatic venous pressure gradient (HPVG), liver stiffness measurement on MRE, and evaluation of histological features of liver biopsy. In some embodiments, magnetic resonance imaging is used to detect one or more of steatohepatitis (NASH-MRI), liver fibrosis (Fibro-MRI), and steatosis. See, for example, U.S. Patent Application Publication Nos. 2016 / 146715 and 2005 / 0215882 (each of which is incorporated herein by reference in its entirety).

[0111] In some embodiments, treatment of NASH can include a reduction in one or more symptoms associated with NASH; a reduction in the amount of hepatic steatosis; a reduction in NAS; a reduction in liver inflammation; a reduction in levels of biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis; and a reduction in fibrosis and / or cirrhosis, a lack of further progression of fibrosis and / or cirrhosis, or a delay in the progression of fibrosis and / or cirrhosis in a subject following administration of one or more doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0112] In some embodiments, treating NASH includes reducing one or more symptoms associated with NASH in a subject. Exemplary symptoms may include one or more of: liver enlargement, fatigue, right upper quadrant pain, abdominal distension, enlarged blood vessels just below the surface of the skin, breast enlargement in men, spleen enlargement, palmar erythema, jaundice, and pruritus. In some embodiments, the subject is asymptomatic. In some embodiments, the subject's overall weight does not increase. In some embodiments, the subject's overall weight decreases. In some embodiments, the subject's body mass index (BMI) does not increase. In some embodiments, the subject's body mass index (BMI) decreases. In some embodiments, the subject's waist-to-hip ratio (WTH) does not increase. In some embodiments, the subject's waist-to-hip ratio (WTH) decreases.

[0113] In some embodiments, the treatment of NASH can be assessed by measuring hepatic steatosis. In some embodiments, the treatment of NASH comprises reducing hepatic steatosis after administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein. In some embodiments, hepatic steatosis is determined by one or more methods selected from the group consisting of ultrasonography, computed tomography (CT), magnetic resonance imaging, magnetic resonance spectroscopy (MRS), magnetic resonance elastography (MRE), transient elastography (TE) (e.g., FIBROSCAN®), measurement of liver size or mass, or by liver biopsy (see, e.g., Di Lascio et al., Ultrasound Med Biol. 2018, 44(8), 1585-1596; Lv et al., J Clin Transl Hepatol. 2018, 6(2), 217-221; Reeder et al., J Magn Reson Imaging. 2011, 34(4), spcone; and de Ledinghen V et al., J Gastroenterol Hepatol. 2016, 31(4), pp. 848-855 (each of which is incorporated herein by reference in its entirety). A subject diagnosed with NASH may have greater than about 5% hepatic steatosis, e.g., greater than about 5% to about 25%, about 25% to about 45%, about 45% to about 65%, or greater than about 65% hepatic steatosis. In some embodiments, a subject with greater than about 5% to about 33% hepatic steatosis has stage 1 hepatic steatosis, a subject with about 33% to about 66% hepatic steatosis has stage 2 hepatic steatosis, and a subject with greater than about 66% hepatic steatosis has stage 3 hepatic steatosis.

[0114] In some embodiments, the amount of hepatic steatosis is determined before administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the amount of hepatic steatosis is determined during or after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a reduction in the amount of hepatic steatosis during or after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof compared to before administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof indicates treatment of NASH. For example, a reduction in the amount of hepatic steatosis of about 1% to about 50%, about 25% to about 75%, or about 50% to about 100% indicates treatment of NASH. In some embodiments, a reduction in the amount of hepatic steatosis by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% indicates treatment of NASH.

[0115] In some embodiments, the presence of liver inflammation is determined by one or more methods selected from the group consisting of biomarkers indicative of liver inflammation and liver biopsy samples from the subject. In some embodiments, the severity of liver inflammation is determined from liver biopsy samples from the subject. For example, liver inflammation in liver biopsy specimens can be assessed as described in Kleiner et al., Hepatology 2005, Vol. 41(6), pp. 1313-1321, and Brunt et al., Am J Gastroenterol 1999, Vol. 94, pp. 2467-2474 (each of which is incorporated herein by reference in its entirety). In some embodiments, the severity of liver inflammation is determined before administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the severity of liver inflammation is determined during or after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a reduction in the severity of liver inflammation during or after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof compared to before administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof indicates treatment of NASH. For example, a reduction in the severity of liver inflammation of about 1% to about 50%, about 25% to about 75%, or about 50% to about 100% indicates treatment of NASH. In some embodiments, a reduction in the severity of liver inflammation of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% indicates treatment of NASH.

[0116] In some embodiments, treating NASH includes treating fibrosis and / or cirrhosis, e.g., reducing the severity of fibrosis, preventing further progression of fibrosis and / or cirrhosis, or delaying the progression of fibrosis and / or cirrhosis. In some embodiments, the presence of fibrosis and / or cirrhosis is determined by one or more methods selected from the group consisting of transient elastography (e.g., FIBROSCAN®), non-invasive markers of liver fibrosis, and histological features of liver biopsies. In some embodiments, the severity (e.g., stage) of fibrosis is determined by one or more methods selected from the group consisting of transient elastography (e.g., FIBROSCAN®), a fibrosis-scoring system, a biomarker of liver fibrosis (e.g., a non-invasive biomarker), and hepatic venous pressure gradient (HVPG). Non-limiting examples of fibrosis scoring systems include the NAFLD fibrosis scoring system (see, e.g., Angulo et al., Hepatology 2007, Vol. 45(4), pp. 846-54), the fibrosis scoring system of Brunt et al., Am. J. Gastroenterol. 1999, Vol. 94, pp. 2467-2474, the fibrosis scoring system of Kleiner et al., Hepatology 2005, Vol. 41(6), pp. 1313-1321, and the Ishak fibrosis scoring system (see, Ishak et al., J. Hepatol. 1995, Vol. 22, pp. 696-699) (the contents of each of which are incorporated herein by reference in their entirety).

[0117] In some embodiments, the severity of fibrosis is determined before administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the severity of fibrosis is determined during or after administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a decrease in the severity of fibrosis during or after administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof compared to before administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof indicates treatment of NASH. In some embodiments, a decrease in the severity of fibrosis, a lack of further progression of fibrosis and / or cirrhosis, or a delay in the progression of fibrosis and / or cirrhosis indicates treatment of NASH. In some embodiments, the severity of fibrosis is determined using a scoring system, such as any of the fibrosis scoring systems described herein; for example, the score can indicate the stage of fibrosis, for example, stage 0 (no fibrosis), stage 1, stage 2, stage 3, and stage 4 (cirrhosis) (see, e.g., Kleiner et al.). In some embodiments, a reduction in the stage of fibrosis is a reduction in the severity of fibrosis. For example, a reduction of 1, 2, 3, or 4 stages is a reduction in the severity of fibrosis. In some embodiments, a reduction in stage, e.g., from stage 4 to stage 3, from stage 4 to stage 2, from stage 4 to stage 1, from stage 4 to stage 0, from stage 3 to stage 2, from stage 3 to stage 1, from stage 3 to stage 0, from stage 2 to stage 1, from stage 2 to stage 0, or from stage 1 to stage 0, indicates treatment of NASH. In some embodiments, the stage of fibrosis is reduced from stage 4 to stage 3, from stage 4 to stage 2, from stage 4 to stage 1, from stage 4 to stage 0, from stage 3 to stage 2, from stage 3 to stage 1, from stage 3 to stage 0, from stage 2 to stage 1, from stage 2 to stage 0, or from stage 1 to stage 0 after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, compared to before administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments,The stage of fibrosis is reduced from Stage 4 to Stage 3, Stage 4 to Stage 2, Stage 4 to Stage 1, Stage 4 to Stage 0, Stage 3 to Stage 2, Stage 3 to Stage 1, Stage 3 to Stage 0, Stage 2 to Stage 1, Stage 2 to Stage 0, or Stage 1 to Stage 0 during the period of administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, compared to before administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the stage of fibrosis is reduced from Stage 4 to Stage 3, Stage 4 to Stage 2, Stage 4 to Stage 1, Stage 4 to Stage 0, Stage 3 to Stage 2, Stage 3 to Stage 1, Stage 3 to Stage 0, Stage 2 to Stage 1, Stage 2 to Stage 0, or Stage 1 to Stage 0 after the period of administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, compared to before administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0118] In some embodiments, the presence of NASH is determined by one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis, or a scoring system thereof. In some embodiments, the severity of NASH is determined by one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis, or a scoring system thereof. The level of a biomarker can be determined, for example, by measuring, quantifying, and monitoring the expression level of the gene or mRNA encoding the biomarker and / or the peptide or protein of the biomarker.Non-limiting examples of biomarkers indicative of one or more liver damage, inflammation, liver fibrosis, and / or cirrhosis, and / or scoring systems thereof, include the aspartate aminotransferase (AST) to platelet ratio index (APRI); the aspartate aminotransferase (AST) to alanine aminotransferase (ALT) ratio (AAR); the FIB-4 score (see, e.g., McPherson et al., Gut 2010, Vol. 59(9), pp. 1265-9 (which is incorporated herein by reference in its entirety); hyaluronic acid; pro-inflammatory cytokines; a panel of biomarkers consisting of α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma glutamyl transpeptidase (GGT) combined with a subject's age and sex to generate a measure of fibrotic and necroinflammatory activity in the liver (e.g., FIBROTEST®, FIBROSURE®), a panel of biomarkers consisting of bilirubin, gamma glutamyl transferase, hyaluronic acid, α2-macroglobulin combined with a subject's age and sex (e.g., HEPASCORE®; see, e.g., Adams et al., Clin. Chem. 2005, 51(10), 1867-1873), and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1, hyaluronic acid, and α2-macroglobulin (e.g., FIBROSPECT®); a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1 (TIMP-1), amino-terminal propeptide of type III procollagen (PIIINP), and hyaluronic acid (HA) (e.g., Enhanced Liver Fibrosis (ELF) score, see, e.g., Lichtinghagen R et al., J Hepatol. 2013 Aug;59(2):236-42, which is incorporated herein by reference in its entirety).In some embodiments, the presence of fibrosis is determined by measuring the FIB-4 score, a panel of biomarkers consisting of α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma glutamyl transpeptidase (GGT) in combination with the subject's age and sex to generate a measure of fibrosis and necroinflammatory activity in the liver (e.g., FIBROTEST®, FIBROSURE®), a panel of biomarkers consisting of bilirubin, gamma glutamyl transferase, hyaluronic acid, α2-macroglobulin in combination with the subject's age and sex (e.g., HEPASCORE®; see, e.g., Adams et al., Clin. Chem. 2005, 51(10), pp. 1867-1873), and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase-1, hyaluronic acid, and α2-macroglobulin (e.g., FIBROSPECT®); and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase-1 (TIMP-1), amino-terminal propeptide of type III procollagen (PIIINP), and hyaluronic acid (HA) (e.g., Enhanced Liver Fibrosis (ELF) score). In some embodiments, the level of aspartate aminotransferase (AST) is not increased. In some embodiments, the level of aspartate aminotransferase (AST) is decreased. In some embodiments, the level of alanine aminotransferase (ALT) is not increased. In some embodiments, the level of alanine aminotransferase (ALT) is decreased. In some embodiments, the "level" of an enzyme refers to the concentration of the enzyme, e.g., the concentration in the blood. For example, AST or ALT levels can be expressed as units / L.

[0119] In some embodiments, the severity of fibrosis is assessed using a panel of biomarkers consisting of α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, and gamma glutamyl transpeptidase (GGT) combined with the subject's age and sex to generate the FIB-4 score, a measure of fibrosis and necroinflammatory activity in the liver (e.g., FIBROTEST®, FIBROSURE®); a panel of biomarkers consisting of bilirubin, gamma glutamyl transferase, hyaluronic acid, and α2-macroglobulin combined with the subject's age and sex (e.g., HEPASCORE®; see, e.g., Adams et al., Clin. Chem. 2005, 51(10), pp. 1867-1873), which is incorporated herein by reference in its entirety, and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase-1, hyaluronic acid, and α2-macroglobulin (e.g., FIBROSPECT®); and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase-1 (TIMP-1), amino-terminal propeptide of type III procollagen (PIIINP), and hyaluronic acid (HA) (e.g., Enhanced Liver Fibrosis (ELF) score).

[0120] In some embodiments, liver inflammation is determined by measuring the level of liver inflammation biomarkers, for example, the level of pro-inflammatory cytokines. Non-limiting examples of biomarkers indicating liver inflammation include interleukin-(IL)6, interleukin-(IL)1β, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, monocyte chemoattractant protein (MCP)-1, C-reactive protein (CRP), PAI-1, and collagen isoforms, such as Col1a1, Col1a2, and Col4a1 (see, for example, Neuman et al., Can. J. Gastroenterol. Hepatol. 2014, Vol. 28(11), pp. 607-618, and U.S. Patent No. 9,872,844, each of which is incorporated herein by reference in its entirety). Liver inflammation can also be assessed by measuring changes in macrophage infiltration, for example, changes in CD68 expression levels. In some embodiments, liver inflammation can be determined by measuring or monitoring serum or circulating levels of one or more of interleukin-(IL)6, interleukin-(IL)1β, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, monocyte chemoattractant protein (MCP)-1, and C-reactive protein (CRP).

[0121] In some embodiments, the level of one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis is determined in a sample from the subject before administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the level of one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis is determined during or after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a decrease in the level of one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis during or after administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof compared to before administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof indicates treatment of NASH. For example, a decrease of at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% in the level of one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis indicates treatment of NASH. In some embodiments, the reduction in the level of one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis following administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99%.In some embodiments, the reduction in the level of one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis during administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99%. In some embodiments, the reduction in the level of one or more biomarkers indicative of one or more of liver damage, inflammation, liver fibrosis, and / or cirrhosis after a period of administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99%.

[0122] In some embodiments, treating NASH reduces serum bile acid levels in a subject. In some embodiments, serum bile acid levels are determined, for example, by an ELISA enzyme assay or a total bile acid measurement assay such as that described in Danese et al., PLoS One. 2017, Vol. 12(6): e0179200, which is incorporated herein by reference in its entirety. In some embodiments, serum bile acid levels can be reduced by, for example, 10% to 40%, 20% to 50%, 30% to 60%, 40% to 70%, 50% to 80%, or more than 90% of the serum bile acid levels prior to administration of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the NASH is NASH with accompanying cholestasis. Cholestasis blocks the release of bile, including bile acids, from the liver. Bile acids can cause hepatocellular damage (see, e.g., Perez MJ, Briz O. World J. Gastroenterol. 2009, vol. 15(14), pp. 1677-1689), which can lead to or increase the development of fibrosis (e.g., cirrhosis) and increase the risk of hepatocellular carcinoma (see, e.g., Sorrentino P et al., Dig. Dis. Sci. 2005, vol. 50(6), pp. 1130-1135, and Satapathy SK and Sanyal AJ. Semin. Liver Dis. 2015, vol. 35(3), pp. 221-235, each of which is incorporated herein by reference in its entirety). In some embodiments, the treatment of NASH includes treatment of pruritus. In some embodiments, the treatment of NASH with associated cholestasis includes treatment of pruritus. In some embodiments, subjects with NASH with associated cholestasis have pruritus.

[0123] Exemplary biomarkers for NASH are shown in Table 7.

[0124] [Table 7]

[0125] References for Table 7 1 McPherson et al., Gut. 2010, vol. 59(9), p. 1265-1269. 2 Adams, et al. Clin Chem. 2005, vol. 51(10), p. 1867-1873. 3 Lichtinghagen, et al. J Hepatol. 2013, vol. 59(2), p. 236-242. 4 Neuman, et al. Can J Gastroenterol Hepatol. 2014, vol. 28(11), p. 607-618. 5 US Patent No. 9,872,844

[0126] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may exhibit a higher free fraction in plasma. In some embodiments, the free fraction is greater than about 0.2%, such as greater than about 0.4%, for example greater than about 0.6%, for example greater than about 0.8%, for example greater than about 1.0%, for example greater than about 1.25%, for example greater than about 1.5%, for example greater than about 1.75%, for example greater than about 2.0%, for example greater than about 2.5%, for example greater than about 3%, for example greater than about 4%, for example greater than about 5%, for example greater than about 7.5%, for example greater than about 10%, or for example greater than about 20%.

[0127] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may be excreted in the urine, in some embodiments, the fraction of the compound excreted in the urine is greater than about 0.2%, such as greater than about 0.4%, for example greater than about 0.6%, for example greater than about 0.8%, such as greater than about 1.0%, for example greater than about 2%, such as greater than about 3%, for example greater than about 5%, such as greater than about 7.5%, for example greater than about 10%, such as greater than about 15%, for example greater than about 20%, for example greater than about 30%, or for example greater than about 50%.

[0128] After absorption from the intestine, some compounds of formula (I) or pharmaceutically acceptable salts thereof can circulate through the enterohepatic circulation. In some embodiments, the fraction of the compound circulating through the enterohepatic circulation is greater than about 0.1%, such as greater than about 0.2%, for example greater than about 0.3%, for example greater than about 0.5%, for example greater than about 1.0%, for example greater than about 1.5%, for example greater than about 2%, for example greater than about 3%, for example greater than about 5%, for example greater than about 7%, for example greater than about 10%, for example greater than about 15%, for example greater than about 20%, for example greater than about 30%, or for example greater than about 50%.

[0129] Some compounds of Formula (I) or pharmaceutically acceptable salts thereof may cause renal excretion of bile salts, in some embodiments, the fraction of circulating bile acids excreted via the renal route is greater than about 1%, such as greater than about 2%, for example greater than about 5%, for example greater than about 7%, for example greater than about 10%, for example greater than about 15%, for example greater than about 20%, or for example greater than about 25%.

[0130] Some compounds of Formula (I) or pharmaceutically acceptable salts thereof may exhibit improved or optimal permeability. Permeability can be measured in Caco2 cells, and values ​​are expressed as Papp (apparent permeability) values ​​in cm / s. In some embodiments, the permeability is at least about 0.1 x 10 -6 cm / s, e.g., about 0.2 × 10 -6 cm / s, e.g., about 0.4 × 10 -6 cm / s, e.g., about 0.7 × 10 -6 cm / s, e.g., about 1.0 × 10 -6 cm / s, e.g., about 2×10 -6 cm / s, e.g., about 3 × 10 -6 cm / s, e.g., about 5 × 10 -6 cm / s, e.g., about 7×10 -6 cm / s, e.g., about 10 × 10 -6 cm / s, e.g., about 15 × 10 -6 Over cm / s.

[0131] Some compounds of Formula (I) or pharmaceutically acceptable salts thereof may exhibit improved or optimal bioavailability. In some embodiments, the oral bioavailability is greater than about 5%, e.g., greater than about 7%, e.g., greater than about 10%, e.g., greater than about 15%, e.g., greater than about 20%, e.g., greater than about 30%, e.g., greater than about 40%, e.g., greater than about 50%, e.g., greater than about 60%, e.g., greater than about 70%, or e.g., greater than about 80%. In other embodiments, the oral bioavailability is between about 10% and about 90%, e.g., between about 20% and about 80%, e.g., between about 30% and about 70%, or e.g., between about 40% and about 60%.

[0132] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may be substrates for relevant transporters in the kidney.

[0133] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may produce concentrations of bile acids in the intestine, liver and serum that do not cause adverse gastrointestinal effects.

[0134] Some compounds of formula (I) or pharmaceutically acceptable salts thereof can reduce the concentration of bile acids in the liver without causing gastrointestinal disorders such as diarrhea.

[0135] As used herein, the terms "treatment," "treat," and "treating" refer to the regression, alleviation, delay in onset, or inhibition of progression of a disease or disorder as described herein, or one or more symptoms thereof. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual (e.g., taking into account a history of the condition and / or taking into account genetic or other predisposing factors) before the onset of symptoms. Treatment may also be continued after symptoms have resolved, e.g., to prevent or delay their recurrence.

[0136] Suitable pharmaceutically acceptable salts of the compounds of the present invention are, for example, base addition salts of compounds of the present invention that are sufficiently acidic, such as alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., calcium or magnesium salts), ammonium salts, or salts with organic bases that provide physiologically acceptable cations, such as salts with methylamine, dimethylamine, trimethylamine, piperidine, morpholine, or tris-(2-hydroxyethyl)amine.

[0137] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may have chiral centers and / or centers of geometric isomeric form (E- and Z-isomers). It should be understood that the present invention encompasses all such optical isomers, diastereoisomers, and geometric isomers that possess ASBT and / or LBAT inhibitory activity. The present invention also encompasses any and all tautomers of compounds of formula (I) or pharmaceutically acceptable salts thereof that possess ASBT and / or LBAT inhibitory activity. Certain compounds of formula (I) or pharmaceutically acceptable salts thereof may exist in solvated forms, such as, for example, hydrated forms, as well as unsolvated forms. It should be understood that the present invention encompasses all such solvated forms that possess ASBT and / or LBAT inhibitory activity.

[0138] In another aspect, the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. The excipients may include, for example, fillers, binders, disintegrants, glidants, and lubricants. In general, the pharmaceutical composition may be prepared in a conventional manner using conventional excipients.

[0139] Examples of suitable fillers include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose (such as lactose monohydrate), sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, dry starch, hydrolyzed starch, and pregelatinized starch.

[0140] Examples of suitable binders include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (such as sucrose, glucose, dextrose, lactose, and sorbitol), polyethylene glycol, waxes, natural and synthetic gums (such as gum arabic and gum tragacanth), sodium alginate, cellulose derivatives (such as hydroxypropyl methylcellulose (or hypromellose), hydroxypropyl cellulose, and ethyl cellulose), and synthetic polymers (such as acrylic acid and methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid / polymethacrylic acid copolymers, and polyvinylpyrrolidone (povidone)).

[0141] Examples of suitable disintegrants include, but are not limited to, dry starch, modified starches (such as (partially) pregelatinized starch, sodium starch glycolate, and sodium carboxymethyl starch), alginic acid, cellulose derivatives (such as sodium carboxymethylcellulose, hydroxypropylcellulose, and low-substituted hydroxypropylcellulose (L-HPC)), and cross-linked polymers (such as carmellose, croscarmellose sodium, carmellose calcium, and cross-linked PVP (crospovidone)).

[0142] Examples of suitable glidants and lubricants include, but are not limited to, talc, magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, colloidal silica, aqueous silicon dioxide, synthetic magnesium silicate, micronized silicon oxide, starch, sodium lauryl sulfate, boric acid, magnesium oxide, waxes (such as carnauba wax), hydrogenated oils, polyethylene glycol, sodium benzoate, polyethylene glycol, and mineral oil.

[0143] The pharmaceutical composition may be conventionally coated with one or more coating layers. An enteric coating layer or a coating layer for delayed or targeted release of the compound of formula (I) or a pharmaceutically acceptable salt thereof is also contemplated. The coating layer may include one or more coating agents, and may optionally include a plasticizer and / or a pigment (or colorant).

[0144] Examples of suitable coating agents include, but are not limited to, cellulosic polymers (such as ethyl cellulose, hydroxypropyl methylcellulose (or hypromellose), hydroxypropyl cellulose, cellulose acetate phthalate, cellulose acetate succinate, hydroxypropyl methylcellulose acetate succinate, and hydroxypropyl methylcellulose phthalate), vinyl polymers (such as polyvinyl alcohol), and polymers based on acrylic acid and its derivatives (such as copolymers of acrylic acid and methacrylic acid, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid / polymethacrylic acid copolymers, etc.).

[0145] Examples of suitable plasticizers include, but are not limited to, triethyl citrate, glyceryl triacetate, tributyl citrate, diethyl phthalate, acetyl tributyl citrate, dibutyl phthalate, dibutyl sebacate, and polyethylene glycol.

[0146] Examples of suitable pigments include, but are not limited to, titanium dioxide, iron oxide (such as yellow, brown, red, or black iron oxide), and barium sulfate.

[0147] The pharmaceutical composition may be in a form suitable for oral administration, injection (including intravenous, subcutaneous, intramuscular, and intravascular injection), topical administration for rectal administration, hi a preferred embodiment, the pharmaceutical composition is in a form suitable for oral administration, such as a tablet or capsule.

[0148] The dosage required for therapeutic or prophylactic treatment will depend on the route of administration, the severity of the disease, the age and weight of the patient, and other factors normally considered by the attending physician when determining the regimen and dosage level appropriate for a particular patient.

[0149] The amount of compound to be administered will vary depending on the patient being treated and can range from about 1 μg / kg body weight to about 50 mg / kg body weight per day. A unit dosage form, such as a tablet or capsule, will typically contain about 1 to about 250 mg of active ingredient, e.g., about 1 to about 100 mg, or, e.g., about 1 to about 50 mg, or, e.g., about 1 to about 20 mg, e.g., about 2.5 mg, or about 5 mg, or about 10 mg, or about 15 mg. The daily dose can be administered as a single dose or divided into one, two, three, or more unit doses. The daily dose of an orally administered bile acid modulator is preferably about 0.1 to about 250 mg, more preferably about 1 to about 100 mg, e.g., about 1 to about 5 mg, e.g., about 1 to about 10 mg, e.g., about 1 to about 15 mg, or, e.g., about 1 to about 20 mg.

[0150] In another aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use as a medicament. The present invention also relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as a medicament.

[0151] In another aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of any of the diseases described herein. The present invention also relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing any of the diseases described herein. The present invention also relates to a method of treating or preventing any of the diseases described herein in a subject, such as a human, comprising administering to a subject in need of such treatment or prevention a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0152] Combination therapy In one aspect of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with at least one other therapeutically active agent, for example, one, two, three, or more other therapeutically active agents. The compound of formula (I) or a pharmaceutically acceptable salt thereof and the at least one other therapeutically active agent may be administered simultaneously, sequentially, or separately. Therapeutically active agents suitable for combination with the compound of formula (I) include, but are not limited to, known active agents useful for treating any of the above-mentioned conditions, disorders, and diseases.

[0153] In one embodiment, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with another ASBT inhibitor. Suitable ASBT inhibitors include WO 93 / 16055, WO 94 / 18183, WO 94 / 18184, WO 96 / 05188, WO 96 / 08484, WO 96 / 16051, WO 97 / 33882, WO 98 / 03818, WO 98 / 07449, WO 98 / 40375, WO 99 / 35135, WO 99 / 64409, WO 99 / 64410, WO 00 / 47568, WO 00 / 61568, WO 00 / 38725, WO 00 / 38726, WO 00 / 38727, WO 00 / 38728, WO 00 / 38729, WO 01 / 66533, WO 01 / 68096, WO 02 / 32428, WO 02 / 50051, WO 03 / 020710, WO 03 / 022286, WO 03 / 022825, WO 03 / 022830, WO 03 / 061663, WO 03 / 091232, WO 03 / 106482, WO 2004 / 006899, WO 2004 / 076430, WO 2007 / 009655, WO 2007 / 009656, WO 2011 / 137135, WO 2019 / 234077, WO 2020 / 161216, WO 2020 / 161217, WO 2021 / 110883, WO 2021 / 110884, WO 2021 / 110885, WO 2021 / 110886, WO 2021 / 110887, DE 19825804, EP 864582, EP 489423, EP 549967, EP 573848, EP 624593, EP 624594, EP 624595, EP 624596, EP 0864582, EP 1173205, EP 1535913, and EP 3210977 (all of which are incorporated herein by reference in their entirety).Specific examples of suitable ASBT inhibitors include 1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-1'-phenyl-1'-[N'-(carboxymethyl)carbamoyl]methyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine (elobixibat) and 1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxypropyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine (odebixibat).

[0154] In another embodiment, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a bile acid binder (also called a bile acid sequestrant or resin), such as colesevelam, cholestyramine, or colestipol. In a preferred embodiment of such a combination, the bile acid binder is formulated for colonic release. Examples of such formulations are disclosed, for example, in WO 2017 / 138877, WO 2017 / 138878, WO 2019 / 032026, and WO 2019 / 032027 (all of which are incorporated herein by reference in their entireties).

[0155] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a DPP-IV inhibitor, such as a gliptin, for example, sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, and dutogliptin, or a pharmaceutically acceptable salt thereof.

[0156] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an HMG CoA reductase inhibitor, such as fluvastatin, lovastatin, pravastatin, simvastatin, atorvastatin, pitavastatin, cerivastatin, mevastatin, rosuvastatin, bervastatin, or dalvastatin, or a pharmaceutically acceptable salt thereof.

[0157] In another embodiment, the compounds of formula (I) or a pharmaceutically acceptable salt thereof are administered in combination with a cholesterol absorption inhibitor, for example, ezetimibe or a pharmaceutically acceptable salt thereof.

[0158] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a PPARα agonist, such as a fibrate, for example, clofibrate, bezafibrate, ciprofibrate, clinofibrate, clofibrate, fenofibrate, gemfibrozil, lonifibrate, and simfribrate, or a pharmaceutically acceptable salt thereof.

[0159] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a PPARγ agonist, for example a thiazolidinedione, for example pioglitazone, rosiglitazone, and lobeglitazone, or a pharmaceutically acceptable salt thereof.

[0160] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a dual PPAR α / γ agonist, such as a glitazar, e.g., saroglitazar, aleglitazar, muraglitazar, or tesaglitazar, or a pharmaceutically acceptable salt thereof.

[0161] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a dual PPARα / δ agonist, such as elafibranor.

[0162] In yet another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a pan-PPAR agonist (i.e., a PPAR agonist with activity across all subtypes: alpha, gamma, and delta), such as IVA337.

[0163] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a farnesoid X receptor (FXR) modulator, e.g., an FXR agonist, such as cafestol, chenodeoxycholic acid, 6α-ethyl-chenodeoxycholic acid (obeticholic acid; INT-747), fexaramine, tropifexor, cilofexor, and MET409.

[0164] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a TGR5 receptor modulator, for example, a TGR5 agonist, for example, 6α-ethyl-23(S)-methylcholic acid (INT-777).

[0165] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a dual FXR / TGR5 agonist, such as INT-767.

[0166] In another embodiment, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with ursodeoxycholic acid (UDCA). In yet another embodiment, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with norursodeoxycholic acid (norUDCA).

[0167] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an FGF19 modulator, such as NGM282.

[0168] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an FGF21 agonist, for example, BMS-986036.

[0169] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an integrin inhibitor, such as PLN-74809 and PLN-1474.

[0170] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a CCR2 / CCR5 inhibitor, such as cenicriviroc.

[0171] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a caspase protease inhibitor, such as emricasan.

[0172] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a galectin-3 inhibitor, such as GR-MD-02.

[0173] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a stearoyl-CoA desaturase (SCD) inhibitor, for example, aramchol (arachidylamide cholanic acid).

[0174] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an apoptosis signal-regulating kinase 1 (ASK1) inhibitor, such as selonsertib.

[0175] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a LOXL2 inhibitor, for example, simtuzumab.

[0176] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an ACC inhibitor, such as GS-0976.

[0177] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a thyroid hormone receptor beta agonist, such as MGL3196.

[0178] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a GLP-1 agonist, such as liraglutide.

[0179] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a glucagon-like peptide and a dual agonist of the glucagon receptor, such as SAR425899.

[0180] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a mitochondrial pyruvate transporter inhibitor, such as MSDC-0602K.

[0181] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an antioxidant, for example, vitamin E.

[0182] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an SGLT1 inhibitor, an SGLT2 inhibitor, or a dual inhibitor of SGLT1 and SGLT2. Examples of such compounds are dapagliflozin, sotagliflozin, canagliflozin, empagliflozin, LIK066, and SGL5213.

[0183] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a diacylglycerol O-acyltransferase 2 (DGAT2) inhibitor, such as DGAT2RX and PF-06865571.

[0184] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a fatty acid synthase (FASN) inhibitor, such as TVB-2640.

[0185] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an AMP-activated protein kinase (AMPK) activator, such as PXL-770.

[0186] In another embodiment, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a glucocorticoid receptor antagonist (GR), a mineralocorticoid receptor antagonist (MR), or a dual GR / MR antagonist. Examples of such compounds are MT-3995 and CORT-118335.

[0187] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a cannabinoid receptor 1 (CB1) antagonist, such as IM102.

[0188] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with Klotho-beta (KLB) and a fibroblast growth factor receptor (FGFR) activator, such as MK-3655 (formerly known as NGM-313).

[0189] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a chemokine (cc motif) ligand 24 (CCL24) inhibitor, such as CM101.

[0190] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an A3 antagonist, such as PBF-1650.

[0191] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a P2x7 receptor antagonist, for example SGM 1019.

[0192] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a P2Y13 receptor agonist, such as CER-209.

[0193] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a sulfated oxysterol, for example, Dur-928.

[0194] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a leukotriene D4 (LTD4) receptor antagonist, such as MN-001.

[0195] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a type 1 natural killer T cell (NKT1) inhibitor, such as GRI-0621.

[0196] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an anti-lipopolysaccharide (LPS) compound, such as IMM-124E.

[0197] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a VAP1 inhibitor, such as, for example, BI1467335.

[0198] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an A3 adenosine receptor agonist, such as CF-102.

[0199] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a SIRT-1 activator, such as NS-20.

[0200] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a nicotinic acid receptor 1 agonist, such as ARI-3037MO.

[0201] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a TLR4 antagonist, for example JKB-121.

[0202] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a ketohexokinase inhibitor, such as PF-06835919.

[0203] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an adiponectin receptor agonist, such as ADP-335.

[0204] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an autotaxin inhibitor, such as PAT-505 and PF8380.

[0205] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a chemokine (cc motif) receptor 3 (CCR3) antagonist, such as bertilimumab.

[0206] In another embodiment, the compounds of formula (I) or pharmaceutically acceptable salts thereof are administered in combination with chloride channel agonists such as cobiprostone and lubiprostone.

[0207] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a heat shock protein 47 (HSP47) inhibitor, such as ND-L02-s0201.

[0208] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a sterol regulatory region binding protein (SREBP) transcription factor inhibitor, such as CAT-2003 and MDV-4463.

[0209] In another embodiment, the compounds of formula (I) or a pharmaceutically acceptable salt thereof are administered in combination with a biguanidine, such as metformin.

[0210] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with insulin.

[0211] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a glycogen phosphorylase inhibitor and / or a glucose-6-phosphatase inhibitor.

[0212] In another embodiment, the compounds of formula (I) or pharmaceutically acceptable salts thereof are administered in combination with a sulfonylurea such as glipizide, glibenclamide, and glimepiride.

[0213] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a meglitinide, such as repaglinide, nateglinide, and ormiglitinide.

[0214] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a glucosidase inhibitor such as, for example, acarbose or miglitol.

[0215] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a squalene synthase inhibitor, such as TAK-475.

[0216] In another embodiment, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a PTPB1 inhibitor such as trodusquemine, eltiprotafib, JTT-551, and claramine.

[0217] Preparation of compounds The compounds of the present invention can be prepared as free acids or pharmaceutically acceptable salts thereof by the processes described below. Throughout the following descriptions of such processes, it will be understood that suitable protecting groups will be added, where appropriate, and subsequently removed from various reactants and intermediates in a manner readily understood by one skilled in the art of organic synthesis. Conventional procedures for using such protecting groups, and examples of suitable protecting groups, can be found, for example, in Greene's Protective Groups in Organic Synthesis, 4th Edition, by PGM Wutz and TW Greene, John Wiley & Sons, Hoboken, 2006.

[0218] General method All solvents used were of analytical grade. Commercially available anhydrous solvents were routinely used in the reactions. Starting materials were either available from commercial sources or prepared according to literature procedures. 3,3-Dibutyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 8-hydroxy-7-(methylthio)-5-phenyl-3,3-dipropyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide may be prepared as described in WO 02 / 50051 (Methods 26 and 117, respectively). 7-Bromo-3,3-diethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3,3-diethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide may be prepared as described in WO 2019 / 234077 (intermediates 56 and 60, respectively). Room temperature refers to 20-25° C. The composition of the solvent mixture is given as a volume percentage or volume ratio.

[0219] LCMS: Instrument name: Agilent 1290 infinity II. Method A: Mobile phase: A: 0.1% HCOOH:ACN (95:5) in H2O, B:ACN; Flow rate: 1.5 mL / min; Cell: ZORBAX XDB C-18 (50×4.6 mm), 3.5 μm. Method B: Mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; flow rate: 1.2 mL / min; cartridge: XBridge C8 (50×4.6 mm), 3.5 μm. Method C: Mobile phase: A: 0.1% HCOOH:ACN (95:5) in water, B:ACN; Flow rate: 1.5 mL / min; Cell: ATLANTIS dC18 (50×4.6 mm), 5 μm. Method D: Mobile phase: A: 10 mM NH4OAc in water, B: ACN; flow rate: 1.2 mL / min; cartridge: Zorbax Extend C18 (50×4.6 mm), 5 μm. Method E: Mobile phase: A: 0.1% TFA:ACN in water (95:5), B: 0.1% TFA in ACN; Flow rate: 1.5 mL / min; Cell: XBridge C8 (50×4.6 mm), 3.5 μm. Method F: Mobile phase: A: 0.1% TFA in water, B: 0.1% TFA in ACN; flow rate: 0.8 mL / min; cell: ZORBAX ECLIPSE PLUS C18 (50×2.1 mm), 1.8 μm. Method G: Mobile phase: A: 0.1% TFA in water, B: 0.1% TFA in ACN; flow rate: 0.8 mL / min; cell: Acquity UPLC BEH C18 (2.1×50 mm), 1.7 μm. Method H: Mobile phase: A: 10 mM NH4OAc, B: 100% ACN; flow rate: 0.8 mL / min; cell: Acquity UPLC BEH C18 (2.1×50) mm; 1.7 μm. Method I: Mobile phase: A: 0.1% HCOOH:ACN (95:5) in water, B:ACN; Flow rate: 0.8 mL / min; Cell: ZORBAX ECLIPSE PLUS C18 (2.1×50) mm, 1.8 μm. Method J: Mobile phase: A: 0.1% TFA in water, B: ACN; Flow rate: 1.0 mL / min; Cell: Zorbax Extend C18 (50×4.6 mm), 5 μM. Method K: Mobile phase: A: 0.1% TFA in water, B: 0.1% TFA in ACN; flow rate: 1.5 mL / min; column: XBridge C8 (50 x 4.6 mm), 3.5 μM. Method L: Mobile phase: A: 0.1% TFA in water, B: 100% ACN; flow rate: 1.5 mL / min; column: XBridge C8 (50 x 4.6 mm), 3.5 μM.

[0220] UPLC: Equipment name: Waters Acquity I Class Method A: Mobile phase: A: 0.1% HCOOH in water, B: 0.1% HCOOH in ACN; flow rate: 0.8 mL / min; column: Acquity UPLC HSS T3 (2.1 × 50) mm; 1.8 μm.

[0221] Device name: Shimadzu Nexera X2 LC / 2020 MSD Method B: Mobile phase: A: 0.1% HCOOH in water, B: ACN; flow rate: 0.8 mL / min; column: Acquity UPLC BEH C18 (2.1 × 50) mm; 1.7 μm.

[0222] HPLC: Instrument name: Agilent 1260 Infinity II series instrument (using % with UV detection (max plot) as follows). Method A: Mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; Flow rate: 1.0 mL / min; Column: XBridge C8 (50 x 4.6 mm, 3.5 μm). Method B: Mobile phase: A: 0.1% TFA in water, B: 0.1% TFA in ACN; flow rate: 2.0 mL / min; column: XBridge C8 (50 x 4.6 mm, 3.5 μm). Method C: Mobile phase: A: 10 mM NH4OAc in Milli-Q water, B: ACN; flow rate: 1.0 ml / min; column: Phenomenex Gemini C18 (150 x 4.6 mm, 3.0 μm). Method D: Mobile phase: A: 0.1% TFA in water, B: ACN; Flow rate: 1.0 mL / min; Column: ATLANTIS dC18 (250 x 4.6 mm, 5.0 μm). Method E: Mobile phase: A: 0.1% TFA in water, B: ACN, flow rate: 2.0 mL / min; column: X-Bridge C8 (50 × 4.6 mm, 3.5 μm).

[0223] Chiral SFC: Equipment name: PIC SFC 10 (for analysis) The CO2 to co-solvent ratio ranges from 60:40 to 80:20 Method A: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 mL / min; column: YMC Amylose-SA (250 x 4.6 mm, 5 μm). Method B: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 mL / min; column: Chiralpak AD-H (250 x 4.6 mm, 5 μm). Method C: Mobile phase: 20 mM ammonia in methanol; flow rate: 3 mL / min; column: YMC Cellulose-SC (250 x 4.6 mm, 5 μm). Method D: Mobile phase: methanol; flow rate: 3 mL / min; column: Lux A1 (250 x 4.6 mm, 5 μm). Method E: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 5 mL / min; column: Lux C4. Method F: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 3 mL / min; column: YMC Cellulose-SC. Method G: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 3 mL / min; column: Lux A1. Method H: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 mL / min; column: Lux A1 (250 x 4.6 mm, 5 μm). Method I: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 3 mL / min; column: Chiral CCS (250 x 4.6 mm, 5 μm). Method J: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 5 mL / min; column: YMC Cellulose-SC AD-H (250 x 4.6 mm, 5 μm). Method K: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 4 mL / min; column: (R,R)-Whelk-01 (250 x 4.6 mm, 5 μm). Method L: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 mL / min; column: Chiralcel OX-H (250 x 4.6 mm, 5 μm). Method M: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 5 mL / min; column: YMC Cellulose-SC (250 x 4.6 mm, 5 μm). Method N: Mobile phase: methanol, flow rate: 5 mL / min; column: Chiralcel OX-H (250 x 4.6 mm, 5 μm). Method O: Mobile phase: 0.1% isopropylamine in IPA:methanol (1:1), flow rate: 3 mL / min; Column: Chiralpak AS-H (250 x 4.6 mm, 5 μm). Method P: Mobile phase: 0.5% isopropylamine in methanol, flow rate: 3 mL / min; column: Chiralpak AS-H (250 x 4.6 mm, 5 μm). Method Q: Mobile phase: IPA, flow rate: 3 mL / min; Column: Lux A1 (250 x 4.6 mm, 5 μm). Method R: Mobile phase: 0.1% isopropylamine in IPA:methanol (1:1), flow rate: 3 mL / min; Column: Lux A1 (250 x 4.6 mm, 5 μm). Method S: Mobile phase: 0.5% isopropylamine in methanol, flow rate: 3 mL / min; column: Chiralpak OX-H (250 x 4.6 mm, 5 μm). Method T: Mobile phase: 0.5% isopropylamine in IPA, flow rate: 4 mL / min; column: YMC Cellulose-SB (250 x 4.6 mm, 5 μm). Method U: Mobile phase: 0.5% isopropylamine in IPA, flow rate: 3 mL / min; column: Chiralpak AS-H (250 x 4.6 mm, 5 μm).

[0224] Preparative HPLC: Instrument name: Agilent 1290 Infinity II Method A: Mobile phase: A: 0.1% TFA in water; mobile phase; B: 0.1% TFA in CAN; flow rate: 2.0 mL / min; column: X-Bridge C8 (50 × 4.6 mm, 3.5 μM). Method B: Mobile phase: A: 10mM NH4OAc in water; B: ACN; flow rate: 35mL / min; column: X select C18 (30 x 150mm, 5μm). Method C: Mobile phase: A: 10 mM NH4HCO3 in water; B: ACN; Flow rate: 1.0 mL / min; Column: XBridge C8 (50 x 4.6 mm, 3.5 μm). Method D: Mobile phase: A: 0.1% HCOOH in water; B: ACN; flow rate: 1.0 mL / min; column: X-select C18 (30 × 150 mm, 5 μm).

[0225] Chiral preparative SFC: Device name: PIC SFC 100 and PSC SFC 400 The CO2 to co-solvent ratio ranges from 60:40 to 80:20 Method A: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 mL / min; column: YMC Amylose-SA (250 x 30 mm, 5 μm). Method B: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 mL / min; column: Chiralpak AD-H (250 x 30 mm, 5 μm). Method C: Mobile phase: 20 mM ammonia in methanol; flow rate: 3 mL / min; column: YMC Cellulose-SC (250 x 30 mm, 5 μm). Method D: Mobile phase: methanol; flow rate: 3 mL / min; column: Chiral CCS (250 x 30 mm, 5 μm). Method E: Mobile phase: methanol; flow rate: 3 mL / min; column: Lux A1 (250 x 30 mm, 5 μm). Method F: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 mL / min; column: Lux A1 (250 x 30 mm, 5 μm). Method G: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 3 mL / min; column: Chiral CCS (250 x 30 mm, 5 μm). Method H: Mobile phase: 0.5% isopropylamine in IPA, flow rate: 5 mL / min; column: YMC Amylose-SC (250 x 30 mm, 5 μm). Method J: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 mL / min; column: Chiralcel OX-H (250 x 30 mm, 5 μm). Method K: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 5 mL / min; column: YMC Cellulose-SC (250 x 30 mm, 5 μm). Method L: Mobile phase: methanol; flow rate: 5 mL / min; column: Chiralcel OX-H (250 x 30 mm, 5 μm). Method M: Mobile phase: 0.5% isopropylamine in methanol, flow rate: 3 mL / min; column: Lux A1 (250 x 30 mm, 5 μm).

[0226] Abbreviation ACN Acetonitrile DCM dichloromethane DBAD Di-tert-butyl azodicarboxylate DMF Dimethylformamide IPA Isopropyl Alcohol LCMS Liquid Chromatography Mass Spectrometry HPLC High-Performance Liquid Chromatography PE Petroleum Ether SFC Supercritical Fluid Chromatography TFA trifluoroacetic acid THF tetrahydrofuran TLC thin layer chromatography UPLC Ultra High Performance Liquid Chromatography

[0227] The present invention will now be illustrated by the following examples, which are not intended to limit the invention in any way. All citations and references are incorporated by reference. [Example]

[0228] Intermediate 1 2-(((2-amino-4-bromo-5-methoxyphenyl)thio)methyl)-2-methylhexanoic acid

[0229] [ka]

[0230] To a stirred solution of 5-bromo-6-methoxybenzo[d]thiazol-2-amine (63 g, 0.243 mol) in water (630 mL) was added KOH (218.2 g, 3.89 mol), and the reaction mixture was stirred at 120 °C for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to room temperature. Then, a solution of 2-(bromomethyl)-2-methylhexanoic acid (70.5 g, 6.31 mol) in THF (210 mL) was added dropwise, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to 0 °C and acidified with concentrated HCl (pH ∼2). The reaction mixture was extracted with EtOAc (2 × 350 mL), and the combined organic layers were washed with water (150 mL) and brine (150 mL). The organic portion was dried over anhydrous NaSO and concentrated in vacuo. The resulting crude product was directly carried to the next step without further purification. Yield: 75 g (crude, brown gum). LCMS: (Method A) 376.1(M + ), 378.0(M + +2), Rt.2.44 minutes, 92.97% (maximum).

[0231] Intermediate 2 7-Bromo-3-butyl-8-methoxy-3-methyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0232] [ka]

[0233] To a stirred solution of 2-(((2-amino-4-bromo-5-methoxyphenyl)thio)methyl)-2-methylhexanoic acid (Intermediate 1; 75.0 g, 0.199 mol) in EtOAc (750 mL) at 0 °C, triethylamine (60.4 g, 0.59 mol) and 1-propanephosphonic anhydride solution (50% in EtOAc, 95.1 g, 0.29 mol) were added dropwise. The reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by UPLC), the reaction mixture was quenched with water (150 mL), and the aqueous layer was extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with brine (150 mL) and dried over anhydrous NaSO. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 10-12% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 63% (45 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 9.62 (s, 1H), 7.33 (s, 1H), 7.13 (s, 1H), 3.83 (s, 3H), 3.17 (s, 2H), 1.46-1.44 (m, 2H), 1.22 (s, 3H), 1.17-1.14 (m, 4H), 0.79 (t, J = 6.8 Hz, 3H). LCMS: (Method A) 360.0 (M + +2), Rt. 2.64 minutes, 97.14% (max).

[0234] Intermediate 3 7-Bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-3-methyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0235] [ka]

[0236] To a solution of 7-bromo-3-butyl-8-methoxy-3-methyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 2; 5 g, 14 mmol) in 1-bromo-4-fluorobenzene (50 mL), dry K2CO3 (3.9 g, 28 mmol), CuI (0.26 g, 1.4 mmol), and tris[2-(2-methoxyethoxy)ethyl]amine (0.9 g, 2.8 mmol) were added, and the reaction mixture was heated at 135 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo. The resulting residue was then partitioned between water (25 mL) and EtOAc (25 mL). The aqueous portion was extracted with EtOAc (2 × 100 mL), and the combined organic layers were washed with ice-cold water (100 mL) and brine (100 mL). The organic portion was dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The resulting residue was triturated with petroleum ether. The resulting solid was then filtered off and dried to give the title compound. Yield: 85% (5.5 g, light brown solid). LCMS: (Method E) 451.9(M + +H), Rt.3.26 min, 81.86% (max).

[0237] Intermediate 4 7-Bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-3-methyl-2,3,4,5-tetrahydro-1,5-benzothiazepine

[0238] [ka]

[0239] To a solution of 7-bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-3-methyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 3; 5.2 g, 11.5 mmol) in THF (50 mL) at 0 °C, borane dimethyl sulfide (1 M in THF, 58 mL, 58 mmol) was added, and the reaction mixture was heated at 75 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0 °C, methanol (60 mL) was added, and the reaction mixture was heated at 60 °C for 2 h. The reaction mixture was then cooled to room temperature and concentrated in vacuo, and the resulting residue was partitioned between water (50 mL) and EtOAc (50 mL). The aqueous layer was extracted with EtOAc (2 × 100 mL), and the combined organic layers were washed with ice-cold water (100 mL) and brine (100 mL). The organic portion was dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give the title compound. Yield: 5.3 g (crude, colorless gum). LCMS: (Method E) 439.9(M + +H), Rt.3.55 minutes, 87.61% (maximum).

[0240] Intermediate 5 7-Bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-3-methyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0241] [ka]

[0242] To a solution of 7-bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-3-methyl-2,3,4,5-tetrahydro-1,5-benzothiazepine (Intermediate 4; 5.3 g, 12.1 mmol) in a mixture of THF and water (8:2, 55 mL) was added Oxone (37.16 g, 120.8 mmol), and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the aqueous layer was extracted with EtOAc (2 × 100 mL), and the combined organic layers were washed with ice-cold water (100 mL) and brine (100 mL). The organic portion was dried over anhydrous NaSO and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 15% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 75% (4.3 g, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.45 (s, 1H), 7.21 (s, 1H), 7.12-7.05 (m, 4H), 3.93 (s, 3H), 3.85-3.61 (m, 2H), 3.29 (s, 2H), 1.52-1.39 (m, 1H), 1.37-1.26 (m, 1H), 1.25-1.03 (m, 4H), 0.98 (s, 3H), 0.81-0.74 (m, 3H). LCMS: (Method E) 470.1 (M + ), Rt. 3.21 minutes, 98.04% (max).

[0243] Intermediate 6 3-Butyl-5-(4-fluorophenyl)-8-hydroxy-3-methyl-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0244] [ka]

[0245] To a solution of 7-bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-3-methyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 5; 4.3 g, 9.14 mmol) in DMF (43 mL) was added sodium thiomethoxide (3.2 g, 45.7 mmol), and the reaction mixture was stirred at 80 °C for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with ice-cold water (25 mL), and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with ice-cold water (50 mL) and brine (50 mL) and dried over anhydrous NaSO. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 30% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 90% (3.5 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.57 (s, 1H), 7.31 (s, 1H), 7.04-6.98 (m, 2H), 6.92-6.88 (m, 2H), 6.71 (s, 1H), 3.81-3.65 (m, 2H), 3.34-3.20 (m, 2H), 2.20 (s, 3H), 1.55-1.39 (m, 1H), 1.38-1.05 (m, 5H), 0.99 (s, 3H), 0.81-0.77 (m, 3H). LCMS: (Method E) 424.2 (M + +H), Rt. 2.78 minutes, 98.08% (max).

[0246] Separation of enantiomers (S)-3-Butyl-5-(4-fluorophenyl)-8-hydroxy-3-methyl-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and (R)-3-butyl-5-(4-fluorophenyl)-8-hydroxy-3-methyl-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0247] [ka]

[0248] The two enantiomers of racemic 3-butyl-5-(4-fluorophenyl)-8-hydroxy-3-methyl-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (2.8 g, 6.61 mmol) were separated by chiral SFC (Method M). The material was concentrated under vacuum at 40° C. The first eluting fraction corresponded to enantiomer 1, and the second eluting fraction corresponded to enantiomer 2. The absolute configurations of the two enantiomers are unknown.

[0249] Enantiomer 1: Yield: 42% (1.2 g, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.56 (s, 1H), 7.30 (s, 1H), 7.04-6.98 (m, 2H), 6.92-6.88 (m, 2H), 6.71 (s, 1H), 3.80-3.50 (m, 2H), 3.20-3.15 (m, 2H), 2.21 (s, 3H), 1.58-1.36 (m, 1H), 1.35-1.03 (m, 5H), 0.99 (s, 3H), 0.80 (t, J = 9.20 Hz, 3H). LCMS: (Method E) 424.2 (M + +H), Rt. 2.79 min, 93.0% (max). HPLC: (Method B) Rt. 5.56 min, 93.77% (max). Chiral SFC: (Method M) Rt. 1.70 min, 99.78% (max).

[0250] Enantiomer 2: Yield: 44% (1.25 g, white solid). 1H NMR (400 MHz, DMSO-d6): δ 10.57 (s, 1H), 7.30 (s, 1H), 7.03-6.99 (m, 2H), 6.92-6.91 (m, 2H), 6.71 (s, 1H), 4.00-3.40 (m, 2H), 3.24-3.16 (m, 2H), 2.21 (s, 3H), 1.62-1.38 (m, 1H), 1.38-1.10 (m, 5H), 0.99 (s, 3H), 0.80 (t, J = 8.40 Hz, 3H). LCMS: (Method E) 424.2 (M + +H), Rt. 2.79 min, 98.86% (max). HPLC: (Method B) Rt. 5.56 min, 97.15% (max). Chiral SFC: (Method M) Rt. 2.51 min, 99.72% (max).

[0251] Intermediate 7 Methyl 3-((3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate (individual diastereomers)

[0252] [ka]

[0253] Diastereomers 1 and 2 To a stirred solution of enantiomer 1 of 3-butyl-5-(4-fluorophenyl)-8-hydroxy-3-methyl-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 6; 400 mg, 0.94 mmol) in DMF (4 mL), CsCO (615 mg, 1.83 mol) and methyl-oxirane-2-carboxylate (867 mg, 8.49 mmol) were added portionwise, and the reaction mixture was stirred at room temperature for 72 h. The reaction mixture was quenched with dilute HCl (1.5 N, 5 mL) and diluted with water (5 mL). The aqueous layer was extracted with EtOAc (2 × 50 mL), and the combined organic layers were washed with water (10 mL) and brine (10 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 35% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 50% (250 mg, white gum). 1 H NMR (400 MHz, DMSO-d6): δ 7.34 (s, 1H), 7.07-6.99 (m, 4H), 6.71 (s, 1H), 5.90 (d, J = 6.0 Hz, 1H), 4.48 (d, J = 4.8 Hz, 1H), 4.27 (s, 2H), 3.69 (s, 5H), 3.40-3.20 (m, 2H), 2.19 (s, 3H), 1.52-1.38 (m, 1H), 1.38-1.02 (m, 5H), 0.99 (s, 3H), 0.79 (t, J = 7.20 Hz, 3H). LCMS: (Method E) 526.0 (M + +H), Rt. 2.95 min, 97.92% (max). HPLC: (Method B) Rt. 5.50 min, 96.34% (max).

[0254] The two diastereomers (210 mg, 0.39 mmol) were separated by chiral SFC (Method G). The material was concentrated under vacuum at 40° C. The first eluting fraction corresponded to diastereomer 1, and the second to diastereomer 2.

[0255] Diastereomers 3 and 4 Following the same procedure, starting from 400 mg of enantiomer 2 of intermediate 6, a mixture of diastereomers 3 and 4 of the title compound was obtained. Yield: 50% (250 mg, yellow gum). 1 H NMR (400 MHz, DMSO-d6): δ 7.34 (s, 1H), 7.08-6.99 (m, 4H), 6.72 (s, 1H), 5.89 (d, J = 7.6 Hz, 1H), 4.49-4.47 (m, 1H), 4.27-4.26 (m, 2H), 3.69-3.67 (m, 5H), 3.23-3.20 (m, 2H), 2.19 (s, 3H), 1.55-1.40 (m, 1H), 1.40-1.05 (m, 5H), 0.99 (s, 3H), 0.79 (t, J = 9.60 Hz, 3H). LCMS: (Method E) 526.0 (M + +H), Rt. 2.95 min, 97.93% (max). HPLC: (Method B) Rt. 5.50 min, 96.46% (max).

[0256] The two diastereomers (210 mg, 0.39 mmol) were separated by chiral SFC (Method G). The material was concentrated under vacuum at 40° C. The first eluting fraction corresponded to diastereomer 3, and the second to diastereomer 4.

[0257] Diastereomer 1: Yield: 38% (80 mg, pale yellow solid). 1H NMR (400 MHz, DMSO-d6): δ 7.34 (s, 1H), 7.08-6.99 (m, 4H), 6.71 (s, 1H), 5.89 (d, J = 8.0 Hz, 1H), 4.48 (d, J = 6.0 Hz, 1H), 4.27 (t, J = 4.0 Hz, 2H), 3.69 (s, 5H), 3.31-3.22 (m, 2H), 2.19 (s, 3H), 1.52-1.38 (m, 1H), 1.30-1.10 (m, 5H), 0.99 (s, 3H), 0.79 (t, J = 9.20 Hz, 3H). LCMS: (Method G) 526.2 (M + +H), Rt. 2.32 minutes, 99.80% (maximum). HPLC: (Method B) Rt. 5.58 minutes, 99.40% (maximum).キラルSFC: (Method G) Rt. 2.68 points, 99.38% (maximum). Sesame 2: Yield: 38% (80 mg, thin yellow solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.34 (s, 1H), 7.08-6.96 (m, 4H), 6.72 (s, 1H), 5.89 (d, J = 7.6 Hz, 1H), 4.48 (d, J = 6.8 Hz, 1H), 4.27 (d, J = 5.6 Hz, 2H), 3.69 (s, 5H), 3.31-3.22 (m, 2H), 2.19 (s, 3H), 1.55-1.40 (m, 1H), 1.24-1.11 (m, 5H), 0.99 (s, 3H), 0.79 (t, J = 9.20 Hz, 3H). LCMS: (Method G) 526.2 (M + +H), Rt. 2.32 minutes, 98.65% (maximum). HPLC: (Method B) Rt. 5.50 minutes, 99.46% (maximum).キラルSFC: (Method G) Rt. 4.02 points, 99.75% (maximum). Sesame 3: Yield: 40% (85 mg, thin yellow solid). 1H NMR (400 MHz, DMSO-d6): δ 7.35 (s, 1H), 7.08-6.99 (m, 4H), 6.72 (s, 1H), 5.89 (d, J = 7.6 Hz, 1H), 4.49-4.48 (m, 1H), 4.28-4.26 (m, 2H), 3.75-3.50 (m, 5H), 3.23-3.17 (m, 2H), 2.19 (s, 3H), 1.55-1.35 (m, 1H), 1.32-1.06 (m, 5H), 1.00 (s, 3H), 0.90-0.70 (m, 3H). LCMS: (Method E) 526.0 (M + +H), Rt. 2.98 min, 97.37% (max). HPLC: (Method B) Rt. 5.58 min, 96.35% (max). Chiral SFC: (Method G) Rt. 2.36 min, 99.90% (max). Diastereomer 4: Yield: 38% (80 mg, pale yellow solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.35 (s, 1H), 7.07-6.99 (m, 4H), 6.72 (s, 1H), 5.89 (d, J = 6.0 Hz, 1H), 4.50-4.47 (m, 1H), 4.30-4.23 (m, LCMS: (Method E) 526.1 (M + +H), Rt. 2.96 min, 97.20% (max). HPLC: (Method B) Rt. 5.51 min, 99.09% (max). Chiral SFC: (Method G) Rt. 3.31 min, 99.48% (max). The absolute configurations of the four diastereomers are unknown.

[0258] Intermediate 8 2-(((2-amino-5-methoxyphenyl)thio)methyl)-2-ethylhexanoic acid

[0259] [ka]

[0260] To a stirred solution of 6-methoxybenzo[d]thiazol-2-amine (270 g, 1.498 mol) in water (2700 mL) was added KOH (1345 g, 23.96 mol), and the reaction mixture was stirred at 120 °C for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to room temperature. Then, a solution of 2-(bromomethyl)-2-ethylhexanoic acid (533 g, 2.25 mol) in THF (1000 mL) was added dropwise, and the resulting reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to 0 °C and acidified with concentrated HCl (pH ∼2). The reaction mixture was extracted with EtOAc (2 × 4000 mL), and the combined organic layers were washed with water (1000 mL) and brine (1000 mL). The organic portion was then dried over anhydrous NaSO and concentrated in vacuo to give the crude material, which was directly carried on to the next step without further purification. Yield: 590 g (crude, brown gum). LCMS: (Method A) 312.1(M + +H), Rt.2.24 min, 97.34% (maximum).

[0261] Intermediate 9 3-Butyl-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0262] [ka]

[0263] To a stirred solution of 2-(((2-amino-5-methoxyphenyl)thio)methyl)-2-ethylhexanoic acid (Intermediate 8; 590 g, 1.89 mol) in EtOAc (2500 mL) at 0 °C, triethylamine (530 mL, 3.78 mol) and 1-propanephosphonic anhydride solution (50% in EtOAc; 785 g, 2.46 mol) were added dropwise, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), water (2000 mL) was added to the reaction mixture, and the aqueous layer was extracted with EtOAc (2 x 2000 mL). The combined organic layers were washed with brine (800 mL), dried over anhydrous NaSO, and concentrated in vacuo. The crude material was purified by washing with methanol to give the title compound. Yield: 48% (265 g, off-white solid). 1 H NMR (300 MHz, DMSO-d6): δ 9.53 (s, 1H), 7.04-7.01 (m, 2H), 6.87-6.86 (m, 1H), 3.72 (s, 3H), 2.50 (s, 2H), 1.68-1.66 (m, 4H), 1.50-1.48 (m, 4H), 0.79-0.72 (m, 6H). LCMS: (Method A) 294.3 (M + +H), Rt. 2.68 minutes, 99.47% (max).

[0264] Intermediate 10 7-Bromo-3-butyl-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0265] [ka]

[0266] To a stirred solution of 3-butyl-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 9; 265 g, 0.903 mol) in a 1:1 mixture of DCM and acetonitrile (2650 mL), N-bromosuccinimide (209 g, 1.17 mol) was added portionwise, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated. The resulting crude material was treated with cold acetonitrile and stirred for 30 minutes. The resulting precipitate was filtered off, washed with cold acetonitrile (2 x 100 mL), and dried under vacuum to give the title compound. Yield: 179 g (79%, crude, brown solid). 1 H NMR (300 MHz, DMSO-d6): δ 9.61 (s, 1H), 7.33 (s, 1H), 7.10 (s, 1H), 3.82 (s, 3H), 2.98 (s, 2H), 1.70-1.68 (m, 4H), 1.48-1.45 (m, 4H), 0.84-0.82 (m, 6H). LCMS: (Method A) 372.0 (M + +H), Rt. 2.83 minutes, 99.20% (max).

[0267] Intermediate 11 7-Bromo-3-butyl-3-ethyl-5-(4-fluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3-butyl-3-ethyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0268] [ka]

[0269] To a stirred solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 10; 15 g, 40.2 mmol) in 1-fluoro-4-iodobenzene (50 mL), copper(I) iodide (1.58 g, 0.8 mmol) and K2CO3 (11 g, 80.5 mmol) were added, and the reaction mixture was purged with nitrogen for 20 minutes to degas. Then, tris[2-(2-methoxyethoxy)ethyl]amine (1.3 mL, 4.0 mmol) was added under a nitrogen atmosphere, and the resulting reaction mixture was heated at 135 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through Celite, and the Celite pad was washed with EtOAc (200 mL). The filtrate was washed with water (100 mL) and brine (75 mL) and dried over anhydrous Na2SO4. The resulting crude material was purified by Isolera column chromatography (eluent: 5% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 64% (12.2 g, off-white solid). LCMS: (Method E) 467.1 (M for 7-bromo substituted compound) + +2) and 514.1 (M + +H), Rt.3.33 minutes, 92.83% (maximum).

[0270] Intermediate 12 7-Bromo-3-butyl-3-ethyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3-butyl-3-ethyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine

[0271] [ka]

[0272] To a stirred solution of a mixture of 7-bromo-3-butyl-3-ethyl-5-(4-fluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3-butyl-3-ethyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 11; 12 g, 25.7 mmol) in THF (100 mL) at 0 °C, borane dimethyl sulfide (2 M in THF; 38 mL, 77 mmol) was added dropwise, and the reaction mixture was refluxed at 65 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0 °C, quenched with methanol (20 mL), and heated at 65 °C for 2 hours. The resulting reaction mixture was then cooled to room temperature and concentrated in vacuo. The residue was diluted with water (100 mL) and the aqueous layer was extracted with DCM (2 x 100 mL). The combined organic layers were then washed with water (50 mL) and brine (50 mL) and dried over anhydrous NaSO. The organic portion was concentrated in vacuo and the resulting crude product was directly carried to the next step without further purification. Yield: 10 g (crude, black gum). LCMS: (Method E) 451.8 (M for 7-bromo substituted compound) + +H) and 499.7 (M + +H), Rt.3.78 min, 75.13% (max).

[0273] Intermediate 13 7-Bromo-3-butyl-3-ethyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 3-butyl-3-ethyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0274] [ka]

[0275] To a stirred solution of a mixture of 7-bromo-3-butyl-3-ethyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3-butyl-3-ethyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine (Intermediate 12; 10 g, 26.6 mmol) in THF (100 mL) and water (60 mL) was added oxone (81 g, 26.6 mmol) at 0 °C. The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a Buchner funnel, the filtrate was extracted with EtOAc (2 × 200 mL), and the combined organic layers were washed with water (100 mL) and brine (100 mL), dried over anhydrous NaSO, and concentrated in vacuo. The resulting crude product was purified by Isolera column chromatography (eluent: 15% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 54% (7 g, white solid). LCMS: (Method E) 486.0 (M for 7-bromo substituted compound + +2) and 532.0 (M + +H), Rt.2.87 min, 91.53% (max).

[0276] Intermediate 14 3-Butyl-3-ethyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0277] [ka]

[0278] To a stirred solution of a mixture of 7-bromo-3-butyl-3-ethyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 3-butyl-3-ethyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 13; 3 g, 6.2 mmol) in DMF (16 mL) was added sodium thiomethoxide (2.1 g, 31 mmol) at room temperature, and the reaction mixture was stirred at 65 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and quenched with water (25 mL). The aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous NaSO, and concentrated in vacuo. The resulting crude product was purified by Isolera column chromatography (eluent: 10% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 77% (2.13 g, brown solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.49 (s, 1H), 7.28 (s, 1H), 7.06-6.96 (m, 4H), 6.61 (s, 1H), 3.62 (bs, 2H), 3.21 (s, 2H), 2.17 (s, 3H), 1.61-1.25 (m, 4H), 1.20-1.01 (m, 4H), 0.81-0.74 (m, 6H). LCMS: (Method A) 438.1 (M + +H), Rt. 2.78 minutes, 87.79 % (max.).

[0279] Intermediate 15 Ethyl 1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylate

[0280] [ka]

[0281] To a stirred solution of 3-butyl-3-ethyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 14; 200 mg, 0.45 mmol) in DMF (5 mL), triphenylphosphine (180 mg, 0.68 mmol), ethyl (1-hydroxymethyl)cyclopropanecarboxylate (99 mg, 0.68 mmol), and DBAD (210 mg, 0.91 mmol) were added, and the reaction mixture was stirred at room temperature for 4 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water (10 mL), and the aqueous layer was extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with water (15 mL) and brine (15 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 11% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 93% (240 mg, colorless gum). LCMS: (Method E) 564.1(M + +H), Rt.3.33 minutes, 95.41% (maximum).

[0282] Intermediate 16 Methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoate

[0283] [ka]

[0284] To a stirred solution of 3-butyl-3-ethyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 14; 0.5 g, 1.10 mmol) in THF (10 mL) at 0 °C, methyl 3-hydroxy-2,2-dimethylpropanoate (0.15 g, 1.10 mmol) and triphenylphosphine (0.43 g, 1.60 mmol) were added, and the reaction mixture was stirred for 5 min. DBAD (0.50 g, 2.20 mmol) was then added, and the reaction mixture was stirred at room temperature for 24 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with water (10 mL), and the aqueous layer was extracted with EtOAc (2 × 15 mL). The combined organic layers were washed with water (2 × 15 mL) and brine (15 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The crude material obtained was purified by Isolera column chromatography (eluent: 10% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 47% (0.30 g, white solid). LCMS: (Method E) 552.1(M + +H), Rt.3.32 min, 85.43% (max).

[0285] Intermediate 17 Methyl 3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxy-2-methylpropanoate

[0286] [ka]

[0287] To a stirred solution of 3,3-diethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (3.0 g, 9.4 mmol) in DMF (10 mL) was added CsCO (6.11 g, 18.8 mmol), and the reaction mixture was stirred at room temperature for 15 minutes. Methyl-2-methylglycidate (3.27 g, 28.2 mmol) was then added, and the reaction mixture was stirred at room temperature for 72 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (15 mL) and diluted with water (10 mL). The aqueous layer was extracted with EtOAc (2 × 15 mL), and the combined organic layers were washed with water (15 mL) and brine (15 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 10% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 29.35% (1.5 g, colorless solid). LCMS: (Method E) 508.2(M + +H), Rt.2.72 minutes, 99.88% (maximum).

[0288] Intermediate 18 Methyl 3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoate

[0289] [ka]

[0290] To a stirred solution of methyl 3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxy-2-methylpropanoate (Intermediate 17; 300 mg, 0.59 mmol) in DCM (10 mL) at −10° C., diethylaminosulfur trifluoride (0.09 g, 0.70 mmol) was added, and the reaction mixture was stirred for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with water (5 mL), and the aqueous layer was extracted with EtOAc (2×5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 20% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 43% (130 mg, off-white solid). LCMS: (Method E) 510.3(M + +H), Rt.2.89 min, 98.90% (max).

[0291] Separation of enantiomers Methyl (R)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoate and methyl (S)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoate

[0292] [ka] The two enantiomers of racemic methyl 3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoate (100 mg, 0.19 mmol) were separated by chiral SFC (Method B). The material was concentrated under vacuum at 40 °C. The first eluting fraction corresponded to enantiomer 1, and the second eluting fraction corresponded to enantiomer 2. The absolute configurations of the two enantiomers are unknown.

[0293] Enantiomer 1: Yield: 30% (30 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.36 (s, 1H), 7.22 (t, J = 8.4 Hz, 2H), 6.99 (d, J = 7.2 Hz, 2H), 6.86 (t, J = 7.6 Hz, 1H), 6.71 (s, 1H), 4.50-4.39 (m, 2H), 3.76 (s, 3H), 3.69 (bs, 2H), 3.26 (s, 2H), 2.16 (s, 3H), 1.60-1.52 (m, 5H), 1.38-1.32 (m, 2H), 0.75 (t, J = 7.2 Hz, 6H). LCMS: (Method E) 510.0 (M + +H), Rt. 3.04 min, 96.42% (max). HPLC: (Method B) Rt. 5.90 min, 97.43% (max). Enantiomer 1: Yield: 30% (30 mg, white solid). 1H NMR (400 MHz, DMSO-d6): 7.35 (s, 1H), 7.22 (t, J = 8.4 Hz, 2H), 6.99 (d, J = 7.2 Hz, 2H), 6.86 (t, J = 7.6 Hz, 1H), 6.70 (s, 1H), 4.50-4.42 (m, 2H), 3.70 (s, 3H), 3.60 (m, 2H), 3.26 (s, 2H), 2.16 (s, 3H), 1.65-1.52 (m, 5H), 1.37-1.32 (m, 2H), 0.75 (t, J = 7.6 Hz, 6H). LCMS: (Method E) 510.0 (M + +H), Rt. 3.04 min, 98.16% (max). HPLC: (Method B) Rt. 5.90 min, 99.29% (max).

[0294] Intermediate 19 Methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate

[0295] [ka]

[0296] To a stirred solution of 3-butyl-3-ethyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 14; 750 mg, 1.71 mmol) in DMF (6 mL), CsCO (1.11 g, 3.42 mmol) was added, and the reaction mixture was stirred at room temperature for 15 minutes. Methyl oxirane-2-carboxylate (0.42 mL, 5.14 mmol) was then added dropwise, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (1.5 N, 1 mL) and diluted with water (1 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 20-30% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 50% (300 mg, colorless gum). 1 HNMR: (400 MHz, DMSO-d6): δ 7.34 (s, 1H), 7.17-7.07 (m, 4H), 6.61 (s, 1H), 5.89 (d, J = 6.0 Hz, 1H), 4.49-4.46 (m, 1H), 4.25 (t, J = 2.8 Hz, 2H), 3.68 (s, 3H), 3.62 (bs, 2H), 3.29 (s, 2H), 2.15 (s, 3H), 1.40-1.33 (m, 4H), 1.20-1.10 (m, 4H), 0.78-0.72 (m, 6H). LCMS: (Method E) 540.2 (M + +H), Rt. 2.90 minutes, 94.13% (max).

[0297] Intermediate 20 Methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoate

[0298] [ka]

[0299] To a stirred solution of methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate (Intermediate 19; 100 mg, 0.18 mmol) in DMF (2 mL) at 0° C., sodium hydride (60%, 3.71 mg, 0.09 mmol) was added, and the reaction mixture was stirred at room temperature for 15 minutes. Then, a solution of methyl iodide (0.04 mL, 0.55 mmol) in DMF (1 mL) was added dropwise, and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (1.5 N, 1 mL) and diluted with water (1 mL). The aqueous layer was extracted with EtOAc (2×10 mL). The combined organic layers were washed with water (10 mL) and brine (10 mL) and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated in vacuo. The crude material obtained was purified by Isolera column chromatography (eluent: 20-30% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 57% (60 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.32 (s, 1H), 7.07 (d, J = 6.80 Hz, 4H), 6.60 (s, 1H), 4.38-4.27 (m, 3H), 3.70 (s, 3H), 3.68 (s, 2H), 3.41 (s, 3H), 3.28 (s, 2H), 2.15 (s, 3H), 1.37-1.32 (m, 4H), 1.16-1.09 (m, 4H), 0.77-0.72 (m, 6H). LCMS: (Method E) 554.3 (M + +H), Rt. 3.01 min, 98.12% (max).

[0300] Intermediate 21 7-Bromo-3,3-dibutyl-5-(4-fluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3,3-dibutyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0301] [ka]

[0302] To a stirred solution of a mixture of 7-bromo-3,3-dibutyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (3 g, 7.49 mmol) in 4-fluoroiodobenzene (30 mL) was added copper(I) iodide (0.14 g, 0.74 mmol) and K2CO3 (2.07 g, 14.9 mmol), and the reaction mixture was purged with nitrogen for 20 minutes to degas. Then, tris[2-(2-methoxyethoxy)ethyl]amine (0.49 g, 1.49 mmol) was added under a nitrogen atmosphere, and the resulting reaction mixture was heated at 135 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through Celite, and the Celite pad was washed with EtOAc (25 mL). The filtrate was washed with water (15 mL) and brine (15 mL) and dried over anhydrous Na2SO4. The resulting crude material was purified by Isolera column chromatography (eluent: 20% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 95% (3.5 g, pale yellow solid). LCMS: (Method E) 494.0 (M for 7-bromo substituted compound + ) and 541.9 (M + +H), Rt.3.50 minutes, 96.61% (maximum).

[0303] Intermediate 22 7-Bromo-3,3-dibutyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3,3-dibutyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine

[0304] [ka]

[0305] To a stirred solution of a mixture of 7-bromo-3,3-dibutyl-5-(4-fluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3,3-dibutyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 21; 3.5 g, 7.07 mmol) in THF (35 mL) at 0° C., borane dimethyl sulfide (2 M in THF; 5.3 mL, 10.61 mmol) was added dropwise, and the reaction mixture was refluxed at 65° C. for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0° C., quenched with methanol (10 mL), and heated at 65° C. for 2 hours. The resulting reaction mixture was then cooled to room temperature, concentrated in vacuo, and the residue was partitioned between water (50 mL) and EtOAc (50 mL). The aqueous layer was extracted with DCM (2 x 50 mL), and the combined organic layers were washed with water (25 mL) and brine (25 mL). The organic portion was dried over anhydrous NaSO and concentrated in vacuo to give the crude product. The crude product was directly used in the next step without further purification. Yield: 3.6 g (crude, pale yellow gum). LCMS: (Method E) 482.0 (M for 7-bromo substituted compound + +2H) and 527.9 for the 7-iodo substituted compound (M + +H), Rt. 3.86 min, 81.04% (combined for bromo and iodo substituted compounds) (max).

[0306] Intermediate 23 7-Bromo-3,3-dibutyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 3,3-dibutyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0307] [ka]

[0308] To a stirred solution of a mixture of 7-bromo-3,3-dibutyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3,3-dibutyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine (Intermediate 22; 3.6 g, 7.49 mmol) in acetic acid (36 mL), sodium tungstate (360 mg, 0.01 mmol) and hydrogen peroxide (30% in HO; 2.6 mL, 22.47 mmol) were added at 0 ° C., and the resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a Buchner funnel, and the filtrate was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with water (25 mL) and brine (25 mL) and dried over anhydrous NaSO. The organic portion was concentrated in vacuo, and the resulting crude material was purified by Isolera column chromatography (eluent: 12% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 79% (3.4 g, off-white solid). LCMS: (Method A) 512.2 (M for 7-bromo substituted compound + +H) and 560.2 for the 7-iodo substituted compound (M + +H);Rt.3.40 minutes, 70.63% (maximum).

[0309] Intermediate 24 3,3-Dibutyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0310] [ka]

[0311] To a stirred solution of a mixture of 7-bromo-3,3-dibutyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 3,3-dibutyl-5-(4-fluorophenyl)-7-iodo-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 23; 1.6 g, 3.12 mmol) in DMF (16 mL) was added sodium thiomethoxide (1.09 g, 15.6 mmol) at room temperature, and the reaction mixture was stirred at 65 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and quenched with water (15 mL). The aqueous layer was extracted with EtOAc (2 × 25 mL), and the combined organic layer was washed with brine (10 mL). The organic portion was dried over anhydrous NaSO and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 30% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 90% (1.3 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.48 (s, 1H), 7.28 (d, J = 4.4 Hz, 1H), 7.08-7.01 (m, 4H), 6.59 (s, 1H), 3.80-3.67 (m, 2H), 3.22 (s, 2H), 2.16 (s, 3H), 1.36-1.33 (m, 4H), 1.12-1.03 (m, 8H), 0.79-0.77 (m, 6H). LCMS: (Method E) 466.0 (M + +H), Rt. 3.23 minutes, 88.86% (max).

[0312] Intermediate 25 Methyl 3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate

[0313] [ka]

[0314] To a stirred solution of 3,3-dibutyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 24; 400 mg, 0.8602 mmol) in DMF (4 mL) was added CsCO (560 mg, 1.72 mmol), and the reaction mixture was stirred at room temperature for 15 minutes. Methyl oxirane-2-carboxylate (0.2 mL, 2.58 mmol) was then added dropwise, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (1.5 N, 5 mL), and the aqueous layer was extracted with EtOAc (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 20-30% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound in 44% yield (215 mg, colorless gum). LCMS: (Method E) 568.1(M + +H), Rt.3.18 minutes, 93.04% (maximum).

[0315] Intermediate 26 Methyl 3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoate

[0316] [ka]

[0317] To a stirred solution of methyl 3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate (Intermediate 25; 125 mg, 0.22 mmol) in DMF (2.5 mL) at 0 °C, sodium hydride (60%, 8.80 mg, 0.22 mmol) was added, and the reaction mixture was stirred for 5 minutes. Then, methyl iodide (0.08 mL, 1.32 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (1.5 N, 5 mL), and the aqueous layer was extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with water (10 mL) and brine (10 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 20-30% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 38% (50 mg, white solid). LCMS: (Method E) 582.2(M + +H), Rt.3.29 min, 97.76% (max).

[0318] Intermediate 27 Methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate

[0319] [ka]

[0320] To a solution of 3-butyl-3-ethyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 14; 750 mg, 1.71 mmol) in DMF (10 mL), CsCO (1.11 g, 3.42 mmol) and methyl oxirane-2-carboxylate (0.52 g, 5.14 mmol) were added, and the reaction mixture was stirred at room temperature for 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with water (10 mL), and the aqueous layer was extracted with EtOAc (2 × 15 mL). The combined organic layers were washed with water (15 mL) and brine (15 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 45% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 50% (300 mg, colorless gum). LCMS: (Method E) 540.2(M + +H), Rt.2.90 minutes, 94.11% (max).

[0321] Intermediate 28 Methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoate

[0322] [ka]

[0323] To a suspension of NaH (60%, 21 mg, 0.55 mmol) in dry DMF (2 mL) at 0 °C, a solution of methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate (Intermediate 27; 300 mg, 0.55 mmol) in DMF (5 mL) was added, and the reaction mixture was stirred for 5 min. Then, ethyl iodide (260 mg, 1.67 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (1.5 N, 5 mL) at 0 °C, and the aqueous layer was extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with water (15 mL) and brine (15 mL) and dried over anhydrous Na SO . The organic portion was filtered and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 25% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 31% (98 mg, colorless gum). LCMS: (Method E) 568.3(M + +H), Rt.3.09 min, 65.63% (max).

[0324] Intermediate 29 Methyl 3-((3,3-dibutyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate

[0325] [ka]

[0326] To a solution of 3,3-dibutyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (1.5 g, 3.35 mmol) in DMF (10 mL) was added CsCO (2.18 g, 6.70 mmol) and methyl oxirane-2-carboxylate (1.02 g, 10.05 mmol), and the reaction mixture was stirred at room temperature for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with water (50 mL), and the aqueous layer was extracted with EtOAc (2 × 30 mL). The combined organic layers were washed with water (50 mL) and brine (50 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 45% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 45% (600 mg, colorless gum). LCMS: (Method E) 550.1(M + +H), Rt.3.20 minutes, 98.81% (maximum).

[0327] Intermediate 30 Methyl 3-((3,3-dibutyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoate

[0328] [ka]

[0329] To a suspension of NaH (60%, 35 mg, 0.55 mmol) in dry DMF (3 mL) at 0 °C, a solution of methyl 3-((3,3-dibutyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate (Intermediate 29; 500 mg, 0.91 mmol) in DMF (7 mL) was added, and the reaction mixture was stirred for 10 min. Then, ethyl iodide (426 mg, 2.73 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (1.5 N, 10 mL) at 0 °C, and the aqueous layer was extracted with EtOAc (2 × 15 mL). The combined organic layers were washed with water (20 mL) and brine (20 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 25% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 30% (300 mg, colorless gum). LCMS: (Method E) 578.3(M + +H), Rt.3.28 minutes, 71.90% (maximum).

[0330] Intermediate 31 7-Bromo-3,3-diethyl-5-(4-fluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0331] [ka]

[0332] To a stirred solution of 7-bromo-3,3-diethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (20 g, 0.06 mol) in 4-bromofluorobenzene (200 mL), copper(I) iodide (1.10 g, 0.006 mol) and K2CO3 (16.05 g, 0.12 mol) were added, and the reaction mixture was purged with nitrogen for 20 minutes to degas it. Then, tris[2-(2-methoxyethoxy)ethyl]amine (3.75 g, 0.01 mol) was added under a nitrogen atmosphere, and the resulting reaction mixture was heated at 135 °C for 3 days. After completion of the reaction (monitored by TLC and UPLC), the reaction mixture was filtered through Celite, and the Celite pad was washed with EtOAc (100 mL). The filtrate was concentrated in vacuo, and the resulting residue was partitioned between water (100 mL) and EtOAc (50 mL). The aqueous layer was extracted with EtOAc (2×100 mL), and the combined organic layers were washed with water (70 mL) and brine (70 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 2-4% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 70% (18.0 g, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.36 (s, 1H), 7.22-7.14 (m, 5H), 3.90 (s, 3H), 3.15 (s, 2H), 1.50-1.45 (m, 4H), 0.78 (t, J = 9.6 Hz, 6H). LCMS: (Method E) 440.1 (M + +2), Rt. 2.97 min, 96.42% (max). HPLC: (Method E) Rt. 5.97 min, 94.16% (max).

[0333] Intermediate 32 7-Bromo-3,3-diethyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine

[0334] [ka]

[0335] To a stirred solution of 7-bromo-3,3-diethyl-5-(4-fluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 31; 18.0 g, 0.04 mol) in THF (90 mL) at 0 °C, borane dimethyl sulfide (2 M in THF, 102 mL, 0.21 mol) was added dropwise, and the reaction mixture was refluxed at 70 °C for 48 h. After completion of the reaction (monitored by UPLC), the reaction mixture was cooled to 0 °C and quenched with methanol (50 mL). The resulting solution was heated at 70 °C for 2 h, then cooled to room temperature and concentrated in vacuo. The resulting residue was partitioned with water (100 ml) and EtOAc (50 mL), and the aqueous layer was extracted with EtOAc (2 × 100 mL). The combined organic layers were washed with water (70 mL) and brine (70 mL) and dried over anhydrous Na SO . The organic portion was concentrated under vacuum and the resulting crude was carried on to the next step without further purification. Yield: 21 g (crude, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.12-6.95 (m, 5H), 6.88 (s, 1H), 3.81 (s, 3H), 3.70-3.45 (m, 2H), 2.78 (s, 2H), 1.45-1.18 (m, 4H), 0.70 (t, J = 7.6 Hz, 6H). LCMS: (Method E) 424.9 (M + +2), Rt. 3.44 min, 86.38% (max). HPLC: (Method E) Rt. 6.97 min, 94.07% (max).

[0336] Intermediate 33 7-Bromo-3,3-diethyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0337] [ka]

[0338] To a stirred solution of 7-bromo-3,3-diethyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine (Intermediate 32; 21.0 g, 0.05 mol) in THF (147 mL), water (63 mL), oxone (152.1 g, 0.49 mol) were added at 0° C., and the reaction mixture was stirred at room temperature for 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered. The filtrate was diluted with water (100 ml), and the aqueous layer was extracted with EtOAc (2×100 mL). The combined organic layers were washed with water (70 mL) and brine (70 mL), dried over anhydrous NaSO, and concentrated in vacuo. The crude material obtained was purified by Isolera column chromatography (eluent: 10% MeOH / DCM; silica gel: 230-400 mesh), and the resulting product was triturated with petroleum ether (2×100 mL) to give the title compound. Yield: 93% (21.0 g, brown solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.44 (s, 1H), 7.25-7.02 (m, 5H), 3.92 (s, 3H), 3.66 (s, 2H), 3.33 (s, 2H), 1.60-1.42 (m, 2H), 1.42-1.28 (m, 2H), 0.71 (t, J = 9.6 Hz, 6H). LCMS: (Method E) 458.1 (M + +2), Rt. 2.94 minutes, 95.31% (max).

[0339] Intermediate 34 3,3-Diethyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0340] [ka]

[0341] To a stirred solution of 7-bromo-3,3-diethyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 33; 10.0 g, 0.02 mol) in DMF (100 mL) was added sodium thiomethoxide (7.67 g, 0.11 mol) at room temperature, and the resulting mixture was stirred at 70 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with water (100 mL), and the aqueous layer was extracted with EtOAc (2 × 100 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 36-50% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 88% (7.90 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.54 (s, 1H), 7.30 (s, 1H), 7.09-6.99 (m, 2H), 6.99-6.91 (m, 2H), 6.64 (s, 1H), 3.72-3.50 (m, 2H), 3.25-3.18 (m, 2H), 2.18 (s, 3H), 1.65-1.46 (m, 2H), 1.43-1.25 (m, 2H), 0.74 (t, J = 7.20 Hz, 6H). LCMS: (Method I) 410.0 (M + +H), Rt. 2.46 min, 92.37% (max). HPLC: (Method E) Rt. 5.33 min, 94.90% (max).

[0342] Intermediate 35 Methyl 3-((3,3-diethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoate

[0343] [ka]

[0344] To a stirred solution of 3,3-diethyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 35; 300 mg, 0.73 mmol) in THF (10 mL), methyl-3-hydroxy-2,2-dimethylpropanoate (96 mg, 0.73 mmol), triphenylphosphine (288 mg, 1.09 mmol), and DBAD (336 mg, 1.46 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water (10 mL), and the aqueous layer was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with water (25 mL) and brine (25 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 10-20% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound in 78% yield (300 mg, white gum). LCMS: (Method E) 524.0(M + +H), Rt.3.17 minutes, 93.5% (maximum).

[0345] Intermediate 36 Methyl 2-hydroxy-3-((7-(methylthio)-1,1-dioxide-5-phenyl-3,3-dipropyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)propanoate

[0346] [ka]

[0347] To a stirred solution of 8-hydroxy-7-(methylthio)-5-phenyl-3,3-dipropyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (500 mg, 1.19 mmol) in DMF (10 mL) was added CsCO (777 mg, 2.38 mmol) and methyl oxirane-2-carboxylate (365 mg, 3.57 mmol), and the reaction mixture was stirred at room temperature for 4 days. After completion of the reaction (monitored by TLC; approximately 50% conversion), the reaction mixture was quenched with dilute HCl (1.5 N, 10 mL), and the aqueous layer was extracted with ethyl acetate (2 × 10 mL). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 30-40% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 24% (150 mg, yellow gum). LCMS: (Method K) 521.9(M + +H), Rt.3.03 minutes, 95.10% (maximum).

[0348] Example 1 3-((3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid (individual diastereomers)

[0349] [ka]

[0350] Diastereomer 1 To a stirred solution of methyl 3-((3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoate diastereomer 1 (Intermediate 7; 80 mg, 0.15 mmol) in 1,4-dioxane (1 mL), aqueous HCl (6 N, 3 mL) was added, and the reaction mixture was heated at 80° C. for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water (5 mL), and the aqueous layer was extracted with EtOAc (2×20 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 10-20% MeOH / DCM; silica gel: 230-400 mesh) to afford the title compound. Starting from 80-85 mg of diastereomers 2, 3 and 4 of intermediate 7, respectively, and following the same procedure, diastereomers 2, 3 and 4 of the title compound were obtained. Diastereomer 1: Yield: 32% (25 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.36 (s, 1H), 7.07-6.99 (m, 4H), 6.73 (s, 1H), 4.33 (d, J = 6.8 Hz, 1H), 4.12 (s, 2H), 3.69-3.52 (m, 2H), 3.31-3.22 (m, 2H), 2.20 (s, 3H), 1.55-1.38 (m, 1H), 1.30-1.10 (m, 5H), 0.99 (s, 3H), 0.76 (t, J = 8.4 Hz, 3H). LCMS: (Method E) 512.0 (M + +H), Rt. 2.83 min, 98.75% (max). HPLC: (Method B) Rt. 5.10 min, 98.25% (max). Chiral SFC: (Method G) Rt. 2.59 min, 99.42% (max).

[0351] Diastereomer 2: Yield: 32% (25 mg, white solid). 1H NMR (400 MHz, DMSO-d6): δ 7.35 (s, 1H), 7.06-6.98 (m, 4H), 6.72 (s, 1H), 4.35 (d, J = 9.2 Hz, 1H), 4.07-4.04 (m, 2H), 3.62 (m, 2H), 3.21 (m, 2H), 2.20 (s, 3H), 1.55-1.39 (m, 1H), 1.35-1.05 (m, 5H), 1.03 (s, 3H), 0.79 (t, J = 8.4 Hz, 3H). LCMS: (Method E) 512.0 (M + +H), Rt. 2.82 points, 99.25% (maximum). HPLC: (Method B) Rt. 5.11 points, 98.71% (maximum).キラルSFC: (Method G) Rt. 4.19 points, 99.14% (maximum).

[0352] Semen 3: Yield: 49% (40 mg, Semen solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.36 (s, 1H), 7.07-6.98 (m, 4H), 6.72 (s, 1H), 4.33-4.30 (m, 1H), 4.21-4.15 (m, 2H), 3.88-3.44 (m, 2H), 3.22 (s, 2H), 2.20 (s, 3H), 1.46-1.33 (m, 1H), 1.32-1.10 (m, 5H), 1.04 (s, 3H), 0.80 (t, J = 7.2 Hz, 3H). LCMS: (Method E) 512.0 (M + +H), Rt. 2.83 points, 98.66% (maximum). HPLC: (Method B) Rt. 5.10 points, 98.19% (maximum).キラルSFC: (Method G) Rt. 2.50 points, 99.54% (maximum).

[0353] Semen 4: Yield: 52% (40 mg, Semen solid). 1H NMR (400 MHz, DMSO-d6): δ 7.36 (s, 1H), 7.07-6.98 (m, 4H), 6.73 (s, 1H), 4.33 (d, J = 8.4 Hz, 1H), 4.17-4.11 (m, 2H), 3.90-3.45 (m, LCMS: (Method H) 512.2 (M + +H), Rt. 2.06 min, 96.13% (max). HPLC: (Method B) Rt. 5.10 min, 96.16% (max). Chiral SFC: (Method G) Rt. 3.88 min, 99.19% (max). The absolute configurations of the four diastereomers are unknown.

[0354] Example 2 1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid

[0355] [ka]

[0356] To a stirred solution of ethyl 1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylate (Intermediate 15; 240 mg, 0.42 mmol) in a mixture of 1,4-dioxane and water (7:3, 10 mL), lithium hydroxide (36 mg, 0.85 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH 4) and diluted with ice-cold water (2 mL). The aqueous layer was extracted with EtOAc (2 × 10 mL), and the combined organic layers were washed with water (10 mL) and brine (10 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was triturated with hexane and then filtered to give the title compound. Yield: 22% (50 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.42 (s, 1H), 7.27 (s, 1H), 7.05-7.07 (m, 4H), 6.63 (s, 1H), 4.19 (s, 2H), 3.84-3.54 (bs, 2H), 3.27 (s, 2H), 2.16 (s, 3H), 1.59-1.36 (m, 4H), 1.35-0.98 (m, 8H), 0.79-0.69 (m, 6H). LCMS: (Method E) 536.2 (M + +H), Rt. 2.92 min, 98.33% (max). HPLC: (Method B) Rt. 5.91 min, 96.58% (max).

[0357] Example 3 (S)-1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid and (R)-1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid

[0358] [ka]

[0359] The two enantiomers of racemic 1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid (Example 2; 40 mg, 0.074 mmol) were separated by chiral SFC (Method O). The material was concentrated under vacuum at 40° C. The first eluting fraction corresponded to Enantiomer 1, and the second eluting fraction to Enantiomer 2. The absolute configurations of the two enantiomers are unknown.

[0360] Enantiomer 1: Yield: 32% (13 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.40 (s, 1H), 7.28 (s, 1H), 7.06 (d, J = 8.0 Hz, 4H), 6.63 (s, 1H), 4.19 (s, 2H), 3.65 (bs, 2H), 3.27 (s, LCMS: (Method E) 536.2 (M) ++H), Rt. 2.91 min, 97.46% (max). HPLC: (Method E) Rt. 5.99 min, 96.04% (max). Chiral SFC: (Method O) Rt. 2.74 min, 97.81% (max).

[0361] Enantiomer 2: Yield: 10% (4 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.41 (s, 1H), 7.28 (s, 1H), 7.06 (d, J = 8.4 Hz, 4H), 6.63 (s, 1H), 4.19 (s, 2H), 3.66 (bs, 2H), 3.32 (s, 2H), 2.16 (s, 3H), 1.51-1.36 (m, 4H), 1.31-1.24 (m, 3H), 1.11-1.07 (m, 6H), 0.80-0.70 (m, 6H). LCMS: (Method E) 536.1 (M + +H), Rt. 3.07 min, 95.83% (max). HPLC: (Method E) Rt. 5.995 min, 95.29% (max). Chiral SFC: (Method O) Rt. 3.70 min, 99.39% (max).

[0362] Example 4 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid

[0363] [ka]

[0364] To a stirred solution of methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoate (Intermediate 16; 300 mg, 0.54 mmol) in a mixture of 1,4-dioxane and water (7:3, 10 mL), lithium hydroxide (45 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH 4) and diluted with ice-cold water (2 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), and the combined organic layers were washed with water (5 mL) and brine (5 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The crude material obtained was purified by Isolera column chromatography (eluent: 9% MeOH / DCM; silica gel: 230-400 mesh), and the residue obtained was repurified by preparative HPLC (Method A) to give the title compound. Yield: 41% (130 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.43 (s, 1H), 7.27 (s,1H), 7.07-7.05 (m, 4H), 6.26 (s, 1H), 4.05 (s, 2H), 3.67 (bs, 2H), 3.34-3.28 (m, 2H), 2.15 (s, 3H), 1.50-1.34 (m, 4H), 1.25 (s, 6H), 1.22-1.04 (m, 4H), 0.78-0.72 (m, 6H). LCMS: (Method E) 538.1 (M + +H), Rt. 3.13 min, 97.56% (max). HPLC: (Method B) Rt. 6.04 min, 96.19% (max).

[0365] Example 5 (S)-3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid and (R)-3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid

[0366] [ka]

[0367] The two enantiomers of racemic 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid (Example 4; 100 mg, 0.18 mmol) were separated by SFC (Method H). The material was concentrated under vacuum at 40° C. The first eluting fraction corresponded to Enantiomer 1, and the second to Enantiomer 2. The absolute configurations of the two enantiomers are unknown.

[0368] Enantiomer 1: Yield: 33% (33 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.43 (s, 1H), 7.27 (s, 1H), 7.07 (t, J = 8.00 Hz, 4H), 6.63 (s, 1H), 4.06 (s, 2H), 3.66 (s, 2H), 3.28 (s, 2H), 2.16 (s, 3H), 1.31-1.33 (m, 10H), 1.02-1.04 (m, 4H), 0.73-0.75 (m, 6H). LCMS: (Method H) 538.5 (M + +H), Rt. 2.33 min, 96.81% (max). HPLC: (Method B) Rt. 6.05 min, 95.30% (max). Chiral SFC: (Method H) Rt. 3.35 min, 99.93% (max).

[0369] Enantiomer 2: Yield: 26% (26 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.40 (s, 1H), 7.27 (s, 1H), 7.05-7.07 (m, 4H), 6.63 (s, 1H), 4.06 (s, 1H), 3.65 (s, 2H), 3.28 (s, 2H), 2.16 (s, 3H), 1.31-1.33 (m, 11H), 1.02-1.04 (m, 4H), 0.73-0.75 (m, 6H). LCMS: (Method H) 538.5 (M + + H), Rt. 2.33 min, 97.41% (max). HPLC: (Method B) Rt. 6.02 min, 95.58% (max). Chiral SFC: (Method H) Rt. 2.74 min, 99.96% (max).

[0370] Example 6 3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid

[0371] [ka]

[0372] To a stirred solution of 3,3-diethyl-8-(2-hydroxy-2-methoxypropoxy)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 18; 30 mg, 0.05 mmol) in a mixture of 1,4-dioxane and water (2:3, 5 mL), lithium hydroxide (4.9 mg, 0.11 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH 4) and diluted with ice-cold water (2 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), and the combined organic layers were washed with water (5 mL) and brine (5 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The crude material obtained was purified by Isolera column chromatography (eluent: 9% MeOH / DCM; silica gel: 230-400 mesh), and the residue obtained was repurified by preparative HPLC (Method A) to give the title compound. Yield: 68% (20 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): 13.53 (bs, 1H), 7.35 (s, 1H), 7.22 (t, J = 8.4 Hz, 2H), 6.99 (d, J = 7.2 Hz, 2H), 6.84 (t, J = 7.6 Hz, 1H), 6.72 (s, 1H), 4.49-4.38 (m, 2H), 3.70 (s, 2H), 3.26 (s, 2H), 2.17 (s, 3H), 1.62-1.57 (m, 5H), 1.37-1.32 (m, 2H), 0.75 (t, J = 7.2 Hz, 6H). LCMS: (Method E) 496.0 (M + +H), Rt. 2.84 min, 99.03% (max). HPLC: (Method B) Rt. 5.25 min, 97.49% (max).

[0373] Example 7 (S)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid and (R)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid

[0374] [ka]

[0375] Enantiomer 1 To a stirred solution of methyl 3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoate enantiomer 1 (Intermediate 18; 30 mg, 0.07 mmol) in a mixture of 1,4-dioxane and water (2:3, 5 mL), lithium hydroxide (5 mg, 0.11 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH 4) and diluted with ice-cold water (2 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), and the combined organic layers were washed with water (5 mL) and brine (5 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 9% MeOH DCM; silica gel: 230-400 mesh) to afford the title compound. Starting from 30 mg of enantiomer 2 of intermediate 18 and following the same procedure, enantiomer 2 of the title compound was obtained.

[0376] Enantiomer 1: Yield: 77% (22 mg, white solid). 1H NMR (400 MHz, DMSO-d6): 13.54 (bs, 1H), 7.36 (s, 1H), 7.22 (dd, J = 7.2, 8.8 Hz, 2H), 6.96 (d, J = 7.2 Hz, 2H), 6.84 (t, J = 7.2 Hz, 1H), 6.72 (s, 1H), 4.47-4.38 (m, 2H), 3.67 (s, 2H), 3.26 (s, 2H), 2.17 (s, 3H), 1.62-1.57 (m, 5H), 1.37-1.32 (m, 2H), 0.75 (t, J = 7.2 Hz, 6H). LCMS: (Method E) 496.2 (M + +H), Rt. 2.64 points, 98.33% (maximum). HPLC: (Method B) Rt. 5.35 points, 98.75% (maximum).キラルSFC (Method G): Rt. 1.80 points, 98.82% (maximum).

[0377] Enatoplast 2: Yield: 77.54% (22 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): 13.52 (bs, 1H), 7.36 (s, 1H), 7.22 (t, J = 8.4 Hz, 2H), 6.96 (d, J = 7.6 Hz, 2H), 6.84 (t, J = 7.6 Hz, 1H), 6.72 (s, 1H), 4.47-4.38 (m, 2H), 3.68 (s, 2H), 3.27 (s, 2H), 2.17 (s, 3H), 1.62-1.57 (m, 5H), 1.37-1.32 (m, 2H), 0.75 (t, J = 7.2 Hz, 6H). LCMS: (Method E) 496.2 (M + +H), Rt. 2.64 points, 98.23% (maximum). HPLC: (Method B) Rt. 5.35 points, 98.42% (maximum).キラルSFC (Method G): Rt. 2.58 points, 98.47% (maximum). 2つのエナンチオマーのJueku configuration is unknown.

[0378] (Example 8) 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid

[0379] [ka]

[0380] To a stirred solution of methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoate (Intermediate 20; 60 mg, 0.10 mmol) in 1,4-dioxane (1.5 mL), lithium hydroxide (5.19 mg, 0.217 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, 2 mL), and the aqueous layer was extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude was purified by preparative HPLC (Method A) to give the title compound. Yield: 27% (16 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 13.20 (s, 1H), 7.33 (s, 1H), 7.07 (d, J = 7.20 Hz, 4H), 6.62 (s, 1H), 4.36-4.09 (m, 3H), 3.65 (bs, 2H), 3.34 (s, 2H), 3.29 (s, 3H), 2.16 (s, 3H), 1.52-1.37 (m, 4H), 1.11-0.99 (m, 4H), 0.77-0.72 (m, 6H). LCMS: (Method E) 540.0 (M + +H), Rt. 2.97 min, 98.12% (max). HPLC: (Method B) Rt. 5.59 min, 99.49% (max).

[0381] Example 9 3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid

[0382] [ka]

[0383] To a stirred solution of methyl 3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoate (Intermediate 26; 50 mg, 0.08 mmol) in a mixture of 1,4-dioxane and water (3 mL), lithium hydroxide (7.22 mg, 0.17 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, 2 mL), and the aqueous layer was extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude was purified by preparative HPLC (Method A) to give the title compound. Yield: 24% (12 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 13.06 (s, 1H), 7.32 (s, 1H), 7.09 (d, J = 6.8 Hz, 4H), 6.59 (s, 1H), 4.36-4.33 (m, 1H), 4.27-4.23 (m, 1H), 4.12 (bs, 1H), 3.64 (bs, 2H), 3.40 (s, 3H), 3.29 (s, 2H), 2.15 (s, 3H), 1.37-1.34 (m, 4H), 1.24-1.02 (m, 8H), 0.76 (t, J = 6.80 Hz, 6H). LCMS: (Method E) 568.1 (M ++H), Rt. 3.15 min, 98.26% (max). HPLC: (Method B) Rt. 6.07 min, 97.56% (max).

[0384] Example 10 (S)-3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid and (R)-3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid

[0385] [ka]

[0386] The two enantiomers of racemic 3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid (Example 9; 30 mg, 5.29 mmol) were separated by chiral SFC (Method H). The material was concentrated under vacuum at 40° C. The first eluting fraction corresponded to Enantiomer 1, and the second eluting fraction to Enantiomer 2. The absolute configurations of the two enantiomers are unknown.

[0387] Enantiomer 1: Yield: 16% (5.0 mg, white solid). 1H NMR (400 MHz, DMSO-d6): δ 13.12 (s, 1H), 7.33 (s, 1H), 7.09 (d, J = 6.8 Hz, 4H), 6.59 (s, 1H), 4.35-4.33 (m, 1H), 4.27-4.23 (m, 1H), 4.15-4.13 (m, 1H), 3.62 (bs, 2H), 3.40 (s, 3H), 3.38 (s, 2H), 2.15 (s, 3H), 1.41-1.37 (m, 6H), 1.35-1.03 (m, 6H), 0.76 (t, J = 6.40 Hz, 6H). LCMS: (Method E) 568.2 (M + +H), Rt. 2.78 points, 94.49% (maximum). HPLC: (Method B) Rt. 6.09 points, 97.7% (maximum).キラルSFC: (Method H) Rt. 3.55 points, 99.58% (maximum).

[0388] Enatoplast 2: Yield: 26% (8 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 13.05 (s, 1H), 7.33 (s, 1H), 7.09 (d, J = 6.8 Hz, 4H), 6.59 (s, 1H), 4.36-4.33 (m, 1H), 4.27-4.23 (m, 1H), 4.15-4.13 (m, 1H), 3.62 (bs, 2H), 3.40 (s, 3H), 3.29 (s, 2H), 2.14 (s, 3H), 1.41-1.30 (m, 6H), 1.30-1.02 (m, 6H), 0.76 (t, J = 6.80 Hz, 6H). LCMS: (Method E) 568.2 (M + + H), Rt. 2.78 points, 87.94% (max). HPLC: (Method B) Rt. 6.09 points, 90.72% (max).キラルSFC: (Method H) Rt. 4.32 points, 95.74% (maximum).

[0389] (Example 11) 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoic acid

[0390] [ka]

[0391] To a stirred solution of methyl 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoate (Intermediate 28; 97 mg, 0.17 mmol) in a mixture of 1,4-dioxane and water (7:3, 4 mL), lithium hydroxide (14 mg, 0.34 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH 3) and diluted with ice-cold water (2 mL). The aqueous layer was extracted with EtOAc (2 × 10 mL), and then the combined organic layers were washed with water (10 mL) and brine (10 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was triturated with hexane, filtered and dried under vacuum to give the title compound. Yield: 40% (20 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.91 (bs, 1H), 7.37 (s, 1H), 7.08-7.06 (m, 4H), 6.62 (s, 1H), 4.35-4.33 (m, 1H), 4.28-4.24 (m, 2H), 3.72-3.68 (m, 2H), 3.57-3.52 (m, 2H), 3.29-3.21 (m, 2H), 2.16 (s, 3H), 1.59-1.33 (m, 4H), 1.22-0.95 (m, 7H), 0.79-0.68 (m, 6H). LCMS: (Method E) 554.3 (M ++H), Rt. 2.87 min, 99.83% (max). HPLC: (Method B) Rt. 5.89 min, 97.41% (max).

[0392] Example 12 3-((3,3-dibutyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoic acid

[0393] [ka]

[0394] To a stirred solution of methyl 3-((3,3-dibutyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoate (Intermediate 30; 50 mg, 0.08 mmol) in a mixture of 1,4-dioxane and water (5:1, 3 mL), lithium hydroxide (7.2 mg, 0.17 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, 2 mL), and the aqueous layer was extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL) and dried over anhydrous NaSO. The organic portion was filtered and concentrated in vacuo. The resulting crude was purified by preparative HPLC (Method A) to give the title compound. Yield: 51% (25 mg, white solid). 1H NMR (400 MHz, DMSO-d6): δ 7.38 (s, 1H), 7.23 (t, J = 8.4 Hz, 2H), 7.01 (d, J = 8.0 Hz, 2H), 6.88 (t, J = 6.8 Hz, 1H), 6.67 (s, 1H), 4.36-4.34 (m, 1H), 4.27-4.23 (m, 2H), 3.70-3.67 (m, 2H), 3.56 (s, 2H), 3.28 (s, 2H), 2.15 (s, 3H), 1.43-1.36 (m, 2H), 1.33-1.24 (m, 3H), 1.17-1.03 (m, 10H), 0.77 (t, J = 6.80 Hz, 6H). LCMS: (Method E) 564.3 (M + +H), Rt. 3.08 min, 99.82% (max). HPLC: (Method B) Rt. 6.39 min, 97.45% (max).

[0395] Example 13 3-((3,3-diethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid

[0396] [ka]

[0397] To a solution of methyl 3-((3,3-diethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoate (Intermediate 35; 300 mg, 0.57 mmol) in a mixture of 1,4-dioxane and water (7:3, 10 mL), lithium hydroxide (48 mg, 1.14 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH 3) and diluted with ice-cold water (2 mL). The aqueous layer was extracted with EtOAc (2 × 10 mL), and then the combined organic layers were washed with water (20 mL) and brine (20 mL). The organic portion was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was triturated with hexane and then filtered to give the title compound. Yield: 15% (45 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.43 (s, 1H), 7.28 (s, 1H), 7.09-7.02 (m, 4H), 6.66 (s, 1H), 4.07 (s, 2H), 3.69 (s, 2H), 3.27 (s, 2H), 2.17 (s, 3H), 1.55-1.49 (m, 2H), 1.38-1.31 (m, 2H), 1.25 (s, 6H), 0.74 (t, J = 7.6 Hz, 6H). LCMS: (Method E) 510.2 (M + +H), Rt. 2.78 min, 98.4% (max). HPLC: (Method B) Rt. 5.55 min, 96.41% (max).

[0398] Example 14 2-Hydroxy-3-((7-(methylthio)-1,1-dioxide-5-phenyl-3,3-dipropyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)propanoic acid

[0399] [ka]

[0400] To a stirred solution of methyl 2-hydroxy-3-((7-(methylthio)-1,1-dioxide-5-phenyl-3,3-dipropyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)propanoate (Intermediate 36; 150 mg, 0.29 mmol) in 1,4-dioxane (5 mL), 6 N HCl (3 mL) was added at room temperature, and the reaction mixture was stirred at 100° C. for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water (10 mL), and the aqueous layer was extracted with ethyl acetate (2×10 mL). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 40-50% EtOAc / PE; silica gel: 230-400 mesh) to give the title compound. Yield: 46% (70 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.81 (s, 1H), 7.35 (s, 1H), 7.23 (t, J = 8.4 Hz, 2H), 7.00 (d, J = 7.6 Hz, 2H), 6.87 (t, J = 7.2 Hz, 1H), 6.68 (s, 1H), 5.80-5.40 (m, 1H), 4.42-4.32 (m, 1H), 4.32-4.15 (m, 2H), 3.88-3.58 (m, 2H), 3.28 (s, 2H), 2.15 (s, 3H), 1.52-1.35 (m, 2H), 1.32-1.12 (m, 6H), 0.70 (t, J = 6.80 Hz, 6H). LCMS: (Method K) 507.9 (M + +H), Rt. 2.69 min, 97.77% (max). HPLC: (Method E) Rt. 5.32 min, 95.61% (max).

[0401] Biological assays IBAT(h / m) assay protocol Ten thousand cells (human or mouse IBAT-overexpressing cells) were seeded into 96-well plates (Corning CLS3809) in 200 μL of MEM-alpha medium (Gibco 12571-063) supplemented with 10% FBS (Gibco 10438026) containing puromycin (Gibco A1113803) (10 μg / mL) and incubated at 37°C in 5% CO for 48 hours. After incubation, the medium was decanted from the wells, and the cells were washed twice with 300 μL of basal MEM-alpha medium (without FBS). After each decantation of basal MEM-alpha medium, the plate was tapped against a paper towel to ensure maximum removal of residual medium. Dilutions of test inhibitors (10 μM highest test concentration, 3-fold serial dilutions, 10-point) prepared in DMSO (Sigma D2650) were added to an incubation mixture containing 0.25 μM 3H-taurocholic acid (ARC ART-1368) and 5 μM cold taurocholic acid (Sigma T4009) (maintaining a final DMSO concentration of 0.2%). Then, 50 μL of incubation mixture containing the test inhibitor was added to the wells (in duplicate), and the plate was incubated at 37°C in a CO2 incubator for 20 min. After incubation, the reaction was stopped by placing the plate on an ice-water mixture for 2–3 min, and then the incubation mixture was completely aspirated from the wells. Wells were washed twice with 250 μL of cold unlabeled 1 mM taurocholate dissolved in HEPES (Gibco 15630080) buffered (10 mM) HBSS (Gibco 14175079), pH 7.4. After each wash, plates were tapped against a paper towel to ensure maximum removal of blocking buffer. 100 μL of MicroScint-20 (PerkinElmer 6013621) was added to the wells and kept at room temperature overnight, after which the plates were read in a PerkinElmer TopCount NXT™ Microplate Scintillation and Luminescence Counter under the 3H Test protocol (set for a read time of 120 seconds per well).

[0402] LBAT(h / m) assay protocol 20,000 cells (human or mouse LBAT-overexpressing cells) were seeded in 100 μL of MEM-alpha medium (Gibco 12571-063) supplemented with 10% FBS (Gibco 10438026) containing Geneticin (Gibco 10131-027) (1 mg / mL) in a 96-well plate (Corning CLS3809) and incubated for 24 hours at 37°C in 5% CO. After incubation, the medium was decanted from the wells, and the cells were washed twice with 300 μL of basal MEM-alpha medium (without FBS). After each decantation of basal MEM-alpha medium, the plate was tapped against a paper towel to ensure maximum removal of residual medium. For human LBAT, incubation mixtures were prepared by adding dilutions of test inhibitors (10-point, 3-fold serial dilutions in DMSO (Sigma D2650)) to MEM-alpha (without FBS) containing 0.3 μM H-taurocholic acid (ARC ART-1368) and 7.5 μM cold taurocholic acid (Sigma T4009) to maintain a final DMSO concentration of 0.2%. For mouse LBAT, incubation mixtures were prepared by adding dilutions of test inhibitors (10-point, 3-fold serial dilutions in DMSO) to MEM-alpha (without FBS) containing 0.3 μM H-taurocholic acid and 25 μM cold taurocholic acid to maintain a final DMSO concentration of 0.2%. Next, 50 μL of incubation mixture containing the test inhibitor was added to the wells (in duplicate), and the plate was incubated for 20 minutes at 37°C in a CO2 incubator. After incubation, the reaction was stopped by placing the plate on an ice-water mixture for 2-3 minutes, and then the incubation mixture was completely aspirated from the wells. The wells were washed twice with 250 μL of chilled unlabeled 1 mM taurocholate dissolved in HEPES (Gibco 15630080)-buffered (10 mM) HBSS (Gibco 14175079), pH 7.4. After each wash, the plate was gently tapped against a paper towel to ensure maximum removal of the blocking buffer. 100 μL of MicroScint-20 (PerkinElmer 6013621) was added to the wells and kept at room temperature overnight, after which the plates were read in a PerkinElmer TopCount NXT™ Microplate Scintillation and Luminescence Counter under the 3H Test protocol (normal plate orientation, set to a read time of 120 seconds per well).

[0403] Bidirectional permeability assay (Caco-2 cells) Caco-2 cells (Evotec) were seeded at a density of 70,000 cells / well in Millicell® 24-well cell culture insert plates and maintained in an incubator (37°C, 5% CO2, 95% RH) for 21 days with medium changes every other day. Stock solutions (10 mM) of the test compounds, atenolol (a low-permeability marker), propranolol (a high-permeability marker), and digoxin (a substrate for the P-gp transport pathway) were prepared in dimethyl sulfoxide (DMSO). Intermediate stock solutions (1 mM) were prepared by diluting 10 μL of the 10 mM master stock solution with 90 μL of neat DMSO. Working stock solutions (10 μM) were prepared by diluting 50 μL of the 1 mM solution with 4950 μL of FaSSIF buffer. After adding the compounds to FaSSIF, the samples were sonicated for 2 hours and centrifuged at 4000 RPM for 30 minutes at 37°C. 4 mL of the resulting supernatant was used directly in the assay. The final DMSO concentration in transport experiments was 1%. On the day of the assay, Caco-2 monolayers were washed twice with transport buffer (HBSS, pH 7.4) and pre-incubated in an incubator for 30 minutes (37°C, 5% CO2, 95% RH). The electrical resistance of the monolayers was measured using a Millicell®-ERS system: 350 ohm.cm 2 Monolayers with transepithelial electrical resistance (TEER) values ​​above 1000 were selected for the assay. Assays were performed in the absorptive (A2B) and secretory (B2A) directions. Transport experiments were initiated by adding compound-containing transport assay buffer (FaSSIF buffer prepared in HBSS) to the donor compartment (apical chamber AB; basolateral chamber BA) in duplicate wells (n = 2). Drug-free HBSS buffer (pH 7.4) containing 1% bovine serum albumin (BSA) was introduced into the receiver compartment (AB - basolateral; BA - apical). The volumes of the apical and basolateral compartments were 0.4 and 0.8 mL, respectively. After addition of the dosing solution, the plate was incubated at 37 °C in an incubator for 120 min. After 120 min, donor and receiver samples were taken and matrix-matched to the opposing buffer (1:1, 30 μL test sample + 30 μL blank buffer). The dosed sample was matrix-matched to the opposing buffer (1:1, 30 μL test sample + 30 μL blank buffer). Samples were processed by adding acetonitrile containing an internal standard (60 μL test sample + 200 μL acetonitrile containing internal standard - tolbutamide, 500 ng / mL). The samples were vortexed and centrifuged at 4000 rpm for 10 minutes. The resulting supernatant (100 μL) was diluted with 100 μL water and transferred to a new 96-well plate. Compound concentrations in the samples, where applicable, were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS / MS) using discovery-grade bioanalytical methods. The mean apparent permeability (P) of the test compounds, atenolol, propranolol, and digoxin app , ×10 -6 The velocity (cm / sec) was calculated as follows:

[0404]

number

[0405] (where dq / dt = transport rate (rate of transport of compound in receiver compartment), C0 = initial concentration in donor compartment, A = effective filtration membrane surface area).

[0406] HepaRG-based assay protocol A cryopreserved vial of differentiated HepaRG cells (Biopredic International HPR116080) is thawed in HepaRG Thawing / Plating / General Purpose Medium (Biopredic International ADD670C) supplemented with 200 mM Glutamax (Gibco 35050061) according to the protocol provided by Biopredic International. 70,000 cells per well are seeded in 100 μL of HepaRG Thawing / Plating / General Purpose Medium supplemented with 200 mM Glutamax in a 96-well plate (Corning CLS3809) and incubated at 37°C in 5% CO for 24 hours. After incubation, the seeding medium is replaced with HepaRG Maintenance / Metabolism Medium (Biopredic International ADD620C) and replenished with fresh HepaRG Maintenance / Metabolism Medium every 48 hours for 6 days. After 7 days of incubation post-seeding, the incubation medium is decanted from the wells and the cells are washed twice with 250 μL of William's E Basal Media (Gibco 12551032). After each decantation of William's E Basal Media, the plate is tapped against a paper towel to ensure maximum removal of residual medium. Incubation mixtures are prepared by adding dilutions of test inhibitors (3-fold serial dilutions in DMSO (Sigma D2650)) to William's E medium (basal) containing 0.3 μM 3H-taurocholic acid (ARC ART-1368) and 7.5 μM cold taurocholic acid (Sigma T4009) (maintaining a final DMSO concentration of 0.2%). Then, 50 μL of incubation mixture containing the test inhibitor is added to the wells (in duplicate), and the plate is incubated for 30 min at 37°C in a 5% CO2 incubator. After incubation, the reaction is stopped by placing the plate on an ice-water mixture for 2-3 min, and then the incubation mixture is completely aspirated from the wells. Wash the wells twice with 250 μL of cold unlabeled 1 mM taurocholate dissolved in HEPES (Gibco 15630080) buffered (10 mM) HBSS (Gibco 14175079), pH 7.4. After each wash, tap the plate against a paper towel to ensure maximum removal of blocking buffer. 100 μL of MicroScint-20 (PerkinElmer 6013621) is added to the wells and kept at room temperature overnight, after which the plate is read in a PerkinElmer TopCount NXT™ Microplate Scintillation and Luminescence Counter under the 3H Test protocol (set to a read time of 120 seconds per well in normal plate orientation).

[0407] Preparation of test compound dilutions All test compounds were prepared in powder form at room temperature. 10 mM DMSO stock solutions of test compounds were prepared, aliquoted, and stored at -20°C. From the 10 mM DMSO stock solutions of the compounds, 3-fold serial dilutions in DMSO were prepared to obtain a total of 10 dilutions of test compounds. 0.5 μL of this DMSO dilution was added to 250 μL of FBS-free basal medium containing 3H-taurocholate and cold taurocholate to prepare the incubation mixture.

[0408] Bioavailability test Eight- to nine-week-old male mice (C57BL / 6 or CD1) or Wistar rats were used. Two groups of three animals were used for each test compound. One group received a single intravenous dose of 1 mg / kg (vehicle 100% DMSO) via the tail vein, and the other group received a single oral dose of 10 mg / kg via gavage needle. The oral dose group was fasted overnight. Blood samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after intravenous administration and at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after oral administration. Blood samples were drawn from the saphenous vein. 0.2% EDTA was used as an anticoagulant. Samples were analyzed using a LC-MS / MS system by a discovery-grade bioanalytical method developed for the estimation of test compounds in plasma.

[0409] result Biological data for the example compounds is shown in Table 8 below.

[0410] [Table 8]

[0411] PD model: Evaluation of test compounds on total bile acid levels in male C57BL / 6 mice Eight- to nine-week-old C57BL / 6N Tac mice were used to test the effects of bile acid modulators on bile acid levels. After completing the quarantine and acclimation period, animals were randomly assigned based on body weight to x experimental groups: (i) vehicle control, and (ii) test compound y mg / kg orally once daily. Animals were treated with the test compound for 7 days. On day 5 of the study, animals were individually housed in new cages. On day 7, feces were collected from each cage, followed by blood collection from each animal via the retro-orbital route. Animals were euthanized, and liver and terminal ileum were collected from each animal for further analysis. Body weight and food intake were measured twice weekly. Serum lipid profiles were analyzed using serum samples on day 7. Serum total bile acids were measured using serum samples on day 7. Fecal bile excretion was measured using fecal samples on day 7. Hepatic expression of CYP7A1 and SHP will be quantified in liver samples on day 7. Hepatic triglycerides and total cholesterol will be analyzed in liver samples on day 7.

[0412] Urinary bile acid model: Evaluation of test compounds on urinary bile acid levels in male C57BL / 6 mice Eight- to nine-week-old C57BL / 6N Tac mice were used to test the effects of bile acid modulators on bile acid levels. After completing the quarantine and acclimation period, animals were randomly assigned based on body weight to x experimental groups: (i) vehicle control, and (ii) test compound y mg / kg orally once daily. Animals were treated with the test compound for 7 days. On day 6 of the study, animals were transferred to metabolic cages. On day 7, feces and urine were collected from each metabolic cage, followed by blood collection from each animal via the retro-orbital route. Animals were euthanized, and kidneys were collected from each animal for further analysis. Body weights were measured twice weekly. Serum total bile acids were measured in serum samples on day 7. Fecal bile acid excretion was measured in fecal samples on day 7. Urinary bile acid excretion was measured in samples on day 7. Renal expression of ASBT, OSTa, OSTAb, and MRP2 will be quantified in day 7 samples.

Claims

1. (S)-3-(((S)-3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; (S)-3-(((R)-3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; (R)-3-(((S)-3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; (R)-3-(((R)-3-butyl-5-(4-fluorophenyl)-3-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; 1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (S)-1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (R)-1-(((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; (S)-3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; (R)-3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; 3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid; (S)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid; (R)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoro-2-methylpropanoic acid; 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid; 3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid; (S)-3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid; (R)-3-((3,3-dibutyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-methoxypropanoic acid; 3-((3-butyl-3-ethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoic acid; 3-((3,3-dibutyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-ethoxypropanoic acid; 3-((3,3-diethyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; and 2-hydroxy-3-((7-(methylthio)-1,1-dioxide-5-phenyl-3,3-dipropyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)propanoic acid; or a pharmaceutically acceptable salt thereof.

2. 10. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 and one or more pharmaceutically acceptable excipients.

3. A pharmaceutical comprising the compound of claim 1.

4. 4. The pharmaceutical composition according to claim 3, for use in the treatment or prevention of cardiovascular diseases or disorders of fatty acid metabolism or disorders of glucose utilization, such as hypercholesterolemia; disorders of fatty acid metabolism; type 1 and type 2 diabetes mellitus; complications of diabetes, such as cataracts, small and large vessel disease, retinopathy, neuropathy, nephropathy, and delayed wound healing, tissue ischemia, diabetic foot, arteriosclerosis, myocardial infarction, acute coronary syndrome, unstable angina, stable angina, stroke, peripheral arterial occlusive disease, cardiomyopathy, heart failure, heart rhythm disorders, and vascular restenosis; diabetes-related diseases, such as insulin resistance (disorders of glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, obesity, dyslipidemia, hyperlipidemia, such as hypertriglyceridemia, metabolic syndrome (syndrome X), atherosclerosis, and hypertension; and increased high density lipoprotein levels.

5. 4. The medicament according to claim 3, for use in the treatment or prevention of gastrointestinal diseases or disorders, such as constipation (including chronic constipation, functional constipation, chronic idiopathic constipation (CIC), intermittent / sporadic constipation, constipation secondary to diabetes mellitus, constipation secondary to stroke, constipation secondary to chronic kidney disease, constipation secondary to multiple sclerosis, constipation secondary to Parkinson's disease, constipation secondary to systemic sclerosis, drug-induced constipation, irritable bowel syndrome with constipation (IBS-C), mixed irritable bowel syndrome (IBS-M), pediatric functional constipation, and opioid-induced constipation); Crohn's disease; primary bile acid malabsorption; irritable bowel syndrome (IBS); inflammatory bowel disease (IBD); ileal inflammation; and reflux disease and its complications, such as Barrett's esophagus, bile reflux esophagitis, and bile reflux gastritis.

6. Liver diseases or disorders, e.g., inherited metabolic disorders of the liver; congenital abnormalities of bile acid synthesis; congenital abnormalities of bile duct course; biliary atresia; biliary atresia after Kasai operation; biliary atresia after liver transplantation; neonatal hepatitis; neonatal cholestasis; inherited forms of cholestasis; cerebrotendinous xanthomatosis; secondary defects of BA synthesis; Zellweger syndrome; liver disease associated with cystic fibrosis; alpha-1 antitrypsin deficiency; Alagille syndrome (ALGS); Byler syndrome; primary defects of bile acid (BA) synthesis; progressive familial intrahepatic cholestasis (PFIC), e.g., PFIC-1, PFIC-2, PFIC-3, and non-specific PFIC, PFIC after bile diversion, and PFIC after liver transplantation; benign recurrent intrahepatic cholestasis (BRIC), e.g., BRIC BRIC 1, BRIC 2, and non-specific BRIC, post-bile diversion BRIC, and post-liver transplant BRIC; autoimmune hepatitis; primary biliary cirrhosis (PBC); liver fibrosis; non-alcoholic fatty liver disease (NAFLD); non-alcoholic steatohepatitis (NASH); portal hypertension; cholestasis; cholestasis of Down syndrome; drug-induced cholestasis; intrahepatic cholestasis of pregnancy (jaundice of pregnancy); intrahepatic cholestasis; extrahepatic cholestasis; parenteral nutrition-associated cholestasis (PNAC); hypophospholipid-associated cholestasis; lymphedema cholestatic syndrome 1 (LCS1); primary sclerosing cholangitis (PSC); immunoglobulin G4-associated cholangitis; primary biliary cholangitis; cholelithiasis (gallstones); biliary lithiasis (biliary lithiasis); choledocholithiasis; gallstone pancreatitis; Caroli disease; malignant tumors of the bile duct; malignant tumors causing obstruction of the biliary tree; bile duct stenosis; AIDS cholangiopathy; ischemic cholangiopathy; pruritus due to cholestasis or jaundice; pancreatitis; chronic autoimmune liver disease leading to progressive cholestasis; hepatic steatosis; alcoholic hepatitis; acute fatty liver; fatty liver of pregnancy; drug-induced hepatitis; iron overload; congenital bile acid metabolism disorder type 1 (BAS disorder type 1); drug-induced liver injury (DILI); hepatic fibrosis; congenital hepatic fibrosis; cirrhosis; Langerhans cell histiocytosis (LCH); neonatal ichthyotic sclerosing cholangitis (NISCH); erythropoietic protoporphyria (EPP); idiopathic adult bile duct paucity (IAD); idiopathic neonatal hepatitis (INH); nonsyndromic intrahepatic bile duct paucity (NS) PILBD); autosomal recessive intrahepatic cholestasis (North American Indian childhood cirrhosis) (NAIC);4. The pharmaceutical composition of claim 3 for use in the treatment or prevention of cholestasis caused by hepatic sarcoidosis; amyloidosis; necrotizing enterocolitis; serum bile acid-induced toxicity, such as cardiac rhythm disorders (e.g., atrial fibrillation) in the setting of an abnormal serum bile acid profile, cardiomyopathy associated with cirrhosis ("cholecardia"), and skeletal muscle wasting associated with cholestatic liver disease; polycystic liver disease; viral hepatitis (including hepatitis A, B, C, D, and E); hepatocellular carcinoma (hepatocellular carcinoma); cholangiocarcinoma; bile acid-related gastrointestinal cancer; and tumors and neoplasms of the liver, biliary tract, and pancreas; or for use in augmenting corticosteroid therapy in liver disease.

7. 4. The pharmaceutical of claim 3, for use in the treatment or prevention of hyperabsorption syndromes (including abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicron retention disease (CRD), and sitosterolemia); hypervitaminosis and osteopetrosis; hypertension; glomerular hyperfiltration; polycystic kidney disease (PKD), such as autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD); and pruritus in renal failure; or for use in protection against kidney damage associated with liver or metabolic disease.

Images