RU2785867C9 - Benzothia(di)azepines and their use as bile acid modulators - Google Patents

Abstract

FIELD: medicine; therapy.

SUBSTANCE: group of inventions relates to the field of medicine, namely to therapy; it is intended for the treatment of cardiovascular diseases, disorders of fatty acid metabolism and glucose use, gastrointestinal diseases, and liver diseases. Derivatives of 1,5-benzothiazepine and 1,2,5-benzothiadiazepine of the formula (I) are presented. In another implementation, a pharmaceutical composition is provided, containing these compounds.

EFFECT: group of inventions provides new derivatives of 1,5-benzothiazepine and 1,2,5-benzothiadiazepine of the formula (I), as well as a pharmaceutical composition containing these compounds, which are bile acid modulators having inhibitory activity relatively to an apical sodium-dependent bile acid transporter (ASBT) and/or a liver bile acid transporter (LBAT).

15 cl, 7 tbl, 110 ex

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to Indian Application No. 201811021016, filed June 5, 2018; according to the application of Sweden No. 1850915-8, filed on July 18, 2018; and Indian Application No. 201911000892, filed January 8, 2019, the disclosures of which are hereby incorporated by reference in their entirety.

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

The present invention relates to 1,5-benzothiazepine and 1,2,5-benzothiadiazepine derivatives of formula (I). These compounds are bile acid modulators having inhibitory activity against apical sodium dependent bile acid transporter (ASBT) and/or liver bile acid transporter (LBAT). The present invention also relates to pharmaceutical compositions containing these compounds and to the use of these compounds for the treatment of cardiovascular diseases, disorders of fatty acid metabolism and glucose utilization, gastrointestinal diseases and liver diseases.

BACKGROUND OF THE INVENTION

Bile acids are physiological detergents that play an important role in the absorption and transport of lipids, nutrients and vitamins in the intestine. 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 that are 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 the gallbladder to contract and thereby release bile acids into the small intestine, where they activate the absorption of fat-soluble vitamins and cholesterol. When they enter the ileum, bile acids are reabsorbed from the intestine and secreted into the portal blood for return to the liver via the portal venous circulation. Thus, more than 90% of bile acids are recycled and returned to the liver. These bile acids are then carried across the sinusoidal membranes of the hepatocytes and re-secreted across the canalicular membranes into the bile. In this last passage, 75-90% of the bile acids are taken up by the hepatocytes, completing one round of the enterohepatic circulation. The remaining portion of the bile acids cleared in the liver enters the systemic circulation, where free bile acids are filtered by the renal glomeruli, effectively enter the proximal renal tubules, and are excreted back into the systemic circulation. Notably, most of the bile acids secreted across the canalicular membranes into the bile originate from the recirculation pool and less than 10% come from new synthesis in the liver. A small proportion of bile acids that are not reabsorbed in the ileum enters the large intestine. In the intestinal lumen, primary bile acids are converted to secondary bile acids by water by the action of intestinal bacteria, many via single or double dehydroxylation reactions of the steroid nucleus. Bile acids that are not absorbed in the intestines are then excreted in the faeces.

In general, an efficient transport system helps to maintain a high constant pool of bile acids, which provides high enough conjugated bile acids in the intestine to stimulate lipid absorption as well as reduce the bacterial load in the small intestine. The system also minimizes fecal and urinary bile acid loss and protects the intestinal and hepatobiliary compartments by removing potentially cytotoxic detergents (reviewed in Kosters and Karpen (Xenobiotica 2008, vol. 38, p. 1043-1071); Chiang (J. Lipid Res. 2009, vol. 50, p. 1955-1966), and Dawson (Handb. Exp. Pharmacol. 2011, vol. 201, p. 169-203)).

It has been established that the regulation of the size of the pool of bile acids plays a key role in cholesterol homeostasis by converting cholesterol into bile acid in the liver, which is the main route of excretion of cholesterol from the body. The liver plays an important role in removing endogenous and xenobiotic compounds from the body. Normal hepatobiliary secretion and enterohepatic circulation are necessary for the elimination of endogenous compounds such as cholesterol and bilirubin and their metabolites from the body, which maintains lipid and bile acid homeostasis. (Kosters and Karpen, Xenobiotica 2008, vol. 38, p. 1043-1071).

The reabsorption of bile acids in the ileum can be suppressed by compounds that inhibit the apical sodium-dependent bile acid transporter (ASBT). Bile acid reuptake suppression has been reported to be useful in the treatment of various diseases including dyslipidemia, diabetes, obesity, constipation, cholestatic liver disease, non-alcoholic steatohepatitis and other liver diseases. A number of ASBT inhibitor compounds have been disclosed over the past decades, see e.g. 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 2015/13713 864582, EP 489423, EP 549967, EP 573848, EP 624593, EP 624594, EP 624595, EP 624596, EP 0864582, EP 1173205 and EP 1535913.

Despite a number of ASBT inhibitor compounds that have been previously described, there is still a need for bile acid modulating compounds that have an optimized profile of activity, selectivity and bioavailability.

DETAILED DESCRIPTION OF THE INVENTION

Certain 1,5-benzothiazepine and 1,2,5-benzothiadiazepine derivatives have been found to be potent inhibitors of apical sodium-dependent bile acid transporter (ASBT) and/or hepatic bile acid transporter (LBAT) and may be useful in the treatment of diseases in which inhibition of bile acid circulation is desirable.

The first object of the present invention is a compound of formula (I)

wherein

M is selected from the group consisting of -CH ₂ - and -NR ⁷ -;

R ¹ and R ² are all independently C _1-4 alkyl;

R ³ is selected from the group consisting of hydrogen, halogen, hydroxy, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, cyano, nitro, amino, N- (C _1-4 alkyl)amino, N, N -di(C _1-4 alkyl)amino, N- (aryl-C _1-4 alkyl)amino, C _1-6 alkylcarbonylamino, C _3-6 cycloalkylcarbonylamino, N- (C _1-4 alkyl)aminocarbonyl, N, N -di(C _1-4 alkyl)aminocarbonyl, C _1-4 alkyloxycarbonylamino group, C _3-6 cycloalkyloxycarbonylamino group, C _1-4 alkylsulfonamide group and C _3-6 cycloalkylsulfonamide group;

n is an integer equal to 1, 2 or 3;

R ⁴ is selected from the group consisting of hydrogen, halogen, cyano, C _1-4 alkyl, C _3-6 cycloalkyl, C _1-4 alkoxy, C _3-6 cycloalkyloxy, C _1-4 alkylthio, C _3-6 cycloalkylthio, amino , N -(C _1-4 alkyl)amino and N,N -di(C _1-4 alkyl)amino;

one of R ⁵ and R ⁶ is carboxy and the other of R ⁵ and R ⁶ is selected from the group consisting of hydrogen, fluorine, C _1-4 alkyl and C _1-4 haloalkyl;

R ⁷ is selected from the group consisting of hydrogen and C _1-4 alkyl; And

R ⁸ is selected from the group consisting of hydrogen and C _1-4 alkyl;

or a pharmaceutically acceptable salt thereof.

In some embodiments, R ¹ and R ² are all independently C _2-4 alkyl. In other embodiments, R ¹ is n-butyl and R ² is C _2-4 alkyl. In a preferred embodiment, both R ¹ and R ² are n-butyl. In another preferred embodiment, R ¹ is n-butyl and R ² is ethyl. In yet another preferred embodiment, both R ¹ and R ² are ethyl.

In some embodiments, R ³ is selected from the group consisting of hydrogen, bromine, hydroxy, methoxy, amino, t-butoxycarbonylamino, methylsulfonamide, and cyclopropylsulfonamide. In a preferred embodiment, n is 1, i.e. the phenyl ring is substituted with only one R ³ substituent. In another preferred embodiment, R ³ is in the para position.

In some embodiments, R ⁴ is selected from the group consisting of hydrogen, bromine, ethyl, cyclopropyl, methoxy, methylthio, and dimethylamino.

In some embodiments, R ⁵ is selected from the group consisting of hydrogen and fluorine.

In some embodiments, R ⁶ is a carboxy group.

In some embodiments, R ⁷ is hydrogen or methyl.

In some embodiments, R ⁸ is hydrogen.

In a preferred embodiment, the compound of formula (I) is a compound of formula (I-a):

wherein

R ¹ and R ² are all independently ethyl or n-butyl;

R ⁴ represents fluorine, chlorine, bromine, methylthio or ethylthio; And

R ⁵ represents hydrogen or fluorine;

or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the compound of formula (I) is a compound of formula (I-b):

wherein

R ¹ and R ² are all independently ethyl or n-butyl;

R ⁴ represents chlorine or methylthio; And

R ⁵ represents hydrogen or fluorine;

or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the compound of formula (I) is a compound of formula (I-c):

wherein

R ⁴ represents chlorine or methylthio;

or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the compound of formula (I) is a compound of formula (I-d):

wherein

R ⁴ represents chlorine, methylthio, ethylthio or dimethylamino;

or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the compound of formula (I) is a compound of formula (Id) wherein R ⁴ is chloro or methylthio;

or a pharmaceutically acceptable salt thereof

In another preferred embodiment, the compound of formula (I) is a compound of formula (I-e):

wherein

R ⁴ represents chlorine or methylthio; And

R ⁵ represents hydrogen or fluorine;

or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the compound of formula (I) is a compound of formula (I-f):

wherein

R ¹ and R ² are all independently ethyl or n-butyl;

R ⁴ represents chlorine or methylthio; And

R ⁵ represents hydrogen or fluorine;

or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the compound of formula (I) is a compound of formula (I-g):

wherein

R ¹ and R ² are all independently ethyl or n-butyl;

R ⁴ represents chlorine or methylthio; And

R ⁵ represents hydrogen or fluorine;

or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the compound of formula (I) is a compound of formula (I-h):

wherein

R ¹ is n-butyl;

R ² is ethyl or n-butyl;

R ³ is selected from the group consisting of N -(C _1-4 alkyl)amino group, N, N -di(C _1-4 alkyl)amino group, N -(aryl-C _1-4 alkyl)amino group, C _1-6 alkylcarbonylamino group , a C _1-4 alkyloxycarbonylamino group, a C _1-4 alkylsulfonamide group, and a C _3-6 cycloalkylsulfonamide group;

R ⁴ is halo or C _1-4 alkylthio; And

R ⁵ represents hydrogen or fluorine;

or a pharmaceutically acceptable salt thereof.

In another preferred embodiment, the compound of formula (I) is a compound of formula (I-i):

wherein

M is CH ₂ or NH;

R ¹ is n-butyl;

R ² is ethyl or n-butyl;

R ⁵ represents hydrogen or fluorine;

or a pharmaceutically acceptable salt thereof.

In a preferred embodiment, the compound of formula (I) is selected from the group consisting of:

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid;

(E) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(E) -3-((5-(4-((tert-butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid;

(R)- (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(S)- (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(E) -3-((3,3-dibutyl-7-cyclopropyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid;

(E) -3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-(cyclopropanesulfonamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-(methylsulfonamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid;

(Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid;

(E) -3-((3,3-dibutyl-7-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid;

(E) -3-((3-butyl-3-ethyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid;

(E) -3-((3,3-dibutyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid;

(E) -3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid;

(R)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid;

(S) -(E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(E) -3-((3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid ;

( E )-3-((5-(4-(benzylamino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid;

( E )-3-((7-bromo-5-(4-((tert-butoxycarbonyl)amino)phenyl)-3-butyl-3-ethyl-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-((methoxycarbonyl)amino)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro- 1,5-benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-(dimethylamino)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid;

(Z) -3-((5-(4-((tert-butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((5-(4-((butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-(3,3-dimethylbutanamido)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro -1,5-benzothiazepin-8-yl)oxy)acrylic acid;

(Z) -3-((3,3-dibutyl-5-(4-isobutyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-isobutyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid;

(Z) -3-((3,3-dibutyl-5-(4-(cyclopentanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-dibutyl-5-(4-(cyclopropanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((3,3-dibutyl-5-(4-(cyclopentanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydrobenzo1-,5- thiazepin-8-yl)hydroxy)acrylic acid;

(E) -3-((3-butyl-5-(4-(cyclopentanecarboxamido)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1 ,5-benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-butyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid;

(R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-dibutyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid;

(R) -(Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(S)- (Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid;

(R)- (E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(S)- (E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(Z) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-dibutyl-7-chloro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) -2-fluoroacrylic acid;

(E) -3-((3,3-diethyl-7-iodo-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid;

(E) -3-((7-bromo-3,3-diethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid;

(Z) -3-((7-bromo-3,3-diethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) -2-fluoroacrylic acid;

(S)- (Z) -3-((3-butyl-3-ethyl-7-iodine-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(R)- (Z) -3-((3-butyl-3-ethyl-7-iodine-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid;

(S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid;

(R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3-butyl-5-(4-(tert-butylcarbamoyl)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro -1,5-benzothiazepin-8-yl)oxy)acrylic acid;

(Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid;

(R)- (Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(S)- (Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((5-(4-(benzylamino)phenyl)-3,3-diethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro-1 ,5-benzothiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3,3-dibutyl-5-(4-(isopropylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid;

(Z) -3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-propionamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid;

(E) -3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid;

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)-2-methacrylic acid;

(E) -3-((7-bromo-3,3-dibutyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl) hydroxy)acrylic acid;

(E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 8-yl)hydroxy)acrylic acid;

(S)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)acrylic acid;

(R)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)acrylic acid;

(Z) -3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro-1 ,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid;

(E) -3-((3,3-dibutyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid;

(E) -3-((3,3-dibutyl-7-cyano-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid;

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)oxy)but-2-enoic acid;

(Z)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)acrylic acid;

(E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)acrylic acid;

(S)- (E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid;

(R)- (E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro1, 2,5-benzothiadiazepin-8-yl)oxy)acrylic acid;

(E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid;

(S)- (E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid;

(R)- (E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid;

(Z) -3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)-2-fluoroacrylic acid;

(Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(S)- (Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(R)- (Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid;

(R)- (E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(S)- (E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid;

(Z) -3-((3,3-dibutyl-5-(4-(dimethylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-dibutyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((7-bromo-3-butyl-3-ethyl-2-methyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3,3-dibutyl-5-(3,4-difluorophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

(Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid;

(S)- (Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; And

(R)- (Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid;

or a pharmaceutically acceptable salt thereof.

When used in the present invention, the term "halogen" represents fluorine, chlorine, bromine and iodine.

When used in the present invention, the term "C _1-6 alkyl" is a linear or branched alkyl group containing from 1 to 6 carbon atoms, and the term "C _1-4 alkyl" is a linear or branched alkyl group containing from 1 to 4 carbon atoms. Examples of C _1-4 alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl.

When used in the present invention, the term "C _1-4 haloalkyl" is a linear or branched C _1-4 alkyl group as defined in the present invention, where one or more hydrogen atoms are replaced by halogen. Examples of C _1-4 haloalkyl include chloromethyl, fluoroethyl and trifluoromethyl.

When used in the present invention, the terms "C _1-4 alkoxy" and "C _1-4 alkylthio" represent a linear or branched C _1-4 alkyl group attached to the rest of the molecule through an oxygen or sulfur atom, respectively.

When used in the present invention, the term "C _3-6 cycloalkyl" is a monocyclic saturated hydrocarbon ring containing from 3 to 6 carbon atoms. Examples of C _3-6 cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term "aryl" represents an aromatic monocyclic ring of 6 carbon atoms or an aromatic bicyclic ring system of 10 carbon atoms. Examples of aryl include phenyl, naphthyl and azulenyl.

The term "amino group" is a -NH ₂ group. When used in the present invention, the terms " N -(C _1-4 alkyl)amino group" and " N,N -di(C _1-4 alkyl)amino group" are an amino group in which one or both hydrogen atoms are replaced, respectively, by a linear or branched C _1-4 alkyl group. Examples of the N -(C _1-4 alkyl)amino group include a methylamino group, an ethylamino group and a tert-butylamino group, and examples of the N,N -di-(C _1-4 alkyl)amino group include a dimethylamino group and a diethylamino group.

When used in the present invention, the term " N -(aryl-C _1-4 alkyl)amino group" is an amino group in which the hydrogen atom is replaced by an aryl-C _1-4 alkyl group. Examples of the N- (aryl-C _1-4 alkyl)amino group include a benzylamino group and a phenylethylamino group. The term "C _1-6 alkylcarbonylamino group" is an amino group in which a hydrogen atom is replaced by a C _1-6 alkylcarbonyl group. Examples of the C _1-6 alkanoylamino group include an acetylamino group and a t-butylcarbonylamino group. The term "C _1-4 alkyloxycarbonylamino group" is an amino group in which a hydrogen atom is replaced by a C _1-4 alkyloxycarbonyl group. An example of a C _1-4 alkyloxycarbonylamino group is t-butoxycarbonylamino group. The terms "C _1-4 alkylsulfonamide group" and "C _3-6 cycloalkylsulfonamide group" represent an amino group in which a hydrogen atom is replaced by a C _1-4 alkylsulfonyl or C _3-6 cycloalkylsulfonyl group, respectively.

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

When used in the present invention, the term "about" in the present invention is a value or parameter that includes (and describes) options that refer to that value or parameter. For example, a description indicating "about 20" includes the indication "20". Numeric ranges include limit values. Typically, the term "about" is the specified value of the variable and all values of the variable that deviate from the value by the amount of experimental error for the specified value (e.g., within a 95% confidence interval of the mean) or within 10% of the specified value, depending on that which is more.

The 1,5-benzothiazepines and 1,2,5-benzothiadiazepines of formula (I), or pharmaceutically acceptable salts thereof, are inhibitors of apical sodium-dependent bile acid transporter (ASBT inhibitors), hepatic bile acid transporter (LBAT inhibitors), or apical sodium- dependent bile acid transporter and hepatic bile acid transporter (dual ASBT/LBAT inhibitors). Therefore, they are useful for treating or preventing pathological conditions, disorders and diseases in which inhibition of bile acid circulation is desirable, such as cardiovascular diseases, disorders of fatty acid metabolism and glucose utilization, gastrointestinal diseases and liver diseases.

Cardiovascular disease and disorders of fatty acid metabolism and glucose use include, but are not limited to, hypercholesterolemia; metabolic disorders of fatty acids; diabetes mellitus type 1 and type 2; complications of diabetes, including cataracts, micro- and macrovascular diseases, retinopathy, neuropathy, nephropathy and slow wound healing, tissue ischemia, diabetic foot, arteriosclerosis, myocardial infarction, acute coronary syndrome, unstable angina, stable angina, stroke, peripheral artery occlusive disease, cardiomyopathy, heart failure, heart rate disturbances and vascular restenosis; diabetes-related diseases such as insulin resistance (impaired glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood fatty acids or glycerol, obesity, dyslipidemia, hyperlipidemia, including hypertriglyceridemia, metabolic syndrome (syndrome X), atherosclerosis, and hypertension; and an increase in high-density lipoprotein content.

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), functional constipation in children and opioid-induced constipation); Crohn's disease; primary malabsorption of bile acids; irritable bowel syndrome (IBS); inflammatory bowel disease (IBD); inflammation of the ileum; and reflux disease and its complications such as Barrett's esophagus, biliary reflux esophagitis and biliary reflux gastritis.

The liver disease defined in the present invention is any disease of the liver and associated organs such as the pancreas, portal vein, liver parenchyma, intrahepatic biliary tree, extrahepatic biliary tree and gallbladder. In some cases, the liver disease is a bile acid dependent liver disease. Diseases and disorders of the liver include, but are not limited to, hereditary metabolic disorder in the liver; congenital defects in the synthesis of bile acids; congenital anomalies of the bile ducts; atresia of the bile ducts; atresia of the bile ducts after the Kasai operation; bile duct atresia after liver transplantation; congenital giant cell hepatitis; cholestasis of newborns; hereditary forms of cholestasis; cerebrotendon xanthomatosis; secondary defect in BA synthesis (bile acids); Zellweger's syndrome; cystic fibrosis-associated liver disease; alpha-1 antitrypsin deficiency; Alagille syndrome (ALGS); Byler's syndrome; primary defect in bile acid synthesis (BA); progressive familial intrahepatic cholestasis (PFIC), including PFIC-1, PFIC-2, PFIC-3 and non-specific PFIC, PFIC after bile diversion and PFIC after liver transplantation; benign recurrent intrahepatic cholestasis (BRIC), including BRIC1, BRIC2 and non-specific BRIC, BRIC after bile diversion and BRIC after liver transplantation; autoimmune hepatitis; primary biliary cirrhosis (PBC); liver fibrosis; non-alcoholic fatty liver disease (NAFLD); non-alcoholic steatohepatitis (NASH); portal hypertension; cholestasis; cholestasis in Down syndrome; drug-induced cholestasis; intrahepatic cholestasis of pregnancy (jaundice during pregnancy); intrahepatic cholestasis; extrahepatic cholestasis; associated with parenteral nutrition cholestasis (PNAC); associated with a low content of phospholipids cholestasis; Aagenes syndrome 1 (LSC1); primary sclerosing cholangitis (PSC); associated with immunoglobulin G4 cholangitis; primary biliary cholangitis; cholelithiasis (gallstones); biliary lithiasis; choledocholithiasis; gallstone pancreatitis; Caroli disease; malignant tumor of the bile ducts; a malignant tumor causing obstruction of the biliary tree; strictures of the bile ducts; AIDS-associated cholangiopathy; ischemic cholangiopathy; pruritis due to cholestasis or jaundice; pancreatitis; chronic autoimmune liver disease leading to progressive cholestasis; fatty hepatosis; alcoholic hepatitis; acute fatty infiltration of the liver; fatty infiltration of the liver of pregnant women; medicinal hepatitis; iron overload disorders; congenital defect in bile acid synthesis type 1 (BAS type 1); drug-induced liver injury (DILI); liver fibrosis; congenital fibrosis of the liver; cirrhosis of the liver; histiocytosis of Langerhans cells (LCH); ichthyosis of newborns - sclerosing cholangitis (NISCH); erythropoietic protoporphyria (EPP); adult idiopathic ductopenia (IAD); idiopathic congenital giant cell hepatitis (INH); non-syndromic interlobular bile duct insufficiency (NS PILBD); North American Indian Infantile Cirrhosis (NAIC); sarcoidosis of the liver; amyloidosis; necrotizing enterocolitis; toxicities caused by serum bile acids, including cardiac arrhythmias (eg, atrial fibrillation) in cases of an abnormal serum bile acid profile, cardiomyopathy associated with cirrhosis of the liver ("cholecardia"), and skeletal muscle weakness associated with cholestatic liver disease; viral hepatitis (including hepatitis A, hepatitis B, hepatitis C, hepatitis D and hepatitis E); hepatocellular carcinoma (hepatoma); cholangiocarcinoma; associated with bile acids gastrointestinal cancers; and cholestasis caused by tumors and neoplasms in the liver, biliary tract and pancreas.

Other diseases that can be treated or prevented by the compounds of formula (I) or their pharmaceutically acceptable salts include hyperabsorption syndromes (including abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicron retention disease (CRD) and sitosterolemia); hypervitaminosis and osteopetrosis; hypertension; glomerular hyperfiltration; and pruritis in renal failure.

Bile acid transport in the human body is regulated by the action of members of the SLC10 family of solute transport proteins, specifically the Na ⁺ -taurocholate co-transport polypeptide (NTCP, also referred to as the liver bile acid transporter (LBAT); gene symbol SLC10A1 ), which is expressed in the sinusoidal membrane of hepatocytes, and 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 renal tubule cells, epithelium liver, large cholangiocytes and gallbladder epithelial cells. In the liver, bile acids are efficiently removed from the portal blood by the hepatic bile acid transporter (LBAT) and secreted via canalicular membranes by the bile salt excretion pump (BSEP; gene symbol ABCB11 ). The reabsorption of bile acids in the ileum is carried out by the apical sodium-dependent bile acid transporter (ASBT), where it is commonly referred to as the ileal bile acid transporter (IBAT). Both LBAT and ASBT act as electrogenic sodium-solute co-transporters that move two or more Na ⁺ ions per solute molecule.

Xenobiotics and endobiotics, including bile acids, enter the liver from the portal blood and are secreted into the bile via specific carrier proteins with individualized substrate specificities. Glycine- and taurine-conjugated bile acids exist in an anionic form and are unable to cross membranes by diffusion, and are therefore entirely dependent on membrane transport proteins for entry into or out of hepatocytes (Kosters and Karpen, Xenobiotica 2008, vol. 38, p. 1043-1071). For ASBT and LBAT, glycine and taurine conjugated bile salts are preferred over their non-conjugated counterparts and have a greater affinity for trihydroxy bile salts. No non-bile acid substrates have yet been identified for ASBT, but LBAT has also been shown to tolerate various steroid sulfates, hormones, and xenobiotics.

In terms of drug inhibition requirements, LBAT has not been as thoroughly characterized as ASBT. Dong et al. identified FDA-approved drugs that inhibit human LBAT and compared LBAT and ASBT in terms of inhibition requirements. Conducted a series of LBAT inhibition studies using FDA-approved drugs in conjunction with an iterative computer model. The screening identified 27 drugs as novel LBAT inhibitors, including irbesartan (Ki=11.9 μM) and ezetimibe (Ki=25.0 μM). It has been found that a common pharmacophore feature is that two hydrophobes and one hydrogen bond acceptor are important for LBAT inhibition. Of the 72 drugs tested in vitro, only 31 drugs inhibited LBAT, and 51 drugs (i.e., more than half) inhibited ASBT. Therefore, since inhibitor overlap was observed, ASBT was unexpectedly more inhibitory drug than LBAT, and this may be due to the fact that LBAT has fewer pharmacophore features (Dong et al., Mol. Pharm. 2013, vol. 10 , pp. 1008-1019).

Vaz et al. reported the identification of LBAT deficiency as a new neonatal metabolic disorder with a relatively "mild" clinical phenotype. The detection of LBAT deficiency confirmed that this transporter is the main system for the transfer of conjugated bile salts to the liver, but also showed that auxiliary transporters are also able to maintain the enterohepatic cycle in its absence (Vaz et al., Hepatology 2015, vol. 61, p. 260 -267). These data support the hypothesis that LBAT inhibition is a safe mechanism of action, since hepatocytes still have the ability to absorb the required amount of bile acids.

Liu et al. described the identification of a new type of hypercholanemia that is associated with the homozygosity of the p.Ser267Phe mutation in SLC10A1 (LBAT). the allele frequency of this mutation in the SLC10A1 gene is different for different populations, with the highest frequency observed in South China (8% and 12% in the Chinese cities of Han and Dai, respectively) and in Vietnam (11%). This "latent" hypercholanemia is thought to affect 0.64% of the population in southern Han, 1.44% of the population in the Chinese city of Dai, and 1.21% of the population in Vietnam. There was also an increase in the content of conjugated and non-conjugated BA in the serum of homozygous individuals. Liu et al. suggested that these findings are most likely due to reduced transfer of BA from the portal circulation to hepatocytes. These data support the hypothesis that the physiological function of the enterohepatic circulation is not only to recirculate bile acids, but also to remove bile acids from the circulation to ensure homeostasis (Karpen and Dawson, Hepatology 2015, vol. 61, p. 24-27 ). Alternatively, the liver may synthesize increased amounts of bile acids to compensate for the decreased enterohepatic circulation in homozygous carriers. Since LBAT also transports non-conjugated bile acids, the increase in non-conjugated bile acids in this study was not unexpected (Liu et al., Scientific Reports 2017, 7: 9214, p. 1-7).

LBAT has been found to be inactivated in some forms of cholestatic liver disease and cholestasis, whereas ASBT has been found to be inactivated in various gastrointestinal disorders such as Crohn's disease, primary bile acid malabsorption, inflammatory bowel disease, and ileum inflammation, but is upregulated in cholestasis . LBAT also acts as a cellular receptor for the entry of hepatitis B virus (HBV) and hepatitis D virus (HDV), which are the main cause of liver disease and hepatocellular carcinoma.

The effect of ASBT inhibition on lowering plasma cholesterol and increasing insulin resistance, as well as on reducing the load of the liver with bile acids in cholestatic liver disease, has been studied. In addition, ASBT inhibition has been found to restore insulin levels and normoglycemia, and it has been shown that ASBT inhibition is a promising treatment for type 2 diabetes mellitus. ASBT inhibitors have also been used to treat functional constipation.

Since ASBT is expressed primarily in the ileum (where it is often presented as IBAT), ASBT inhibitors are not required to be available systemically. On the other hand, ASBT is also expressed in proximal tubular cells of the kidney. Therefore, systemically available ASBT inhibitors can also inhibit bile acid reuptake in the kidneys. It is hypothesized that this leads to increased levels of bile acids in the urine and increased excretion of bile acids from the body in the urine. Therefore, systemically available ASBT inhibitors that act not only in the ileum but also in the kidney are expected to result in greater reductions in bile acids than systemically available ASBT inhibitors that act only in the ileum.

Compounds having high activity in inhibiting ASBT 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).

Bile duct atresia is a rare liver disease in children that involves complete or partial blockage (or even absence) of the large bile ducts. This blockage or absence causes cholestasis, which leads to a buildup of bile acids that damages the liver. In some embodiments, the accumulation of bile acids occurs in the extrahepatic biliary tree. In some embodiments, the accumulation of bile acids occurs in the intrahepatic biliary tree. The current standard of care is the Kasai operation, which is an operation that removes blocked bile ducts and directly connects often the small intestine to the liver. There are currently no approved drugs for this disorder.

The present invention relates to 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 subjected to Kasai surgery prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a compound of formula (I), or a pharmaceutically acceptable salt thereof, is administered to the subject prior to undergoing Kasai surgery. In some embodiments, treatment of biliary atresia results in a decrease in serum bile acids in the subject. In some embodiments, serum bile acids are determined, for example, using an ELISA enzyme assay or total bile acid assays as described in Danese et al., PLoS One. 2017, vol. 12(6): e0179200, which is hereby incorporated by reference in its entirety. In some embodiments, serum bile acids may decrease, for example, by 10%-40%, 20%-50%, 30%-60%, 40%-70%, 50%-80% or more than 90% compared with the content of bile acids in the serum before the introduction of the compounds of formula (I) or its pharmaceutically acceptable salt. In some embodiments, treatment of biliary atresia includes treatment of pruritis.

PFIC is a rare genetic disorder that is estimated to affect one in every 50,000-100,000 babies born worldwide and cause progressive, life-threatening liver disease.

One manifestation of PFIC is pruritis, which often leads to a severe deterioration in quality of life. In some cases, PFIC leads to cirrhosis and liver failure. Current treatments include partial bile diversion (PEBD) and liver transplantation, but these options can lead to significant postoperative risk, as well as psychological and social problems.

Three alternative gene defects have been identified that correlate with three distinct subtypes of PFIC, known as types 1, 2, and 3.

PFIC type 1, often referred to as "Byler's disease", is caused by impaired bile secretion due to mutations in the ATP8B1 gene, which codes for a protein that helps maintain the proper balance of fats, known as phospholipids, in cell membranes in the bile ducts. An imbalance of these phospholipids is associated with cholestasis and elevated bile acids in the liver. Subjects with type 1 PFIC usually develop cholestasis in the first months of life, if left untreated, progress to cirrhosis and end-stage liver disease before the end of the first decade of life.

Type 2 PFIC, sometimes referred to as Byler's syndrome, is caused by a defect in bile salt secretion due to mutations in the ABCB11 gene, which codes for a protein known as the bile salt excretion pump, which removes bile acids from the liver. Subjects with type 2 PFIC often develop liver failure in the early years of life and are at increased risk of developing a type of liver cancer known as hepatocellular carcinoma.

Type 3 PFIC, which is commonly seen in the early years of childhood with progressive cholestasis, is caused by mutations in the ABCB4 gene, which codes for a transporter that moves phospholipids across cell membranes.

In addition, mutations in the TJP2 gene, NR1H4 gene, or Myo5b gene have been suggested to be the cause of PFIC. In addition, some subjects with PFIC do not have a mutation in any of the ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b genes. In these cases, the cause of the pathological condition is unknown.

Exemplary mutations in the ATP8B1 gene or resulting protein are shown in Tables 1 and 2, where the numbering is based on the wild-type human ATP8B1 protein (eg, SEQ ID NO: 1) or gene (eg, SEQ ID NO: 2). Exemplary mutations in the ABCB11 gene or resulting protein are shown in Tables 4 and 5, where the numbering is based on the wild-type human ABCB11 protein (eg, SEQ ID NO: 3) or gene (eg, SEQ ID NO: 4).

As may be known to those skilled in the art, the amino acid position in a protein reference sequence that corresponds to the position of a particular amino acid in SEQ ID NO: 1 or 3 can be determined by aligning the protein reference sequence to SEQ ID NO: 1 or 3 (e.g., using software such as ClustalW2). Alterations in these residues (referred to herein as "mutations") may include substitutions of one or more amino acids, sequence insertions or flanks, and sequence deletions or flanks. As may be known to those skilled in the art, the position of a nucleotide in a gene reference sequence that corresponds to the position of a particular nucleotide in SEQ ID NO: 2 or 4 can be determined by aligning the gene reference sequence to SEQ ID NO: 2 or 4 (e.g., using software such as ClustalW2). Changes in these residues (referred to in the present invention as "mutations") substitutions of one or more nucleotides, insertions in sequences or flanks thereof, and deletions from sequences or flanks thereof. 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 hereby incorporated by reference in its entirety.

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

MSTERDSETT FDEDSQPNDE VVPYSDDETE DELDDQGSAV EPEQNRVNRE AEENREPFRK ECTWQVKAND RKYHEQPHFM NTKFLCIKES KYANNAIKTY KYNAFTFIPM NLFEQFKRAA NLYFLALLIL QAVPQISTLA WYTTLVPLLV VLGVTAIKDL VDDVARHKMD KEINNRTCEV IKDGRFKVAK WKEIQVGDVI RLKKNDFVPA DILLLSSSEP NSLCYVETAE LDGETNLKFK MSLEITDQYL QREDTLATFD GFIECEEPNN RLDKFTGTLF WRNTSFPLDA DKILLRGCVI RNTDFCHGLV IFAGADTKIM KNSGKTRFKR TKIDYLMNYM VYTIFVVLIL LSAGLAIGHA YWEAQVGNSS WYLYDGEDDT PSYRGFLIFW GYIIVLNTMV PISLYVSVEV IRLGQSHFIN WDLQMYYAEK DTPAKARTTT LNEQLGQIHY IFSDKTGTLT QNIMTFKKCC INGQIYGDHR DASQHNHNKI EQVDFSWNTY ADGKLAFYDH YLIEQIQSGK EPEVRQFFFL LAVCHTVMVD RTDGQLNYQA ASPDEGALVN AARNFGFAFL ARTQNTITIS ELGTERTYNV LAILDFNSDR KRMSIIVRTP EGNIKLYCKG ADTVIYERLH RMNPTKQETQ DALDIFANET LRTLCLCYKE IEEKEFTEWN KKFMAASVAS TNRDEALDKV YEEIEKDLIL LGATAIEDKL QDGVPETISK LAKADIKIWV LTGDKKETAE NIGFACELLT EDTTICYGED INSLLHARME NQRNRGGVYA KFAPPVQESF FPPGGNRALI ITGSWLNEIL LEKKTKRNKI LKLKFPRTEE ERRMRTQSKR RLEAKKEQRQ KNFVDLACEC SAVICCRVTP KQKAMVVDLV KRYKKAITLA IGDGANDVNM IKTAHIGVGI SGQEGMQAVM SSDYSFAQFR YLQRLLLVHG RWSYIRMCKF LRYFFYKNFA FTLVHFWYSF FNGYSAQTAY EDWFITLYNV LYTSLPVLLM GLLDQDVSDK LSLRFPGLYI VGQRDLLFNY KRFFVSLLHG VLTSMILFFI PLGAYLQTVG QDGEAPSDYQ SFAVTIASAL VITVNFQIGL DTSYWTFVNA FSIFGSIALY FGIMFDFHSA GIHVLFPSAF QFTGTASNAL RQPYIWLTII LAVAVCLLPV VAIRFLSMTI WPSESDKIQK HRKRLKAEEQ WQRRQQVFRR GVSTRRSAYA FSHQRGYADL ISSGRSIRKK RSPLDAIVAD GTAEYRRTGD S

Canonical DNA sequence for ATP8B1 (SEQ ID NO: 2)

ATG AGT ACA GAA AGA GAC TCA GAA ACG ACA TTT GAC GAG GAT TCT CAG CCT AAT GAC GAA GTG GTT CCC TAC AGT GAT GAT GAA ACA GAA GAT GAA CTT GAT GAC CAG GGG TCT GCT GTT GAA CCA GAA CAA AAC CGA GTC AAC AGG GAA GCA GAG GAG AAC CGG GAG CCA TTC AGA AAA GAA TGT ACA TGG CAA GTC AAA GCA AAC GAT CGC AAG TAC CAC GAA CAA CCT CAC TTT ATG AAC ACA AAA TTC TTG TGT ATT AAG GAG AGT AAA TAT GCG AAT AAT GCA ATT AAA ACA TAC AAG TAC AAC GCA TTT ACC TTT ATA CCA ATG AAT CTG TTT GAG CAG TTT AAG AGA GCA GCC AAT TTA TAT TTC CTG GCT CTT CTT ATC TTA CAG GCA GTT CCT CAA ATC TCT ACC CTG GCT TGG TAC ACC ACA CTA GTG CCC CTG CTT GTG GTG CTG GGC GTC ACT GCA ATC AAA GAC CTG GTG GAC GAT GTG GCT CGC CAT AAA ATG GAT AAG GAA ATC AAC AAT AGG ACG TGT GAA GTC ATT AAG GAT GGC AGG TTC AAA GTT GCT AAG TGG AAA GAA ATT CAA GTT GGA GAC GTC ATT CGT CTG AAA AAA AAT GAT TTT GTT CCA GCT GAC ATT CTC CTG CTG TCT AGC TCT GAG CCT AAC AGC CTC TGC TAT GTG GAA ACA GCA GAA CTG GAT GGA GAA ACC AAT TTA AAA TTT AAG ATG TCA CTT GAA ATC ACA GAC CAG TAC CTC CAA AGA GAA GAT ACA TTG GCT ACA TTT GAT GGT TTT ATT GAA TGT GAA GAA CCC AAT AAC AGA CTA GAT AAG TTT ACA GGA ACA CTA TTT TGG AGA AAC ACA AGT TTT CCT TTG GAT GCT GAT AAA ATT TTG TTA CGT GGC TGT GTA ATT AGG AAC ACC GAT TTC TGC CAC GGC TTA GTC ATT TTT GCA GGT GCT GAC ACT AAA ATA ATG AAG AAT AGT GGG AAA ACC AGA TTT AAA AGA ACT AAA ATT GAT TAC TTG ATG AAC TAC ATG GTT TAC ACG ATC TTT GTT GTT CTT ATT CTG CTT TCT GCT GGT CTT GCC ATC GGC CAT GCT TAT TGG GAA GCA CAG GTG GGC AAT TCC TCT TGG TAC CTC TAT GAT GGA GAA GAC GAT ACA CCC TCC TAC CGT GGA TTC CTC ATT TTC TGG GGC TAT ATC ATT GTT CTC AAC ACC ATG GTA CCC ATC TCT CTC TAT GTC AGC GTG GAA GTG ATT CGT CTT GGA CAG AGT CAC TTC ATC AAC TGG GAC CTG CAA ATG TAC TAT GCT GAG AAG GAC ACA CCC GCA AAA GCT AGA ACC ACC ACA CTC AAT GAA CAG CTC GGG CAG ATC CAT TAT ATC TTC TCT GAT AAG ACG GGG ACA CTC ACA CAA AAT ATC ATG ACC TTT AAA AAG TGC TGT ATC AAC GGG CAG ATA TAT GGG GAC CAT CGG GAT GCC TCT CAA CAC AAC CAC AAC AAA ATA GAG CAA GTT GAT TTT AGC TGG AAT ACA TAT GCT GAT GGG AAG CTT GCA TTT TAT GAC CAC TAT CTT ATT GAG CAA ATC CAG TCA GGG AAA GAG CCA GAA GTA CGA CAG TTC TTC TTC TTG CTC GCA GTT TGC CAC ACA GTC ATG GTG GAT AGG ACT GAT GGT CAG CTC AAC TAC CAG GCA GCC TCT CCC GAT GAA GGT GCC CTG GTA AAC GCT GCC AGG AAC TTT GGC TTT GCC TTC CTC GCC AGG ACC CAG AAC ACC ATC ACC ATC AGT GAA CTG GGC ACT GAA AGG ACT TAC AAT GTT CTT GCC ATT TTG GAC TTC AAC AGT GAC CGG AAG CGA ATG TCT ATC ATT GTA AGA ACC CCA GAA GGC AAT ATC AAG CTT TAC TGT AAA GGT GCT GAC ACT GTT ATT TAT GAA CGG TTA CAT CGA ATG AAT CCT ACT AAG CAA GAA ACA CAG GAT GCC CTG GAT ATC TTT GCA AAT GAA ACT CTT AGA ACC CTA TGC CTT TGC TAC AAG GAA ATT GAA GAA AAA GAA TTT ACA GAA TGG AAT AAA AAG TTT ATG GCT GCC AGT GTG GCC TCC ACC AAC CGG GAC GAA GCT CTG GAT AAA GTA TAT GAG GAG ATT GAA AAA GAC TTA ATT CTC CTG GGA GCT ACA GCT ATT GAA GAC AAG CTA CAG GAT GGA GTT CCA GAA ACC ATT TCA AAA CTT GCA AAA GCT GAC ATT AAG ATC TGG GTG CTT ACT GGA GAC AAA AAG GAA ACT GCT GAA AAT ATA GGA TTT GCT TGT GAA CTT CTG ACT GAA GAC ACC ACC ATC TGC TAT GGG GAG GAT ATT AAT TCT CTT CTT CAT GCA AGG ATG GAA AAC CAG AGG AAT AGA GGT GGC GTC TAC GCA AAG TTT GCA CCT CCT GTG CAG GAA TCT TTT TTT CCA CCC GGT GGA AAC CGT GCC TTA ATC ATC ACT GGT TCT TGG TTG AAT GAA ATT CTT CTC GAG AAA AAG ACC AAG AGA AAT AAG ATT CTG AAG CTG AAG TTC CCA AGA ACA GAA GAA GAA AGA CGG ATG CGG ACC CAA AGT AAA AGG AGG CTA GAA GCT AAG AAA GAG CAG CGG CAG AAA AAC TTT GTG GAC CTG GCC TGC GAG TGC AGC GCA GTC ATC TGC TGC CGC GTC ACC CCC AAG CAG AAG GCC ATG GTG GTG GAC CTG GTG AAG AGG TAC AAG AAA GCC ATC ACG CTG GCC ATC GGA GAT GGG GCC AAT GAC GTG AAC ATG ATC AAA ACT GCC CAC ATT GGC GTT GGA ATA AGT GGA CAA GAA GGA ATG CAA GCT GTC ATG TCG AGT GAC TAT TCC TTT GCT CAG TTC CGA TAT CTG CAG AGG CTA CTG CTG GTG CAT GGC CGA TGG TCT TAC ATA AGG ATG TGC AAG TTC CTA CGA TAC TTC TTT TAC AAA AAC TTT GCC TTT ACT TTG GTT CAT TTC TGG TAC TCC TTC TTC AAT GGC TAC TCT GCG CAG ACT GCA TAC GAG GAT TGG TTC ATC ACC CTC TAC AAC GTG CTG TAC ACC AGC CTG CCC GTG CTC CTC ATG GGG CTG CTC GAC CAG GAT GTG AGT GAC AAA CTG AGC CTC CGA TTC CCT GGG TTA TAC ATA GTG GGA CAA AGA GAC TTA CTA TTC AAC TAT AAG AGA TTC TTT GTA AGC TTG TTG CAT GGG GTC CTA ACA TCG ATG ATC CTC TTC TTC ATA CCT CTT GGA GCT TAT CTG CAA ACC GTA GGG CAG GAT GGA GAG GCA CCT TCC GAC TAC CAG TCT TTT GCC GTC ACC ATT GCC TCT GCT CTT GTA ATA ACA GTC AAT TTC CAG ATT GGC TTG GAT ACT TCT TAT TGG ACT TTT GTG AAT GCT TTT TCA ATT TTT GGA AGC ATT GCA CTT TAT TTT GGC ATC ATG TTT GAC TTT CAT AGT GCT GGA ATA CAT GTT CTC TTT CCA TCT GCA TTT CAA TTT ACA GGC ACA GCT TCA AAC GCT CTG AGA CAG CCA TAC ATT TGG TTA ACT ATC ATC CTG GCT GTT GCT GTG TGC TTA CTA CCC GTC GTT GCC ATT CGA TTC CTG TCA ATG ACC ATC TGG CCA TCA GAA AGT GAT AAG ATC CAG AAG CAT CGC AAG CGG TTG AAG GCG GAG GAG CAG TGG CAG CGA CGG CAG CAG GTG TTC CGC CGG GGC GTG TCA AAG CGC TCG CCG CTT GAT GCC ATC GTG GCG GAT GGC ACC GCG GAG TAC AGG CGC ACC GGG GAC AGC TGA

Table 1 Typical ATP8B1 mutations

Amino acid position 3 (e.g. T3K) ²⁷ Amino acid position 23 (e.g. P23L) ⁵ Amino acid position 45 (eg N45T) ^5,8,9 Amino acid position 46 (e.g. R46X) ^A.25 Amino acid position 62 (e.g. C62R) ²⁸ Amino acid position 63 (e.g. T63T) ⁴¹ Amino acid position 70 (eg D70N) ^1.6 Amino acid position 71 (e.g. R71H) ⁴³ Amino acid position 78 (e.g. H78Q) ¹⁹ Amino acid position 82 (e.g. T82T) ⁴¹ Amino acid position 92 (e.g. Y92Y) ⁴¹ Amino acid position 93 (e.g. A93A) ⁶ Amino acid position 96 (e.g. A96G) ²⁷ Amino acid position 114 (e.g. E114Q) ⁸ Amino acid position 127 (eg L127P ⁶ , L127V ³⁶ ) Amino acid position 177 (e.g. T177T) ⁶ Amino acid position 179 (e.g. E179X) ²⁹ Δ Amino acid positions 185-282 ⁴⁴ Amino acid position 197 (eg G197Lfs*10) ²² Amino acid position 201 (e.g. R201S ²⁷ , R201H ³⁵ ) Amino acid position 203 (e.g. K203E ^5.8 , K203R ⁹ , K203fs ²⁵ ) Amino acid position 205 (e.g. N205fs ⁶ , N205Kfs*2 ³⁵ ) Amino acid position 209 (e.g. P209T) ⁴ Amino acid position 217 (e.g. S217N) ⁴³ Amino acid position 232 (e.g. D232D) ³⁰ Amino acid position 233 (e.g. G233R) ³⁸ Amino acid position 243 (e.g. L243fs*28) ³³ Amino acid position 265 (e.g. C265R) ²⁵ Amino acid position 271 (e.g. R271X ¹³ , R271R ³⁰ ) Amino acid position 288 (e.g. L288S) ⁶ Amino acid position 294 (e.g. L294S) ⁴³ Amino acid position 296 (e.g. R296C) ¹¹ Amino acid position 305 (eg F305I) ²⁸ Amino acid position 306 (e.g. C306R) ²³ Amino acid position 307 (e.g. H307L) ³⁵ Amino acid position 308 (e.g. G308V ¹ , G308D ⁶ , G308S ³⁵ ) Amino acid position 314 (e.g. G314S) ¹³ Amino acid position 320 (e.g. M320Vfs*13) ¹¹ Amino acid position 337 (e.g. M337R) ¹⁸ Amino acid position 338 (e.g. N338K) ¹⁸ Amino acid position 340 (e.g. M340V) ¹⁸ Amino acid position 344 (eg I344F) ^6.20 Amino acid position 349 (e.g. I349T) ⁴¹ Amino acid position 358 (eg G358R) ²⁸ Amino acid position 367 (e.g. G367G) ⁴¹ Amino acid position 368 (e.g. N368D) ⁴¹ Amino acid position 393 (eg I393V) ²⁷ Amino acid position 403 (e.g. S403Y) ⁶ Amino acid position 407 (e.g. S407N) ⁴⁰ Amino acid position 412 (e.g. R412P) ⁶ Amino acid position 415 (e.g. Q415R) ²⁷ Amino acid position 422 (e.g. D422H) ³⁵ Amino acid position 429 (e.g. E429A) ⁶ Amino acid position 446 (eg G446R) ^4.11 Amino acid position 453 (e.g. S453Y) ⁶ Amino acid position 454 (e.g. D454G) ⁶ Amino acid position 455 (e.g. K455N) ⁴³ Amino acid position 456 (e.g. T456M ^3.6 , T456K ³⁵ ) Amino acid position 457 (e.g. G457G ⁶ , G457fs*6 ³³ ) Amino acid position 469 (e.g. C469G) ⁴¹ Amino acid position 478 (e.g. H478H) ⁴¹ Amino acid position 500 (e.g. Y500H) ⁶ Amino acid position 525 (e.g. R525X) ⁴ Δ Amino acid position 529 ⁶ Amino acid position 535 (e.g. H535L ⁶ , H535N ⁴¹ ) Amino acid position 553 (e.g. P553P) ⁴³ Amino acid position 554 (e.g. D554N ^1.6 , D554A ³⁵ ) Δ Amino acid positions 556-628 ⁴⁴ Δ Amino acid positions 559-563 ³⁵ Amino acid position 570 (e.g. L570L) ⁴¹ Amino acid position 577 (eg I577V) ¹⁹ Amino acid position 581 (e.g. E581K) ³⁵ Amino acid positions 554 and 581 (e.g. D554A+E581K) ³⁵ Amino acid position 585 (e.g. E585X) ²¹ Amino acid position 600 (e.g. R600W ^2,4 , R600Q ⁶ ) Amino acid position 602 (e.g. R602X) ^3.6 Amino acid position 628 (e.g. R628W) ⁶ Amino acid position 631 (e.g. R631Q) ²⁸ Δ Amino acid positions 645-699 ⁴ Amino acid position 661 (eg I661T) ^1,4,6 Amino acid position 665 (e.g. E665X) ^4.6 Amino acid position 672 (e.g. K672fs ⁶ , K672Vfs*1 ³⁵ ) Amino acid position 674 (e.g. M674T) ¹⁹ Amino acid positions 78 and 674 (e.g. H78Q/M674T) ¹⁹ Amino acid position 684 (e.g. D684D) ⁴¹ Amino acid position 688 (e.g. D688G) ⁶ Amino acid position 694 (eg I694T ⁶ , I694N ¹⁷ ) Amino acid position 695 (e.g. E695K) ²⁷ Amino acid position 709 (e.g. K709fs ⁶ , K709Qfs*41 ¹³ ) Amino acid position 717 (e.g. T717N) ⁴ Amino acid position 733 (e.g. G733R) ⁶ Amino acid position 757 (eg Y757X) ⁴ Amino acid position 749 (e.g. L749P) ²¹ Amino acid position 792 (eg P792fs) ⁶ Δ Amino acid position 795-797 ⁶ Amino acid position 809 (eg I809L) ²⁷ Amino acid position 814 (e.g. K814N) ²⁸ Amino acid position 833 (e.g. R833Q ²⁷ , R833W ⁴¹ ) Amino acid position 835 (for example, K835Rfs*36) ³⁵ Amino acid position 845 (e.g. K845fs) ²⁵ Amino acid position 849 (e.g. R849Q) ²⁴ Amino acid position 853 (e.g. F853S, F853fs) ⁶ Amino acid position 867 (e.g. R867C ¹ , R867fs ⁶ , R867H ²³ ) Amino acid position 885 (e.g. K885T) ⁴¹ Amino acid position 888 (e.g. T888T) ⁴¹ Amino acid position 892 (e.g. G892R) ⁶ Amino acid position 912 (e.g. G912R) ³⁵ Amino acid position 921 (e.g. S921S) ⁴¹ Amino acid position 924 (e.g. Y924C) ²⁸ Amino acid position 930 (e.g. R930X ⁶ , R930Q ²⁸ ) Amino acid position 941 (e.g. R941X) ³⁵ Amino acid position 946 (e.g. R946T) ⁴¹ Amino acid position 952 (e.g. R952Q ^5,9,15 , R952X ⁶ ) Amino acid position 958 (e.g. N958fs) ⁶ Amino acid position 960 (e.g. A960A) ⁴¹ Δ Amino acid position 971 ⁴³ Amino acid position 976 (e.g. A976E ⁴¹ , A976A ⁴³ ) Amino acid position 981 (e.g. E981K) ²⁰ Amino acid position 994 (e.g. S994R) ⁴ Amino acid position 1011 (for example, L1011fs*18) ³³ Amino acid position 1012 (e.g. S1012I) ¹⁰ Amino acid position 1014 (e.g. R1014X) ^6.11 Amino acid position 1015 (eg F1015L) ²⁷ Amino acid position 1023 (e.g. Q1023fs) ⁶ Amino acid position 1040 (eg G1040R) ^1.6 Amino acid position 1044 (e.g. S0144L) ³⁴ Amino acid position 1047 (e.g. L1047fs) ⁶ Amino acid position 1050 (for example, I1050K) ³¹ Amino acid position 1052 (e.g. L1052R) ²⁸ Amino acid position 1095 (e.g. W1095X) ¹¹ Amino acid position 1098 (e.g. V1098X) ³⁵ Amino acid position 1131 (e.g. Q1131X) ⁴⁴ Amino acid position 1142 (e.g. A1142Tfs*35) ⁴³ Amino acid position 1144 (eg Y1144Y) ⁴³ Amino acid position 1150 (for example, I1150T) ⁴¹ Amino acid position 1152 (e.g. A1152T) ³⁰ Amino acid position 1159 (e.g. P1159P) ^25.43 Amino acid position 1164 (e.g. R1164X) ⁶ Amino acid position 1193 (e.g. R1193fs*39) ³³ Amino acid position 1197 (e.g. V1197L) ⁴¹ Amino acid position 1208 (e.g. A1208fs) ⁶ Amino acid position 1209 (eg Y1209Lfs*28) ⁴ Amino acid position 1211 (eg F1211L) ²⁷ Amino acid position 1219 (e.g. D1219H ⁵ , D1219G ²⁷ ) Amino acid position 1223 (e.g. S1223S) ⁴¹ Amino acid position 1233 (e.g. P1233P) ⁴¹ Amino acid position 1241 (eg G1241fs) ⁶ Amino acid position 1248 (e.g. T1248T) ⁴³ Splicing site mutation IVS3+1_+3delGTG ⁶ Splicing site mutation IVS3-2A>G ⁶ IVS6+5T>G ^17.25 Splicing site mutation IVS8+1G>T ⁶ IVS9-G>A ²⁶ IVS12+1G>A ²⁵ Splicing site mutation IVS17-1G>A ⁶ Splicing site mutation IVS18+2T>C ⁶ Splicing site mutation IVS20-4CT>AA Splicing site mutation IVS21+5G>A ⁶ Splicing site mutation IVS23-3C>A ⁶ Splicing site mutation IVS26+2T>A ⁶ g.24774-42062del ⁴ c.-4C>G ⁴¹ c.145C>T ¹² c.181-72G> ^A9 c.182-5T>A ⁴¹ c.182-72G> ^A41 c.246A> ^G9 c.239G>A ³⁹ c.279+1_279+3delGTG ⁴⁶ c.280-2A> ^G46 c.625_62715delinsACAGTAAT ⁴⁶ c.554+122C>T ⁹ c.555-3T>C ²⁷ c.625+5 G>T ⁴ Amino acid position 209 (eg P209T) and c.625+5 G>T ⁴ c.628-30G>A ⁴¹ c.628-31C>T ⁴¹ c.698+1G>T ⁴⁶ c.698+20C>T ⁴¹ c.782-1G>A ⁴⁶ c.782-34G>A ⁴¹ Δ795-797 ¹⁴ c.782 -1G>A ⁴ c.852A>C ²⁷ c.941-1G>A ⁴⁶ c.1014C>T ⁹ c.1029+35G>A ⁹ c.1221-8C.G ⁴¹ 1226delA ¹⁶ c.1429+1G>A ⁴⁶ c.1429+2T>G ¹³ c.1429+49G>A ⁴¹ c.1430-42A>G ⁴¹ c.1493T>C ¹² c.1587_1589delCTT ⁴⁶ c.1630+2T>G ²⁷ c.1631-10T>A ⁴¹ c.1637-37T>C ⁴¹ 1660 G>A ¹⁴ 1798 C>T ¹⁴ 1799 G>A ¹⁴ c.1819-39_41delAA ⁹ c.1819+1G>A ³¹ c.1820-27G>A ⁴¹ c.1918+8C>T ²⁷ c.1933-1G>AK46 c.2097+2T>C ³² c.2097+60T>G ⁴¹ c.2097+89T>C ⁴¹ c.2097+97T>G ⁴¹ c.2210-114T> ^C9 2210delA ¹⁶ c.2210-45_50dupATAAAA ⁹ c.2285+29C.T ⁴¹ c.2285+32A>G ⁴¹ c.2286-4_2286-3delinsAA ⁴⁶ c.2418+5G>A ⁴⁶ c.2707+3G>C ²⁷ c.2707+9T>G ⁴¹ c.2707+43A>G ⁴¹ c.2709-59T>C ⁴¹ c.2931+9A> ^G41 c.2931+59T>A ⁴¹ c.2932-3C>A ⁴⁶ c.2932+59T>A ⁹ c.2937A>C ²⁷ c.3016-9C>A ³¹ c.3033-3034del ¹⁹ 3122delTCCTA/
insACATCGATGTTGATGTTAGG ⁴⁵ 3318 G>A ¹⁴ c.3400+2T>A ⁴⁶ c.3401-175C>T ⁹ c.3401-167C>T ⁹ c.3401-108C>T ⁹ c.3531+8G>T ^9.15 c.3532-15C>T ⁹ ∆ Phe ex 15 ⁴ Ex1_Ex13del ⁶ Ex2_Ex6del ³³ Ex12_Ex14del ²⁷ Skipped exon 24 ⁴⁵ del5'UTR-ex18 ¹¹ c.*11C>T ⁴¹ c.*1101+366G > A ⁷ g.92918del565 ³¹ Preceding GC exon 16 (e.g. resulting in a 4 bp deletion) ⁴² Reading frame shift from the end of the 5' exon 16 ⁴² deletion 5' 1.4 kb ⁴⁶

Table 2 Selected ATP8B1 Mutations Associated with PFIC-1

Amino acid position 23 (e.g. P23L) ⁵ Amino acid position 78 (e.g. H78Q) ¹⁹ Amino acid position 93 (e.g. A93A) ⁶ Amino acid position 96 (e.g. A96G) ²⁷ Amino acid position 127 (e.g. L127P) ⁶ Amino acid position 197 (eg G197Lfs*10) ²² Amino acid position 205 (eg N205fs) ⁶ Amino acid position 209 (e.g. P209T) ⁴ Amino acid position 233 (e.g. G233R) ³⁸ Amino acid position 243 (e.g. L243fs*28) ³³ Amino acid position 288 (e.g. L288S) ⁶ Amino acid position 296 (e.g. R296C) ¹¹ Amino acid position 308 (e.g. G308V ^1.6 ) Amino acid position 320 (e.g. M320Vfs*13) ¹¹ Amino acid position 403 (e.g. S403Y) ⁶ Amino acid position 407 (e.g. S407N) ⁴⁰ Amino acid position 412 (e.g. R412P) ⁶ Amino acid position 415 (e.g. Q415R) ²⁷ Amino acid position 429 (e.g. E429A) ⁶ Amino acid position 446 (eg G446R) ⁴ Amino acid position 456 (e.g. T456M) ^3.6 Amino acid position 457 (e.g. G457G ⁶ , G457fs*6 ³³ ) Amino acid position 500 (e.g. Y500H) ⁶ Amino acid position 525 (e.g. R525X) ⁴ Δ Amino acid position 529 ⁶ Amino acid position 535 (e.g. H535L) ⁶ Amino acid position 554 (eg D554N) ^1.6 Amino acid position 577 (eg I577V) ¹⁹ Amino acid position 585 (e.g. E585X) ²¹ Amino acid position 600 (e.g. R600W) ⁴ Amino acid position 602 (e.g. R602X) ^3.6 Amino acid position 661 (eg I661T) ^4.6 Amino acid position 665 (e.g. E665X) ^4.6 Δ Amino acid positions 645-699 ⁴ Amino acid position 672 (e.g. K672fs) ⁶ Amino acid position 674 (e.g. M674T) ¹⁹ Amino acid positions 78 and 674 (e.g. H78Q/M674T) ¹⁹ Amino acid position 688 (e.g. D688G) ⁶ Amino acid position 694 (for example, I694N) ¹⁷ Amino acid position 695 (e.g. E695K) ²⁷ Amino acid position 709 (e.g. K709fs) ⁶ Amino acid position 717 (e.g. T717N) ⁴ Amino acid position 733 (e.g. G733R) ⁶ Amino acid position 749 (e.g. L749P) ²¹ Amino acid position 757 (eg Y757X) ⁴ Amino acid position 792 (eg P792fs) ⁶ Amino acid position 809 (eg I809L) ²⁷ Amino acid position 853 (e.g. F853S, F853fs) ⁶ Amino acid position 867 (e.g. R867fs) ⁶ Amino acid position 892 (e.g. G892R) ⁶ Amino acid position 930 (eg R930X ⁶ , R952Q ¹⁵ ) Amino acid position 952 (e.g. R952X) ⁶ Amino acid position 958 (e.g. N958fs) ⁶ Amino acid position 981 (e.g. E981K) ²⁰ Amino acid position 994 (e.g. S994R) ⁴ Amino acid position 1014 (e.g. R1014X) ^6.11 Amino acid position 1015 (eg F1015L) ²⁷ Amino acid position 1023 (e.g. Q1023fs) ⁶ Amino acid position 1040 (eg G1040R) ^1.6 Amino acid position 1047 (e.g. L1047fs) ⁶ Amino acid position 1095 (e.g. W1095X) ¹¹ Amino acid position 1208 (e.g. A1208fs) ⁶ Amino acid position 1209 (eg Y1209Lfs*28) ⁴ Amino acid position 1211 (eg F1211L) ²⁷ Amino acid position 1219 (e.g. D1219H ⁵ , D1219G ²⁷ ) Splicing site mutation IVS3+1_+3delGTG ⁶ Splicing site mutation IVS3-2A>G ⁶ IVS6+5T>G ¹⁷ Splicing site mutation IVS8+1G>T ⁶ IVS9-G>A ²⁶ Splicing site mutation IVS17-1G>A ⁶ Splicing site mutation IVS18+2T>C ⁶ Splicing site mutation IVS21+5G>A ⁶ g.24774-42062del ⁴ c.145C>T ¹² c.239G>A ³⁹ c.625+5 G>T ⁴ Amino acid position 209 (eg P209T) and c.625+5 G>T ⁴ c.782 -1G>A ⁴ c.1493T>C ¹² c.1630+2T>G ²⁷ 1660 G>A ¹⁴ c.2707+3G>C ²⁷ c.2097+2T>C ³² c.3033-3034del ¹⁹ 3318 G>A ¹⁴ c.3158+8G>T ¹⁵ ∆ Phe ex 15 ⁴ Ex1_Ex13del ⁶ Ex2_Ex6del ³³ Ex12_Ex14del ²⁷ del5'UTR-ex18 ¹¹ c.*1101+366G > A ⁷ Preceding GC exon 16 (e.g. resulting in a 4 bp deletion) ⁴² Reading frame shift from the end of the 5' exon 16 ⁴²

^A Mutation in 'X' is an early stop codon

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¹⁶ Mizuochi et al., Clin Chim Acta. 2012, vol. 413(15-16), p. 1301-1304.

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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.

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⁴⁰ Jericho et al., Journal of Pediatric Gastroenterology and Nutrition 2015, vol. 60(3), p. 368-374.

⁴¹ van der Woerd et al., PLoS One. 2013, vol. 8(11): e80553.

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

Canonical sequence of ABCB11 proteins (SEQ ID NO: 3) - Uniprot ID O95342

MSDSVILRSI KKFGEENDGF ESDKSYNNDK KSRLQDEKKG DGVRVGFFQL FRFSSSTDIW LMFVGSLCAF LHGIAQPGVL LIFGTMTDVF IDYDVELQEL QIPGKACVNN TIVWTNSSLN QNMTNGTRCG LLNIESEMIK FASYYAGIAV AVLITGYIQI CFWVIAAARQ IQKMRKFYFR RIMRMEIGWF DCNSVGELNT RFSDDINKIN DAIADQMALF IQRMTSTICG FLLGFFRGWK LTLVIISVSP LIGIGAATIG LSVSKFTDYE LKAYAKAGVV ADEVISSMRT VAAFGGEKRE VERYEKNLVF AQRWGIRKGI VMGFFTGFVW CLIFLCYALA FWYGSTLVLD EGEYTPGTLV QIFLSVIVGA LNLGNASPCL EAFATGRAAA TSIFETIDRK PIIDCMSEDG YKLDRIKGEI EFHNVTFHYP SRPEVKILND LNMVIKPGEM TALVGPSGAG KSTALQLIQR FYDPCEGMVT VDGHDIRSLN IQWLRDQIGI VEQEPVLFST TIAENIRYGR EDATMEDIVQ AAKEANAYNF IMDLPQQFDT LVGEGGGQMS GGQKQRVAIA RALIRNPKIL LLDMATSALD NESEAMVQEV LSKIQHGHTI ISVAHRLSTV RAADTIIGFE HGTAVERGTH EELLERKGVY FTLVTLQSQG NQALNEEDIK DATEDDMLAR TFSRGSYQDS LRASIRQRSK SQLSYLVHEP PLAVVDHKST YEEDRKDKDI PVQEEVEPAP VRRILKFSAP EWPYMLVGSV GAAVNGTVTP LYAFLFSQIL GTFSIPDKEE QRSQINGVCL LFVAMGCVSL FTQFLQGYAF AKSGELLTKR LRKFGFRAML GQDIAWFDDL RNSPGALTTR LATDASQVQG AAGSQIGMIV NSFTNVTVAM IIAFSFSWKL SLVILCFFPF LALSGATQTR MLTGFASRDK QALEMVGQIT NEALSNIRTV AGIGKERRFI EALETELEKP FKTAIQKANI YGFCFAFAQC IMFIANSASY RYGGYLISNE GLHFSYVFRV ISAVVLSATA LGRAFSYTPS YAKAKISAAR FFQLLDRQPP ISVYNTAGEK WDNFQGKIDF VDCKFTYPSR PDSQVLNGLS VSISPGQTLA FVGSSGCGKS TSIQLLERFY DPDQGKVMID GHDSKKVNVQ FLRSNIGIVS QEPVLFACSI MDNIKYGDNT KEIPMERVIA AAKQAQLHDF VMSLPEKYET NVGSQGSQLS RGEKQRIAIA RAIVRDPKIL LLDEATSALD TESEKTVQVA LDKAREGRTC IVIAHRLSTI QNADIIAVMA QGVVIEKGTH EELMAQKGAY YKLVTTGSPI S

Canonical DNA sequence for ABCB11 (SEQ ID NO: 4)

ATG TCT GAC TCA GTA ATT CTT CGA AGT ATA AAG AAA TTT GGA GAG GAG AAT GAT GGT TTT GAG TCA GAT AAA TCA TAT AAT AAT GAT AAG AAA TCA AGG TTA CAA GAT GAG AAG AAA GGT GAT GGC GTT AGA GTT GGC TTC TTT CAA TTG TTT CGG TTT TCT TCA TCA ACT GAC ATT TGG CTG ATG TTT GTG GGA AGT TTG TGT GCA TTT CTC CAT GGA ATA GCC CAG CCA GGC GTG CTA CTC ATT TTT GGC ACA ATG ACA GAT GTT TTT ATT GAC TAC GAC GTT GAG TTA CAA GAA CTC CAG ATT CCA GGA AAA GCA TGT GTG AAT AAC ACC ATT GTA TGG ACT AAC AGT TCC CTC AAC CAG AAC ATG ACA AAT GGA ACA CGT TGT GGG TTG CTG AAC ATC GAG AGC GAA ATG ATC AAA TTT GCC AGT TAC TAT GCT GGA ATT GCT GTC GCA GTA CTT ATC ACA GGA TAT ATT CAA ATA TGC TTT TGG GTC ATT GCC GCA GCT CGT CAG ATA CAG AAA ATG AGA AAA TTT TAC TTT AGG AGA ATA ATG AGA ATG GAA ATA GGG TGG TTT GAC TGC AAT TCA GTG GGG GAG CTG AAT ACA AGA TTC TCT GAT GAT ATT AAT AAA ATC AAT GAT GCC ATA GCT GAC CAA ATG GCC CTT TTC ATT CAG CGC ATG ACC TCG ACC ATC TGT GGT TTC CTG TTG GGA TTT TTC AGG GGT TGG AAA CTG ACC TTG GTT ATT ATT TCT GTC AGC CCT CTC ATT GGG ATT GGA GCA GCC ACC ATT GGT CTG AGT GTG TCC AAG TTT ACG GAC TAT GAG CTG AAG GCC TAT GCC AAA GCA GGG GTG GTG GCT GAT GAA GTC ATT TCA TCA ATG AGA ACA GTG GCT GCT TTT GGT GGT GAG AAA AGA GAG GTT GAA AGG TAT GAG AAA AAT CTT GTG TTC GCC CAG CGT TGG GGA ATT AGA AAA GGA ATA GTG ATG GGA TTC TTT ACT GGA TTC GTG TGG TGT CTC ATC TTT TTG TGT TAT GCA CTG GCC TTC TGG TAC GGC TCC ACA CTT GTC CTG GAT GAA GGA GAA TAT ACA CCA GGA ACC CTT GTC CAG ATT TTC CTC AGT GTC ATA GTA GGA GCT TTA AAT CTT GGC AAT GCC TCT CCT TGT TTG GAA GCC TTT GCA ACT GGA CGT GCA GCA GCC ACC AGC ATT TTT GAG ACA ATA GAC AGG AAA CCC ATC ATT GAC TGC ATG TCA GAA GAT GGT TAC AAG TTG GAT CGA ATC AAG GGT GAA ATT GAA TTC CAT AAT GTG ACC TTC CAT TAT CCT TCC AGA CCA GAG GTG AAG ATT CTA AAT GAC CTC AAC ATG GTC ATT AAA CCA GGG GAA ATG ACA GCT CTG GTA GGA CCC AGT GGA GCT GGA AAA AGT ACA GCA CTG CAA CTC ATT CAG CGA TTC TAT GAC CCC TGT GAA GGA ATG GTG ACC GTG GAT GGC CAT GAC ATT CGC TCT CTT AAC ATT CAG TGG CTT AGA GAT CAG ATT GGG ATA GTG GAG CAA GAG CCA GTT CTG TTC TCT ACC ACC ATT GCA GAA AAT ATT CGC TAT GGC AGA GAA GAT GCA ACA ATG GAA GAC ATA GTC CAA GCT GCC AAG GAG GCC AAT GCC TAC AAC TTC ATC ATG GAC CTG CCA CAG CAA TTT GAC ACC CTT GTT GGA GAA GGA GGA GGC CAG ATG AGT GGT GGC CAG AAA CAA AGG GTA GCT ATC GCC AGA GCC CTC ATC CGA AAT CCC AAG ATT CTG CTT TTG GAC ATG GCC ACC TCA GCT CTG GAC AAT GAG AGT GAA GCC ATG GTG CAA GAA GTG CTG AGT AAG ATT CAG CAT GGG CAC ACA ATC ATT TCA GTT GCT CAT CGC TTG TCT ACG GTC AGA GCT GCA GAT ACC ATC ATT GGT TTT GAA CAT GGC ACT GCA GTG GAA AGA GGG ACC CAT GAA GAA TTA CTG GAA AGG AAA GGT GTT TAC TTC ACT CTA GTG ACT TTG CAA AGC CAG GGA AAT CAA GCT CTT AAT GAA GAG GAC ATA AAG GAT GCA ACT GAA GAT GAC ATG CTT GCG AGG ACC TTT AGC AGA GGG AGC TAC CAG GAT AGT TTA AGG GCT TCC ATC CGG CAA CGC TCC AAG TCT CAG CTT TCT TAC CTG GTG CAC GAA CCT CCA TTA GCT GTT GTA GAT CAT AAG TCT ACC TAT GAA GAA GAT AGA AAG GAC AAG GAC ATT CCT GTG CAG GAA GAA GTT GAA CCT GCC CCA GTT AGG AGG ATT CTG AAA TTC AGT GCT CCA GAA TGG CCC TAC ATG CTG GTA GGG TCT GTG GGT GCA GCT GTG AAC GGG ACA GTC ACA CCC TTG TAT GCC TTT TTA TTC AGC CAG ATT CTT GGG ACT TTT TCA ATT CCT GAT AAA GAG GAA CAA AGG TCA CAG ATC AAT GGT GTG TGC CTA CTT TTT GTA GCA ATG GGC TGT GTA TCT CTT TTC ACC CAA TTT CTA CAG GGA TAT GCC TTT GCT AAA TCT GGG GAG CTC CTA ACA AAA AGG CTA CGT AAA TTT GGT TTC AGG GCA ATG CTG GGG CAA GAT ATT GCC TGG TTT GAT GAC CTC AGA AAT AGC CCT GGA GCA TTG ACA ACA AGA CTT GCT ACA GAT GCT TCC CAA GTT CAA GGG GCT GCC GGC TCT CAG ATC GGG ATG ATA GTC AAT TCC TTC ACT AAC GTC ACT GTG GCC ATG ATC ATT GCC TTC TCC TTT AGC TGG AAG CTG AGC CTG GTC ATC TTG TGC TTC TTC CCC TTC TTG GCT TTA TCA GGA GCC ACA CAG ACC AGG ATG TTG ACA GGA TTT GCC TCT CGA GAT AAG CAG GCC CTG GAG ATG GTG GGA CAG ATT ACA AAT GAA GCC CTC AGT AAC ATC CGC ACT GTT GCT GGA ATT GGA AAG GAG AGG CGG TTC ATT GAA GCA CTT GAG ACT GAG CTG GAG AAG CCC TTC AAG ACA GCC ATT CAG AAA GCC AAT ATT TAC GGA TTC TGC TTT GCC TTT GCC CAG TGC ATC ATG TTT ATT GCG AAT TCT GCT TCC TAC AGA TAT GGA GGT TAC TTA ATC TCC AAT GAG GGG CTC CAT TTC AGC TAT GTG TTC AGG GTG ATC TCT GCA GTT GTA CTG AGT GCA ACA GCT CTT GGA AGA GCC TTC TCT TAC ACC CCA AGT TAT GCA AAA GCT AAA ATA TCA GCT GCA CGC TTT TTT CAA CTG CTG GAC CGA CAA CCC CCA ATC AGT GTA TAC AAT ACT GCA GGT GAA AAA TGG GAC AAC TTC CAG GGG AAG ATT GAT TTT GTT GAT TGT AAA TTT ACA TAT CCT TCT CGA CCT GAC TCG CAA GTT CTG AAT GGT CTC TCA GTG TCG ATT AGT CCA GGG CAG ACA CTG GCG TTT GTT GGG AGC AGT GGA TGT GGC AAA AGC ACT AGC ATT CAG CTG TTG GAA CGT TTC TAT GAT CCT GAT CAA GGG AAG GTG ATG ATA GAT GGT CAT GAC AGC AAA AAA GTA AAT GTC CAG TTC CTC CGC TCA AAC ATT GGA ATT GTT TCC CAG GAA CCA GTG TTG TTT GCC TGT AGC ATA ATG GAC AAT ATC AAG TAT GGA GAC AAC ACC AAA GAA ATT CCC ATG GAA AGA GTC ATA GCA GCT GCA AAA CAG GCT CAG CTG CAT GAT TTT GTC ATG TCA CTC CCA GAG AAA TAT GAA ACT AAC GTT GGG TCC CAG GGG TCT CAA CTC TCT AGA GGG GAG AAA CAA CGC ATT GCT ATT GCT CGG GCC ATT GTA CGA GAT CCT AAA ATC TTG CTA CTA GAT GAA GCC ACT TCT GCC TTA GAC ACA GAA AGT GAA AAG ACG GTG CAG GTT GCT CTA GAC AAA GCC AGA GAG GGT CGG ACC TGC ATT GTC ATT GCC CAT CGC TTG TCC ACC ATC CAG AAC GCG GAT ATC ATT GCT GTC ATG GCA CAG GGG GTG GTG ATT GAA AAG GGG ACC CAT GAA GAA CTG ATG GCC CAA AAA GGA GCC TAC TAC AAA CTA GTC ACC ACT GGA TCC CCC ATC AGT TGA

Table 3. Typical ABCB11 mutations

Amino acid position 1 (e.g. M1V) ⁹ Amino acid position 4 (e.g. S4X) ^A.64 Amino acid position 8 (e.g. R8X) ⁸⁸ Amino acid position 19 (eg G19R) ⁵⁶ Amino acid position 24 (e.g. K24X) ³⁵ Amino acid position 25 (eg S25X) ^5.14 Amino acid position 26 (e.g. Y26Ifs*7) ³⁸ Amino acid position 36 (e.g. D36D) ²⁷ Amino acid position 38 (e.g. K38Rfs*24) ⁷³ Amino acid position 43 (e.g. V43I) ⁵⁷ Amino acid position 49 (e.g. Q49X) ⁷³ Amino acid position 50 (e.g. L50S, L50W) ⁵⁷ Amino acid position 52 (e.g. R52W ²⁶ , R52R ²⁸ ) Amino acid position 56 (e.g. S56L) ⁵⁸ Amino acid position 58 (e.g. D58N) ⁶² Amino acid position 62 (e.g. M62K) ⁹ Amino acid position 66 (e.g. S66N) ¹⁷ Amino acid position 68 (e.g. C68Y) ⁴¹ Amino acid position 50 (e.g. L50S) ^5.7 Amino acid position 71 (e.g. L71H) ⁷³ Amino acid position 74 (e.g. I74R) ⁷¹ Amino acid position 77 (e.g. P77A) ⁷³ Amino acid position 87 (e.g. T87R) ⁶⁷ Amino acid position 90 (eg F90F) ^7.27 Amino acid position 93 (e.g. Y93S ¹³ , Y93X ⁸⁸ ) Amino acid position 96 (e.g. E96X) ⁸⁸ Amino acid position 97 (e.g. L97X) ³⁹ Amino acid position 101 (e.g. Q101Dfs*8) ⁹ Amino acid position 107 (e.g. C107R) ³⁶ Amino acid position 112 (for example, I112T) ⁹ Amino acid position 114 (eg W114R) ^2.9 Amino acid position 123 (e.g. M123T) ⁶⁷ Amino acid position 127 (e.g. T127Hfs*6) ⁵ Amino acid position 129 (e.g. C129Y) ²⁵ Amino acid position 130 (e.g. G130G) ⁷⁷ Amino acid position 134 (eg I134I) ²⁸ Amino acid position 135 (e.g. E135K ^7.13 , E135L ¹⁷ ) Amino acid position 137 (e.g. E137K) ⁷ Amino acid position 157 (e.g. Y157C) ⁵ Amino acid position 161 (e.g. C161X) ³⁹ Amino acid position 164 (e.g. V164Gfs*7 ³⁰ , V164I ⁸⁵ ) Amino acid position 167 (e.g. A167S ⁴ , A167V ⁷ , A167T ^9.17 ) Amino acid position 181 (e.g. R181I) ³⁵ Amino acid position 182 (for example, I182K) ⁹ Amino acid position 183 (e.g. M183V ⁸ , M183T ⁹ ) Amino acid position 185 (e.g. M185I) ⁷³ Amino acid position 186 (e.g. E186G) ^2,7,22 Amino acid position 188 (e.g. G188W) ⁷³ Amino acid position 194 (e.g. S194P) ⁷ Amino acid position 198 (e.g. L198P) ⁷ Amino acid position 199 (eg N199Ifs*15X) ⁸⁸ Amino acid position 206 (eg I206V) ²⁸ Amino acid position 212 (e.g. A212T) ⁷³ Amino acid position 217 (e.g. M217R) ⁸⁸ Amino acid position 225 (e.g. T225P) ⁵⁷ Amino acid position 226 (e.g. S226L) ⁹ Amino acid position 232 (e.g. L232Cfs*9) ⁹ Amino acid position 233 (e.g. L233S) ⁸⁶ Amino acid position 238 (eg G238V) ^2.7 Amino acid position 242 (e.g. T242I) ^5.7 Amino acid position 245 (for example, I245Tfs*26) ⁵⁷ Amino acid position 256 (e.g. A256G) ⁹ Amino acid position 260 (eg G260D) ⁷ Amino acid position 269 (e.g. Y269Y) ²⁷ Amino acid position 277 (e.g. A277E) ⁷⁷ Amino acid position 283 (e.g. E283D) ⁷³ Amino acid positions 212 and 283 (e.g. A212T+E283D) ⁷³ Amino acid position 284 (e.g. V284L ^7.39 , V284A ⁷ , V284D ²³ ) Amino acid position 297 (e.g. E297G ^1,2,5,7 , E297K ⁷ ) Amino acid position 299 (e.g. R299K) ²⁸ Amino acid position 303 (e.g. R303K ⁸ , R303M ⁶³ R303fsX321 ⁸³ ) Amino acid position 304 (eg Y304X) ²⁶ Amino acid position 312 (e.g. Q312H) ⁷ Amino acid position 313 (e.g. R313S) ^5.7 Amino acid position 314 (e.g. W314X) ⁵⁷ Amino acid position 318 (e.g. K318Rfs*26) ²⁹ Amino acid position 319 (e.g. G319G) ⁷ Amino acid position 327 (eg G327E) ^5.7 Amino acid position 330 (e.g. W330X) ²⁴ Amino acid position 336 (eg C336S) ^2.7 Amino acid position 337 (eg Y337H) ^21.27 Amino acid position 342 (e.g. W342G) ⁵⁰ Amino acid position 354 (e.g. R354X) ⁹ Amino acid position 361 (e.g. Q361X ⁵⁷ , Q361R ⁷⁴ ) Amino acid position 366 (e.g. V366V ²⁸ , V366D ⁵⁷ ) Amino acid position 368 (for example, V368Rfs*27) ⁵ Amino acid position 374 (eg G374S) ³ Amino acid position 380 (e.g. L380Wfs*18) ⁵ Amino acid position 382 (e.g. A382G) ⁸⁸ Δ Amino acid positions 382-388 ⁵ Δ Amino acid positions 383-389 ⁵⁷ Amino acid position 387 (e.g. R387H) ⁹ Amino acid position 390 (eg A390P) ^5.7 Amino acid position 395 (e.g. E395E) ²⁸ Amino acid position 404 (e.g. D404G) ⁹ Amino acid position 410 (eg G410D) ^5.7 Amino acid position 413 (e.g. L413W) ^5.7 Amino acid position 415 (e.g. R415X) ⁴² Amino acid position 416 (eg I416I) ²⁷ Amino acid position 420 (e.g. I420T) ⁹ Amino acid position 423 (e.g. H423R) ¹³ Amino acid position 432 (eg R432T) ^1,2,7 Amino acid position 436 (e.g. K436N) ⁴⁰ Amino acid position 440 (e.g. D440E) ⁸⁸ Amino acid position 444 (e.g. V444A) ² Amino acid position 454 (e.g. V454X) ⁴⁹ Amino acid position 455 (e.g. G455E) ⁹ Amino acid position 457 (e.g. S457Vfs*23) ⁸⁸ Amino acid position 461 (e.g. K461E) ^2.7 Amino acid position 462 (e.g. S462R) ⁸⁸ Amino acid position 463 (e.g. T463I) ^5.7 Amino acid position 466 (e.g. Q466K) ^5.7 Amino acid position 470 (e.g. R470Q ^5.7 , R470X ⁹ ) Amino acid position 471 (eg Y472X) ⁵ Amino acid position 472 (e.g. Y472C ^5.27 , Y472X ¹⁴ ) Amino acid position 473 (e.g. D473Q ³⁵ , D473V ⁸⁸ ) Amino acid position 475 (e.g. C475X) ²⁹ Amino acid position 481 (e.g. V481E) ^5.7 Amino acid position 482 (e.g. D482G) ^2,5,7 Amino acid position 484 (e.g. H484Rfs*5) ⁹ Amino acid position 487 (e.g. R487H ² , R487P ⁵ ) Amino acid position 490 (eg N490D) ^5.7 Amino acid position 493 (e.g. W493X) ⁸ Amino acid position 496 (e.g. D496V) ⁸⁸ Amino acid position 498 (eg I498T) ^2.7 Amino acid position 499 (e.g. G499E) ⁷³ Amino acid position 501 (e.g. V501G) ⁶⁸ Amino acid position 504 (e.g. E504K) ⁷⁹ Amino acid position 510 (e.g. T510T) ⁷ Amino acid position 512 (eg I512T) ^5.7 Amino acid position 515 (e.g. N515T ^5.7 , N515D ⁶⁴ ) Amino acid position 516 (eg I516M) ¹⁷ Amino acid position 517 (e.g. R517H) ^5.7 Amino acid position 520 (e.g. R520X) ⁵ Amino acid position 523 (e.g. A523G) ¹³ Amino acid position 528 (e.g. I528Sfs*21 ⁵ , I528X ⁹ , I528T ⁷³ ) Amino acid position 535 (e.g. A535A ⁷ , A535X ⁸⁹ ) Amino acid position 540 (eg F540L) ⁴⁶ Amino acid position 541 (e.g. I541L ^5.7 , I541T ^5.17 ) Amino acid position 546 (e.g. Q546K ³⁹ , Q546H ⁷³ ) Amino acid position 548 (eg F548Y) ^5.7 Amino acid position 549 (e.g. D549V) ⁹ Amino acid position 554 (e.g. E554K) ²¹ Amino acid position 556 (e.g. G556R) ⁶⁷ Amino acid position 558 (e.g. Q558H) ²³ Amino acid position 559 (e.g. M559T) ⁵⁷ Amino acid position 562 (e.g. G562D ^5.7 , G562S ⁷³ ) Amino acid position 570 (e.g. A570T ^2,5,7 , A570V ²⁶ ) Amino acid position 575 (e.g. R575X ^2.5 , R575Q ²¹ ) Amino acid position 580 (e.g. L580P) ⁵⁷ Amino acid position 586 (e.g. T586I) ⁷ Amino acid position 587 (e.g. S587X) ⁷³ Amino acid position 588 (e.g. A588V ^5.7 , A588P ⁷³ ) Amino acid position 591 (e.g. N591S) ^2.7 Amino acid position 593 (e.g. S593R) ^2.7 Amino acid position 597 (e.g. V597V ⁹ , V597L ¹³ ) Amino acid position 603 (e.g. K603K) ⁵⁵ Amino acid position 609 (e.g. H609Hfs*46) ²⁶ Amino acid position 610 (for example, I610Gfs*45 ⁹ , I610T ⁵⁷ ) ⁹ Amino acid position 615 (e.g. H615R) ²⁶ Amino acid position 616 (e.g. R616G ²⁸ , R616H ⁷³ ) Amino acid position 619 (e.g. T619A) ²⁸ Amino acid position 623 (e.g. A623A) ²⁸ Amino acid position 625 (e.g. T625Nfs*5) ²⁶ Amino acid position 627 (for example, I627T) ⁷ Amino acid position 628 (e.g. G628Wfs*3) ⁷⁰ Amino acid position 636 (e.g. E636G) ² Amino acid position 648 (e.g. G648Vfs*6 ⁵ , G648V ⁵⁰ ) Amino acid position 655 (e.g. T655I) ⁷ Amino acid position 669 (for example, I669V) ²⁶ Amino acid position 676 (e.g. D676Y) ¹¹ Amino acid position 677 (e.g. M677V) ^7.13 Amino acid position 679 (e.g. A679V) ⁵⁸ Amino acid position 685 (e.g. G685W) ⁶⁰ Amino acid position 696 (e.g. R696W ²⁷ , R696Q ⁵⁸ ) Amino acid position 698 (e.g. R698H ^7.9 , R698K ⁶¹ , R698C ⁸⁸ ) Amino acid position 699 (e.g. S699P) ⁹ Amino acid position 701 (e.g. S701P) ⁵⁸ Amino acid position 702 (e.g. Q702X) ⁸⁹ Amino acid position 709 (e.g. E709K) ⁷ Amino acid position 710 (e.g. P710P) ⁷ Amino acid position 712 (e.g. L712L) ²⁸ Amino acid position 721 (e.g. Y721C) ⁸⁸ Amino acid position 729 (e.g. D724N) ³⁹ Amino acid position 731 (e.g. P731S) ²³ Amino acid position 740 (eg P740Qfs*6) ⁷³ Amino acid position 758 (e.g. G758R) ⁵ Amino acid position 766 (eg G766R) ^5.24 Amino acid position 772 (eg Y772X) ⁵ Amino acid position 804 (e.g. A804A) ⁷ Amino acid position 806 (e.g. G806D ⁴⁴ , G806G ⁵⁵ ) Amino acid position 809 (e.g. S809F) ⁸¹ Amino acid position 817 (e.g. G817G) ⁸⁸ Amino acid position 818 (e.g. Y818F) ⁷ Amino acid position 824 (e.g. G824E) ⁴² Amino acid position 825 (e.g. G825G) ⁷³ Amino acid position 830 (e.g. R830Gfs*28) ⁷³ Amino acid position 832 (e.g. R832C ^7.26 , R832H ⁴¹ ) Amino acid position 842 (e.g. D842G) ² Amino acid position 848 (e.g. D848N) ⁷³ Amino acid position 855 (e.g. G855R) ¹¹ Amino acid position 859 (e.g. T859R) ^5.7 Amino acid position 865 (e.g. A865V) ²⁷ Amino acid position 866 (e.g. S866A) ⁵⁷ Amino acid position 868 (e.g. V868D) ⁷³ Amino acid position 869 (e.g. Q869P) ⁷³ Amino acid position 875 (e.g. Q875X) ⁷³ Amino acid position 877 (e.g. G877R) ⁵⁶ Amino acid position 879 (e.g. I879R) ⁸⁸ Amino acid position 893 (e.g. A893V) ⁵⁷ Amino acid position 901 (e.g. S901R ¹⁷ , S901I ⁷³ ) Amino acid position 903 (e.g. V903G) ⁵⁷ Δ Amino acid position 919 ¹² Amino acid position 923 (e.g. T923P) ^2.7 Amino acid position 926 (e.g. A926P) ^2.7 Amino acid position 928 (e.g. R928X ¹⁵ , R928Q ⁴⁰ ) Amino acid position 930 (e.g. K930X ⁵ , K930Efs*79 ^5.10 , K930Efs*49 ²⁶ ) Amino acid position 931 (e.g. Q931P) ²⁷ Amino acid position 945 (e.g. S945N) ⁵⁷ Amino acid position 948 (e.g. R948C) ^5,7,26 Amino acid position 958 (e.g. R958Q) ²⁸ Amino acid position 969 (e.g. K969K) ⁸⁸ Δ Amino acid positions 969-972 ⁵ Amino acid position 973 (e.g. T973I) ⁵⁷ Amino acid position 976 (e.g. Q976R ⁵⁸ , Q976X ⁸⁸ ) Amino acid position 979 (e.g. N979D) ^5.7 Amino acid position 981 (e.g. Y981Y) ²⁸ Amino acid position 982 (eg G982R) ^2,5,7 Amino acid positions 444 and 982 (e.g. V444A+G982R) ³⁸ Amino acid position 995 (e.g. A995A) ²⁸ Amino acid position 1001 (e.g. R1001R) ⁹ Amino acid position 1003 (e.g. G1003R) ²⁴ Amino acid position 1004 (eg G1004D) ^2.7 Amino acid position 1027 (e.g. S1027R) ²⁶ Amino acid position 1028 (e.g. A1028A ^7,10,88 , A1028E ⁸⁸ ) Amino acid position 1029 (e.g. T1029K) ⁵ Amino acid position 1032 (e.g. G1032R) ¹² Amino acid position 1041 (e.g. Y1041X) ⁹ Amino acid position 1044 (e.g. A1044P) ⁸⁸ Amino acid position 1050 (e.g. R1050C) ^2,7,57 Amino acid position 1053 (e.g. Q1053X) ⁵⁷ Amino acid position 1055 (e.g. L1055P) ³⁶ Amino acid position 1057 (e.g. R1057X ² , R1057Q ⁵⁸ ) Amino acid position 1058 (e.g. Q1058Hfs*38 ⁹ , Q1058fs*38 ¹⁷ , Q1058X ⁷³ ) Amino acid position 1061 (for example, I1061Vfs*34) ⁹ Amino acid position 1083 (eg C1083Y) ⁴⁷ Amino acid position 1086 (e.g. T1086T) ²⁸ Amino acid position 1090 (eg R1090X) ^2.5 Amino acid position 1099 (for example, L1099Lfs*38) ²⁶ Amino acid position 1100 (for example, S1100Qfs*38) ¹³ Amino acid position 1110 (eg A1110E) ^5.7 Amino acid position 1112 (e.g. V1112F) ⁷⁰ Amino acid position 1116 (e.g. G1116R ⁷ , G1116F ^9.17 , G1116E ³⁶ ) Amino acid position 1120 (e.g. S1120N) ⁸⁸ Amino acid position 1128 (e.g. R1128H ^2.7 , R1128C ^5,7,13 ) Amino acid position 1131 (e.g. D1131V) ²⁷ Amino acid position 1144 (e.g. S1144R) ⁷ Amino acid position 1147 (e.g. V1147X) ⁵ Amino acid position 1153 (e.g. R1153C ^2,5,7 , R1153H ⁵ ) Amino acid position 1154 (eg S1154P) ^5.7 Amino acid position 1162 (e.g. E1162X) ³⁹ Δ Amino acid position 1165 ⁸⁸ Amino acid position 1164 (for example, V1164Gfs*7) Amino acid position 1173 (e.g. N1173D) ⁵⁷ Amino acid position 1175 (e.g. K1175T) ⁵⁸ Amino acid position 1186 (e.g. E1186K) ⁷ Amino acid position 1192 (e.g. A1192Efs*50) ⁹ Amino acid position 1196 (e.g. Q1196X) ⁸⁸ Amino acid position 1197 (e.g. L1197G) ⁷ Amino acid position 1198 (e.g. H1198R) ²⁷ Amino acid position 1204 (e.g. L1204P) ⁸⁸ Amino acid position 1208 (e.g. Y1208C) ⁷³ Amino acid position 1210 (e.g. T1210P ^5.7 , T1210F ⁵⁷ ) Amino acid position 1211 (eg N1211D) ⁷ Amino acid position 1212 (e.g. V1212F) ³⁶ Amino acid position 1215 (e.g. Q1215X) ⁵ Amino acid position 1221 (e.g. R1221K) ⁵³ Amino acid position 1223 (e.g. E1223D) ⁷ Amino acid position 1226 (e.g. R1226P) ⁷³ Amino acid position 1228 (eg A1228V) ⁷ Amino acid position 1231 (e.g. R1231W ^5.7 , R1231Q ^5.7 ) Amino acid position 1232 (e.g. A1232D) ¹⁷ Amino acid position 1235 (eg R1235X) ^5.12 Amino acid position 1242 (eg L1242I) ^5.7 Amino acid position 1243 (e.g. D1243G) ⁶⁷ Amino acid position 1249 (e.g. L1249X) ⁷³ Amino acid position 1256 (e.g. T1256fs*1296) ⁸³ Amino acid position 1268 (e.g. R1268Q) ^2.7 Amino acid position 1276 (e.g. R1276H) ³⁰ Amino acid position 1283 (e.g. A1283A ²⁸ , A1283V ⁸⁸ ) Amino acid position 1292 (eg G1292V) ⁷³ Amino acid position 1298 (eg G1298R) ⁵ Amino acid position 1302 (e.g. E1302X) ⁵ Amino acid position 1311 (eg Y1311X) ⁵⁷ Amino acid position 1316 (e.g. T1316Lfs*64) ¹⁵ Amino acid position 1321 (e.g. S1321N) ⁵⁷ intron 4 ((+3)A>C) ¹ IVS4-74A>T ⁸⁹ Splicing site mutation 3' intron 5 c.3901G>A ⁵ Splicing site mutation 5; intron 7 c.6111G>A ⁵ Splicing site mutation IVS7+1G>A ¹⁴ IVS7+5G>A ⁴⁰ IVS8+1G>C ⁷⁶ Splicing site mutation 5' intron 9 c.9081delG ⁵ Splicing site mutation 5' intron 9 c.9081G>T ⁵ Splicing site mutation 5' intron 9 c.9081G>A ⁵ Splicing site mutation IVS9+1G>T ¹⁴ Splicing site mutation 3' intron 13 c.143513_1435-8del ⁵ Splicing site mutation IVS13del-13^-8 ¹⁴ Splicing site mutation 3' intron 16 c.20128T>G ⁵ Splicing site mutation IVS16-8T>G ¹⁴ Splicing site mutation 5' intron 18 c.21781G>T ⁵ Splicing site mutation 5' intron 18 c.21781G>A ⁵ Splicing site mutation 5' intron 18 c.21781G>C ⁵ Splicing site mutation 3' intron 18 c.21792A>G ⁵ Splicing site mutation IVS18+1G>A ¹⁴ Splicing site mutation 5' intron 19 c.2343+1G>T ⁵ Splicing site mutation 5' intron 19 c.2343+2T>C ⁵ Splicing site mutation IVS19+2T>C ¹⁴ Splicing site mutation IVS19+1G>A ²² Splicing site mutation 3' intron 21 c.26112A>T ⁵ IVS22+3A>G ⁸⁹ IVS 23-8 GA ³⁶ IVS24+5G>A ⁵¹ Splicing site mutation 5' intron 24 c.32131delG ⁵ IVS35-6C>G ⁸⁹ Putative splicing mutation 1198-1G>C ¹⁷ Putative splicing mutation 1810-3C>G ¹⁷ Putative splicing mutation 2178+1G>A ¹⁷ Putative splicing mutation 2344-1G>T ¹⁷ Putative splicing mutation c.2611-2A>T ³⁹ Putative splicing mutation 3213+1_3213+2delinsA ¹⁷ c.-24C>A ^44.78 c,76 13 G>T ⁹ c.77-19T>A ⁵² c.90_93delGAAA ¹⁸ c.124G>A ⁶⁹ c.150 +3 A>C ¹⁰ 174C>T ⁵⁴ c.245T>C ⁸⁷ c.249_250insT ¹⁸ 270T>C ⁵⁴ 402C>T ⁵⁴ 585G>C ⁵⁴ c.611+1G>A ⁷⁰ c.611+4A>G ³⁶ c.612-15_-6del10bp ⁵⁵ c.625A>C ³¹ c.627+5G>T ³¹ c.625A>C/ c.627+5G>T ³¹ 696G>T ⁵⁴ c. 784+1G>C ⁴⁹ 807T> ^C54 c.886C>T ³¹ c.890A> ^G59 c.908+1G>A ⁵⁷ c.908+5G>A ⁵⁵ c.908delG ⁵⁹ c.909-15A> ^G66 957A> ^G54 c.1084-2A> ^G57 1145 1bp deletion ⁹⁰ 1281C>T ^54.57 c.1309-165C > T ¹⁹ c.1434+174G > A ¹⁹ c.1434+70C > T ¹⁹ c.1530C>A ⁵⁷ c.1587-1589delCTT ³¹ c.1621A>C ^33.59 c.1638+32T>C ⁶⁶ c.1638+80C>T ⁶⁶ 1671C>T ⁵⁴ 1791G>T ⁵⁴ 1939delA ¹⁴ c.2075+3A>G ⁵³ c.2081T>A ³¹ c.2093G>A ⁶⁵ 2098 delA ¹⁶ c.2138-8T> ^G67 2142A> ^G54 c.2178+1G>T ^36.39 c.2179-17C> ^A66 p.2344-157T> ^G66 c.2344-17T> ^C66 c.2417G>A ⁷⁸ c.2541delG ⁸⁷ c.2620C>T ^32.33 c.2815-8A>G ⁵⁵ c.3003A>G ³⁷ c.3084A>G ^48.54 c.3213 +4 A>G ^9.37 c.3213 +5 G>A ⁹ c.3268C>T ⁷⁵ 3285A> ^G54 c.3382C>T ⁷⁵ 3435A> ^G54 c.3491delT ⁷² c.3589C>T ⁵⁷ c.3765(+1 +5)del5 ⁴² c.3766-34A> ^{G66 c.3767-3768insC6} c.3770delA ⁶⁷ c.3826C>T ⁷² c.3846C>T ⁵⁷ c.3929delG ⁶⁷ c.*236A> ^G66 1145delC ⁸ Ex13_Ex17del ⁸²

Table 4 Selected ABCB11 Mutations Associated with PFIC-2

Amino acid position 1 (e.g. M1V) ⁹ Amino acid position 4 (e.g. S4X) ⁶⁴ Amino acid position 19 (eg G19R) ⁵⁶ Amino acid position 25 (e.g. S25X) ¹⁴ Amino acid position 26 (e.g. Y26Ifs*7) ³⁸ Amino acid position 50 (e.g. L50S) ^7.57 Amino acid position 52 (e.g. R52W) ²⁶ Amino acid position 58 (e.g. D58N) ⁶² Amino acid position 62 (e.g. M62K) ⁹ Amino acid position 66 (e.g. S66N) ¹⁷ Amino acid position 68 (e.g. C68Y) ⁴¹ Amino acid position 93 (e.g. Y93S) ¹³ Amino acid position 101 (e.g. Q101Dfs*8) ⁹ Amino acid position 107 (e.g. C107R) ³⁶ Amino acid position 112 (for example, I112T) ⁹ Amino acid position 114 (eg W114R) ^2.9 Amino acid position 129 (e.g. C129Y) ²⁵ Amino acid position 135 (e.g. E135K ¹³ , E135L ¹⁷ ) Amino acid position 167 (e.g. A167V ⁷ , A167T ^9.17 ) Amino acid position 182 (for example, I182K) ⁹ Amino acid position 183 (e.g. M183V ⁸ , M183T ⁹ ) Amino acid position 225 (e.g. T225P) ⁵⁷ Amino acid position 226 (e.g. S226L) ⁹ Amino acid position 232 (e.g. L232Cfs*9) ⁹ Amino acid position 233 (e.g. L233S) ⁸⁶ Amino acid position 238 (eg G238V) ^2.7 Amino acid position 242 (e.g. T242I) ⁷ Amino acid position 245 (for example, I245Tfs*26) ⁵⁷ Amino acid position 256 (e.g. A256G) ⁹ Amino acid position 260 (eg G260D) ⁵⁷ Amino acid position 284 (e.g. V284L) ⁷ Amino acid position 297 (e.g. E297G) ^2.7 Amino acid position 303 (e.g. R303K ⁸ , R303M ⁶³ , R303fsX321 ⁸³ ) Amino acid position 304 (eg Y304X) ²⁶ Amino acid position 312 (e.g. Q312H) ⁷ Amino acid position 313 (e.g. R313S) ⁷ Amino acid position 314 (e.g. W314X) ⁵⁷ Amino acid position 318 (e.g. K318Rfs*26) ²⁹ Amino acid position 327 (e.g. G327E) ⁷ Amino acid position 330 (e.g. V330X) ²⁴ Amino acid position 336 (eg C336S) ^2.7 Amino acid position 337 (e.g. Y337H) ²¹ Amino acid position 342 (e.g. W342G) ⁵⁰ Amino acid position 354 (e.g. R354X) ⁹ Amino acid position 361 (e.g. Q361X) ⁵⁷ Amino acid position 366 (e.g. V366D) ⁵⁷ Amino acid position 386 (eg G386X) ³⁴ Δ Amino acid positions 383-389 ⁵⁷ Amino acid position 387 (e.g. R387H) ⁹ Amino acid position 390 (e.g. A390P) ⁷ Amino acid position 410 (eg G410D) ⁷ Amino acid position 413 (e.g. L413W) ⁷ Amino acid position 415 (e.g. R415X) ⁴² Amino acid position 420 (e.g. I420T) ⁹ Amino acid position 454 (e.g. V454X) ⁴⁹ Amino acid position 455 (e.g. G455E) ⁹ Amino acid position 461 (e.g. K461E) ^2.7 Amino acid position 463 (e.g. T463I) ⁷ Amino acid position 466 (e.g. Q466K) ⁷ Amino acid position 470 (eg R470Q ⁷ , R470X ⁹ ) Amino acid position 472 (e.g. Y472X ¹⁴ , Y472C ²⁷ ) Amino acid position 475 (e.g. C475X) ²⁹ Amino acid position 481 (e.g. V481E) ⁷ Amino acid position 482 (eg D482G) ^2.7 Amino acid position 484 (e.g. H484Rfs*5) ⁹ Amino acid position 487 (e.g. R487H ² , R487P ⁸⁴ ) Amino acid position 490 (e.g. N490D) ⁷ Amino acid position 493 (e.g. W493X) ⁸ Amino acid position 498 (for example, I498T) ⁷ Amino acid position 501 (e.g. V501G) ⁶⁸ Amino acid position 512 (for example, I512T) ⁷ Amino acid position 515 (e.g. N515T ⁷ , N515D ⁶⁴ ) Amino acid position 516 (eg I516M) ¹⁷ Amino acid position 517 (e.g. R517H) ⁷ Amino acid position 520 (e.g. R520X) ⁵⁷ Amino acid position 523 (e.g. A523G) ¹³ Amino acid position 528 (eg I528X) ⁹ Amino acid position 540 (eg F540L) ⁴⁶ Amino acid position 541 (eg I541L ⁷ , I541T ¹⁷ ) Amino acid position 548 (eg F548Y) ⁷ Amino acid position 549 (e.g. D549V) ⁹ Amino acid position 554 (e.g. E554K) ²¹ Amino acid position 559 (e.g. M559T) ⁵⁷ Amino acid position 562 (eg G562D) ⁷ Amino acid position 570 (e.g. A570T ⁷ , A570V ²⁶ ) Amino acid position 575 (e.g. R575X ² , R575Q ²¹ ) Amino acid position 588 (e.g. A588V) ⁷ Amino acid position 591 (e.g. N591S) ^9.17 Amino acid position 593 (e.g. S593R) ^2.7 Amino acid position 597 (e.g. V597V ⁹ , V597L ¹³ ) Amino acid positions 591 and 597 (e.g. N591S+V597V) ⁹ Amino acid position 603 (e.g. K603K) ⁵⁵ Amino acid position 609 (e.g. H609Hfs*46) ²⁶ Amino acid position 610 (for example, I610Gfs*45) ⁹ Amino acid position 615 (e.g. H615R) ²⁶ Amino acid position 625 (e.g. T625Nfs*5) ²⁶ Amino acid position 627 (for example, I627T) ⁷ Amino acid position 636 (e.g. E636G) ² Amino acid position 669 (for example, I669V) ²⁶ Amino acid position 698 (e.g. R609H) ⁹ Amino acid positions 112 and 698 (e.g. I112T+R698H) ⁹ Amino acid position 699 (e.g. S699P) ⁹ Amino acid position 766 (eg G766R) ²⁴ Amino acid position 806 (e.g. G806G) ⁵⁵ Amino acid position 824 (e.g. G824E) ⁴² Amino acid position 832 (e.g. R832C ^7.26 , R832H ⁴¹ ) Amino acid position 842 (e.g. D842G) ² Amino acid position 859 (e.g. T859R) ⁷ Amino acid position 865 (e.g. A865V) ⁴⁵ Amino acid position 877 (e.g. G877R) ⁵⁶ Amino acid position 893 (e.g. A893V) ⁵⁷ Amino acid position 901 (e.g. S901R) ¹⁷ Amino acid position 903 (e.g. V903G) ⁵⁷ Δ Amino acid position 919 ¹² Amino acid position 928 (e.g. R928X) ^15.21 Amino acid position 930 (e.g. K930Efs*79 ¹⁰ , K930Efs*49 ²⁶ ) Amino acid position 948 (e.g. R948C) ^7.26 Amino acid position 979 (e.g. N979D) ⁷ Amino acid position 982 (eg G982R) ^2.7 Amino acid positions 444 and 982 (e.g. V444A+G982R) ³⁸ Amino acid position 1001 (e.g. R1001R) ⁹ Amino acid position 1003 (e.g. G1003R) ²⁴ Amino acid position 1004 (eg G1004D) ^2.7 Amino acid position 1027 (e.g. S1027R) ²⁶ Amino acid position 1028 (e.g. A1028A) ¹⁰ Amino acid position 1032 (e.g. G1032R) ¹² Amino acid position 1041 (e.g. Y1041X) ⁹ Amino acid position 1050 (e.g. R1050C) ⁵⁷ Amino acid position 1053 (e.g. Q1053X) ⁵⁷ Amino acid position 1055 (e.g. L1055P) ³⁶ Amino acid position 1057 (e.g. R1057X) ² Amino acid position 1058 (e.g. Q1058Hfs*38 ⁹ , Q1058fs*38 ¹⁷ ) Amino acid position 1061 (for example, I1061Vfs*34) ⁹ Amino acid position 1083 (eg C1083Y) ⁴⁷ Amino acid position 1090 (eg R1090X) ² Amino acid position 1099 (for example, L1099Lfs*38) ²⁶ Amino acid position 1100 (for example, S1100Qfs*38) ¹³ Amino acid position 1110 (eg A1110E) ⁷ Amino acid position 1116 (e.g. G1116R ⁷ , G1116F ^9.17 , G1116E ³⁶ ) Amino acid position 1128 (e.g. R1128C) ^7.13 Amino acid position 1131 (e.g. D1131V) ²⁷ Amino acid position 1144 (e.g. S1144R) ⁷ Amino acid position 1153 (e.g. R1153C ^2.7 , R1153H ^7.26 ) Amino acid position 1154 (e.g. S1154P) ⁷ Amino acid position 1173 (e.g. N1173D) ⁵⁷ Amino acid position 1192 (e.g. A1192Efs*50) ⁹ Amino acid position 1198 (e.g. H1198R) ²⁷ Amino acid position 1210 (e.g. T1210P ⁷ , T1210F ⁵⁷ ) Amino acid position 1211 (eg N1211D) ⁷ Amino acid position 1212 (e.g. V1212F) ³⁶ Amino acid position 1231 (eg R1231W ⁷ , R1223Q ⁷ ) Amino acid position 1232 (e.g. A1232D) ¹⁷ Amino acid position 1235 (e.g. R1235X) ¹² Amino acid position 1242 (e.g. L1242I) ⁷ Amino acid position 1256 (e.g. T1256fs*1296) ⁸³ Amino acid position 1268 (e.g. R1268Q) ^2.7 Amino acid position 1302 (e.g. E1302X) ⁵⁷ Amino acid position 1311 (eg Y1311X) ⁵⁷ Amino acid position 1316 (e.g. T1316Lfs*64) ¹⁵ intron 4 ((+3)A>C) ¹ Splicing site mutation IVS7+1G>A ¹⁴ IVS8+1G>C ⁷⁶ Splicing site mutation IVS9+1G>T ¹⁴ Splicing site mutation IVS13del-13^-8 ¹⁴ Splicing site mutation IVS16-8T>G ¹⁴ Splicing site mutation IVS18+1G>A ¹⁴ Splicing site mutation IVS19+2T>C ¹⁴ IVS 23-8 GA ³⁶ IVS24+5G>A ⁵¹ Putative splicing mutation 1198-1G>C ¹⁷ Putative splicing mutation 1810-3C>G ¹⁷ Putative splicing mutation 2178+1G>A ¹⁷ Putative splicing mutation 2344-1G>T ¹⁷ Putative splicing mutation 3213+1_3213+2delinsA ¹⁷ c.-24C>A ⁷⁸ p.76 13 G>T ⁹ c.77-19T>A ⁵² c.90_93delGAAA ¹⁸ c.124G>A ⁶⁹ c.150 +3 A>C ¹⁰ c.249_250insT ¹⁸ c.611+1G>A ⁸⁴ c.611+4A>G ³⁶ c.612-15_-6del10bp ⁵⁵ c.625A>C ³¹ c.627+5G>T ³¹ c.625A>C/ c.627+5G>T ³¹ c.886C>T ³¹ c.890A> ^G59 c.908+1G>A ⁵⁷ c.908+5G>A ⁵⁵ c.908delG ⁵⁹ 1273 1bp deletion ⁹¹ c.1084-2A> ^G57 c.1445A> ^G59 c.1587-1589delCTT ³¹ c.1621A> ^C59 1939delA ¹⁴ c.2081T>A ³¹ 2098 delA ¹⁶ c.2343+1 G>T ⁸⁰ c.2178+1G>T ³⁶ c.2417G>A ⁷⁸ c.2620C>T ³² c.2815-8A>G ⁵⁵ c.3003A>G ³⁷ c.3213 +4 A>G ^9.37 c.3213 +5 G>A ⁹ c.3268C>T ⁷⁵ c.3382C>T ⁷⁵ c.3765(+1 +5)del5 ^{42 c.3767-3768insC6} 1145delC ⁸ Ex13_Ex17del ⁸²

^A Mutation in 'X' is an early stop codon

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⁹¹ U.S. Patent No. 9,295,677

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

Methods are provided for treating PFIC (e.g., PFIC-1 and PFIC-2) in a subject, which comprises analyzing a sample taken from the subject to determine if the subject has a PFIC-associated mutation (e.g., ATP8B1, ABCB11, ABCB4 mutation). , TJP2, NR1H4, or Myo5b) and administering (eg, specific or selective administration) a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, to a subject who has a PFIC-associated mutation. In some embodiments, the mutation is ATP8B1 or ABCB11. For example, a mutation listed in any of tables 1-4. In some embodiments, the mutation in ATP8B1 is selected from the group consisting of L127P, G308V, T456M, D554N, F529del, I661T, E665X, R930X, R952X, R1014X, and G1040R. In some embodiments, the mutation in ABCB11 is selected from the group consisting of A167T, G238V, V284L, E297G, R470Q, R470X, D482G, R487H, A570T, N591S, A865V, G982R, R1153C, and R1268Q.

Also provided are methods for treating PFIC (e.g., PFIC-1 and PFIC-2) in a subject in need thereof, the method comprising: (a) detecting a PFIC-associated mutation (e.g., ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b mutations) , at 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, methods for treating PFIC may include administering a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, to a subject who has a PFIC-associated mutation (e.g., an ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b mutation). In some embodiments, the mutation is ATP8B1 or ABCB11. For example, a mutation listed in any of tables 1-4. In some embodiments, the mutation in ATP8B1 is selected from the group consisting of L127P, G308V, T456M, D554N, F529del, I661T, E665X, R930X, R952X, R1014X, and G1040R. In some embodiments, the mutation in ABCB11 is selected from the group consisting of A167T, G238V, V284L, E297G, R470Q, R470X, D482G, R487H, A570T, N591S, A865V, G982R, R1153C, and R1268Q.

In some embodiments, the subject is determined to have a mutation associated with PFIC in the subject or in a biopsy sample from the subject using any of the tests known in the art, including next generation sequencing (NGS). In some embodiments, a subject is determined to have a PFIC-associated mutation using a regulatory agency-approved, e.g., FDA-approved test or assay to detect a PFIC-associated mutation in the subject, or a sample taken from the subject for a biopsy, or by performing any from non-limiting examples of assays described in the present invention. Additional diagnostic techniques for PFIC are described in Gunaydin, M. et al., Hepat Med. 2018, vol. 10, p. 95-104, which is hereby incorporated by reference in its entirety.

In some embodiments, PFIC treatment (eg, PFIC-1 or PFIC-2) reduces serum bile acids in a subject. In some embodiments, serum bile acid levels are determined, for example, using an enzymatic ELISA or total bile acid assays as described in Danese et al., PLoS One. 2017, vol. 12(6): e0179200, which is hereby incorporated by reference in its entirety. In some embodiments, serum bile acids may decrease, for example, by 10% - 40%, 20% - 50%, 30% - 60%, 40% - 70%, 50% - 80%, or more than 90% % compared with the content of bile acids in the serum before the introduction of the compounds of formula (I) or its pharmaceutically acceptable salt. In some embodiments, the implementation of the treatment of PFIC includes the treatment of pruritis.

Since LBAT is expressed in hepatocytes, LBAT and dual ASBT/LBAT inhibitory substances must have at least some bioavailability and a free fraction in the blood. Since LBAT inhibitory compounds should only be stable when passing from the gut to the liver, it is expected that relatively little systemic exposure to such compounds is sufficient to minimize the potential risk of any side effects in the rest of the body. It is assumed that the inhibition of LBAT and ASBT has at least additive effects in reducing the concentration of intrahepatic bile acids. It is also contemplated that a dual ASBT/LBAT inhibitor may be able to reduce bile acids without causing diarrhea, as is sometimes seen with ASBT inhibitors.

Compounds having high LBAT inhibitory activity and sufficient bioavailability are believed to be particularly suitable for the treatment of hepatitis. Compounds with dual ASBT/LBAT inhibition and sufficient bioavailability are believed to be particularly relevant to non-alcoholic steatohepatitis (NASH).

NASH is a common and severe chronic liver disease that resembles alcoholic liver disease but occurs in people who drink little or no alcohol. In patients with NASH, the accumulation of fat in the liver, known as non-alcoholic fatty liver disease (NAFLD) or steatosis, and other factors such as high LDL cholesterol and insulin resistance cause chronic inflammation of the liver and can lead to progressive tissue scarring, known like fibrosis and cirrhosis, eventually leading to liver failure and death. Patients with NASH were found to have a significantly higher total fasting serum bile acid concentration than healthy subjects ((2.2-2.4-fold increase in NASH) and at all times after meals (1.7-fold increase in NASH). -2.2-fold).This is caused by an increase in glycine- and taurine-conjugated primary and secondary bile acids.NASH patients exhibit greater variability in fasting and postprandial bile acid profiles.These results indicate that NASH patients experience more fasting and postprandial bile acid exposure, including more hydrophobic and cytotoxic secondary compounds Increased bile acid exposure may be involved in liver damage and the pathogenesis of NAFLD and NASH (Ferslew et al., Dig Dis Sci. 2015, vol. 60, p. 3318- 3328) Therefore, inhibition of ASBT and/or LBAT is likely to be beneficial in the treatment of NASH.

NAFLD is characterized by fatty liver in the absence of secondary causes of fatty liver, including excess alcohol consumption, other known liver disease, or long-term steatogenic treatment (Chalasani et al., Hepatology 2018, vol. 67(1), p. 328-357). NAFLD can be subdivided into non-alcoholic fatty liver (NAFL) and non-alcoholic steatohepatitis (NASH). According to Chalasani et al. NAFL is defined as the presence of ≥ 5% fatty liver without evidence of hepatocellular damage in the form of hepatocyte swelling. NASH is defined as the presence of ≥ 5% fatty liver and inflammation with hepatocyte involvement (eg, swelling) with or without the presence of any liver fibrosis. NASH is also commonly associated with liver inflammation and liver fibrosis, which can progress to cirrhosis, end-stage liver disease, and hepatocellular carcinoma. Although hepatic fibrosis is not always seen in NASH, the severity of fibrosis, if present, can be associated with long-term consequences.

A variety of approaches are used to assess and determine if a subject has NAFLD, and if so, the severity of the disease, including determining whether NAFLD is NAFL or NASH. In some embodiments, the implementation of the severity of NAFLD can be assessed using NAS. In some embodiments, NAFLD treatment can be assessed using NAS. In some embodiments, 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 5 for a simplified NAS diagram adapted from a Kleiner publication.

Table 5. Example of NAFLD (NAS) Activity Scores at the Fibrosis Stage

Characteristic Degree Index Steatosis <5% 0 5-33% 1 >33-66% 2 >66% 3 Lobular inflammation Absence of foci 0 <2 lesions/200x 1 2-4 hearths/200x 2 >4 outbreaks/200x 3 Swelling degeneration Absence 0 A little 1 Many cells/pronounced swelling 2 Fibrosis Absence 0 Perisinusoidal or periportal 1 Perisinusoidal and portal/periportal 2 Connective fibrosis 3 cirrhosis 4

In some embodiments, NAS is determined non-invasively, such as as described in U.S. Application Publication No. 2018/0140219, which is hereby incorporated by reference in its entirety. In some embodiments, NAS is determined from a sample taken from a subject prior to administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, NAS is determined during or after a time period of administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the NAS score is less during or after the time period of administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof than before administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, indicative of treatment with NAFLD (e.g., NASH). For example, a decrease in NAS by 1, 2, 3, 4, 5, 6, or 7 indicates treatment for NAFLD (eg, NASH). In some embodiments, the NAS following administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof is 7 or less. In some embodiments, the NAS during the time period of 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 implementation of the NAS during the time period of administration of the compounds of formula (I) or its pharmaceutically acceptable salt is 7 or less. In some embodiments, the NAS during the time period of 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 a period of time of administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof is 7 or less. In some embodiments, the NAS after a period of time of administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof is 5 or less, 4 or less, 3 or less, or 2 or less.

Additional techniques for assessing and calculating NASH in a subject include determining one or more of the following: fatty liver (eg, accumulation of fat in the liver); inflammation of the liver; biomarkers indicating one or more of the following: liver damage, liver inflammation, liver fibrosis, and/or cirrhosis (eg, markers and groups of markers for serum). Other examples of physiological indicators of NASH may include liver morphology, liver stiffness, and the size or mass of the subject's liver.

In some embodiments, NASH in a subject is evidenced by accumulation of liver fat and detection of a biomarker indicative of liver damage. For example, elevated serum ferritin and low serum antibody titers may be common features of NASH.

In some embodiments, NASH assessment techniques include magnetic resonance imaging, quantification of steatosis by spectroscopy or proton density of fat fraction (MRI-PDFF), transient elastography (FIBROSCAN®), determination of hepatic venous pressure gradient (HPVG), measurement of stiffness liver using MRE to diagnose significant fibrosis and/or cirrhosis of the liver and assess the histological characteristics of a liver biopsy. In some embodiments, magnetic resonance imaging is used to detect one or more of the following: steatohepatitis (NASH-MRI), liver fibrosis (Fibro-MRI), and steatosis. See, for example, U.S. Application Publication Nos. 2016/146715 and 2005/0215882, which are hereby incorporated by reference in their entirety.

In some embodiments, the implementation of the treatment of NASH may include the reduction of one or more symptoms associated with NASH; reduction of the degree of fatty hepatosis; decrease in NAS; reduction of inflammation of the liver; a decrease in the content of biomarkers indicating one or more of the following: damage, inflammation of the liver, liver fibrosis and/or cirrhosis of the liver; and attenuation of fibrosis and/or cirrhosis, no further progression of fibrosis and/or cirrhosis, or slowing of 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.

In some embodiments, the implementation of the treatment of NASH includes alleviation of one or more symptoms associated with NASH in the subject. Typical symptoms may include one or more of the following: enlarged liver, fatigue, pain in the right upper abdomen, bloating, enlarged blood vessels just under the surface of the skin, male breast enlargement, enlarged spleen, red palms, jaundice, and pruritis. In some embodiments, the subject is asymptomatic. In some embodiments, the subject's total body weight does not increase. In some embodiments, the overall body weight of the subject is reduced. In some embodiments, the body mass index (BMI) of the subject does not increase. In some embodiments, the body mass index (BMI) of the subject decreases. In some embodiments, the subject's waist to hip ratio (WTH) does not increase. In some embodiments, the waist-to-hip ratio (WTH) of the subject is reduced.

In some embodiments, NASH treatment can be assessed by determining fatty liver. In some embodiments, the implementation of the treatment of NASH includes the reduction of fatty liver after administration of the compounds of formula (I) or its pharmaceutically acceptable salt described in the present invention. In some embodiments, fatty liver is determined using one or more techniques selected from the group including ultrasound, 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 liver biopsy (see e.g. Di Lascio et al., Ultrasound Med Biol. 2018, vol. 44(8), p. 1585- 1596; Lv et al., J Clin Transl Hepatol. 2018, vol. 6(2), pp. 217-221; Reeder et al., J Magn Reson Imaging. 2011, vol. 34(4), spcone; and de Lédinghen V, et al., J Gastroenterol Hepatol 2016, vol 31(4), pp 848-855, which are hereby incorporated by reference in their entirety). A subject diagnosed with NASH may have more than about 5% fatty liver, for example, more than about 5% to about 25%, about 25% to about 45%, about 45% to about 65%, or more than about 65% fatty liver . In some embodiments, a subject with greater than about 5% to about 33% fatty liver has stage 1 fatty liver, a subject with from about 33% to about 66% fatty liver has stage 2 fatty liver, and a subject with greater than about 66% fatty hepatosis there is a stage 3 fatty hepatosis.

In some embodiments, the degree of fatty liver is determined prior to administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the degree of fatty liver is determined during or after the time period of administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a decrease in the degree of fatty liver during or after the period of time of administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, compared to the degree before administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, is indicative of NASH treatment. For example, a decrease in the degree of fatty liver, ranging from about 1% to about 50%, from about 25% to about 75%, or from about 50% to about 100% indicates treatment of NASH. In some embodiments, a decrease in the degree of fatty liver, which is 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% indicate NASH treatment.

In some embodiments, the presence of liver inflammation is determined using one or more techniques selected from the group consisting of biomarkers indicative of liver inflammation and biopsy of the subject's liver sample(s). In some embodiments, the severity of liver inflammation is determined from a biopsy of the subject's liver sample(s). For example, inflammation of the liver can be assessed from a biopsy of a liver sample, as described in Kleiner et al., Hepatology 2005, vol. 41(6), p. 1313-1321 and Brunt et al., Am J Gastroenterol 1999, vol. 94, p. 2467-2474, which are hereby incorporated by reference in their entirety. In some embodiments, the severity of liver inflammation is determined prior to 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 the time period of 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 the time period of administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, as compared to the degree prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, is indicative of NASH treatment. 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% is indicative of NASH treatment. 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% indicate NASH treatment.

In some embodiments, treatment of NASH includes treatment of fibrosis and/or cirrhosis, such as reducing the severity of fibrosis, preventing further progression of fibrosis and/or cirrhosis, or slowing the progression of fibrosis and/or cirrhosis. In some embodiments, the presence of fibrosis and/or cirrhosis is determined using one or more techniques selected from the group including transient elastography (eg, FIBROSCAN®), non-invasive liver fibrosis markers, and histological features of a liver biopsy. In some embodiments, the severity (e.g., stage) of fibrosis is determined using one or more techniques selected from the group including transient elastography (e.g., FIBROSCAN®), fibrosis scoring system, liver fibrosis biomarkers (e.g., non-invasive biomarkers), 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), p. 846-54), the fibrosis scoring system described in Brunt et al. al., Am. J. Gastroenterol. 1999, vol. 94, p. 2467-2474, the fibrosis scoring system described in Kleiner et al., Hepatology 2005, vol. 41(6), p. 1313-1321, and the ISHAK Fibrosis Scoring System (see Ishak et al., J. Hepatol. 1995, vol. 22, p. 696-699), the contents of which are incorporated herein by reference in their entirety.

In some embodiments, the severity of fibrosis is determined prior to administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the severity of fibrosis is determined during or after the time period of administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a decrease in the severity of fibrosis during or after the time period of administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, compared to the degree prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, is indicative of NASH treatment. In some embodiments, a reduction in the severity of fibrosis, no further progression of fibrosis and/or cirrhosis, or a slowdown in the progression of fibrosis and/or cirrhosis is indicative of NASH treatment. In some embodiments, the severity of fibrosis is determined using a scoring system, such as any of the fibrosis scoring systems described in the present invention, for example, the score may indicate the stage of fibrosis, for example, stage 0 (no fibrosis), stage 1, stage 2 , stage 3 and stage 4 (cirrhosis) (see, for example, Kleiner et al). In some embodiments, the reduction in fibrosis stage is a reduction in the severity of fibrosis. For example, a reduction of 1, 2, 3 or 4 stages represents a decrease in the severity of fibrosis. In some embodiments, step down, e.g., step 4 to step 3, step 4 to step 2, step 4 to step 1, step 4 to step 0, step 3 to step 2, step 3 to step 1 , stage 3 to stage 0, stage 2 to stage 1, stage 2 to stage 0, or stage 1 to stage 0 indicates NASH treatment. In some embodiments, the fibrosis stage decreases 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 the compound of formula (I) or its pharmaceutically acceptable salt compared to the stage before administration of the compound of formula (I) or its pharmaceutically acceptable salt. In some embodiments, the fibrosis stage decreases 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 step 3 to step 0, step 2 to step 1, step 2 to step 0, or step 1 to step 0 during the time period of administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, compared to the step prior to administration of the compound of formula ( I) or a pharmaceutically acceptable salt thereof. In some embodiments, the fibrosis stage decreases 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 Step 3 to Step 0, Step 2 to Step 1, Step 2 to Step 0, or Step 1 to Step 0 after a period of time for administering a compound of formula (I) or a pharmaceutically acceptable salt thereof compared to a step prior to administering a compound of formula (I) ) or a pharmaceutically acceptable salt thereof.

In some embodiments, the presence of NASH is determined by one or more biomarkers indicating one or more of the following: liver disease, inflammation, liver fibrosis, and/or cirrhosis, or by scoring systems thereof. In some embodiments, the severity of NASH is determined using one or more biomarkers indicating one or more of the following: liver disease, inflammation, liver fibrosis, and/or cirrhosis, or scoring systems thereof. The content of a biomarker can be determined by, for example, measuring, quantifying and monitoring the degree of expression of a gene or mRNA encoding a biomarker and/or a peptide or protein biomarker. Non-limiting examples of biomarkers indicative of one or more of the following: lesion, inflammation of the liver, fibrosis of the liver and/or cirrhosis of the liver and/or its quantitative indicators include aspartate aminotransferase (AST) to platelet count ratio (APRI); the ratio of the amount of aspartate aminotransferase (AST) to the amount of alanine aminotransferase (ALT) (AAR); FIB-4 score, which is based on APRI, alanine aminotransferase (ALT) levels, and subject age (see, for example, McPherson et al., Gut 2010, vol. 59(9), p. 1265-9, which in its entirety the entirety is incorporated into the present invention by reference); hyaluronic acid; pro-inflammatory cytokines; biomarker groups including α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma-glutamyl transpeptidase (GGT) together with the age and sex of the subject to form a measure of the degree of fibrosis and necroinflammatory activity in the liver (for example, FIBROTEST®, FIBROSURE®), biomarker groups, including bilirubin, gamma-glutamyltransferase, hyaluronic acid, α2-macroglobulin, together with the age and sex of the subject (for example, HEPASCORE®; see, for example, Adams et al., Clin. Chem. 2005, vol. 51(10), p. 1867-1873) and groups of biomarkers, including tissue inhibitor of metalloprotease-1, hyaluronic acid and α2-macroglobulin (for example, FIBROSPECT®); biomarker groups including tissue inhibitor of metalloprotease 1 (TIMP-1), amino-terminated type III polypeptide, procollagen (PIIINP) and hyaluronic acid (HA) (e.g. Enhanced Liver Fibrosis (ELF), 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 one or more of the following: FIB-4 score, biomarker groups including α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma-glutamyl transpeptidase (GGT) together with the subject's age and sex to form a measure degree of fibrosis and necroinflammatory activity in the liver (e.g. FIBROTEST®, FIBROSURE®), biomarker groups including bilirubin, gamma-glutamyltransferase, hyaluronic acid, α2-macroglobulin in conjunction with subject age and sex (e.g. HEPASCORE®; see, e.g., Adams et al., Clin. Chem. 2005, vol. 51(10), p. 1867-1873) and biomarker groups including tissue metalloprotease-1 inhibitor, hyaluronic acid, and α2-macroglobulin (eg, FIBROSPECT®); and groups of biomarkers including tissue inhibitor of metalloprotease 1 (TIMP-1), amino-terminated type III polypeptide, procollagen (PIIINP), and hyaluronic acid (HA) (eg, an indicator of enhanced liver fibrosis (ELF)).

In some embodiments, the aspartate aminotransferase (AST) content is not increased. In some embodiments, the implementation of the content of aspartate aminotransferase (AST) is reduced. In some embodiments, the implementation of the content of alanine aminotransferase (ALT) does not increase. In some embodiments, the implementation of the content of alanine aminotransferase (ALT) is reduced. In some embodiments, the implementation of the "content" of the enzyme is the concentration of the enzyme, for example, in the blood. For example, the content of AST or ALT can be represented as U/l.

In some embodiments, the severity of fibrosis is determined using one or more of the following: FIB-4 score, biomarker groups including α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma-glutamyl transpeptidase (GGT) together with the subject's age and sex to form a measure degree of fibrosis and necroinflammatory activity in the liver (e.g. FIBROTEST®, FIBROSURE®), biomarker groups including bilirubin, gamma-glutamyltransferase, hyaluronic acid, α2-macroglobulin in conjunction with subject age and sex (e.g. HEPASCORE®; see, e.g., Adams et al., Clin. Chem. 2005, vol. 51(10), pp. 1867-1873, incorporated herein by reference in its entirety) and biomarker groups including tissue metalloprotease inhibitor-1, hyaluronic acid and α2-macroglobulin (eg FIBROSPECT®); and groups of biomarkers including tissue inhibitor of metalloprotease 1 (TIMP-1), amino-terminated type III polypeptide, procollagen (PIIINP), and hyaluronic acid (HA) (eg, an indicator of enhanced liver fibrosis (ELF)).

In some embodiments, liver inflammation is determined by the content of biomarkers of liver inflammation, for example, pro-inflammatory cytokines. Non-limiting examples of biomarkers indicative of liver inflammation include interleukin-(IL) 6, interleukin-(IL) 1β, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, monocyte chemotactic 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. 9872844, which are incorporated herein by reference in their entirety). Liver inflammation can also be assessed by the change in macrophage infiltration, for example by measuring the change in CD68 expression. In some embodiments, liver inflammation can be determined by measuring or monitoring serum levels or circulating levels of one or more of the following: interleukin-(IL) 6, interleukin-(IL) 1β, tumor necrosis factor (TNF)-α, transforming factor growth (TGF)-β, monocyte chemotactic protein (MCP)-1 and C-reactive protein (CRP).

In some embodiments, the presence of one or more biomarkers indicative of one or more of the following: liver disease, inflammation, liver fibrosis, and/or cirrhosis is determined using a sample taken from the subject prior to administration of the compound of formula (I) or its pharmaceutically acceptable salt . In some embodiments, the presence of one or more biomarkers indicative of one or more of the following: liver disease, inflammation, liver fibrosis, and/or cirrhosis is determined during or after the time period of administration of the compound of formula (I) or its pharmaceutically acceptable salt. In some embodiments, a decrease in one or more biomarkers indicative of one or more of the following: liver injury, inflammation, liver fibrosis, and/or cirrhosis of the liver during or after a time period of administration of a compound of formula (I) or a pharmaceutically acceptable thereof salt compared to the level prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, indicates NASH treatment. For example, a decrease in one or more biomarkers indicative of one or more of the following: liver disease, inflammation, liver fibrosis, and/or cirrhosis of the liver 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% indicates NASH treatment. In some embodiments, the reduction in one or more biomarkers indicative of one or more of the following: liver injury, 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 content of one or more biomarkers indicative of one or more of the following: damage, inflammation of the liver, fibrosis of the liver and/or cirrhosis of the liver during the time period of administration of the compound of formula (I) or its pharmaceutically acceptable salt is at least 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 content of one or more biomarkers indicative of one or more of the following: damage, inflammation of the liver, fibrosis of the liver and/or cirrhosis of the liver after a period of time of administration of the compound of formula (I) or its pharmaceutically acceptable salt 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, NASH treatment reduces serum bile acids in a subject. In some embodiments, serum bile acid levels are determined, for example, using an enzymatic ELISA or total bile acid assays as described in Danese et al., PLoS One. 2017, vol. 12(6): e0179200, which is hereby incorporated by reference in its entirety. In some embodiments, serum bile acids may decrease, for example, by 10% - 40%, 20% - 50%, 30% - 60%, 40% - 70%, 50% - 80%, or more than 90% % compared with the content of bile acids in the serum before the introduction of the compounds of formula (I) or its pharmaceutically acceptable salt. In some embodiments, NASH is NASH with concomitant cholestasis. With cholestasis, the secretion of bile from the liver, including bile acids, is blocked. Bile acids can cause damage to hepatocytes (see, e.g., Perez MJ, Briz O. World J. Gastroenterol. 2009, vol. 15(14), p. 1677-1689), likely leading to fibrosis or increased fibrosis progression ( cirrhosis) and an increased risk of hepatocellular carcinoma (see, for example, Sorrentino P et al., Dig. Dis. Sci. 2005, vol. 50(6), p. 1130-1135 and Satapathy SK and Sanyal AJ Semin Liver Dis 2015, vol 35(3), pp 221-235, which are hereby incorporated by reference in their entirety). In some embodiments, the NASH treatment includes the treatment of pruritis. In some embodiments, treatment of NASH with concomitant cholestasis includes treatment of pruritis. In some embodiments, the NASH subject with concomitant cholestasis has pruritis.

Typical biomarkers for NASH are shown in Table 6.

Table 6 Typical NASH biomarkers

Biomarkers of liver fibrosis Aspartate aminotransferase (AST) to platelet count (APRI) ratio The ratio of aspartate aminotransferase (AST) to alanine aminotransferase (ALT) (AAR) FIB-4 score ¹ Hyaluronic acid Pro-inflammatory cytokines A group including α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma-glutamyl transpeptidase (GGT) together with the subject's age and sex to form a measure of the degree of fibrosis and necroinflammatory activity in the liver (e.g., FIBROTEST®, FIBROSURE®) A group including bilirubin, gamma-glutamyltransferase, hyaluronic acid, α2-macroglobulin in conjunction with the age and sex of the subject (for example, HEPASCORE® ² ) Group comprising tissue metalloprotease inhibitor-1, hyaluronic acid and α2-macroglobulin (eg FIBROSPECT®) A group including tissue inhibitor of metalloprotease 1 (TIMP-1), amino-terminated type III polypeptide, procollagen (PIIINP) and hyaluronic acid (HA) (for example, an indicator of enhanced liver fibrosis (ELF) ³ ) Biomarkers of liver inflammation ^4.5 Interleukin-(IL) 6 Interleukin-(IL) 1β Tumor necrosis factor (TNF)-α Transforming growth factor (TGF)-β Monocytic chemotactic protein (MCP)-1 C-reactive protein (CRP) PAI-1 Collagen isoforms (eg Col1a1, Col1a2 and Col4a1) Change in macrophage infiltration (eg, change in CD68 expression level)

Literature for table 6

¹ McPherson et al., Gut. 2010, vol. 59(9), p. 1265-1269.

² Adams, et al. Clinic 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 U.S. Patent No. 9,872,844

When used in the present invention, the terms "treatment", "treat" are the reversal, alleviation, delay the onset or suppression of the progression of the disease or disorder, or one or more of its symptoms, described in the present invention. In some embodiments, the implementation of the treatment can be carried out after the development of one or more symptoms. In other embodiments, the implementation of the treatment can be carried out in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (eg, based on history of symptoms and/or genetic or other susceptibility factors). Treatment can also be continued after symptoms have resolved, for example to prevent or delay their recurrence.

A suitable pharmaceutically acceptable salt of a compound of the present invention is, for example, a base addition salt of a compound of the present invention that is sufficiently acidic, such as an alkali metal salt (e.g., sodium or potassium salt), an alkaline earth metal salt ( e.g. a calcium or magnesium salt), an ammonium salt or a salt with an organic base which contains a physiologically acceptable cation, for example a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.

Some compounds of formula (I) or their pharmaceutically acceptable salts may contain chiral centers and/or centers of geometric isomerism (E and Z isomers). It is to be understood that the present invention includes all such optical isomers, diastereoisomers and geometric isomers which have ASBT and/or LBAT inhibitory activity. The present invention also relates to any and all tautomeric forms of the compounds of formula (I) or their pharmaceutically acceptable salts, which have inhibitory activity against ASBT and/or LBAT. Certain compounds of formula (I), or their pharmaceutically acceptable salts, may exist in unsolvated as well as solvated forms, such as, for example, hydrated forms. It should be understood that the present invention includes all such solvated forms that have inhibitory activity against ASBT and/or LBAT.

Another object of the present invention is a pharmaceutical composition containing a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. Inert fillers may, for example, include fillers, binders, disintegrators, lubricity agents and lubricants. Typically, pharmaceutical compositions can be prepared in the usual manner using conventional excipients.

Examples of suitable excipients 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 starches, and pregelatinized starch. In some embodiments, the filler is mannitol and/or microcrystalline cellulose.

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 acacia gum and gum tragacanth). ), sodium alginate, cellulose derivatives (such as hydroxypropylmethylcellulose (or hypromellose), hydroxypropylcellulose and ethylcellulose) and synthetic polymers (such as acrylic acid-methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, acrylic acid/methacrylic acid copolymers and polyvinylpyrrolidone (povidone)). In some embodiments, the binder is hydroxypropyl methylcellulose (hypromellose).

Examples of suitable disintegrants include, but are not limited to, dry starch, modified starch (such as (partially) pregelatinized starch, sodium starch glycolate, and sodium carboxymethyl starch), alginic acid, cellulose derivatives (such as sodium carboxymethyl cellulose, hydroxypropyl cellulose, and low substituted hydroxypropyl cellulose (L-HPC)) and cross-linked polymers (such as carmellose, croscarmellose sodium, carmellose calcium and cross-linked PVP (crospovidone)). In some embodiments, the disintegrant is croscarmellose sodium.

Examples of suitable glidants and lubricants include, but are not limited to, talc, magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, colloidal silica, hydrous silica, synthetic magnesium silicate, fine granular silica, starch, sodium lauryl sulfate, peric acid, magnesium oxide, waxes (such as carnauba wax), hydrogenated oil, polyethylene glycol, sodium benzoate, polyethylene glycol, and mineral oil. In some embodiments, the lubricating agent or lubricant is magnesium stearate or colloidal silicon dioxide.

The pharmaceutical composition can usually be coated with one or more layers. Also included are enteric coatings or coating layers for delayed or targeted release of the compounds of formula (I) or their pharmaceutically acceptable salts. The coating layers may contain one or more coating agents and may optionally contain plasticizers and/or pigments (or colorants).

Examples of suitable coating agents include, but are not limited to, cellulose-based polymers (such as ethyl cellulose, hydroxypropyl methyl cellulose (or hypromellose), hydroxypropyl cellulose, cellulose acetate phthalate, cellulose acetate succinate, hydroxypropyl methyl acetate cellulose succinate, and hydroxypropyl methyl cellulose phthalate), vinyl-based polymers (such as polyvinyl alcohol); and polymers based on acrylic acid and their derivatives (such as acrylic acid-methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, acrylic acid/methacrylic acid copolymers). In some embodiments, the coating agent is hydroxypropyl methylcellulose. In other embodiments, the coating agent is polyvinyl alcohol.

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. In some embodiments, the plasticizer is polyethylene glycol.

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

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

The dose required for therapeutic or prophylactic treatment depends on the route of administration, the severity of the disease, the age and body weight of the patient, and other factors usually taken into account by the attending physician in determining the appropriate regimen and dose for a particular patient.

The amount of compound administered varies for the patient being treated and may vary 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, typically contains from about 1 to about 250 mg of the active ingredient, for example, from about 1 to about 100 mg, or, for example, from about 1 to about 50 mg, or, for example, from about 1 to about 20 mg, such as about 2.5 mg, or about 5 mg, or about 10 mg, or about 15 mg. The daily dose may be administered as a single dose or divided into two, three or more single doses. The orally administered daily dose of the bile acid modulator is preferably from about 0.1 to about 250 mg, more preferably from about 1 to about 100 mg, for example, from about 1 to about 5 mg, for example, from about 1 to about 10 mg, for example , from about 1 to about 15 mg, or, for example, from about 1 to about 20 mg.

Another aspect of the present invention is 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 medicine.

Another object of the present invention is a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of any of the diseases specified in the present invention. The present invention also relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment or prevention of any of the diseases referred to in the present invention. The present invention also relates to a method of treating or preventing any of the diseases described in the present invention 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.

Combination Therapy

In one aspect of the present invention, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are 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 at least one other therapeutically active agent may be administered simultaneously, sequentially or separately. Therapeutically active agents that are suitable for combination with the compounds of formula (I) include, but are not limited to, known active agents that are useful in the treatment of any of the above pathological conditions, disorders and diseases.

In one embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with another ASBT inhibitor. Suitable ASBT inhibitors are disclosed in 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 00/38729; WO 01/66533; WO 01/68096; WO 02/32428; WO 02/50051; WO 03/020710; WO 03/022286; Wo 03/031232, Wo 03/106482, Wo 2004/006899, Wo 2004/076430, Wo 2007/009655, Wo 2007/009656, Wo 2011/137135, De 19825804, EP 864582, EP 489423, EP 549967, EP 573848, EP 573848, EP 573848 EP 624593, EP 624594, EP 624595, EP 624596, EP 0864582, EP 1173205 and EP 1535913, all of which are incorporated herein by reference in their entirety.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a bile acid binder (also referred to as a bile acid sequestrant or resin), such as colesevelam, cholestyramine, or cholestipol. In a preferred embodiment of such a combination, the bile acid binder is formulated for release in the colon. Examples of such formulations are disclosed in, for example, WO 2017/138877, WO 2017/138878, WO 2019/032026 and WO 2019/032027, all of which are hereby incorporated by reference in their entirety.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a DPP-IV inhibitor, including gliptins such as sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gozogliptin and dutogliptin or a pharmaceutically acceptable salt thereof.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are 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 form thereof. salt.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a cholesterol absorption inhibitor such as ezetimibe, or a pharmaceutically acceptable salt thereof.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a PPAR alpha agonist, including fibrates such as clofibrate, bezafibrate, ciprofibrate, clinofribrate, clofibride, fenofibrate, gemfibrozil, ronifibrate, and simfibrate, or a pharmaceutically acceptable salt thereof. .

In another embodiment, compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a PPAR gamma agonist, including thiazolidinediones such as pioglitazone, rosiglitazone, and lobeglitazone, or a pharmaceutically acceptable salt thereof.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a dual alpha/gamma PPAR agonist, including glitazars such as saroglitazar, aleglitazar, muraglitazar, or tezaglitazar, or a pharmaceutically acceptable salt thereof.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a dual alpha/delta PPAR agonist such as elafibranor.

In yet another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a pan PPAR agonist (i.e., a PPAR agonist that has activity on all α, γ, and δ subtypes), such as IVA337.

In another embodiment, compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with farnesoid X receptor (FXR) modulators, including FXR agonists such as cafestol, chenodeoxycholic acid, 6α-ethylchenodeoxycholic acid (obeticholic acid; INT-747), fexaramine, tropifexor, cilofexor and MET409.

In another embodiment, compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a TGR5 receptor modulator, including TGR5 agonists such as 6α-ethyl-23(S)-methylcholic acid (INT-777).

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a dual FXR/TGR5 agonist such as INT-767.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with ursodeoxycholic acid (UDCA). In yet another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with norursodeoxycholic acid (nor-UDCA).

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an FGF19 modulator such as NGM282.

In another embodiment, compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an FGF21 agonist such as BMS-986036.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an integrin inhibitor such as PLN-74809 and PLN-1474.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a CCR2/CCR5 inhibitor such as cenicriviroc.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a caspase protease inhibitor such as emricazan.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a galectin-3 inhibitor such as GR-MD-02.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a stearoyl-CoA desaturase (SCD) inhibitor such as aramchol (arachidylamidcholanic acid).

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an inhibitor of apoptosis signal-regulating kinase 1 (ASK1), such as seloncertib.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a LOXL2 inhibitor such as simtuzumab.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an ACC inhibitor such as GS-0976.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a thyroid hormone receptor-β agonist such as MGL3196.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a GLP-1 agonist such as liraglutide.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with dual glucagon-like peptide and glucagon receptor agonists such as SAR425899.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a mitochondrial pyruvate carrier inhibitor such as MSDC-0602K.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an antioxidant such as vitamin E.

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

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a diacylglycerol-O-acyltransferase 2 (DGAT2) inhibitor such as DGAT2RX and PF-06865571.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a fatty acid synthase (FASN) inhibitor such as TVB-2640.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an AMP-activated protein kinase (AMPK) activator, such as PXL-770.

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

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a cannabinoid receptor 1 (CB1) antagonist such as IM102.

In another embodiment, compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with Klothoβ (KLB) and a fibroblast growth factor receptor (FGFR) activator such as MK-3655 (formerly known as NGM-313).

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a chemokine (c-c motif) ligand 24 (CCL24) inhibitor, such as CM101.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an A3 antagonist such as PBF-1650.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a P2x7 receptor antagonist such as SGM 1019.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with P2Y13 receptor agonists such as CER-209.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a sulfonated oxysterol such as Dur-928.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a leukotriene D4 (LTD4) receptor antagonist such as MN-001.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a natural killer T cell type 1 (NKT1) inhibitor, such as GRI-0621.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an anti-lipopolysaccharide (LPS) such as IMM-124E.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a VAP1 inhibitor such as BI1467335.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an A3 adenosine receptor agonist such as CF-102.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a SIRT-1 activator such as NS-20.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a nicotinic acid 1 receptor agonist such as ARI-3037MO.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a TLR4 antagonist such as JKB-121.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a ketohexokinase inhibitor such as PF-06835919.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an adiponectin receptor agonist such as ADP-335.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with an autotoxin inhibitor such as PAT-505 and PF8380.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a chemokine (c-c motif) receptor 3 (CCR3) antagonist, such as bertilimumab.

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

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a heat shock protein 47 (HSP47) inhibitor such as ND-L02-s0201.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a sterol regulatory element binding protein (SREBP) transcription factor inhibitor such as CAT-2003 and MDV-4463.

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

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with insulin.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a glycogen phosphorylase inhibitor and/or a glucose-6-phosphatase inhibitor.

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.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a meglitinide such as repaglinide, nateglinide and ormiglitinide.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a glucosidase inhibitor such as acarbose or miglitol.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a squalene synthase inhibitor such as TAK-475.

In another embodiment, the compounds of formula (I), or pharmaceutically acceptable salts thereof, are administered in combination with a PTPB1 inhibitor such as troduskvemin, ertiprotabib, JTT-551, and claramine.

Getting connections

The compounds of formula (I) can be obtained in the form of the free acid or its pharmaceutically acceptable salt according to the methods described below. With respect to the following description of such procedures, it should be understood that, if appropriate, suitable protecting groups can be introduced and then removed from various reactants and intermediates in a manner known to those skilled in the art of organic synthesis. Conventional techniques for using such protecting groups, as well as examples of suitable protecting groups, are described, for example, in Greene's Protective Groups in Organic Synthesis by PGM Wutz and TW Greene, 4th Edition, John Wiley & Sons, Hoboken, 2006.

General Methods

All solvents used were pure for analysis. Commercially available anhydrous solvents were commonly used to carry out the reactions. Starting materials were purchased from commercial sources or prepared according to literature procedures. 7-Bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3,3-dibutyl-8-hydroxy- 7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide was prepared as described in WO 02/50051 (method 26). 7-Bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide can be prepared as described in WO 96/ 16051 (example 21). 3,3-Dibutyl-7-chloro-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide can be prepared as described in WO 02/08211 ( example 35). 7-Bromo-3,3-dibutyl-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide and 3,3-dibutyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1- the dioxide can be prepared as described in WO 03/022286 (procedures 23 and 24, respectively). Room temperature is 20-25°C. The compositions of solvent mixtures are given in percent by volume or in volume ratios.

LCMS:

Instrument name: Agilent 1290 infinity II.

Method A: Mobile phase: A: 0.1% HCOOH in water: ACN (95:5), B: ACN; flow rate: 1.5 ml/min; column: ZORBAX XDB C-18 (50×4.6 mm) 3.5 μM.

Method B: Mobile phase: A: 10 mM NH ₄ HCO ₃ in water, B: ACN; flow rate: 1.2 ml/min; column: XBridge C8 (50×4.6 mm), 3.5 μM.

Method C: Mobile phase: A: 0.1% HCOOH in water: ACN (95:5), B: ACN; flow rate: 1.5 ml/min; column: ATLANTIS dC18 (50×4.6 mm), 5 μM.

Method D: Mobile phase: A: 10 mM NH ₄ OAc in water, B: ACN; flow rate: 1.2 ml/min; column: Zorbax Extend C18 (50×4.6 mm) 5 μM.

Method E: Mobile phase: A: 0.1% TFA in water: ACN (95:5), B: 0.1% TFA in ACN; flow rate: 1.5 ml/min; column: XBridge C8 (50×4.6 mm), 3.5 μM.

UPLC:

Device 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.

HPLC:

Instrument Name: Agilent 1260 Infinity II Series instruments listed below with % determination by UV detection (maxplot).

Method A: Mobile phase: A: 10 mM NH ₄ HCO ₃ in water, B: ACN; flow rate: 1.0 ml/min; column: XBridge C8 (50×4.6 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 (50×4.6 mm, 3.5 µm).

Method C: Mobile phase: A: 10 mM NH ₄ OAc in Milli-q water, B: ACN; flow rate: 1.0 ml/min; column: Phenomenex Gemini C18 (150×4.6 mm, 3.0 µm).

Chiral SFC:

Instrument name: THAR-SFC 80 and THAR-SFC 200 (analytical)

The ratio of the amount of CO ₂ to the amount of co-solvent in the range from 60:40 to 80:20

Method A: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 ml/min; column: YMC Amylose-SA (250×4.6 mm, 5 µm).

Method B: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 ml/min; column: Chiralpak AD-H (250×4.6 mm, 5 µm).

Method C: Mobile phase: 20 mM ammonia in methanol; flow rate: 3 ml/min; column: YMC Cellulose-SC (250×4.6 mm, 5 µm).

Method D: Mobile phase: methanol; flow rate: 3 ml/min; column: Lux A1 (250×4.6 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 (250×4.6 mm, 5 µm).

Method G: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 3 ml/min; column: Lux A1 (250×4.6 mm, 5 µm).

Method H: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 ml/min; column: Lux A1 (250×4.6 mm, 5 µm).

Method I: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 3 ml/min; column: Chiral CCS (250×4.6 mm, 5 µm).

Prep-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: 10 mM NH ₄ OAc in water; B: ACN; flow rate: 35 ml/min; column: X select C18 (30×150 mm, 5 µm).

Method C: Mobile phase: A: 10 mM NH ₄ HCO ₃ in water; B: ACN; flow rate: 1.0 ml/min; column: XBridge C8 (50×4.6 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).

Chiral preparative SFC:

Instrument name: PIC SFC 100/PIC SFC 400

The ratio of the amount of CO ₂ to the amount of co-solvent in the range from 60:40 to 80:20

Method A: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 ml/min; column: YMC Amylose-SA (250×30 mm, 5 µm).

Method B: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 ml/min; column: Chiralpak AD-H (250×30 mm, 5 µm).

Method C: Mobile phase: 20 mM ammonia in methanol; flow rate: 3 ml/min; column: YMC Cellulose-SC (250×30 mm, 5 µm).

Method D: Mobile phase: methanol; flow rate: 3 ml/min; column: Chiral CCS (250×30 mm, 5 μm).

Method E: Mobile phase: methanol; flow rate: 3 ml/min; column: Lux A1 (250×30 mm, 5 µm).

Method F: Mobile phase: 0.5% isopropylamine in IPA; flow rate: 3 ml/min; column: Lux A1 (250×30 mm, 5 µm).

Method G: Mobile phase: 0.5% isopropylamine in methanol; flow rate: 3 ml/min; column: Chiral CCS (250×30 mm, 5 µm).

Abbreviations

ACN acetonitrile Boc tert-butoxycarbonyl DABCO 1,4-diazabicyclo[2.2.2]octane DCM dichloromethane DIPEA N,N-diisopropylethylamine DMF dimethylformamide HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate IPA isopropyl alcohol LCMS liquid chromatography - mass spectrometry HPLC high performance liquid chromatography PE petroleum ether SFC supercritical liquid chromatography TFA trifluoroacetic acid THF tetrahydrofuran TLC thin layer chromatography XPhos 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl UPLC ultra high performance liquid chromatography

Below, the present invention is described with the help of the presented examples, which in no way limit the present invention. All cited documents and literature referenced in the present invention are incorporated herein by reference in their entirety.

EXAMPLES

Intermediate 1

Ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)acrylate and ethyl-(Z)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of 3,3-dibutyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (4 g, 8 .93 mmol) in dry DMF (50 ml) were added ethyl-(E)-3-bromoacrylate (2.4 g, 13.4 mmol), potassium carbonate (2.46 g, 17.87 mmol) and tetra-butyl bromide ammonium (0.287 g, 0.89 mmol) at room temperature and the reaction mixture was heated at 90°C for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water and extracted with EtOAc (3× 25 ml). The combined organic layer was washed with brine (50 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the title compound as a mixture of (E)- and (Z)-isomers (ratio 1.7:1). This mixture was separated by preparative HPLC (method A) to give a first eluting fraction corresponding to the (Z)-isomer and a second eluting fraction corresponding to the (E)-isomer in an overall yield of 73%.

(E)-isomer: Yield: 39% (1.9 g, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.72 (d, J =16.0 Hz, 1H), 7.48 (s, 1H), 7.32-7.27 (m, 2H ), 7.19-7.16 (m, 2H), 7.02-6.97 (m, 1H), 6.63 (s, 1H), 5.48 (d, J =16.0 Hz, 1H), 4.14-4.07 (m, 2H), 3.75 (bs, 2H), 3.36 (s, 2H), 2.14 (s, 3H), 1.40-1.31 (m, 4H), 1.27-1.08 (m, 11H), 0.75-0.73 (m, 6H). LCMS: (Method C) 546.1 (M+H), Rt. 3.47 min, 97.89% (maximum)

(Z)-isomer: Yield: 34% (1.65 g, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.47 (s, 1H), 7.32-7.28 (m, 2H), 7.20-7.19 (m, 2H), 7 .16-7.14 (m, 1H), 7.01-6.97 (m, 1H), 6.66 (s, 1H), 5.26 (d, J =8.0 Hz, 1H), 4.13-4.08 (m, 2H), 3.75 (bs, 2H), 3.33 (s, 2H), 2.18 (s, 3H), 1.43-1.36 (m, 2H), 1.33-1.30(m, 2H), 1.22(m, 3H), 1.10-0.98(m, 8H), 0.76-0.73(m, 6H) . LCMS: (Method C) 546.1 (M+H), Rt. 3.34 min, 98.32% (maximum).

Intermediate 2

(E) -3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring to a solution of (E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylic acid (Example 1; 1.0 g, 1.93 mmol) in a 1:1 mixture of DCM and AcOH (10 ml) was added dropwise at 0°C HNO ₃ (65%, 0.183 g, 2.90 mmol) in a mixture of DCM and AcOH (5 ml). After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM and the organic layer was washed with water (30 ml) and saturated NaHCO ₃ solution (30 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the title compound. The crude compound was sent to the next step without any further purification. Yield: 92% (1 g, crude brown solid).

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.14 (d, J =9.2 Hz, 2H), 7.82 (d, J =12.0 Hz, 1H), 7.75 (s, 1H), 6.94 (s, 1H), 6.91 (d, J =8.4 Hz, 2H), 5.69 (d, J =12.0 Hz, 1H), 3.78 (s, 2H), 3.20 (s, 2H), 2.35 (s, 3H), 1.38-1.24 (m, 12H), 0.89 (t, J =6.4 Hz, 6H). LCMS: (Method A) 562.0 (M+H), Rt. 2.73 min, 98.4% (maximum). HPLC: (method A) Rt. 5.67 min, 99.3% (maximum).

Intermediate 3

Ethyl-(E)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)acrylate and ethyl-(Z)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (0.5 g , 1.1 mmol) in dry DMF (20 ml) was added ethyl-(E)-3-bromoacrylate (0.59 g, 3.3 mmol), sodium carbonate (0.35 g, 3.3 mmol) and tetra -butyl ammonium bromide (0.035 g, 0.1 mmol) at room temperature and the reaction mixture was heated at 85-90°C for 5 h. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water and extracted with EtOAc (3×15 ml). The combined organic layer was washed with brine (20 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound as a mixture of (E)- and (Z)-isomers (ratio 1:1). The mixture was purified by Isolera column chromatography (eluent: 8-9% EtOAc/PE; Silica gel: 230-400 mesh) to give a first eluting fraction corresponding to the (E)-isomer and a second eluting fraction corresponding to the (Z)-isomer.

(E)-isomer: Yield: 32% (0.28 g, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.80 (d, J= 12.2 Hz, 1H), 7.66 (s, 1H), 7.38-7.25 (m, 4H ), 7.10-7.04 (m, 2H), 5.47 (d, J= 12.3 Hz, 1H), 4.13 (q, J= 7.1 Hz, 2H), 3.81 -3.78(m, 2H), 3.46(s, 2H), 1.54-1.50(m, 1H), 1.43-1.30(m, 3H), 1.21(t , J= 7.08 Hz, 3H), 1.12-0.98 (m, 4H), 0.74-0.71 (m, 6H). LCMS: (Method A) 552.1 (M+ 2), Rt. 3.33 min, 97.8% (maximum).

(Z)-isomer: Yield: 41% (0.23 g, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.60 (s, 1H), 7.37-7.31 (m, 2H), 7.21-7.19 (m, 3H), 7 .07-7.03 (m, 2H), 5.32 (d, J= 6.9 Hz, 1H), 4.11 (q, J= 6.9 Hz, 2H), 3.78-3, 74 (m, 2H), 3.42-3.38 (m, 2H), 1.51-1.29 (m, 4H), 1.22 (t, J= 7.2 Hz, 3H), 1 .08-1.01 (m, 4H), 0.73-0.71 (m, 6H). LCMS: (Method A) 552.1(M+ 2), Rt. 3.18 min, 98.2% (maximum).

Intermediate 4

3,3-Dibutyl-7-cyclopropyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (3 g, 6.1 mmol) in a mixture of toluene and water (30 ml, 9:1) was added 2-cyclopropyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.04 g, 12.1 mmol) and K ₃ PO ₄ (3.86 g, 18.0 mmol) and the reaction mixture was degassed with N ₂ for 10 min. Then Pd(dppf)Cl ₂ DCM (0.25 g, 0.3 mmol) was added and the reaction mixture was heated at 85° C. overnight. After completion of the reaction (monitored by TLC), the reaction mass was filtered through celite, washed with EtOAc and the combined organics were concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (5-8% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 99% (3 g, crude yellow gum).

LCMS: (Method A) 456.2 (M+H), Rt. 3.58 min, 92.56% (maximum).

Intermediate 5

3,3-Dibutyl-7-cyclopropyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 3,3-dibutyl-7-cyclopropyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 4; 3 g, 6.6 mmol) in DCM (30 ml) at 0° C. was added dropwise BBr₃ (3.2 ml, 33.0 mmol) and the reaction mixture was stirred at room temperature for 1 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C. EtOAc (10 ml) and ice-cold water (5 ml) were added dropwise and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was then partitioned between water (15 ml) and DCM (15 ml) and the aqueous layer was extracted with using DCM (3×15 ml). The combined organics were washed with brine (15 ml), dried over anhydrous Na₂SO₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 20-23% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 69% (2 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.04 (s, 1H), 7.34 (s, 1H), 7.18-7.14 (m, 2H), 6.88-6 .86 (m, 2H), 6.80-6.78 (m, 1H), 6.40 (s, 1H), 3.61 (s, 2H), 3.18 (s, 2H), 2, 08-2.04 (m, 1H), 1.41-1.31 (m, 4H), 1.20-1.06 (m, 8H), 0.88-0.86 (m, 2H), 0.79-0.77 (m, 6H), 0.44-0.43 (m, 2H). LCMS: (Method A) 442.1 (M+H), Rt. 3.22 min, 92.53% (maximum).

Intermediate 6

Ethyl-(E)-3-((3,3-dibutyl-7-cyclopropyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylate and ethyl-(Z)-3-((3,3-dibutyl-7-cyclopropyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylate

With stirring to a solution of 3,3-dibutyl-7-cyclopropyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 5; 2 g , 4.5 mmol) in dry toluene (20 ml) was added ethyl-(E)-3-bromoacrylate (1.22 g, 6.8 mmol), Na ₂ CO ₃ (0.96 g, 9.0 mmol) and tetra-butyl ammonium bromide (0.13 g, 0.4 mmol) at room temperature and the reaction mixture was heated at 85° C. overnight. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water and extracted with EtOAc (3×15 ml). The combined organic layer was washed with brine (20 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude material. The resulting material, which contained the (E)- and (Z)-isomers in a 1:1 ratio, was purified by Isolera column chromatography (eluent: 10-12% EtOAc/PE; silica gel: 230-400 mesh). The resulting mixture was further purified by preparative HPLC (method A) to give a first eluting fraction corresponding to the (Z)-isomer and a second eluting fraction corresponding to the (E)-isomer.

(Z)-isomer: Yield: 26% (0.65 g, almost white gummy). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.47 (s, 1H), 7.28-7.24 (m, 3H), 7.07-7.06 (m, 2H), 6 .96-6.93 (m, 1H), 6.42 (s, 1H), 5.24 (d, J = 6.8 Hz, 1H), 3.70 (s, 2H), 3.26 ( s, 2H), 2.12-2.11 (m, 1H), 1.41-1.39 (m, 2H), 1.33-1.30 (m, 2H), 1.10-1, 07 (m, 5H), 1.00-0.95 (m, 4H), 0.76-0.72 (m, 6H), 0.51-0.50 (m, 2H). LCMS: (Method A) 540.3 (M+H), Rt. 3.51 min, 98.18% (maximum).

(E)-isomer: Yield: 12% (0.29 g, almost white gummy). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.81 (d, J =12.3 Hz, 1H), 7.47 (s, 1H), 7.29-7.26 (m, 2H ), 7.13-7.10 (m, 2H), 6.99-6.95 (m, 1H), 6.39 (s, 1H), 5.44 (d, J =12.2 Hz, 1H), 4.14-4.10 (m, 2H), 3.72 (s, 2H), 3.16 (s, 2H), 1.94-1.91 (m, 1H), 1.41 -1.31 (m, 4H), 1.22-1.18 (m, 4H), 1.09-1.07 (m, 6H), 1.01-0.89 (m, 3H), 0 .75-0.74 (m, 6H), 0.43-0.41 (m, 2H). LCMS: (Method A) 540.3 (M+H), Rt. 3.65 min, 97.68% (maximum).

Intermediate 7

3,3-Dibutyl-7-(dimethylamino)-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (0.8 g, 1 .6 mmol) in toluene (8 ml), dimethylamine (2M in THF; 2.4 ml, 4.8 mmol) was added Cs ₂ CO ₃ (1.31 g, 4.0 mmol) and the reaction mixture was degassed with N ₂ within 10 min. Then Pd(OAc) ₂ (0.036 g, 0.16 mmol) and XPhos (0.077 g, 0.16 mmol) were added and the reaction mixture was heated at 90° C. overnight. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and washed with EtOAc. The combined organics were concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 13-15% EtOAc/PE, silica gel: 230-400 mesh) to give the title compound. Yield: 54% (0.4 g, yellow gum).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.27 (s, 1H), 7.23-7.19 (m, 2H), 7.00-6.98 (m, 2H), 6 .86-6.82(m, 1H), 6.32(s, 1H), 3.85(s, 3H), 3.65(s, 2H), 3.20(s, 2H), 2, 66 (s, 6H), 1.40-1.32 (m, 4H), 1.20-1.12 (m, 8H), 0.79-0.72 (m, 6H). LCMS: (Method A) 459.3 (M+H), Rt. 3.40 min, 92.77% (maximum).

Intermediate 8

3,3-Dibutyl-7-(dimethylamino)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3,3-dibutyl-7-(dimethylamino)-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 7; 0.4 g, 0.87 mmol) in DCM (4 ml) at 0°C was added dropwise BBr ₃ (0.42 ml, 4.3 mmol) and the reaction mixture was stirred at room temperature for 2 h. reaction (monitored by TLC), the reaction mass was cooled to 0°C. The reaction was stopped by dropwise addition of methanol (5 ml) and then added ice-cold water (10 ml). The aqueous layer was extracted with DCM (2×15 ml), washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 15-18% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 66% (0.33 g, yellow gum).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.83 (s, 1H), 7.28 (s, 1H), 7.20-7.16 (m, 2H), 6.94-6 .92 (m, 2H), 6.81-6.79 (m, 1H), 6.31 (s, 1H), 3.62 (s, 2H), 3.14 (s, 2H), 2, 66 (s, 6H), 1.34-1.24 (m, 4H), 1.16-1.06 (m, 8H), 0.85-0.77 (m, 6H). LCMS: (Method A) 445.2 (M+H), Rt. 3.10 min, 81.86% (maximum)

Intermediate 9

Ethyl-(E)-3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)acrylate and ethyl-(Z)-3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of 3,3-dibutyl-7-(dimethylamino)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 8; 2 g, 4.5 mmol) in toluene (20 ml) was added ethyl-(E)-3-bromoacrylate (1.22 g, 6.8 mmol), sodium carbonate (0.96 g, 9.0 mmol) and tetra-butyl ammonium bromide (0.13 g, 0.4 mmol) and the resulting mixture was heated at 85°C for 14 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was dissolved in EtOAc (25 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ , filtered and then 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 as a mixture of (E)- and (Z)-isomers. The resulting mixture was further purified by preparative HPLC (method B) to give a first eluting fraction corresponding to the (Z)-isomer and a second eluting fraction corresponding to the (E)-isomer.

(Z)-isomer: Yield: 13% (0.05 g, white gummy solid). ¹ H NMR (300 MHz, DMSO- d ₆ ): δ 7.36 (s, 1H), 7.30-7.25 (m, 2H), 7.14-7.12 (m, 2H), 7 .02 (d, J = 6.9 Hz, 1H), 6.98-6.95 (m, 1H), 6.25 (s, 1H), 5.20 (d, J = 6.9 Hz, 1H), 4.13-4.06(m, 2H), 3.71(s, 2H), 3.26(s, 2H), 2.71(s, 6H), 1.41-1.32 (m, 4H), 1.20 (t, J =7.20 Hz, 3H), 1.10-1.01 (m, 8H), 0.76-0.72 (m, 6H). LCMS: (Method A) 543.3 (M+H), Rt. 3.52 min, 87.35% (maximum).

(E)-isomer: Yield: 20% (0.08 g, white gummy solid). LCMS: (Method A) 543.3 (M+H), Rt. 3.65 min, 95.73% (maximum).

Intermediate 10

Methyl-(Z)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)-2-fluoroacrylate

With stirring, to a solution of 3,3-dibutyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (12.0 g , 26.80 mmol) in DMF (120 mL) at 0°C was added 60% NaH (2.6 g, 53.60 mmol) in portions and the reaction mixture was stirred for 15 min. Then methyl 3-bromo-2,2-difluoropropanoate (10.88 g, 53.60 mmol) was added dropwise and the reaction mixture was heated at 85° C. for 16 h. cooled to 0°C, the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with ice-cold water (150 ml). The aqueous portion was extracted with EtOAc (200 ml). Then the organic layer was washed with brine (150 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 12-13% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 72% (8.2 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.67 (d, J =18.8 Hz, 1H), 7.59 (s, 1H), 7.32-7.30 (m, 2H ), 7.16 (d, J =7.6 Hz, 2H), 7.00 (t, J =7.2 Hz, 1H), 6.66 (s, 1H), 3.79 (s, 3H ), 3.75 (s, 2H), 3.36 (s, 2H), 2.18 (s, 3H), 1.44-1.30 (m, 4H), 1.11-0.99 ( m, 8H), 0.76 (t, J = 6.80 Hz, 6H). LCMS: (Method C) 550.1 (M ⁺ +H), Rt. 3.30 min, 98.40% (maximum).

Intermediate 11

3,3-dibutyl-8-methoxy-5-phenyl-7-vinyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (1.2 g, 24 .2 mmol) in a mixture of 1,4-dioxane and water (13 ml; 12:1) was added pinacoline ester of vinylboronic acid (0.56 g, 36.3 mmol) and potassium carbonate (1.0 g, 72.6 mmol ) and the reaction mixture was degassed with N ₂ for 10 min. Then Pd(dppf) ₂ Cl ₂ (0.09 g, 0.12 mmol) was added and the resulting mixture was heated at 85° C. for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was filtered through celite. The resulting filtrate was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 19% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 85% (0.91 g, brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.28 (s, 1H), 7.20-7.13 (m, 2H), 7.09 (s, 1H), 7.00-6 .90 (m, 2H), 6.87-6.83 (m, 2H), 5.68 (d, J = 17.7 Hz, 1H), 5.34 (d, J = 11.3 Hz, 1H), 3.90 (s, 3H), 3.66-3.80 (m, 2H), 3.30 (s, 2H), 2.00-1.31 (m, 4H), 1.18 -1.04 (m, 8H), 0.79-0.74 (m, 6H). LCMS: (Method C) 442.1 (M+H), Rt. 3.47 min, 91.5% (maximum).

Intermediate 12

3,3-Dibutyl-7-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

To a degassed solution of 3,3-dibutyl-8-methoxy-5-phenyl-7-vinyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 11; 0.6 g, 1.35 mmol) in ethanol (10 ml) was added with Pd/C (60 mg, 10%) and the resulting mixture was stirred for 4 h. After completion of the reaction (monitored by LCMS), the reaction mixture was filtered through celite and washed with ethanol . The resulting filtrate was concentrated in vacuo to give the title compound. Yield: 91.3% (0.55 g, white resinous solid).

LCMS: (Method A) 444.2 (M+H), Rt. 3.63 min, 67.11% (maximum).

Intermediate 13

3,3-Dibutyl-7-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 3,3-dibutyl-7-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 12; 0. 55 g, 1.23 mmol) in DCM (10 ml) at 0°C was added dropwise BBr ₃ (0.59 ml, 6.19 mmol) and the resulting mixture was stirred for 1 h. TLC) the reaction was stopped with methanol (0.5 ml) and then water (15 ml) was added. The aqueous layer was separated, extracted with DCM (2×15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 40% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 76% (0.4 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.96 (s, 1H), 7.35-7.32 (m, 1H), 7.30-7.16 (m, 2H), 6 .93-6.87 (m, 2H), 6.81-6.77 (m, 2H), 5.79-3.55 (m, 2H), 3.20 (s, 2H), 1.40 -1.24 (m, 4H), 1.18-1.14 (m, 10H), 1.10-1.00 (m, 9H). LCMS: (Method A) 430.1 (M+H), Rt. 3.22 min, 53.5% (maximum).

Intermediate 14

Ethyl-(Z)-3-((3,3-dibutyl-7-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylate and ethyl-(E)-3-((3,3-dibutyl-7-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylate

With stirring to a solution of 3,3-dibutyl-7-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 13; 0. 4 g, 0.92 mmol) in a mixture of toluene (5 ml) and DMF (2 ml) was added ethyl-(E)-3-bromoacrylate (0.33 g, 1.86 mmol), sodium carbonate (0.25 g , 2.32 mmol) and tetra-butyl ammonium bromide (0.03 g, 0.09 mmol) and the resulting mixture was heated at 85°C for 14 h. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated in vacuo and the resulting residue was dissolved in EtOAc (25 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ , filtered 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 as a mixture of (E)- and (Z)-isomers.

(Z)-isomer: Yield: 19.5% (96 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.51 (s, 1H), 7.34-7.26 (m, 3H), 7.11-7.13 (m, 2H), 7 .00-6.94 (m, 1H), 6.85 (s, 1H), 5.25 (d, J = 6.8 Hz, 1H), 4.11 (q, J = 6.8 Hz, 2H), 3.81-3.61(m, 2H), 3.37(s, 2H), 1.51-1.32(m, 5H), 1.24-1.20(m, 5H) , 1.07-1.03 (m, 8H), 0.77-0.73 (m, 8H). LCMS: (Method D) 528.2 (M+H), Rt. 3.44 min, 79.78% (maximum).

(E)-isomer: Yield: 24.5% (120 mg, almost white solid). ¹ H NMR(400 MHz, DMSO- d ₆ ): δ 7.80 (d, J =12.0 Hz, 1H), 7.49 (s, 1H), 7.32-7.28 (m, 2H ), 7.17-7.16 (m, 1H), 7.91-6.99 (m, 2H), 6.85 (s, 1H), 5.48 (d, J= 12.0 Hz, 1H), 4.15-4.10 (m, 2H), 3.82-3.71 (m, 2H), 3.37 (s, 2H), 1.49-1.31 (m, 4H) , 1.27-1.20 (m, 6H), 1.09-1.02 (m, 8H), 1.01-0.99 (m, 8H). LCMS: (Method A) 528.2 (M+H), Rt. 3.7 min, 74.4% (maximum).

Intermediate 15

3-butyl-3-ethyl-8-hydroxy-7-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (1 g, 2 .21 mmol) in sodium methoxide (21%, 4.42 ml, 4.42 mmol) was added CuBr (0.1 g, 0.31 mmol) at room temperature and the resulting mixture was heated 6 h at 85°C. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting crude mass was subjected to distribution between EtOAc (25 ml) and water (25 ml). The aqueous layer was extracted with EtOAc (2×25 ml), the combined organic layer was washed with brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 5% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 41% (0.36 g, pale pink solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.71 (s, 1H), 7.29 (s, 1H), 7.19 (t, J =7.6 Hz, 2H), 6, 92 (d, J =8.0 Hz, 2H), 6.80 (t, J =7.6 Hz, 1H), 6.54 (s, 1H), 3.63 (s, 3H), 3, 16 (s, 2H), 2.45 (s, 2H), 1.62-1.51 (m, 2H), 1.40-1.31 (m, 2H), 1.28-1.18 ( m, 4H), 0.78-0.74 (m, 6H). LCMS: (Method A) 404.2 (M+H), Rt. 2.64 min, 94.56% (maximum).

Intermediate 16

Ethyl-(E)-3-((3-butyl-3-ethyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)acrylate and ethyl-(Z)-3-((3-butyl-3-ethyl-7-methoxy-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of 3-butyl-3-ethyl-8-hydroxy-7-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 15; 0.36 g, 0.89 mmol) in DMF (10 ml) was added ethyl-(E)-3-bromoacrylate (0.32 g, 1.78 mmol), potassium carbonate (0.25 g, 1.78 mmol ) and tetra-butyl ammonium bromide (5 mg, 0.016 mmol) and the resulting mixture was heated at 90°C for 24 h. After completion of the reaction (monitored by TLC), the reaction was stopped with water (5 ml) and the aqueous layer was extracted with EtOAc ( 2×15 ml). The combined organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material (1:1 for both diastereoisomers) was further purified by preparative HPLC (method B) to give a first eluting fraction corresponding to the (Z)-isomer and a second eluting fraction corresponding to the (E)-isomer in a total yield of 53 %.

(Z)-isomer: Yield: 0.13 g (almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.49 (s, 1H), 7.29 (t, J =7.6 Hz, 2H), 7.14 (d, J =8.4 Hz, 2H), 7.03-6.91 (m, 2H), 6.55 (s, 1H), 5.16 (d, J =6.8 Hz, 1H), 4.10 (q, J =7.2 Hz, 2H), 3.90-3.72 (m, 2H), 3.63 (s, 3H), 3.31 (s, 2H), 1.42-1.33 (m, 3H), 1.32-1.29(m, 4H), 1.19-1.02(m, 4H), 0.76-0.71(m, 6H). LCMS: (Method C) 502.1 (M+H), Rt. 3.03 min, 99.13% (maximum).

(E)-isomer: Yield: 0.12 g (almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.72 (d, J =12.2 Hz, 1H), 7.54 (s, 1H), 7.32-7.28 (m, 2H ), 7.18-7.16 (m, 2H), 7.01-6.97 (m, 1H), 6.55 (s, 1H), 5.40 (d, J =12.2 Hz, 1H), 4.11 (q, J = 7.0 Hz, 2H), 3.92-3.75 (m, 2H), 3.62 (s, 3H), 3.31 (m, 2H), 1.36-1.31(m, 3H), 1.24-1.20(m, 4H), 1.21-0.95(m, 4H), 0.76-0.71(m, 6H ). LCMS: (Method C) 502.1 (M+H), Rt. 3.12 min, 99.23% (maximum).

Intermediate 17

7-bromo-3,3-dibutyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (0.8 g, 1 .62 mmol) in DCM (20 ml) at 0°C was added dropwise BBr ₃ (0.78 ml, 8.1 mmol) and the reaction mixture was stirred at room temperature for 1 h. After completion of the reaction (monitored by TLC ) the reaction mixture was cooled to 0°C. EtOAc (10 ml) and ice-cold water (5 ml) were added dropwise and the mixture was stirred at room temperature for 1 h. The reaction mixture was then partitioned between water (15 ml) and DCM (15 ml) and the aqueous layer was extracted with DCM (3×15 ml). The combined organics were washed with brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The obtained crude material was sent to the next stage without any further purification. Yield: 90% (700 mg, crude off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.83 (s, 1H), 7.46 (s, 1H), 7.23 (t, J =8.0 Hz, 2H), 7, 07 (s, 1H), 6.99 (d, J =7.6 Hz, 2H), 6.88 (t, J =7.2 Hz, 1H), 3.80-3.72 (m, 2H ), 3.27 (s, 2H), 1.34-1.31 (m, 4H), 1.13-0.99 (m, 8H), 0.75-0.72 (m, 6H). LCMS: (Method A) 480.1 (M ⁺ ), Rt. 3.19 min, 89.95% (maximum).

Intermediate 18

3,3-dibutyl-8-hydroxy-7-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 7-bromo-3,3-dibutyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 17; 0. 9 g, 1.87 mmol) in sodium methoxide (21%, 10 ml, 10.2 mmol) was added CuBr (0.15 g, 1.05 mmol) at room temperature and the resulting mixture was heated at 85°C for 6 hours After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting crude mass was subjected to distribution between EtOAc (25 ml) and water (25 ml). The aqueous layer was extracted with EtOAc (2×25 ml) and the combined organic layer was washed with brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 5% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 78% (0.63 g, light pink gum).

UPLC: (Method A) 432.5 (M+H), Rt. 1.93 min, 90.07% (maximum).

Intermediate 19

Ethyl-(E)-3-((3,3-dibutyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylate and ethyl-(Z)-3-((3,3-dibutyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylate

With stirring to a solution of 3,3-dibutyl-8-hydroxy-7-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 18; 0. 63 g, 1.45 mmol) in DMF (10 ml), ethyl-(E)-3-bromoacrylate (0.52 g, 2.90 mmol) sodium carbonate (0.39 g, 3.62 mmol) and tetra-butyl ammonium bromide (0.05 g, 0.14 mmol) were added and the resulting mixture was heated at 90° C. within 24 hours After completion of the reaction (monitored by TLC), the reaction was stopped with water (5 ml) and the aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na₂SO₄ and concentrated in vacuo. The resulting crude material, which contained the (E)- and (Z)-isomers in a 1:1 ratio, was further purified by preparative HPLC (method B) to obtain a first eluting fraction corresponding to the (Z)-isomer and a second eluting fraction, corresponding to the (E)-isomer with a total yield of 60%.

(Z)-isomer: Yield: 0.23 g (almost white solid); ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.46 (s, 1H), 7.28-7.27 (m, 2H), 7.16-7.14 (m, 2H), 7 .01-7.00 (m, 2H), 6.50 (s, 1H), 5.14 (d, J =6.8 Hz, 1H), 4.09 (q, J =7 Hz, 2H) , 3.75-3.65 (m, 2H), 3.60 (s, 3H), 3.32-3.30 (m, 2H), 1.21-1.19 (m, 4H), 1 .18-1.07 (m, 11H), 0.75-0.73 (m, 6H).

(E)-isomer: Yield: 0.23 g (almost white solid); ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.69 (d, J =12.2 Hz, 1H), 7.51 (s, 1H), 7.32-7.28 (m, 2H ), 7.18-7.16 (m, 2H), 7.01-6.99 (m, 1H), 6.51 (s, 1H), 5.37 (d, J =12.2 Hz, 1H), 4.09 (q, J = 7.0 Hz, 2H), 3.85 (bs, 2H), 3.59 (s, 3H), 3.32 (s, 2H), 1.40- 1.31 (m, 4H), 1.21-1.09 (m, 11H), 1.07-1.00 (m, 6H).

Intermediate 20

5-(4-bromophenyl)-3,3-dibutyl-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3,3-dibutyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (2 g, 4 .47 mol) in dry DMF (20 ml) N-bromosuccinimide (0.875 g, 4.92 mmol) dissolved in dry DMF (20 ml) was added dropwise at 0°C and the reaction mixture was stirred at 0°C for 1 hours After completion of the reaction (monitored by LCMS), the reaction mixture was poured into crushed ice. The resulting solid was filtered off, washed with water (2 x 20 ml) and dried in vacuo to give the title compound. Yield: 89% (2.1 g, brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.65 (bs, 1H), 7.32-7.30 (m, 3H), 6.82-6.81 (m, 2H), 6 .75 (s, 1H), 3.80-3.50 (m, 2H), 3.25-3.15 (m, 2H), 2.23 (s, 3H), 1.46-1.26 (m, 4H), 1.16-1.08 (m, 8H), 0.82-0.80 (m, 6H). UPLC: (Method A) 526.4 (M+H), Rt. 1.42 min, 94.32%.

Intermediate 21

Ethyl-(E)-3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate and ethyl-(Z)-3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3, 4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of 5-(4-bromophenyl)-3,3-dibutyl-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide ( intermediate 20; 0.49 g, 0.93 mmol) in DMF (5 ml) was added ethyl (E)-3-bromoacrylate (0.25 g, 4.92 mmol) and potassium carbonate (0.26 g, 1.86 mmol) at room temperature and the resulting mixture was heated at 100°C for 4 hours. After completion of the reaction (monitored by TLC), the reaction was stopped with water (20 ml) and the aqueous layer was extracted with EtOAc (2×10 ml) . The combined organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material, which contained the (E)- and (Z)-isomers in a 1:1 ratio, was purified by preparative HPLC (method D) to obtain a first eluting fraction corresponding to the (Z)-isomer and a second eluting fraction corresponding to ( E)-isomer, with a total yield of 50%.

(Z)-isomer: Yield: 0.12 g (almost white solid); ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.49 (s, 1H), 7.40 (d, J =8.8 Hz, 2H), 7.24 (d, J =6.8 Hz, 2H), 7.01-6.99 (m, 2H), 6.84 (s, 1H), 5.29 (d, J =6.8 Hz, 1H), 4.11 (q, J =7.2 Hz, 2H), 3.92-3.55 (m, 2H), 3.33 (s, 2H), 2.28 (s, 3H), 1.45-1.05 (m, 14H), 0.80-0.77 (m, 6H). LCMS: (Method C) 624.0 (M ⁺ ), Rt. 3.43 min, 96.50% (maximum).

(E)-isomer: Yield: 0.13 g (white solid); ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.77 (d, J =12.0 Hz, 1H), 7.52 (s, 2H), 7.42 (d, J =8.8 Hz, 2H), 7.06-7.04 (m, 2H), 6.84 (s, 1H), 5.55 (d, J =12.0 Hz, 1H), 4.13 (q, J =7.2 Hz, 2H), 3.92-3.55 (m, 2H), 3.36 (s, 2H), 2.27 (s, 3H), 1.45-1.04 (m, 14H), 0.80-0.77 (m, 6H). LCMS: (Method C) 624.0 (M ⁺ ), Rt. 3.54 min, 97.99% (maximum).

Intermediate 22

7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (1.0 g , 2 mmol) in dry DMF (15 mL) sodium hydride (60% in mineral oil) (0.09 mg, 2.40 mmol) was added at 0°C and the reaction mixture was stirred for 10 min. Then, methyl iodide (0.4 ml, 6 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 30 minutes. After completion of the reaction (monitored by TLC), the reaction was stopped with ice-cold water (2 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The obtained crude material was sent to the next stage without any further purification. Yield: 96% (900 mg, crude brown resinous solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.45 (s, 1H), 7.26 (t, J =8.0 Hz, 2H), 7.18-7.16 (m, 1H ), 7.06-7.04 (m, 2H), 6.93 (t, J = 7.2 Hz, 1H), 3.93 (s, 3H), 3.70-3.61 (m, 2H), 3.10 (s, 2H), 1.51-0.90 (m, 8H), 0.80-0.72 (m, 6H). LCMS: (Method A) 468.1 (M+2), Rt. 3.21 min, 96.82% (maximum).

Intermediate 23

3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 22; 0.9 g, 1.90 mmol) in dry DMF (15 ml) was added sodium thiomethoxide (687 mg, 9.5 mmol) at room temperature and the reaction mixture was stirred at 60°C for 16 h. using LCMS), the reaction was stopped with ice-cold water (2 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 10-15% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 93% (750 mg, pale brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.16-7.12 (m, 3H), 6.85-6.83 (m, 2H), 6.71 (t, J =7, 2 Hz, 1H), 6.60 (s, 1H), 3.74-3.61 (m, 2H), 3.12 (s, 2H), 2.13 (s, 3H), 1.70- 1.08 (m, 8H), 0.80-0.74 (m, 6H). UPLC: (Method A) 420.5 (M+H), Rt. 1.86 min, 91.84% (maximum).

Intermediate 24

Ethyl-(E)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylate and ethyl-(Z)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring to a solution of 3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 23; 0.75 g, 1.79 mmol) in dry DMF (20 ml) was added ethyl-(E)-3-bromoacrylate (480 mg, 2.68 mmol) and potassium carbonate (494 mg, 3.57 mmol) and the reaction mixture was heated at 90°C for 4 hours After completion of the reaction (monitored by TLC), the reaction was stopped with water (5 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material, which contained the (E)- and (Z)-isomers in a 1:1 ratio, was purified by preparative HPLC (method D) to obtain a first eluting fraction corresponding to the (Z)-isomer and a second eluting fraction corresponding to (E)-isomer, with a total yield of 55%.

(Z)-isomer: Yield: 29% (0.21 g, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.75 (d, J =12.4 Hz, 1H), 7.51 (s, 1H), 7.31 (t, J =8.0 Hz, 2H), 7.17 (d, J =8.0 Hz, 2H), 7.00 (t, J =7.2 Hz, 1H), 6.68 (s, 1H), 5.51 ( d, J =12.4 Hz, 1H), 4.12 (q, J =7.2 Hz, 2H), 3.85-3.62 (m, 2H), 3.38 (s, 2H), 2.17 (s, 3H), 1.43-1.00 (m, 11H), 0.76-0.73 (m, 6H). LCMS: (Method C) 518.1 (M+H), Rt. 3.25 min, 99.38% (maximum).

(E)-isomer: Yield: 26% (0.22 g, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.48 (s, 1H), 7.29 (t, J =8.0 Hz, 2H), 7.20 (d, J =6.8 Hz, 1H), 7.13 (d, J =7.6 Hz, 2H), 6.98 (t, J =7.2 Hz, 1H), 6.69 (s, 1H), 5.27 ( d, J = 7.2 Hz, 1H), 4.11 (q, J = 7.2 Hz, 2H), 3.75-3.74 (m, 2H), 3.34 (s, 2H), 2.19 (s, 3H), 1.55-0.98 (m, 11H), 0.76-0.75 (m, 6H). LCMS: (Method C) 518.1 (M+H), Rt. 3.15 min, 98.88% (maximum).

Intermediate 25

3,3-dibutyl-8-hydroxy-5-(4-methoxyphenyl)-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 5-(4-bromophenyl)-3,3-dibutyl-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide ( intermediate 20; 1.5 g, 2.84 mmol) in dry DMF (5 ml) was added sodium methoxide (5 ml, 20% in methanol) and CuBr (0.06 g, 0.284 mmol) at room temperature and the reaction mixture heated at 100°C for 24 hours After completion of the reaction (monitored by LCMS), the reaction mixture was filtered through celite and washed with EtOAc (15 ml). The organic portion was concentrated in vacuo and the resulting crude material was passed on to the next step without any further purification. Yield: 44% (0.600 g, brown resinous solid).

LCMS: (Method C) 477.68 (M+H), Rt. 3.51 min, 76.40% (maximum).

Intermediate 26

Ethyl-(E)-3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate and ethyl-(Z)-3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3, 4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of 3,3-dibutyl-8-hydroxy-5-(4-methoxyphenyl)-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide ( intermediate 25; 0.65 g, 1.36 mmol) in dry DMF (10 mL) was added ethyl (E)-3-bromoacrylate (0.88 g, 4.92 mmol) and potassium carbonate (0.56 g , 4.08 mmol) at room temperature and the reaction mixture was heated at 100°C for 4 h. After completion of the reaction (monitored by TLC), the reaction was stopped with water (5 ml) and the aqueous layer was extracted with ethyl acetate (2×10 ml). The combined organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material, which contained the (E)- and (Z)-isomers in a 1:1 ratio, was purified by preparative HPLC (method D) to obtain a first eluting fraction corresponding to the (Z)-isomer and a second eluting fraction corresponding to (E)-isomer, with a total yield of 51%.

(Z)-isomer: Yield: 25% (0.20 g, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.42 (s, 1H), 7.25 (d, J =8.6 Hz, 2H), 7.12 (d, J =6.8 Hz, 1H), 6.94 (d, J =9.0 Hz, 2H), 6.43 (s, 1H), 5.22 (d, J =6.8 Hz, 1H), 4.10 ( q, J = 7.0 Hz, 2H), 3.82-3.65 (m, 5H), 3.40 (s, 2H), 2.13 (s, 3H), 1.42-0.99 (m, 15H), 0.76-0.73 (m, 6H).

(E)-isomer: Yield: 26% (0.21 g, almost white solid), 1H NMR (400 MHz, DMSO- d ₆ ): δ 7.69 (d, J =12.4 Hz, 1H) , 7.45 (s, 1H), 7.27 (d, J =9.2 Hz, 2H), 6.95 (d, J =8.8 Hz, 2H), 6.41 (s, 1H) , 5.42 (d, J =12.4 Hz, 1H), 4.09 (q, J =6.8 Hz, 2H), 3.82-3.65 (m, 5H), 3.42 ( s, 2H), 2.10 (s, 3H), 1.42-0.97 (m, 15H), 0.79-0.75 (m, 6H).

Intermediate 27

Ethyl-(E)-3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- BBr ₃ (1M in DCM, 0 .3 ml, 0.26 mmol) and the reaction mixture was stirred at room temperature for 1 h. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to 0°C, EtOAc (10 ml) and ice-cold water were added dropwise (5 ml) and the reaction mixture was stirred at room temperature for 1 h. Then the reaction mixture was subjected to distribution between water (15 ml) and DCM (15 ml) and the aqueous layer was extracted with DCM (3×15 ml). The combined organics were washed with brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The obtained crude material was sent to the next stage without any further purification. Yield: 100 mg (crude brown resinous solid).

LCMS: (Method C) 562.2 (M+H), Rt. 3.09 min, 73.17% (maximum).

Intermediate 28

Ethyl-(E)-3-((3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylate

With stirring to a solution of ethyl-(E)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 3; 0.35 g, 0.64 mmol) in a mixture of MeOH and water (10 ml, 4:1 ratio) was added NH ₄ Cl (0.03 g, 5. 08 mmol) and powdered Zn (0.21 g, 3.18 mmol) and the reaction mixture was heated at 65°C for 2 h. After completion of the reaction (monitored by TLC), the reaction was stopped with water (15 ml) and the aqueous layer was extracted with EtOAc (2×25 ml). The combined organic layer was washed with brine (15 ml), dried over anhydrous Na ₂ SO ₄ 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: 52% (0.150 g, white resinous solid).

LCMS: (Method A) 472.1 (M+H), Rt. 3.12 min, 98.24% (maximum).

Intermediate 29

(E) -3-((7-bromo-3-butyl-3-ethyl-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring to a solution of (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid (Example 4; 0.56 g, 1.07 mol) in a 1:1 mixture of DCM and AcOH (10 ml) was added dropwise with HNO ₃ (65%; 0.101 g 1.6 mmol) in a mixture of DCM and AcOH (5 ml) at 0°C. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml) and the organic layer was washed with water (15 ml) and saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the title compound. The crude material was sent to the next step without further purification. Yield: 500 mg (crude brown solid).

LCMS: (Method A) 565.0 (M ⁺ -H), Rt, 2.53 min, 70.98% (maximum).

Intermediate 30

(E) -3-((5-(4-aminophenyl)-7-bromo-3-butyl-3-ethyl-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring to a solution of (E) -3-((7-bromo-3-butyl-3-ethyl-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylic acid (intermediate 29; 0.225 g, 0.39 mol) in THF (10 ml) was added concentrated HCl (0.1 ml) and SnCl ₂ (0.225 g 1.19 mmol ) at room temperature and the reaction mixture was stirred for 12 h at 70°C. After completion of the reaction (monitored by TLC), the reaction was stopped with a saturated solution of NaHCO ₃ (15 ml), filtered through a pad of celite and washed with EtOAc (2×15 ml). The resulting filtrate was washed with water (20 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 160 mg (crude brown solid).

LCMS: (Method A) 537.8 (M ⁺ +H), Rt. 2.17 min, 90.93% (maximum).

Intermediate 31

Ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 1; 0.33 g, 0.59 mmol) in a 1:1 mixture of DCM and AcOH (3 ml) was added HNO ₃ (65%, 0.056 g, 0.89 mmol ) in a mixture of DCM and AcOH (1 ml) at 0°C. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml) and the organic layer was washed with water (15 ml) and saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the title compound. The crude material was sent to the next step without further purification. Yield: 300 mg (crude brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 8.04 (d, J =12.4 Hz, 2H), 7.82 (d, J =16.4 Hz, 1H), 7.57 ( s, 1H), 7.17 (s, 1H), 6.98 (s, 2H), 5.65 (d, J =16.4 Hz, 1H), 4.13 (q, J =9.6 Hz, 2H), 3.74 (s, 2H), 3.37 (s, 2H), 2.49 (s, 3H), 1.31-1.25 (m, 2H), 1.23-1 .14 (m, 10H), 0.83-0.81 (m, 6H). LCMS: (Method D) 591.2 (M ⁺ +H), Rt. 4.04 min, 93.4% (maximum).

Intermediate 32

Methyl-(E)-3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylic acid (intermediate 31; 0.8 g, 1.42 mol) oxalyl chloride (0.24 ml, 2.84 mmol) was added to DCM at 0°C. DMF (0.05 ml) was then added at this temperature and the reaction mixture was stirred for 1 h at 0°C. After completion of the reaction (monitored by TLC), the reaction was stopped with MeOH (10 ml) and diluted with DCM (15 ml). The organic layer was washed with water (15 ml) and saturated NaHCO ₃ solution (15 ml). The organic layer was then dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the title compound. The crude material was sent to the next step without any further purification. Yield: 775 mg (crude brown solid).

LCMS: (Method C) 577.1 (M ⁺ +H), Rt. 3.07 min, 78.21% (maximum).

Intermediate 33

Methyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of methyl-(E)-3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 32; 0.775 g, 1.34 mol) in THF (10 ml) was added concentrated HCl (1 ml) and SnCl ₂ (1.01 g, 5, 37 mmol) at room temperature and the reaction mixture was stirred for 12 h at 70°C. After completion of the reaction (monitored by TLC), the reaction was stopped with a saturated solution of NaHCO ₃ (25 ml), filtered through a pad of celite and washed with EtOAc (15 ml). The resulting filtrate was washed with water (2×15 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 0.7 g (crude brown solid).

LCMS: (Method C) 547.1 (M ⁺ +H), Rt. 2.87 min, 79.4% (maximum).

Intermediate 34

Methyl-(E)-3-((3,3-dibutyl-5-(4-((methoxycarbonyl)amino)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of methyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (intermediate 33, 0.06 g, 0.1 mol) in DCM (5 ml) was added triethylamine (0.03 g, 0.3 mmol) at 0° C. Then methyl chloroformate (0.014 g, 0.15 mmol) was added to the reaction mixture and stirred for 3 hours at RT. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (10 ml) and the organic layer was washed with saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the title compound. The crude material was sent to the next step without any further purification. Yield: 60 mg (crude off-white solid).

LCMS: (Method A) 605.2 (M ⁺ +H), Rt. 2.97 min, 66.2% (maximum).

Intermediate 35

Ethyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 31; 0.4 g, 0.67 mmol) in THF (5 ml) was added concentrated HCl (0.1 ml) and SnCl ₂ (0.512 g, 2.71 mmol) at room temperature and the reaction mixture was stirred for 12 h at 70°C. After completion of the reaction (monitored by TLC), the reaction was stopped with a saturated solution of NaHCO ₃ (15 ml), filtered through a pad of celite and washed with EtOAc (2×15 ml). The resulting filtrate was washed with water (20 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 0.38 g (crude brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.66 (d, J =16.4 Hz, 1H), 7.39 (s, 1H), 7.04 (d, J =11.6 Hz, 2H), 6.59 (d, J =11.2 Hz, 2H), 6.31 (s, 1H), 5.37 (d, J =16.4 Hz, 1H), 5.09 ( s, 2H), 4.10 (q, J =9.2 Hz, 2H), 3.73 (s, 2H), 3.41 (s, 2H), 2.07 (s, 3H), 1, 45-1.36 (m, 4H), 1.19-1.01 (m, 8H), 0.91 (t, J =14.4 Hz, 6H). LCMS: (Method C) 561.2 (M ⁺ +H), Rt. 2.99 min, 89.73% (maximum).

Intermediate 36

Methyl-(E)-3-((3,3-dibutyl-5-(4-(dimethylamino)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-8-yl)oxy)acrylate

To a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-benzothiazepin-8-yl)oxy)acrylate (intermediate 35; 100 mg, 0.18 mmol) in methanol (3 ml) and AcOH (0.5 ml) formaldehyde (37%, 9.2 mg, 0 .18 mmol) and the reaction mixture was stirred for 3 h at ambient temperature. Then NaCNBH ₃ (23 mg, 0.366 mmol) was added and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo. The resulting residue was subjected to distribution between water (10 ml) and EtOAc (15 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with brine (25 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 70% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound (transesterification product in the presence of methanol). Yield: 43% (46 mg, yellow solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.70 (d, J =12.4 Hz, 1H), 7.43 (s, 1H), 7.20 (d, J =9, 2 Hz, 2H), 6.76 (d, J =8.4 Hz, 2H), 6.38 (s, 1H), 5.45 (d, J =12.0 Hz, 1H), 3.76 (s, 2H), 3.64 (s, 3H), 3.43 (s, 2H), 2.89 (s, 6H), 2.08 (s, 3H), 1.48-1.31 ( m, 4H), 1.24-0.84 (m, 8H), 0.74 (t, J =6.8 Hz, 6H). LCMS: (Method A) 575.3 (M ⁺ -H), Rt. 3.39 min, 96.49% (maximum).

Intermediate 37

(Z) -3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid (Example 11; 2.4 g, 4.48 mmol) in a 1:1 mixture of DCM and AcOH (24 ml) was added dropwise with HNO ₃ (65%, 0.42 g , 6.70 mmol) in a mixture of DCM and AcOH (12 ml) at 0°C. Then the reaction mixture was stirred for 1 h. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (25 ml) and the organic layer was washed with water (3×20 ml). The combined organic layer was washed with brine (2×20 ml) and dried over anhydrous Na ₂ SO ₄ . The solvent was evaporated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 2.53 g (crude brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.71 (s, 1H), 8.05 (d, J =8.8 Hz, 2H), 7.70-7.65 (m, 2H ), 7.16 (s, 1H), 7.01-6.86 (m, 2H), 3.86 (s, 2H), 2.67 (s, 2H), 2.38 (s, 3H) , 1.59-1.55 (m, 2H), 1.31-1.14 (m, 10H), 0.84-0.80 (m, 6H). LCMS: (Method E) 579.2 (M ⁺ -H), Rt. 3.26 min, 93.55% (maximum).

Intermediate 38

(Z) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (intermediate 37; 2.53 g, 4.30 mmol) in THF (25.3 ml) was added concentrated HCl (2.53 ml) and SnCl ₂ (3.29 g, 17.40 mmol) at room temperature and the reaction mixture was heated at 80° C. overnight. After completion of the reaction (monitored by TLC), water (20 ml) was added to the reaction mixture and the resulting solid was filtered through a pad of celite, which was washed with EtOAc (50 ml). The organic portion was washed with brine (20 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 6-7% MeOH in DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 89% (2.14 g, off-white solid).

LCMS: (Method C) 548.9 (M-H), Rt. 2.47 min, 95.69% (maximum).

Intermediate 39

Ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (intermediate 35; 0.06 g, 0.1 mol) in DCM (5 ml) was added triethylamine (0.03 ml, 0.21 mmol) at 0° C. Then pivaloyl chloride (0.016 g, 0.12 mmol) was added and the reaction mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml) and the organic part was washed with saturated NaHCO ₃ solution (2×10 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 65 mg (crude off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.16 (s, 1H), 7.72 (d, J =16.4 Hz, 1H), 7.60 (d, J =11.6 Hz, 2H), 7.47 (s, 1H), 7.16 (d, J =12.0 Hz, 2H), 6.60 (s, 1H), 5.48 (d, J =16.4 Hz, 1H), 4.12 (d, J =9.6 Hz, 2H), 3.74 (bs, 2H), 3.38 (s, 2H), 2.27 (s, 3H), 1, 42 (s, 9H), 1.32-1.22 (m, 6H), 1.20-1.18 (m, 6H), 1.11 (t, J =18.4 Hz, 6H). LCMS: (Method A) 645.3 (M ⁺ +H), Rt. 3.21 min, 91.1% (maximum).

Intermediate 40

Ethyl-(E)-3-((5-(4-((butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (intermediate 35; 0.035 g, 0.06 mmol) in DCM (3 ml) was added dropwise triethylamine (0.012 g, 0.12 mmol) at 0°C. Butyl chloroformate (0.011 g, 0.08 mmol) was then added to the reaction mixture, and stirring was continued for 2 hours at RT. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (5 ml) and the DCM layer was washed with saturated NaHCO ₃ solution (20 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound which was passed on to the next step without any further purification. Yield: 41 mg (crude off-white solid). LCMS: (Method A) 660.9 (M ⁺ +H), Rt. 3.32 min, 85.01% (maximum).

Intermediate 41

Ethyl-(E)-3-((3,3-dibutyl-5-(4-(3,3-dimethylbutanamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (Intermediate 35; 0.150 g, 0.26 mmol) in DCM (5 ml) was added dropwise with triethylamine (0.074 g, 0.53 mmol) at 0°C. Then 3,3-dimethylbutanoyl chloride (0.054 g, 0.4 mmol) was added to the reaction mixture, and stirring was continued for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (5 ml) and the organic layer was washed with saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 150 mg (crude off-white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 9.74 (s, 1H), 7.72 (d, J =16.4 Hz, 1H), 7.56 (d, J =10, 8 Hz, 2H), 7.47 (s, 1H), 7.18 (d, J =11.6 Hz, 2H), 6.58 (s, 1H), 5.47 (d, J =16, 4 Hz, 1H), 4.12 (q, J =10.0 Hz, 2H), 3.75 (bs, 2H), 3.39 (s, 2H), 2.17 (s, 3H), 2 .15 (s, 2H), 1.42-1.32 (m, 3H), 1.27-1.08 (m, 6H), 1.02 (s, 9H), 0.98-0.91 (m, 6H), 0.7 (t, J = 8.4 Hz, 6H). LCMS: (method C) 659.3 (M ⁺ +H), Rt. 3.30 min, 93.37% (maximum).

Intermediate 42

7-bromo-3,3-dibutyl-8-methoxy-5-(4-(trifluoromethyl)phenyl)-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

To a short degassed solution of 7-bromo-3,3-dibutyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (2 g, 5.01 mmol) in 4-bromobenzotrifluoride ( 6 ml) was added tris[2-(2-methoxyethoxy)ethyl]amine (0.16 g, 0.5 mmol), CuI (0.19 g, 1.0 mmol) and dry K ₂ CO ₃ (1.38 g, 10.02 mmol) and the reaction mixture was heated for 24 h at 130°C. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was taken up in water (15 ml). The aqueous layer was extracted with EtOAc (2×25 ml) and the combined organic layer was washed with brine (50 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 19% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 73% (2.2 g, yellow gum).

LCMS: (Method C) 544.1 (M+H), Rt. 3.54 min, 89.96% (maximum).

Intermediate 43

7-bromo-3,3-dibutyl-8-methoxy-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-(4-(trifluoromethyl)phenyl)-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate product 42; 2 g, 3.68 mmol) in dry THF (20 ml) at 0°C was added dropwise borane dimethyl sulfide (1M in THF; 7.5 ml, 7.37 mmol) and the reaction mixture was heated for 16 h at 80°C. After completion of the reaction (monitored by TLC), the reaction was stopped with methanol (15 ml) at 0°C and heated for 1 h at 80°C. The reaction mixture was then concentrated in vacuo. The resulting residue was taken up in water (15 ml) and extracted with EtOAc (2×50 ml). The combined organic layer was washed with brine (50 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 4% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 62% (1.21 g, off-white solid).

LCMS: (Method D) 532.1 (M+H), Rt. 3.26 min, 81.24% (maximum).

Intermediate 44

7-bromo-3,3-dibutyl-8-methoxy-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine (intermediate 43; 1 .21 g, 2.26 mmol) in a mixture of acetone and water (12 ml, 3:1) was added oxone (7 g, 2.27 mmol) and the reaction mixture was stirred for 48 h at room temperature. The progress of the reaction was followed by TLC and LCMS, which indicated the formation of sulfoxide and sulfone. The reaction mixture was then filtered through celite to remove excess oxone and the filtrate was diluted with water (30 ml). The aqueous layer was extracted with EtOAc (2×40 ml) and the combined organic layer was washed with brine (40 ml). The organic was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 3% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 68% (0.87 g, pale yellow solid).

LCMS: (Method D) 562.1 (M ⁺ +H), Rt. 4.39 min, 95.43% (maximum).

Intermediate 45

3,3-dibutyl-8-hydroxy-7-(methylthio)-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 44; 0.87 g, 1.54 mmol) NaSMe (0.57 g, 7.73 mmol) was added to DMF (5 ml) and the solution was heated for 4 h at 60°C. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (15 ml) and stirred for 5 minutes. The aqueous layer was extracted with EtOAc (2×15 ml) and the combined organic layer was washed with brine (15 ml). The organic portion was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 71% (0.57 g, yellow gummy).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.95 (s, 1H), 7.45 (d, J =8.0 Hz, 2H), 7.32 (s, 1H), 6 .92-6.85(m, 2H), 6.84(s, 1H), 3.85(s, 2H), 3.21(s, 2H), 2.25(s, 3H), 1, 52-1.39 (m, 2H), 1.26-1.13 (m, 10H), 0.82-0.79 (m, 6H). LCMS: (Method C) 516.1 (M+H), Rt. 3.09 min, 81.04% (maximum).

Intermediate 46

Ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-8-yl)oxy)acrylate

With stirring to a solution of 3,3-dibutyl-8-hydroxy-7-(methylthio)-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1 -dioxide (intermediate 45; 0.15 g, 0.29 mmol) in THF (3 ml) was added DABCO (0.003 g, 0.029 mmol) and ethyl propiolate (0.043 g, 0.43 mmol) and the reaction mixture was stirred for 30 min at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and the residue was subjected to distribution between ice water (10 ml) and EtOAc (10 ml). The aqueous layer was extracted with EtOAc (2×8 ml) and the combined organic layer was washed with brine (8 ml). The organic was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 40% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 50% (0.09 g, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.81 (d, J =12.0 Hz, 1H), 7.56 (s, 1H), 7.52 (d, J =8, 8 Hz, 2H), 7.08-7.04 (m, 3H), 5.61 (d, J =12.0 Hz, 1H), 4.13 (q, J =7.2 Hz, 2H) , 3.82(s, 2H), 3.34(s, 2H), 2.32(s, 3H), 1.53-1.41(m, 2H), 1.33-1.31(m , 1H), 1.27-1.20 (m, 12H), 0.81-0.77 (m, 6H). LCMS: (Method D) 614.2 (M+H), Rt. 4.35 min, 91.98% (maximum).

Intermediate 47

Ethyl-(E)-3-((3,3-dibutyl-5-(4-isobutyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (intermediate 35, 0.125 g, 0.22 mmol) DIPEA (0.11 ml, 0.66 mmol), isobutyric acid (0.023 g, 0.26 mmol) and HATU (0.17 g, 0.44 mmol) at 0°C and then the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (10 ml) and the resulting solid was collected by filtration to give the title compound. The compound was sent to the next step without any further purification. Yield: 130 mg (crude off-white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 9.78 (s, 1H), 7.72 (d, J =16.4 Hz, 1H), 7.57 (d, J =11, 2 Hz, 2H), 7.47 (s, 1H), 7.18 (d, J =10.8 Hz, 2H), 6.58 (s, 1H), 5.47 (d, J =16, 4 Hz, 1H), 4.12 (q, J = 9.2 Hz, 2H), 3.75 (bs, 2H), 3.35-3.34 (m, 1H), 3.39 (s, 2H), 2.16 (s, 3H), 1.44-1.33 (m, 4H), 1.28-1.19 (m, 6H), 1.11-1.05 (m, 11H) , 0.77 (t, J = 8.00 Hz, 6H). LCMS: (Method C) 630.9 (M ⁺ +H), Rt. 3.15 min, 82.1% (maximum).

Intermediate 48

Ethyl-(E)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylate

With stirring to a solution of 3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 23; 1.5 g, 3.57 mmol) in THF (15 ml) was added DABCO (0.04 g, 0.35 mmol) and ethyl propiolate (0.42 g, 4.28 mmol) at 0°C and reaction the mixture was stirred for 1 hour at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (25 ml). The organic layer was washed with water (2×15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was purified using Isolera column chromatography (eluent: 10% EtOAc in hexane; silica gel: 230-400 mesh ) and obtained the desired compound. Yield: 81% (1.5 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.74 (d, J =12.4 Hz, 1H), 7.50 (s, 1H), 7.31 (d, J =8.0 Hz, 1H), 7.28 (s, 1H), 7.16 (d, J = 7.6 Hz, 2H), 6.99 (t, J = 7.2 Hz, 1H), 6.67 ( s, 1H), 5.50 (d, J =12.4 Hz, 1H), 4.12 (q, J =7.2 Hz, 2H), 3.74 (s, 2H), 3.37 ( s, 2H), 2.16 (s, 3H), 1.53-1.44 (m, 1H), 1.42-1.22 (m, 3H), 1.17 (t, J =6, 80 Hz, 3H), 1.09-0.99 (m, 4H), 0.7 (t, J =6.0 Hz, 6H). LCMS: (Method C) 518.1 (M ⁺ +H), Rt. 3.21 min, 98.39% (maximum).

Intermediate 49

Ethyl-(E)-3-((3-butyl-3-ethyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate

With stirring to a solution of ethyl-(E)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-benzothiazepin-8-yl)oxy)acrylate (intermediate 48; 1.3 g, 2.51 mmol) in a 1:1 mixture of DCM and AcOH (15 mL) HNO ₃ (65%, 0.238 g, 3.77 mmol) in a mixture of DCM and AcOH (5 ml). After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml) and the organic layer was washed with water (15 ml) and saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 1.4 g (crude brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 8.05 (d, J =9.2 Hz, 2H), 7.84 (d, J =12.4 Hz, 1H), 7.59 ( s, 1H), 7.20 (s, 1H), 6.99 (bs, 2H), 5.67 (d, J =12.4 Hz, 1H), 4.14 (q, J =6.8 Hz, 2H), 3.84 (bs, 2H), 3.40 (s, 2H), 2.38 (s, 3H), 1.66-1.40 (m, 2H), 1.38-1 .31 (m, 4H), 1.28-1.19 (m, 5H), 0.82 (t, J =6.40 Hz, 6H). LCMS: (Method C) 562.8 (M ⁺ +H), Rt, 3.01 min, 97.69% (maximum).

Intermediate 50

Ethyl-(E)-3-((5-(4-aminophenyl)-3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((3-butyl-3-ethyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 49; 1.4 g, 2.49 mmol) in THF (20 ml) was added at room temperature with concentrated HCl (1 ml) and SnCl ₂ ( 1.88 g, 9.96 mmol) and the reaction mixture was stirred for 12 hours at 70°C. After completion of the reaction (monitored by TLC) the reaction was stopped with a saturated solution of NaHCO ₃ (15 ml) and filtered through a pad of celite. The Celite layer was washed with EtOAc (2×15 ml) and the resulting filtrate was washed with water (2×25 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 1.3 g (crude brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.67 (dd, J =12.2, 1.52 Hz, 1H), 7.41 (d, J =1.4 Hz, 1H), 7.04 (d, J =7.3 Hz, 2H), 6.60 (d, J =7.3 Hz, 2H), 6.31 (s, 1H), 5.38 (dd, J =12 .3, 1.5 Hz, 1H), 5.10 (s, 2H), 4.13-4.08 (m, 2H), 3.73 (bs, 2H), 3.42 (s, 2H) , 2.08 (s, 3H), 1.52-1.38 (m, 4H), 1.36-1.22 (m, 4H), 1.20 (t, J =5.52 Hz, 3H ), 0.76 (t, J = 5.48 Hz, 6H). LCMS: (Method C) 532.8 (M ⁺ +H), Rt. 2.73 min, 91.68% (maximum).

Intermediate 51

Ethyl-(E)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate

With stirring to a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 50; 0.12 g, 0.22 mmol) in DCM (5 ml) was added triethylamine (0.045 g, 0.45 mmol) at 0° C. Then pivaloyl chloride (0.032 g, 0.27 mmol) was added and the reaction mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml) and the organic layer was washed with saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 135 mg (crude off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.17 (s, 1H), 7.73 (d, J =12.2 Hz, 1H), 7.60 (d, J =8.6 Hz, 2H), 7.49 (s, 1H), 7.16 (d, J = 8.0 Hz, 2H), 6.62 (s, 1H), 5.49 (d, J = 12.3 Hz, 1H), 4.12 (q, J = 7.0 Hz, 2H), 3.74 (bs, 2H), 3.38 (s, 2H), 2.17 (s, 3H), 1, 54-1.42 (m, 4H), 1.36 (s, 9H), 1.35-1.08 (m, 4H), 0.79-0.76 (m, 3H), 0.7 ( t, J = 7.2 Hz, 6H). LCMS: (Method A) 616.9 (M ⁺ +H), Rt. 3.08 min, 94.2% (maximum).

Intermediate 52

Ethyl-(E)-3-((3,3-dibutyl-5-(4-(cyclopentanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (intermediate 35; 0.125 g, 0.22 mmol) in DMF (3 mL) DIPEA (0.12 mL, 0.66 mmol) was added at 0°C, cyclopentanecarboxylic acid (0.038 g, 0.33 mmol) and HATU (0.17 g, 0.44 mmol) and the reaction mixture was stirred for 1 hour at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (15 ml) and the aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The obtained crude material was sent to the next stage without any further purification. Yield: 140 mg (crude off-white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 9.83 (s, 1H), 7.72 (d, J =16.4 Hz, 1H), 7.58 (d, J =12, 0 Hz, 2H), 7.47 (s, 1H), 7.18 (d, J =11.6 Hz, 2H), 6.57 (s, 1H), 5.47 (d, J =16, 0 Hz, 1H), 4.12 (q, J = 9.6 Hz, 2H), 3.75 (bs, 2H), 3.36 (s, 2H), 2.55-2.52 (m, 1H), 2.15 (s, 3H), 1.91-1.83 (m, 4H), 1.81-1.72 (m, 4H), 1.69-1.64 (m, 6H) , 1.63-1.41 (m, 6H), 1.36-1.08 (m, 3H), 0.8 (t, J =8.8 Hz, 6H). LCMS: (Method A) 657.2 (M ⁺ +H), Rt. 3.74 min, 93.37% (maximum).

Intermediate 53

Ethyl-(E)-3-((3-butyl-5-(4-(cyclopentanecarboxamido)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro -1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring to a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 50; 0.125 g, 0.23 mmol) in DMF (3 mL) DIPEA (0.12 mL, 0.7 mmol) was added at 0°C ), cyclopentanecarboxylic acid (0.040 g, 0.35 mmol) and HATU (0.178 g, 0.46 mmol) and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (10 ml) and the aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed to the next step without any further purification. Yield: 140 mg (crude off-white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 9.83 (s, 1H), 7.73 (d, J =12.2 Hz, 1H), 7.57 (d, J =8, 7 Hz, 2H), 7.48 (s, 1H), 7.17 (d, J = 7.6 Hz, 2H), 6.58 (s, 1H), 5.48 (d, J = 12, 3 Hz, 1H), 4.12 (q, J = 7.0 Hz, 2H), 3.74 (bs, 2H), 3.33 (s, 2H), 2.77-2.74 (m, 1H), 2.16 (s, 3H), 1.85-1.83 (m, 2H), 1.74-1.72 (m, 4H), 1.69-1.55 (m, 3H) , 1.54-1.44 (m, 3H), 1.38-1.35 (m, 3H), 1.31-1.21 (m, 4H), 0.8 (t, J =6, 3Hz, 6H). LCMS: (Method A) 629.2 (M ⁺ +H), Rt. 3.57 min, 90.68% (maximum).

Intermediate 54

2-(((2-Amino-5-methoxyphenyl)thio)methyl)-2-ethylbutanoic acid

KOH (1688 g, 30.09 mol) was added to a stirred solution of 6-methoxybenzo[d]thiazol-2-amine (339 g, 1.88 mol) in water (3390 ml) and the reaction mixture was stirred for 16 h at 120°C. After completion of the reaction (monitored by LCMS), the mixture was cooled to room temperature. Then 2-(bromomethyl)-2-ethylbutanoic acid (590 g, 2.82 mol; solution in 1500 ml THF) was added dropwise and the mixture was stirred for 16 hours at room temperature. 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 layer was washed with water (1000 ml) and brine (500 ml). The organic portion was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 650 g (crude brown gum).

LCMS: (Method A) 284 (M ⁺ +H), Rt. 1.82 min, 88.77% (maximum).

Intermediate 55

3,3-Diethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring, to a solution of 2-(((2-amino-5-methoxyphenyl)thio)methyl)-2-ethylbutanoic acid (intermediate 54; 650 g, 2.29 mol) in EtOAc (2500 ml) at 0°C by triethylamine (463 g, 4.586 mol) and a solution of 1-propane phosphoric anhydride (50% in EtOAc; 1021 g, 3.211 mol) were added dropwise and the reaction mixture was stirred for 16 h at room temperature. 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 (2×2000 ml). The combined organic layer was washed with brine (50 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by washing with methanol to give the title compound. Yield: 65% (295 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.56 (s, 1H), 7.06-7.03 (m, 1H), 6.97 (s, 1H), 6.87-6, 86 (m, 1H), 3.73 (s, 3H), 2.96 (s, 2H), 1.67-1.66 (m, 2H), 1.56-1.54 (m, 2H) , 0.79-0.77 (m, 6H). LCMS: (Method A) 266.1 (M ⁺ +H), Rt. 2.30 min, 99.45% (maximum).

Intermediate 56

7-Bromo-3,3-diethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring, to a solution of 3,3-diethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 55; 100 g, 0.376 mol) in 1:1 DCM and acetonitrile (1000 ml) was added portionwise N-bromosuccinimide (80 g, 0.452 mol) and the solution was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and the resulting crude material was treated with cold acetonitrile and stirred for 30 minutes. The resulting precipitate was filtered off and washed with cold acetonitrile (2×80 ml) and dried in vacuo to give the title compound. Yield: 180 g (crude brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.63 (s, 1H), 7.34 (s, 1H), 7.11 (s, 1H), 3.83 (s, 3H), 2 .98 (s, 2H), 1.64-1.66 (m, 2H), 1.50-1.52 (m, 2H), 0.76-0.78 (m, 6H). LCMS: (Method A) 344.1 (M ⁺ +H), Rt. 2.47 min, 96.70% (maximum).

Intermediate 57

7-Bromo-3,3-diethyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3,3-diethyl-7-iodo-8-methoxy -5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring, to a solution of 7-bromo-3,3-diethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 56; 180 g, 0.522 mol) in iodobenzene ( 1400 ml) copper(I) iodide (20 g, 0.104 mol) and K ₂ CO ₃ (144 g, 1.044 mol) were added and the solution was purged with nitrogen for 20 min to degas. Tris[2-(2-methoxyethoxy)ethyl]amine (16.8 g, 0.052 mol) was then added under nitrogen atmosphere and the resulting reaction mixture was heated for 40 h at 135°C. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through celite and the celite layer was washed with EtOAc (2000 ml). The filtrate was concentrated in vacuo to give a crude product which was crystallized from cold petroleum ether. The resulting precipitate was filtered off and washed with cold petroleum ether to give a mixture of the desired compounds. Yield: 200 g (crude light yellow solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.37-7.37 (m, 2H), 7.23-7.28 (m, 2H), 7.08-7.09 (m, 2H), 3.89 (s, 2H), 3.48-3.48 (m, 1H), 3.42-3.42 (m, 3H), 3.24 (s, 2H), 3.15 (d, J = 3.28 Hz, 2H), 2.64-2.65 (m, 1H), 1.46-1.48 (m, 4H), 0.77-0.77 (m, 6H ). LCMS: (Method A) 420.1 (M ⁺ +H) 30.8%; 468.1 (M ⁺ +H) 51.8%; Rt. 2.97 and 2.98 min, 82.6% (maximum).

Intermediate 58

7-Bromo-3,3-diethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3,3-diethyl-7-iodine-8-methoxy-5- phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine

With stirring, to a solution of a mixture of 7-bromo-3,3-diethyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3,3-diethyl-7- iodo-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 57; 100 g, 0.2378 mol) in THF (1000 ml) at 0° Borane dimethylsulfide (2M in THF; 356 ml, 0.713 mol) was added dropwise and the reaction mixture was refluxed for 40 h at 75°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C and the reaction was stopped with methanol (200 ml). The resulting solution was heated for 2 h at 65°C, then cooled to room temperature and concentrated in vacuo to give a mixture of the desired compounds. The resulting crude material was passed on to the next step without any further purification. Yield: 120 g (crude dark brown liquid).

LCMS: (Method A) 407.0 (M ⁺ +H), 41.3% and 454.0 (M ⁺ +H) 53.1%; Rt. 3.81 and 3.95 min, 94.4% (maximum).

Intermediate 59

7-Bromo-3,3-diethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3,3-diethyl-7-iodine- 8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of a mixture of 7-bromo-3,3-diethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3,3-diethyl-7-iodine-8 -methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine (intermediate 58; 120 g, 0.295 mol) in THF (600 ml) at room temperature was added water (600 ml) and oxone (907 g, 2.95 mol) and the reaction mixture was stirred for 24 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a Buchner funnel and the filtrate was extracted with EtOAc (2×2500 ml). The combined organic layer was washed with water (1000 ml) and brine (500 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by column chromatography (eluent: 8-10% EtOAc/PE; silica gel: 230-400 mesh) to give a mixture of the title compounds. Yield: 73% (95 g, yellowish solid).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.33-7.34 (m, 2H), 7.19-7.22 (m, 2H), 6.87-6.89 (m, 2H ), 4.26 (t, J = 7.04 Hz, 1H), 3.90-3.92 (m, 3H), 3.67-3.69 (m, 2H), 2.41-2, 43 (m, 1H), 2.11-2.13 (m, 2H), 1.50-1.52 (m, 2H), 1.30-1.31 (m, 2H), 0.71- 0.73(m, 6H). LCMS: (Method A) 440.0 (M ⁺ +H), 486.0 (M ⁺ +H), Rt. 2.98 min and 2.99 min, 80.7% (maximum).

Intermediate 60

3,3-Diethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of a mixture of 7-bromo-3,3-diethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3,3-diethyl -7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 59; 95 g, 0.216 mmol) in DMF (950 ml ) was added sodium thiomethoxide (45.6 g, 0.650 mmol) and the resulting reaction mixture was heated for 16 h at 60°C. After completion of the reaction (monitored by TLC), water (500 ml) was added to the reaction mixture and the aqueous layer was extracted with EtOAc (2×1000 ml). The combined organic layer was washed with brine (50 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by column chromatography (eluent: 15-20% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 59% (50 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.57 (s, 2H), 7.31 (s, 2H), 7.16-7.18 (m, 2H), 6.89 (d , J = 7.68 Hz, 2H), 6.76-6.78 (m, 2H), 6.66 (s, 2H), 3.64 (s, 1H), 3.20 (s, 2H) , 2.18 (s, 3H), 1.53-1.55 (m, 2H), 1.38-1.30 (m, 2H), 0.74-0.76 (m, 6H). LCMS: (Method A) 392.1 (M ⁺ +H), Rt. 2.54 min, 74.77% (maximum).

Intermediate 61

Ethyl-(E)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylate

With stirring, to a solution of 3,3-diethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 60; 0.3 g, 0.76 mmol) in dry THF (6 mL) was added ethyl propiolate (0.11 g, 1.14 mmol) and DABCO (8.6 mg, 0.076 mmol) and the solution was stirred for 30 min. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated. The resulting residue was subjected to distribution between water (10 ml) and EtOAc (15 ml) and the aqueous layer was extracted with EtOAc (2×15 ml). The combined organics were then washed with brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 45% (0.17 g, white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ ¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.76 (dd, J =12.2, 0.8 Hz, 1H), 7.52 (d, J = 0.80 Hz, 1H), 7.32-7.28 (m, 2H), 7.14 (d, J = 7.6 Hz, 2H), 6.98 (t , J = 7.2 Hz, 1H), 6.72 (s, 1H), 5.53 (dd, J = 12.0, 1.2 Hz, 1H), 4.13 (q, J= 0, 8 Hz, 2H), 3.76 (s, 2H), 3.37 (s, 2H), 2.19 (s, 3H), 1.54-1.52 (m, 2H), 1.39- 1.34 (m, 2H), 1.24-1.20 (m, 6H).

Intermediate 62

Ethyl-(E)-3-((3,3-dibutyl-5-(4-butyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of ethyl-(E)-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 35; 0.125 g, 0.22 mmol) in DCM (3 ml) at 0°C was added triethylamine (0.045 g 0.45 mmol) and then butyryl chloride ( 0.029 g 0.27 mmol) and the reaction mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (10 ml) and the organic layer was washed with saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 145 mg (crude off-white solid).

¹ H-NMR (400 MHz, CDCl ₃ ): δ 7.69 (d, J =Hz, 1H), 7.64 (d, J =9.6 Hz, 1H), 7.52 (d, J = 10.8 Hz, 2H), 7.17-7.11 (m, 3H), 6.56 (s, 1H), 5.54 (d, J =16.4 Hz, 1H), 4.21 ( q, J = 9.2 Hz, 2H), 3.77 (bs, 2H), 3.22 (s, 2H), 2.45 (t, J = 9.60 Hz, 2H), 2.16 ( s, 3H), 1.84-1.79 (m, 2H), 1.76-1.68 (m, 3H), 1.53-1.44 (m, 6H), 1.41-1, 20 (m, 9H), 0.99-0.93 (m, 6H). LCMS: (Method A) 631.3 (M ⁺ +H), Rt. 2.86 min, 92.69% (maximum).

Intermediate 63

Methyl-(Z)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylate

To a solution of 3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 23; 0.2 g, 0.5 mmol) in DMF (2 ml) at 0° C. 60% NaH (0.024 g, 1.02 mmol) was added in portions and the solution was stirred for 15 min. Methyl 3-bromo-2,2-difluoropropanoate (0.21 g, 1.02 mmol) was added and the reaction mixture was heated at 85° C. overnight. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C and the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude title compound, which was further triturated with diethyl ether. The resulting compound was dried in vacuo and passed on to the next step without any further purification. Yield: 46% (0.11 g, brown gummy).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.70-7.65 (m, 1H), 7.60 (s, 1H), 7.32-7.30 (m, 2H), 7 .15-7.13(m, 2H), 7.00-6.98(m, 1H), 6.69(s, 1H), 3.80(s, 3H), 3.75(s, 2H ), 3.36 (s, 2H), 2.20 (s, 3H), 1.43-1.41 (m, 4H), 1.18-1.10 (m, 4H), 0.74 ( t, J =4.80 Hz, 6H). LCMS: (Method A) 522.3 (M ⁺ +H), Rt. 3.01 min, 89.38% (maximum).

Intermediate 64

Methyl-(Z)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)-2-fluoroacrylate

With stirring, to a solution of 3,3-diethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 60; 0.2 g, 0.5 mmol) in DMF (2 ml) at 0°C was added 60% NaH (0.024 g, 1.02 mmol) in portions and the mixture was stirred for 15 min. Methyl 3-bromo-2,2-difluoropropanoate (0.21 g, 1.02 mmol) was added and the reaction mixture was heated at 85° C. overnight. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C and the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude title compound, which was further triturated with diethyl ether. The resulting compound was dried in vacuo and passed on to the next step without any further purification. Yield: 25% (0.06 g, off-white solid).

¹ H NMR (300 MHz, DMSO- d ₆ ): δ 7.71-7.65 (m, 1H), 7.61 (s, 1H), 7.31-7.29 (m, 2H), 7 .12-7.09(m, 2H), 6.99-6.94(m, 1H), 6.72(s, 1H), 3.79(s, 3H), 3.83(s, 2H ), 3.35 (s, 2H), 2.21 (s, 3H), 1.56-1.54 (m, 2H), 1.40-1.37 (m, 2H), 0.73 ( t, J = 6.90 Hz, 6H). LCMS: (Method A) 550.2 (M ⁺ +H), Rt. 2.87 min, 93.92% (maximum).

Intermediate 65

3,3-Dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (0.5 g, 1 01 mmol) in a mixture of MeOH and water (10 ml, 4:1) was added NH ₄ Cl (0.86 g, 8.08 mmol) and powdered Zn (0.32 g, 5.06 mmol) and the reaction mixture was heated for 3 hours at 65°C. After completion of the reaction (monitored by TLC), the reaction was stopped with water (15 ml) and the aqueous layer was extracted with EtOAc (2×25 ml). The combined organic layer was washed with brine (15 ml), dried over anhydrous Na ₂ SO ₄ 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: 66% (0.28 g, white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.52 (d, J =2.8 Hz, 1H), 7.30-7.24 (m, 2H), 7.05-7, 03 (m, 2H), 6.99-6.93 (m, 3H), 3.84 (s, 3H), 3.72 (s, 2H), 3.20 (s, 2H), 1.53 -1.41 (m, 4H), 1.28-1.12 (m, 8H), 1.00-0.91 (m, 6H). LCMS: (Method A) 416.2 (M+H), Rt. 3.29 min, 88.66% (maximum).

Intermediate 66

3,3-Dibutyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 65; 0.28 g, 0 .67 mmol) in dry DCM (6 ml) at 0°C was added BBr ₃ (1M, 1.4 ml, 1.34 mmol) and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was cooled and the reaction was stopped with a saturated solution of Na ₂ CO ₃ (6 ml). The organic layer was separated, washed with brine (6 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 74% (200 mg, brown gum).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 9.99 (s, 1H), 7.29 (d, J =2.7 Hz, 1H), 7.18 (t, J =7, 6 Hz, 2H), 6.97-6.89 (m, 4H), 6.81 (t, J = 7.2 Hz, 1H), 3.64 (s, 2H), 3.24 (s, 2H), 1.42-1.39(m, 4H), 1.35-1.24(m, 8H), 1.17-1.14(m, 6H). LCMS: (Method A) 402.2 (M+H), Rt. 2.98 min, 83.09% (maximum).

Intermediate 67

Methyl-(Z)-3-((3,3-dibutyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2 -fluoroacrylate

With stirring, to a solution of 3,3-dibutyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 66; 0.2 g, 0 .49 mmol) in DMF (2 ml) at 0°C was added NaH (60% in mineral oil; 0.03 g, 0.75 mmol) and the reaction mixture was stirred for 15 min. Then methyl 3-bromo-2,2-difluoropropanoate (0.15 g, 0.74 mmol) was added dropwise and the mixture was heated for 16 h at 85°C. After completion of the reaction (monitored by TLC), the reaction was stopped with dilute HCl (3 ml, 1.5 N.) and the aqueous layer was extracted with EtOAc (2×15 ml). The combined organics were washed with brine (10 ml), dried over anhydrous Na ₂ SO ₄ 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: 32% (0.08 g, brown solid).

LCMS: (Method E) 504.2 (M+H), Rt. 2.98 min, 24.81% (maximum).

Intermediate 68

5-Chloro-6-methoxybenzo[d]thiazol-2-amine

Ammonium thiocyanate (5.3 g, 69.8 mmol) was added to a solution of 3-chloro-4-methoxyaniline (10 g, 63.4 mmol) in acetic acid (100 ml) at room temperature with stirring, and then the mixture was stirred for 30 min. Bromine (3.2 ml, 63.4 mmol) dissolved in acetic acid (20 ml) was added dropwise to the reaction mixture at 15° C., and the resulting mixture was stirred for 3 hours at room temperature. After completion of the reaction, the resulting solid was filtered off, washed with acetic acid (20 ml) and then dried in vacuo. The solid was then suspended in water (20 ml) and basified with 10% NaOH solution to approximately pH 10. The solid was filtered off, washed with water (3 x 25 ml) and then dried in vacuo to give the title compound. Yield: 80% (11 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.53 (s, 1H), 7.41 (bs, 2H), 7.36 (s, 1H), 3.82 (s, 3H). LCMS: (Method A) 215.0 (M ⁺ +H), Rt. 1.39 min, 97.22% (maximum).

Intermediate 69

2-(((2-Amino-4-chloro-5-methoxyphenyl)thio)methyl)-2-ethylhexanoic acid

KOH (34 g, 0.596 mol) was added to a stirred solution of 5-chloro-6-methoxybenzo[d]thiazol-2-amine (intermediate 68; 8 g, 0.037 mol) in water (120 ml) and the reaction mixture was stirred in for 16 h at 120°C. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to room temperature. 2-(Bromomethyl)-2-ethylhexanoic acid (13.29 g, 0.0558 mol; solution in 40 ml THF) was added dropwise and the mixture was then stirred for 16 hours at room temperature. 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 (2×25 ml). The combined organic layer was then washed with water (30 ml) and brine (30 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude material. The obtained crude material was sent to the next stage without any further purification. Yield: 21 g (crude brown gum).

UPLC: (Method A) 345.8 (M ⁺ +H), Rt. 1.58 min, 90.21% (maximum).

Intermediate 70

3-Butyl-7-chloro-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring, to a solution of 2-(((2-amino-4-chloro-5-methoxyphenyl)thio)methyl)-2-ethylhexanoic acid (Intermediate 69; 21 g, 0.0607 mol) in EtOAc (130 mL) at 0° C., triethylamine (12.26 g, 0.1214 mol) and a solution of 1-propane phosphoric anhydride (50% EtOAc; 23.16 g, 0.073 mol) were added dropwise and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by UPLC), water (25 ml) was added to the reaction mixture and the aqueous layer was extracted with EtOAc (2×25 ml). The combined organic layer was washed with brine (25 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 10-12% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 55% (11 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.61 (s, 1H), 7.16 (d, J =14.8 Hz, 2H), 3.82 (s, 3H), 2, 98 (s, 2H), 1.51-1.53 (m, 4H), 1.32-1.26 (m, 4H), 0.92-0.91 (m, 6H). LCMS: (Method A) 328.1 (M ⁺ +H), Rt. 2.60 min, 95.79% (maximum). HPLC: (method B) Rt. 5.51 min, 97.62% (maximum).

Intermediate 71

3-Butyl-7-chloro-3-ethyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring, to a solution of 3-butyl-7-chloro-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 70; 11 g, 0.034 mol) in iodobenzene (110 ml) copper(I) iodide (0.640 g, 0.0034 mol) and K ₂ CO ₃ (9.25 g, 0.067 mol) were added and the solution was purged with nitrogen for 20 min to degas. Tris[2-(2-methoxyethoxy)ethyl]amine (2.16 g, 0.0067 mol) was then added under nitrogen atmosphere and the resulting reaction mixture was heated for 40 h at 135°C. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through celite and the celite layer was washed with EtOAc (25 ml). The filtrate was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 3-5% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 86% (11.7 g, pale brown solid).

¹ H NMR (300 MHz, DMSO- d ₆ ): δ 7.40-7.39 (m, 3H), 7.29 (d, J =7.2 Hz, 1H), 7.09 (d, J =6.9 Hz, 2H), 6.96 (s, 1H), 3.91 (s, 3H), 3.16 (s, 2H), 1.57-1.55 (m, 4H), 1 .19 (d, J = 6.9 Hz, 5H), 0.79 (t, J = 6.3 Hz, 7H). LCMS: (Method A) 404.1 (M ⁺ +H), Rt. 3.19 min, 98.20% (maximum).

Intermediate 72

3-Butyl-7-chloro-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine

With stirring to a solution of 3-butyl-7-chloro-3-ethyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 71; 11.7 g, 0.029 mol) in THF (110 ml) at 0°C was added dropwise borane dimethyl sulfide (2M in THF; 73 ml, 0.144 mol) and the reaction mixture was refluxed for 40 h at 75°C. After completion of the reaction (monitored by UPLC), the reaction mixture was cooled to 0°C and the reaction was stopped with methanol (50 ml). The resulting solution was heated for 2 h at 65°C, then cooled to RT and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 8-10% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 90% (10.2 g, colorless liquid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.19 (t, J =7.2 Hz, 2H), 7.08 (s, 1H), 6.93 (s, 2H), 6, 79 (d, J =4.0 Hz, 2H), 3.82 (s, 3H), 2.76 (s, 2H), 1.26-1.24 (m, 9H), 0.76-0 .71 (m, 6H). LCMS: (Method A) 390.2 (M ⁺ +H), Rt. 3.01 min, 99.61% (maximum).

Intermediate 73

3-Butyl-7-chloro-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3-butyl-7-chloro-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine (intermediate 72; 10.2 g, 0 .0261 mol) in 1,4-dioxane (100 ml) at room temperature were added water (100 ml) and oxone (81 g, 0.2615 mol) and the reaction mixture was stirred for 24 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a Buchner funnel and the filtrate was extracted with EtOAc (2×25 ml). The combined organic layer was washed with water (25 ml) and brine (25 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 10-12% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 56% (6.2 g, yellowish solid).

¹ H NMR (300 MHz, DMSO- d ₆ ): δ 7.50 (s, 1H), 7.27 (t, J =7.8 Hz, 2H), 7.07-7.04 (m, 4H ), 3.94 (s, 3H), 3.69 (s, 2H), 3.33 (s, 2H), 1.52-1.37 (m, 8H), 0.77-0.74 ( m, 6H). LCMS: (Method A) 422.1 (M ⁺ +H), Rt. 3.18 min, 98.51% (maximum).

Intermediate 74

3-Butyl-7-chloro-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3-butyl-7-chloro-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 73; 1.1 g, 2.60 mmol) in DCM (11 ml) at 0°C was added BBr ₃ (1M solution in DCM, 13.03 ml, 13.03 mmol) and the solution was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), methanol was added dropwise at 0° C. until gas evolution ceased. The reaction mixture was diluted with DCM (20 ml) and washed with water (2×20 ml) and brine (20 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 30-32% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 94% (1.0 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.83 (s, 1H), 7.50 (s, 1H), 7.23 (t, J =8.0 Hz, 2H), 6, 99 (d, J =7.6 Hz, 2H), 6.97 (s, 1H), 6.88 (t, J =7.2 Hz, 1H), 3.64 (s, 2H), 3, 28 (s, 2H), 1.51-1.34 (m, 4H), 1.12-1.02 (m, 4H), 0.77-0.72 (m, 6H). LCMS: (Method A) 408.2 (M ⁺ +H), Rt. 2.87 min, 93.25% (maximum).

Intermediate 75

Methyl-(Z)-3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)-2-fluoroacrylate

With stirring, to a solution of 3-butyl-7-chloro-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 74; 1.0 g, 2.45 mmol) in DMF (10 mL) at 0°C was added 60% NaH (0.18 g, 7.35 mmol) in portions and the reaction mixture was stirred for 15 min. Then methyl 3-bromo-2,2-difluoropropanoate (0.99 g, 4.90 mmol) was added and the reaction mixture was heated at 70° C. overnight. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C and the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (10 ml). The aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude title compound, which was further triturated with diethyl ether. The resulting compound was dried in vacuo and passed on to the next step without any further purification. Yield: 59% (0.74 g, pale yellow gum).

LCMS: (Method A) 510.1 (M+H), Rt. 3.13 min, 94.43% (maximum).

Intermediate 76

Ethyl-(E)-3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylate

With stirring, to a solution of 3-butyl-7-chloro-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 74; 0.95 g, 2.33 mmol) in THF (10 ml) was added DABCO (0.026 g, 0.23 mmol) and ethyl propiolate (0.28 ml, 2.8 mmol) at 0°C. The reaction mixture was then stirred for 1 hour at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (15 ml) and the aqueous layer was extracted with EtOAc (3×15 ml). The combined organic layer was washed with brine (20 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 8-9% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 84.7% (1.0 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.81 (d, J =16.4 Hz, 1H), 7.70 (s, 1H), 7.39-7.34 (m, 2H ), 7.28-7.26 (m, 2H), 7.09 (t, J =9.6 Hz, 1H), 6.90 (s, 1H), 5.48 (d, J =16, 4 Hz, 1H), 4.13 (q, J = 9.2 Hz, 2H), 3.81 (bs, 2H), 3.46 (s, 2H), 1.55-1.50 (m, 4H), 1.34 (t, J =10.00 Hz, 3H), 1.30-1.15 (m, 4H), 0.92 (t, J =4.40 Hz, 6H). LCMS: (Method A) 506.1(M ⁺ ), Rt. 3.28 min, 97.07% (maximum).

Intermediate 77

Methyl-(Z)-3-((3,3-diethyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate

With stirring, to a solution of methyl-(Z)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 64; 1.0 g, 1.97 mmol) in a 1:1 mixture of DCM and AcOH (15 mL) HNO ₃ (65%; 0.186 g, 2.95 mmol) in a mixture of DCM and AcOH (5 ml). After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml). The organic layer was washed with water (15 ml) and saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title product, which was passed on to the next step without any further purification. Yield: 900 mg (crude brown solid).

LCMS: (Method A), Rt, 2.62 min, 88% (maximum).

Intermediate 78

Methyl-(Z)-3-((5-(4-aminophenyl)-3,3-diethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate

With stirring, to a solution of methyl-(Z)-3-((3,3-diethyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 77; 0.9 g, 1.67 mmol) in THF (15 mL) was added at room temperature with concentrated HCl (1 mL) and SnCl ₂ (1.26 g, 6.69 mmol) and the reaction mixture was stirred for 12 h at 70°C. After completion of the reaction (monitored by TLC), the reaction was stopped with a saturated solution of NaHCO ₃ (15 ml) and filtered through celite. The Celite layer was washed with EtOAc (2×15 ml) and the resulting filtrate was washed with water (2×25 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 650 mg (crude yellow solid).

Intermediate 79

Methyl-(Z)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate

With stirring, to a solution of methyl-(Z)-3-((5-(4-aminophenyl)-3,3-diethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 78; 0.15 g, 0.29 mmol) in DCM (5 ml) was added TEA (0.082 ml, 0.59 mmol) and pivaloyl chloride (0.038 ml, 0.44 mmol) at 0°C and the reaction mixture was stirred for 2 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml) and washed with saturated NaHCO ₃ solution (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 180 mg (crude white gum).

Intermediate 80

Methyl-(Z)-3-((3,3-dibutyl-7-chloro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylate

With stirring to a solution of 3,3-dibutyl-7-chloro-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (300 mg, 0.688 mmol) NaH (60%; 138 mg, 3.44 mmol) was added in DMF (3 ml) at 0° C. in portions and the mixture was stirred for 15 minutes. Then methyl 3-bromo-2,2-difluoropropanoate (419 mg, 2.06 mmol) was added and the reaction mixture was heated for 16 h at 85°C. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C, the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude title compound, which was passed on to the next step without any further purification. Yield: 617 mg (crude pink liquid).

Intermediate 81

3,3-Diethyl-8-hydroxy-7-iodo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3,3-diethyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (obtained by purification using preparative HPLC (method B) of a mixture of 7-brominated and 7-iodinated intermediate 59; 250 mg, 0.52 mmol) in DCM (5 ml) was added dropwise BBr ₃ (1M solution in DCM, 0.80 ml, 0.77 mmol) at -40°C and the resulting reaction mixture was stirred for 4 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C and MeOH was added dropwise until gas evolution ceased. The reaction mixture was diluted with DCM (20 ml) and the organic layer was washed with water (2×15 ml), brine (20 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 45-50% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 82% (200 mg, pale yellow solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.92 (s, 1H), 7.38 (s, 1H), 7.33 (s, 1H), 7.23-7.19 (m , 2H), 6.94-6.81 (m, 3H), 3.63 (m, 1H), 3.25 (s, 2H), 1.55-1.48 (m, 2H), 1, 36-1.23 (m, 2H), 0.73 (t, J =9.6 Hz, 6H). LCMS: (Method A) 472.1 (M ⁺ +H), Rt. 2.49 min, 93.10%.

Intermediate 82

tert-Butyl-(E)-3-((3,3-diethyl-7-iodine-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylate

With stirring to a solution of 3,3-diethyl-8-hydroxy-7-iodo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 81; 100 mg , 0.212 mmol) in THF (3 ml) at 0°C was added DABCO (3 mg, 0.021 mmol) and tert-butyl propiolate (32 mg, 0.2545 mmol) and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (15 ml) and the aqueous layer was extracted with EtOAc (3×15 ml). The combined organic layer was washed with brine (20 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound, which was passed to the next step without any further purification. Yield: 130 mg (crude off-white solid).

LCMS: (Method A) 542.1 (M ⁺ - ^t Bu+H), Rt. 3.03 min, 76.11% (maximum).

Intermediate 83

7-Bromo-3,3-diethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-diethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (obtained by purification using preparative HPLC (method B) of a mixture of 7-brominated and 7-iodinated intermediate 59; 750 mg, 1.71 mmol) in DCM (10 ml) was added dropwise BBr ₃ (1M solution in DCM, 2.60 ml, 2.57 mmol) at -40°C and the resulting reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C and MeOH was added dropwise until gas evolution ceased. The reaction mixture was diluted with DCM (25 ml) and the organic layer was washed with water (2×15 ml), brine (20 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 45-50% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 80% (580 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.89 (s, 1H), 7.48 (s, 1H), 7.24-7.13 (m, 3H), 6.96-6 .83 (m, 3H), 3.64 (m, 1H), 3.32 (s, 2H), 1.53-1.48 (m, 2H), 1.37-1.30 (m, 2H ), 0.73 (t, J = 9.6 Hz, 6H). LCMS: (Method A) 424.0 (M ⁺ ), Rt. 2.44 min, 98.18%.

Intermediate 84

Ethyl-(E)-3-((7-bromo-3,3-diethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylate

With stirring to a solution of 7-bromo-3,3-diethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 83; 200 mg , 0.471 mmol) in THF (6 ml) was added DABCO (5.3 mg, 0.047 mmol) and ethyl propiolate (56 mg, 0.566 mmol) at 0°C, then the reaction mixture was stirred for 1 h at RT. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (15 ml) and the aqueous layer was extracted with EtOAc (3×15 ml). The combined organic layer was washed with brine (20 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude material. The obtained crude material was sent to the next stage without any further purification. Yield: 250 mg (crude brown solid).

LCMS: (Method A) 522.2 (M ⁺ +H), Rt. 2.40 min, 86.64% (maximum).

Intermediate 85

Methyl-(Z)-3-((7-bromo-3,3-diethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylate

With stirring to a solution of 7-bromo-3,3-diethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 83; 200 mg , 0.4713 mmol) in DMF (5 ml) at 0°C was added NaH (60%; 95 mg, 2.356 mmol) in portions and the mixture was stirred for 15 min. Then methyl 3-bromo-2,2-difluoropropanoate (290 mg, 1.414 mmol) was added and the reaction mixture was heated at 80° C. overnight. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (10 ml). The aqueous layer was extracted with EtOAc (2×15 ml) and the combined organic layer was washed with brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude title compound, which was further triturated with diethyl ether. The resulting compound was dried in vacuo and passed on to the next step without any further purification. Yield: 250 mg (crude brown gum). UPLC: (Method A) 528.5 (M ⁺ +2), Rt. 1.88 min, 39.34% (maximum).

Intermediate 86

2-(((2-Amino-5-methoxyphenyl)thio)methyl)-2-ethylhexanoic acid

KOH (1345 g, 23.96 mol) was added to a stirred solution of 6-methoxybenzo[d]thiazol-2-amine (270 g, 1.498 mol) in water (2700 ml) and the reaction mixture was stirred for 16 h at 120° C. 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 for 16 hours at room temperature. 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 layer was washed with water (1000 ml) and brine (1000 ml). The organic portion was then dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give a crude material which was passed on to the next step without any further purification. Yield: 590 g (crude brown gum).

LCMS: (Method A) 312.1 (M ⁺ +H), Rt. 2.24 min, 97.34% (maximum).

Intermediate 87

3-Butyl-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring, to a solution of 2-(((2-amino-5-methoxyphenyl)thio)methyl)-2-ethylhexanoic acid (intermediate 86; 590 g, 1.89 mol) in EtOAc (2500 ml) at 0°C by triethylamine (530 ml, 3.78 mol) and a solution of 1-propane phosphoric anhydride (50% in EtOAc; 785 g, 2.46 mol) were added dropwise and the reaction mixture was stirred for 16 h at room temperature. 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 (2×2000 ml). The combined organic layer was washed with brine (800 ml), dried over anhydrous Na ₂ SO ₄ 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- d ₆ ): 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 min, 99.47% (maximum).

Intermediate 88

7-Bromo-3-butyl-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

To a 1:1 solution of 3-butyl-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 87; 265 g, 0.903 mol) with stirring DCM and acetonitrile (2650 ml) were added N-bromosuccinimide (209 g, 1.17 mol) in portions and the reaction mixture was stirred for 16 hours at room temperature. 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 and washed with cold acetonitrile (2×100 ml) and dried in vacuo to give the title compound. Yield: 179 g (79%, crude brown solid).

¹ H NMR (300 MHz, DMSO- d ₆ ): 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 min, 99.20% (maximum).

Intermediate 89

7-Bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3-butyl-3-ethyl-7-iodine -8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 88; 179 g, 0.483 mol) in iodobenzene (1800 ml) copper(I) iodide (18.5 g, 0.096 mol) and K ₂ CO ₃ (134 g, 0.967 mol) were added and the solution was purged with nitrogen for 20 min to degas. Tris[2-(2-methoxyethoxy)ethyl]amine (15.6 g, 0.04834 mol) was then added under nitrogen atmosphere and the resulting reaction mixture was heated for 40 h at 135°C. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through celite and the celite layer was washed with EtOAc (2000 ml). The filtrate was concentrated in vacuo to give a crude product which was crystallized from cold petroleum ether. The resulting precipitate was filtered off and washed with cold petroleum ether to give a mixture of the desired compounds. Yield: 180 g (83%, crude light yellow solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.42-7.40 (m, 2H), 7.30-7.28 (m, 2H), 7.10-7.09 (m, 2H), 3.90 (s, 3H), 3.15 (s, 2H), 1.58-1.52 (m, 8H), 0.83-0.81 (m, 6H). LCMS: (Method A) 448.0 (M ⁺ +H) 40.8%; 496.0 (M ⁺ +H) 53.98%; Rt. 3.27 and 3.28 min, 94.7% (maximum).

Intermediate 90

7-Bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3-butyl-3-ethyl-7-iodine-8- methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine

With stirring, to a solution of a mixture of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3-butyl-3- ethyl 7-iodo-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 89; 180 g, 0.40178 mol) in THF (1800 ml ) at 0°C was added dropwise borane dimethylsulfide (2M in THF; 602 ml, 1.2053 mol) and the reaction mixture was refluxed for 40 h at 75°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C and the reaction was stopped with methanol (400 ml). The resulting solution was heated at 65°C for 2 h, then cooled to room temperature and concentrated in vacuo to give a mixture of the desired compounds. The resulting crude material was passed on to the next step without any further purification. Yield: 195 g (crude dark brown liquid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.22-7.20 (m, 2H), 7.04-6.93 (m, 1H), 6.88-6.83 (m, 3H), 6.81-6.79(m, 1H), 3.82(s, 3H), 3.63(s, 2H), 2.76(s, 2H), 1.41-1.40 (m, 8H), 0.80-0.78 (m, 6H). LCMS: (Method A) 448.1 (M ⁺ +H), 89.8% and 482.2 (M ⁺ +H) 8.18%; Rt. 3.02 and 3.19 min, 97.98% (maximum).

Intermediate 91

7-Bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3-butyl-3-ethyl- 7-iodine-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of a mixture of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3-butyl-3-ethyl-7 -iodine-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine (intermediate 90; 195 g, 0.4488 mol) in THF (1950 ml) water (1950 ml) was added ) and oxone (1380 g, 4.488 mol) and the reaction mixture was stirred for 24 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a Buchner funnel and the filtrate was extracted with EtOAc (2×2000 ml). The combined organic layer was washed with water (1000 ml) and brine (1000 ml), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The crude material was purified by column chromatography (eluent: 10-12% EtOAc/PE; silica gel: 230-400 mesh) to give a mixture of the title compounds. Yield: 60% (126 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.45 (s, 1H), 7.36-7.32 (m, 2H), 7.25 (s, 1H), 7.16-7 .06 (m, 2H), 6.93 (t, J = 7.20 Hz, 1H), 3.90 (s, 3H), 3.78 (s, 2H), 1.52-1.41 ( m, 2H), 1.36-1.34 (m, 4H), 1.27-1.24 (m, 4H), 1.00-0.80 (m, 6H). LCMS: (Method B) 468.1 (M ⁺ +H) 91.72%, 514.2 (M ⁺ +H) 5.70%, Rt. 1.76 min and 1.88 min, 97.42% (maximum).

Intermediate 92

7-Bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3-butyl-3-ethyl- 8-hydroxy-7-iodine-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of a mixture of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3-butyl -3-ethyl-7-iodine-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 91; 6.5 g, 13, 93 mmol) in DCM (65 ml) at 0°C was added dropwise BBr ₃ (1M solution in DCM; 69.67 ml, 69.67 mmol) and the solution was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C and MeOH was added dropwise until gas evolution ceased. The reaction mixture was diluted with DCM (100 ml) and the organic layer was washed with water (2×50 ml) and brine (100 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was then concentrated in vacuo to give a crude material which was purified by Isolera column chromatography (eluent: 30-32% EtOAc/PE; silica gel: 230-400 mesh) to give a mixture of the title compounds. Yield: 95% (6.0 g, off-white solid).

LCMS: (Method A) 452.1 (M ⁺ ) and 498.0 (M ⁺ -H), Rt. 3.00 and 3.02 min, 92.15% (total maximum).

Intermediate 93

Ethyl-(E)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)acrylate and ethyl-(E)-3-((3-butyl-3-ethyl-7-iodine-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of a mixture of 7-bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3-butyl -3-ethyl-8-hydroxy-7-iodo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 92; 6 g, 13.26 mmol ) in THF (60 ml) at 0°C was added DABCO (0.15 g, 1.32 mmol) and then ethyl propiolate (1.61 ml, 15.91 mmol) and the mixture was stirred at room temperature for 1 h. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water (100 ml) and was extracted with EtOAc (2×50 ml). The combined organic layer was washed with brine (50 ml), dried over anhydrous Na₂SO₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 17-18% EtOAc/PE; silica gel: 230-400 mesh) to give a mixture of the title compounds. Yield: 90% (6.6 g, colorless gum).

LCMS: (method C) 598.1 (M ⁺ + H) and 550.1 (M ⁺ ), Rt. 3.03 min, 93.95% (maximum).

Intermediate 94

(E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid and (E) -3-((3-butyl-3-ethyl-7-iodo-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine -8-yl)hydroxy)acrylic acid

With stirring, to a solution of a mixture of ethyl-(E)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylate and ethyl-(E)-3-((3-butyl-3-ethyl-7-iodo-1,1-dioxydo-5-phenyl-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 93; 6.6 g, 11.98 mmol) in a mixture of 1,4-dioxane and water (60 ml, 4:1) hydroxide was added lithium (1.01 g, 23.97 mmol) and the reaction mixture was stirred at room temperature overnight. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 15 ml) and the aqueous layer was extracted with EtOAc (2×100 ml). The combined organics were washed with water (50 ml) and brine (50 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 3% MeOH/DCM, silica gel: 230-400 mesh) to give a mixture of the title compounds. Yield: 54% (3.38 g, white solid).

LCMS: (method A) 520.9 (bromine, iodine M-H), Rt. 2.90 min, 97.86% (maximum).

Intermediate 95

5-(4-bromophenyl)-3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 23; 2 g, 4.76 mmol) in DMF (15 mL) N-bromosuccinimide (0.93 g, 5.24 mmol) in DMF (5 mL) was added dropwise at -10°C and the reaction mixture was stirred for 1 hour at temperatures below 0°C. After completion of the reaction (monitored by TLC), the reaction mixture was poured onto crushed ice and stirred vigorously for 5 min. The solid that precipitated was filtered off, washed with ice-cold water and dried in vacuo to give the title compound. Yield: 98% (2.37 g, off-white solid).

LCMS: (Method A) 496.1 (M ⁺ -2H), Rt. 2.97 min, 90.53% (maximum).

Intermediate 96

4-(3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepin-5(2H)-yl) benzonitrile

With stirring to a solution of 5-(4-bromophenyl)-3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1- dioxide (Intermediate 95; 2.37 g, 4.75 mmol) in DMF (24 ml) was added zinc cyanide (2.23 g, 19 mmol) and the solution was degassed with N ₂ for 30 min. Then tetrakis(triphenylphosphine)palladium(0) (0.54 g, 0.47 mmol) was added and the solution was heated for 24 h at 100°C. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water and the aqueous layer was extracted with EtOAc (2×50 ml). The combined organic layer was washed with ice-cold water (50 ml), brine (50 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and 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: 52% (1.1 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.84 (s, 1H), 7.54 (d, J =11.6 Hz, 2H), 7.34 (s, 1H), 6, 87-6.85 (m, 2H), 3.51 (bs, 2H), 3.65 (s, 3H), 2.28 (s, 3H), 1.65-1.41 (m, 2H) , 1.42-1.22 (m, 6H), 0.84-0.65 (m, 6H). LCMS: (Method A) 445.1 (M ⁺ +H), Rt. 2.53 min, 96.66% (maximum).

Intermediate 97

4-(3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepin-5(2H)-yl) benzoic acid

With stirring, to a solution of 4-(3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine-5(2H )-yl)benzonitrile (intermediate 96; 0.5 g, 1.12 mmol) in a mixture of ethanol and water (4:1, 20 ml) was added NaOH (0.45 g, 11.2 mmol) and the solution was heated in for 4 days at 100°C. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and acidified with concentrated HCl (pH~2). The aqueous layer was extracted with EtOAc (2×50 ml) and the combined organic layer was washed with ice-cold water (50 ml), brine (50 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 9% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 50% (0.16 g, off-white solid).

LCMS: (Method A) 464.1 (M ⁺ +H), Rt. 2.19 min, 95.59% (maximum).

Intermediate 98

N-(tert-butyl)-4-(3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine -5(2H)-yl)benzamide

With stirring, to a solution of 4-(3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine-5(2H )-yl)-benzoic acid (intermediate 97; 160 mg, 0.34 mmol) triethylamine (0.1 ml, 0.41 mmol), tert-butylamine (104 mg, 1.03 mmol ) and HATU (230 mg, 0.64 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was dissolved in ice-cold water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with ice-cold water (10 ml), brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 35-40 EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 28% (50 mg, almost white solid).

LCMS: (Method A) 517.3 (M ⁺ -H), Rt. 2.56 min, 95.71% (maximum).

Intermediate 99

Ethyl-(E)-3-((3-butyl-5-(4-(tert-butylcarbamoyl)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring to a solution of N-(tert-butyl)-4-(3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-5(2H)-yl)benzamide (Intermediate 98; 50 mg, 0.096 mmol) in THF (2 ml) was added DABCO (1 mg, 0.0096 mmol) and ethyl propiolate (14.18 mg, 0 .14 mmol). The reaction mixture was then stirred for 30 minutes at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was dissolved in ice-cold water (5 ml). The aqueous layer was extracted with EtOAc (2×5 ml), the combined organic layer was washed with ice-cold water (5 ml) and brine (5 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give a crude material which was passed on to the next step without any further purification. Yield: 50 mg (crude brown gum).

LCMS: (Method A) 617.3 (M ⁺ +H), Rt. 2.74 min, 12.07% (maximum).

Intermediate 100

Methyl-(Z)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)-2-fluoroacrylate

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (0.5 g , 1.11 mmol) in DMF (5 ml) at 0°C NaH (60%, 0.08 g, 3.31 mmol) was added in portions and the mixture was stirred for 15 min at 0°C. Methyl 3-bromo-2,2-difluoropropanoate (0.45 g, 2.2 mmol) was added and the reaction mixture was heated for 16 h at 85°C. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C, the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude title compound, which was further triturated with Et ₂ O. The resulting compound was dried in vacuo and passed on to the next step without any further purification. Yield: 90% (0.55 g, colorless gum).

LCMS: (method A) 556.1 (M ⁺ +2), Rt. 3.09 min, 85.5% (maximum).

Intermediate 101

3,3-dibutyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (2.0 g, 4 04 mmol) in dry DMF (20 ml) was added sodium thioethoxide (1.7 g, 20.24 mmol) at room temperature and the reaction mixture was stirred for 16 h at 60°C. After completion of the reaction (monitored by LCMS), the reaction was stopped with ice-cold water (10 ml) and the aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with water (15 ml) and brine (15 ml) and then dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 10-15% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 65% (1.2 g, off-white solid).

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.44 (s, 1H), 7.30 (s, 1H), 7.20 (t, J=10.8 Hz, 2H), 6, 94 (d, J=10.8 Hz, 2H), 6.82 (t, J=9.6 Hz, 1H), 6.69 (s, 1H), 3.65 (bs, 2H), 3, 22 (s, 2H), 2.69 (q, J=9.6 Hz, 2H), 1.40-1.35 (m, 4H), 1.14-1.05 (m, 11H), 0 .59-0.81 (m, 6H). LCMS: (Method A) 462.1 (M ⁺ +H), Rt. 3.18 min, 94.69% (maximum).

Intermediate 102

Ethyl-(E)-3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylate

With stirring, to a solution of 3,3-dibutyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 101; 0.2 g, 0.43 mmol) in dry THF (3 ml) was added ethyl propiolate (0.043 g, 0.52 mmol) and DABCO (5 mg, 0.04 mmol) and then the solution was stirred for 30 min at room temperature . After completion of the reaction (monitored by TLC), the reaction mixture was concentrated and the resulting residue was subjected to distribution between water (10 ml) and EtOAc (15 ml). The aqueous layer was extracted with EtOAc (2×15 ml) and the combined organics were washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 93% (0.23 g, white solid).

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 7.86 (d, J=10.8 Hz, 1H), 7.49 (s, 1H), 7.35-7.32 (m, 2H), 7.20-7.14 (m, 2H), 7.02 (t, J=9.6 Hz, 1H), 6.65 (s, 1H), 5.47 (d, J=16 .0 Hz, 1H), 4.12 (q, J=9.2 Hz, 2H), 3.78 (bs, 2H), 3.47 (s, 2H), 2.76 (q, J=10 .8 Hz, 2H), 1.40-1.33 (m, 3H), 1.28 (t, J=7.60 Hz, 4H), 1.23-1.18 (m, 11H), 0 .87-0.62 (m, 6H).

Intermediate 103

4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepin-5(2H)-yl)benzonitrile

With stirring, to a solution of 5-(4-bromophenyl)-3,3-dibutyl-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide ( intermediate 20; 4.4 g, 8.36 mmol) in DMF (40 ml) zinc cyanide (4.9 g, 41.01 mmol) was added at room temperature and then the mixture was degassed with N ₂ for 30 min. Then tetrakis(triphenylphosphine)palladium(0) (0.96 g, 0.83 mmol) was added and the mixture was heated for 24 h at 100°C. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (25 ml) and the aqueous layer was extracted with EtOAc (2×50 ml). The combined organic layer was washed with ice-cold water (30 ml), brine (30 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 90-100% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 48% (1.9 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.83 (s, 1H), 7.54 (d, J =11.6 Hz, 2H), 7.34 (s, 1H), 6, 92-6.71(m, 3H), 3.62(bs, 2H), 3.21(s, 2H), 2.27(s, 3H), 1.65-1.35(m, 3H) , 1.29-1.14 (m, 9H), 0.82 (t, J = 8.8 Hz, 6H). LCMS: (Method A) 473.2 (M ⁺ +H), Rt, 2.67 min, 87.18% (maximum).

Intermediate 104

4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepin-5(2H)-yl)benzoic acid

With stirring, to a solution of 4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine-5(2H)- yl)benzonitrile (intermediate 103; 1.5 g, 3.17 mmol) in a mixture of ethanol and water (4:1, 20 ml) was added NaOH (1.9 g, 47.67 mmol) and the reaction mixture was heated for 72 h at 100°C. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and acidified with concentrated HCl at 0°C to pH ~2. The aqueous layer was extracted with EtOAc (2×50 ml) and the combined organic layer was washed with ice-cold water (50 ml) and brine (50 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 9% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 58% (0.76 g, off-white solid).

LCMS: (Method A) 492.1 (M ⁺ +H), Rt. 2.37 min, 38.32% (maximum).

Intermediate 105

N-(tert-butyl)-4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine-5 (2H)-yl)benzamide

With stirring, to a solution of 4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine-5(2H)- yl)benzoic acid (intermediate 104, 150 mg, 0.3 mmol) in DMF (3 ml) at room temperature was added triethylamine (92.4 mg, 0.91 mmol), tert-butylamine (44.6 mg, 0 .61 mmol) and HATU (232 mg, 0.61 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between cold water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with ice-cold water (10 ml) and brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 35-40% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 42% (70 mg, pale yellow solid).

LCMS: (Method A) 547.2 (M ⁺ +H), Rt. 2.78 min, 87.33% (maximum).

Intermediate 106

Ethyl-(E)-3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro -1,5-benzothiazepin-8-yl)oxy)acrylate

With stirring to a solution of N-(tert-butyl)-4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-5(2H)-yl)benzamide (intermediate 105; 70 mg, 0.12 mmol) in THF (2 mL) DABCO (1.4 mg, 0.012 mmol) and ethyl propiolate (18.9 mg, 0.19 mmol). The reaction mixture was then stirred for 30 minutes at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (2 ml) and EtOAc (2 ml). The aqueous layer was extracted with EtOAc (2×3 ml). The combined organic layer was washed with ice-cold water (3 ml) and brine (3 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo 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: 66% (55 mg, pale yellow gum).

LCMS: (Method A) 645.3 (M ⁺ +H), Rt. 3.14 min, 94.44% (maximum).

Intermediate 107

4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepin-5(2H)-yl)-N -isopropylbenzamide

With stirring, to a solution of 4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine-5(2H)- yl)benzoic acid (intermediate 104; 0.23 g, 0.46 mmol) in DMF (3 ml) was added triethylamine (0.19 ml, 1.4 mmol), isopropylamine (0.55 g, 0.93 mmol ) and HATU (0.36 g, 0.93 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with ice-cold water (10 ml) and brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 50% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 28% (70 mg, pale brown solid).

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.65-7.63 (m, 2H), 7.15 (s, 1H), 6.92 (d, J =8.4 Hz, 2H), 6.47 (s, 1H), 5.81 (d, J =10.4 Hz, 1H), 4.32-4.28 (m, 1H), 3.65 (bs, 2H), 3.15 (s, 2H), 2.33 (s, 3H), 1.41-1.21 (m, 18H), 0.87-0.74 (m, 6H). LCMS: (Method A) 533.3 (M ⁺ +H), Rt. 2.58 min, 84.35% (maximum).

Intermediate 108

Ethyl-(E)-3-((3,3-dibutyl-5-(4-(isopropylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of 4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine-5(2H)- yl)-N-isopropylbenzamide (intermediate 107; 70 mg, 0.12 mmol) in THF (5 mL) was added DABCO (1.5 mg, 0.013 mmol) and ethyl propiolate (19.3 mg, 0.19 mmol) and the reaction mixture was stirred for 30 min at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (2 ml) and EtOAc (2 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organic layer was washed with ice-cold water (5 ml) and brine (5 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified using Isolera column chromatography (eluent: 70% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound Yield: 50% (50 mg, off-white solid).

LCMS: (Method A) 631.3 (M ⁺ +H), Rt. 2.96 min, 70.99% (maximum).

Intermediate 109

Methyl-(Z)-3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)-2-fluoroacrylate

With stirring, to a solution of 3,3-dibutyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 101; 0.4 g, 0.86 mmol) in DMF (5 mL) at 0°C NaH (60%, 0.06 g, 2.6 mmol) was added in portions and the mixture was stirred for 15 min. Methyl 3-bromo-2,2-difluoropropanoate (0.35 g, 1.7 mmol) was then added and the reaction mixture was heated for 16 h at 85°C. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C, the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was triturated with Et ₂ O. The resulting compound was dried in vacuo and passed on to the next step without any further purification. Yield: 73% (0.36 g, off-white solid).

LCMS: (Method A) 564.1 (M ⁺ +H), Rt. 3.47 min, 54.37% (maximum).

Intermediate 110

3-butyl-3-ethyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of a mixture of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3-butyl -3-ethyl-7-iodine-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 91; 1.2 g, 2, 3 mmol) in dry DMF (12 ml) was added sodium thioethoxide (0.98 g, 11.6 mmol) at room temperature and the reaction mixture was stirred for 16 h at 100°C. After completion of the reaction (monitored by LCMS), the reaction was stopped with ice-cold water (10 ml) and the aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with water (15 ml) and brine (15 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 10-15% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 72% (0.72 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.48 (s, 1H), 7.38 (s, 1H), 7.31 (d, J =3.2 Hz, 2H), 7, 22-7.17(m, 2H), 6.96-6.91(m, 1H), 6.79-6.72(m, 1H), 3.65(bs, 2H), 3.26( s, 2H), 2.70 (q, J =7.2 Hz, 2H), 1.34-1.30 (m, 4H), 1.15-1.07 (m, 7H), 0.79 (t, J = 7.20 Hz, 6H). LCMS: (Method A) 434.2 (M ⁺ +H), Rt. 2.92 min, 98.63% (maximum).

Intermediate 111

Methyl-(Z)-3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylate

With stirring, to a solution of 3-butyl-3-ethyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 110; 0.4 g, 0.92 mmol) in DMF (4 mL) NaH (60%, 0.07 g, 2.7 mmol) was added portionwise at 0°C and the mixture was stirred at this temperature for 15 min. Then methyl 3-bromo-2,2-difluoropropanoate (0.37 g, 1.8 mmol) was added and the reaction mixture was heated for 16 h at 60°C. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C, the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 15-20% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 47% (0.23 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.68 (s, 1H), 7.63-7.62 (m, 1H), 7.30 (t, J =8.0 Hz, 2H ), 7.18-7.15 (m, 2H), 6.99 (t, J =7.6 Hz, 1H), 6.69 (s, 1H), 3.88 (s, 3H), 3 .78 (s, 2H), 3.38 (s, 2H), 2.71 (q, J = 7.2 Hz, 2H), 1.43-1.31 (m, 4H), 1.17- 1.03 (m, 7H), 0.72 (t, J =4.40 Hz, 6H). LCMS: (Method A) 536.2 (M ⁺ +H), Rt. 3.17 min, 89.63% (maximum).

Intermediate 112

Ethyl-(E)-3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylate

With stirring, to a solution of 3-butyl-3-ethyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 110; 0.3 g, 0.69 mmol) in dry THF (3 ml) was added ethyl propiolate (0.081 g, 0.83 mmol) and DABCO (8 mg, 0.06 mmol) and the reaction mixture was stirred for 30 min at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between water (10 ml) and EtOAc (10 ml). The aqueous layer was extracted with EtOAc (2×15 ml) and the combined organics were then washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 15-20% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 95% (0.36 g, colorless gum).

LCMS: (Method A) 532.3 (M ⁺ +H), Rt. 3.34 min, 95.17% (maximum).

Intermediate 113

Methyl-(E)-3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)-2-methylacrylate

With stirring to a solution of 3,3-dibutyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 1,1-dioxide (0.5 g, 0.88 mmol) in dry DMF (4 ml) was added methyl-(E)-3-bromo-2-methylacrylate (0.32 g, 1.76 mmol) and carbonate potassium (0.37 g, 2.64 mmol) and the reaction mixture was heated for 16 h at 50°C. After completion of the reaction (monitored by TLC), the reaction mixture 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: 68% (0.4 g, brown solid).

Intermediate 114

(E) -3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1, 2,5-Benzothiadiazepin-8-yl)oxy)-2-methacrylic acid

With stirring, to a solution of methyl-(E)-3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4, 5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methylacrylate (intermediate 113; 0.2 g, 0.29 mol) in a mixture of 1,4-dioxane and water (2: 1, 6 ml) lithium hydroxide (0.03 g, 0.59 mmol) was added at 0° C. and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1 ml, 1.5 N.) and the aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with water (8 ml) and brine (8 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was passed on to the next step without any further purification. Yield: 0.2 g (crude, off-white gum).

LCMS: (Method E) 653.2 (M ⁺ +H), Rt. 3.02 min, 89.63% (maximum)

Intermediate 115

7-Bromo-3,3-dibutyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide

With stirring, to a solution of a mixture of 7-bromo-3,3-dibutyl-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1 ,1-dioxide (1.0 g, 1.62 mmol) with DCM (20 ml) BBr ₃ (1M solution in DCM; 1.94 ml, 1.94 mmol) was added dropwise at 0°C and the reaction mixture was stirred within 15 min. The progress of the reaction was followed by UPLC, which indicated the formation of a PMB-deprotected product. BBr ₃ (1M solution in DCM; 3.89 ml, 3.89 mmol) was added again at 0° C. and the reaction mixture was stirred for 48 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C and the reaction was stopped with MeOH (10 ml). The reaction mixture was then concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 16% (130 mg, brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.54 (s, 1H), 7.38-7.26 (m, 4H), 7.09 (d, J =4.0 Hz, 2H ), 6.95 (t, J = 8.0 Hz, 2H), 4.00 (s, 2H), 1.49-1.45 (m, 2H), 1.39-1.31 (m, 2H), 1.23-1.14 (m, 3H), 1.09-0.94 (m, 5H), 0.73 (t, J =8.0 Hz, 6H).

Intermediate 116

tert-Butyl-(E)-3-((7-bromo-3,3-dibutyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 8-yl)hydroxy)acrylate

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (intermediate 115; 0.1 g, 0.20 mmol) in THF (5 ml) was added DABCO (0.002 g, 0.02 mmol) and tert-butyl propiolate (0.04 g, 0.31 mmol) and the reaction mixture was stirred for 15 min at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (20 ml). The organic layer was washed with water (2×10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 15% EtOAc in hexanes; silica gel: 230-400 mesh) to give the title compound. Yield: 87% (0.11 g, brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.78 (bs, 1H), 7.65 (d, J =12.0 Hz, 1H), 7.51 (s, 1H), 7, 43 (t, J =8.0 Hz, 2H), 7.33 (d, J =6.8 Hz, 2H), 7.18 (t, J= 8.0 Hz, 1H), 6.84 ( s, 1H), 5.18 (d, J= 12.0 Hz, 1H), 4.02 (bs, 2H), 1.51-1.35 (m, 13H), 1.26-1.01 (m, 4H), 0.90-0.80 (m, 4H), 0.80-0.60 (m, 6H). LCMS: (Method A) 605.2 (M ⁺ -2H), Rt. 3.78 min, 90.48% (maximum).

Intermediate 117

Ethyl 2-aminobutanoate hydrochloride

With stirring, to a solution of 2-aminobutanoic acid (100 g, 0.97 mol) in ethanol (750 ml) was added thionyl chloride (78 ml, 1.07 mol) at 0°C. Then the reaction mixture was heated for 16 h at 80°C. After completion of the reaction, the reaction mixture was concentrated in vacuo to give the crude title compound, which was used in the next step without any further purification. Yield: 93% (152 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 8.66 (bs, 3H), 4.25-4.16 (m, 2H), 3.98-3.85 (m, 1H), 1 .84 (t, J =7.2 Hz, 2H), 1.23 (t, J =6.8 Hz, 3H), 0.92 (t, J =7.6 Hz, 3H).

Intermediate 118

Ethyl-(E)-2-(benzylideneamino)butanoate

With stirring, to a solution of ethyl 2-aminobutanoate hydrochloride (intermediate 117; 152 g, 0.91 mol) in DCM (900 ml) was added triethylamine (152 ml, 1.09 mol) at 0°C over 30 min. Magnesium sulfate (98 g, 0.82 mol) was added portionwise to the reaction mixture at 0°C. Then, benzaldehyde (84 ml, 0.82 mol) was added to the reaction mixture at 0° C. over 20 minutes, and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and the filtrate was concentrated in vacuo. The resulting crude material was dissolved in petroleum ether (1000 ml) and filtered again through celite. The filtrate was then concentrated in vacuo to give the title compound. This crude material was sent to the next step without any further purification. Yield: 90% (180 g, pale brown liquid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 8.40 (s, 1H), 7.79-7.76 (m, 2H), 7.49-7.47 (m, 3H), 4 .16-4.10 (m, 2H), 3.98-3.95 (m, 1H), 1.92-1.89 (m, 1H), 1.79-1.74 (m, 1H) , 1.19 (t, J =7.2 Hz, 3H), 0.85 (t, J =7.2 Hz, 3H).

Intermediate 119

Ethyl-(E)-2-(benzylideneamino)-2-ethylhexanoate

Ethyl-(E)-2-(benzylideneamino)butanoate (intermediate 118; 180 g, 0.82 mol) in DMF (800 ml). The reaction mixture was then stirred for 1.5 hours at room temperature. n-Butyl iodide (93 ml, 0.82 mol) was added to the reaction mixture at 0° C., and the mixture was stirred for 1 hour at room temperature. After completion of the reaction (monitored by TLC), the reaction was stopped with 2-propanol (100 ml) at 0°C and then diluted with water (1000 ml). The aqueous layer was extracted with petroleum ether (1000 ml). The organic layer was washed with brine (200 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was passed on to the next step without any further purification. Yield: 88% (200 g, yellow liquid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 8.34 (s, 1H), 7.80-7.77 (m, 2H), 7.47-7.44 (m, 3H), 4 .16 (q, J = 7.0 Hz, 2H), 2.51-1.79 (m, 4H), 1.31-1.18 (m, 7H), 0.88-0.84 (m , 6H).

Intermediate 120

Ethyl 2-amino-2-ethylhexanoate

Dilute HCl (1000 ml, 1.5 N .) at 0°C and the reaction mixture was vigorously stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the organic layer was separated and the aqueous layer was washed with EtOAc (2×100 ml). The aqueous layer was then basified (pH ~8.5) with solid sodium bicarbonate (200 g) and extracted with EtOAc (2 x 200 ml). The organic layer was washed with water (2×15 ml). The combined organics were dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the title compound. The crude material was sent to the next step without any further purification. Yield: 80% (110 g, pale yellow liquid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 4.08 (q, J =7.1 Hz, 2H), 1.68-1.00 (m, 13H), 0.85 (t, J =7.2 Hz, 3H), 0.77 (t, J =7.4 Hz, 3H).

Intermediate 121

2-Amino-2-ethyl- N -phenylhexanamide

With stirring, to a solution of aniline (48.3 ml, 534 mmol) in THF (250 ml) at -78°C was added dropwise n -BuLi (2.6M in hexanes; 205 ml, 534 mmol) over 30 min and reaction the mixture was stirred for 45 min at -25°C to -30°C. Ethyl 2-amino-2-ethylhexanoate (intermediate 120; 50 g, 267 mmol) in THF (250 ml) was then added to the reaction mixture at -78°C and the reaction mixture was stirred for 2 h at -78°C. After completion of the reaction (monitored by TLC), the reaction was stopped with water (500 ml) at -78°C. The reaction mixture was extracted with EtOAc (2×250 ml) and the organic layer was washed with water (2×15 ml). The organic portion was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude title compound. The crude product was dissolved in petroleum ether (1000 ml). The organic portion was washed with 30% methanol in water (2×250 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was passed on to the next step without any further purification. Yield: 66 g (crude brown liquid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.64 (d, J =8.4 Hz, 2H), 7.30 (t, J =7.4 Hz, 2H), 7.05 ( t, J = 7.4 Hz, 1H), 6.55 (d, J = 8.5 Hz, 1H), 1.76-1.07 (m, 10H), 0.86-0.77 (m , 6H).

Intermediate 122

2-Ethyl-N1-phenylhexane-1,2-diamine

With stirring, to a solution of 2-amino-2-ethyl- N -phenylhexanamide (intermediate 121; 66 g, 0.28 mol) in THF (600 ml) was added borane dimethyl sulfide (2M in THF, 253 ml, 0.51 mol) at 0°C and the reaction mixture was heated for 16 h at 70°C. After completion of the reaction (monitored by TLC), the reaction was stopped with methanol (300 ml) at 0°C. Then the reaction mixture was heated for 2 h at 70°C. The reaction mixture was concentrated in vacuo and the resulting residue was dissolved in EtOAc (1000 ml). The organic layer was washed with water (2×150 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 40% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 82% (50 g, brown liquid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.04 (t, J =7.2 Hz, 2H), 6.61 (d, J =8.4 Hz, 2H), 6.49 ( t, J =7.2 Hz, 1H), 5.15 (t, J =4.8 Hz, 1H), 2.79 (d, J =5.6 Hz, 2H), 1.39-1, 17(m, 10H), 0.88-0.79(m, 6H).

Intermediate 123

1,2-bis(2,4-dibromo-5-methoxyphenyl)disulfan

With stirring, to a solution of 3-methoxybenzenethiol (100 g, 0.7 mol) in methanol (1000 ml) was added dropwise bromine (73 ml, 1.4 mol) at 0°C and the reaction mixture was stirred for 24 h at room temperature . The reaction mixture was evaporated in vacuo and the resulting crude material was diluted with EtOAc (2000 ml) and washed with water (2×500 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was dissolved in glacial acetic acid (600 ml), bromine (20 ml) was added dropwise at room temperature, and the reaction mixture was stirred for 2 hours at room temperature. The resulting solid was filtered off, triturated with DCM and dried in vacuo to give the pure title compound. Yield: 37% (78 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.69 (s, 2H), 7.17 (s, 2H), 3.84 (s, 6H).

Intermediate 124

2,4-Dibromo-5-methoxybenzenesulfonyl chloride

With stirring to a suspension of 1,2-bis(2,4-dibromo-5-methoxyphenyl)disulfan (intermediate 123; 20.0 g, 33.67 mmol) and potassium nitrate (17.02 g, 168.35 mmol) in acetonitrile (200 ml) sulfuryl chloride (13.6 ml, 168.35 mmol) was added dropwise at 0° C. and the reaction mixture was stirred for 24 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into crushed ice and the resulting solid was filtered off. The solid was washed with water and dried in vacuo to give the pure title compound. Yield: 91% (22.5 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 8.05 (s, 1H), 7.66 (s, 1H), 4.01 (s, 3H).

Intermediate 125

2,4-Dibromo-5-methoxy- N- (3-((phenylamino)methyl)heptan-3-yl)benzenesulfonamide

To a stirred solution of 2-ethyl-N1-phenylhexane-1,2-diamine (intermediate 122; 4.9 g, 22.34 mmol) in THF (10 ml) was added 2,4-dibromo-5-methoxybenzenesulfonyl chloride (intermediate product 124; 10.5 g, 28.91 mmol) and triethylamine (9.3 ml, 67.02 mmol) at 0° C. and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (50 ml). The organic layer was washed with water (2×15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and 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: 59% (7.2 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 8.01 (s, 1H), 7.60 (s, 1H), 7.50 (s, 1H), 7.03 (t, J =8 .1 Hz, 2H), 6.54-6.46 (m, 3H), 4.80 (t, J = 5.1 Hz, 1H), 3.86 (s, 3H), 3.07-2 .96 (m, 2H), 1.66-1.41 (m, 4H), 1.15-0.95 (m, 4H), 0.78-0.69 (m, 6H).

Intermediate 126

7-Bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide

With stirring, to a solution of 2,4-dibromo-5-methoxy- N- (3-((phenylamino)methyl)heptan-3-yl)benzenesulfonamide (intermediate 125; 7.2 g, 13.1 mmol) in DMF ( 50 ml) was added potassium carbonate (3.62 g, 26.2 mmol) and powdered copper (834 mg, 13.1 mmol) and the reaction mixture was heated for 24 h at 150°C. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and washed with EtOAc (25 ml). The filtrate was concentrated in vacuo and 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: 83% (5.1 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.43-7.30 (m, 4H), 7.15-7.13 (m, 2H), 7.03-7.01 (m, 2H), 4.00-3.60 (m, 5H), 1.62-1.34 (m, 4H), 1.08-0.95 (m, 4H), 0.74-0.71 ( m, 6H). LCMS: (Method A) 467.0 (M ⁺ ), Rt. 3.06 min, 95.31% (maximum).

Intermediate 127

7-Bromo-3-butyl-3-ethyl-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1- dioxide

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (intermediate product 126; 20.0 g, 42.7 mmol) in N -methyl-2-pyrrolidone (100 ml) were added Cs ₂ CO ₃ (27.8 g, 85.5 mmol) and p-methoxybenzyl bromide (7.98 ml, 39.5 mmol) at 0°C and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (200 ml) and the organic layer was washed with water (2×50 ml). The organic part was dried over anhydrous Na ₂ SO ₄ 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: 64% (16 g, white solid).

LCMS: (method A) 587.2 (M ⁺ ), Rt. 3.51 min, 92.94% (maximum).

Intermediate 128

3-Butyl-3-ethyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1, 1-dioxide

With stirring to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 1,1-dioxide (intermediate 127; 16.0 g, 27.2 mmol) in DMF (120 ml) was added sodium thiomethoxide (9.5 g, 136.1 mmol) and the reaction mixture was heated for 16 h at 60 °C After completion of the reaction (monitored by LCMS), the reaction mixture was diluted with EtOAc (200 ml) and the organic layer was washed with water (2×50 ml). The organic part was dried over anhydrous Na ₂ SO ₄ 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: 65% (9.2 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.37 (bs, 1H), 7.31-7.22 (m, 5H), 7.01-6.65 (m, 6H), 4 .32-4.13 (m, 2H), 4.10-3.90 (m, 2H), 3.74 (s, 3H), 2.15 (s, 3H), 1.62-1.34 (m, 4H), 1.08-0.98 (m, 4H), 0.74-0.65 (m, 6H). LCMS: (Method E) 541.2 (M ⁺ +H), Rt. 2.86 min, 93.67% (maximum).

Intermediate 129

tert-Butyl-(E)-3-((3-butyl-3-ethyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4, 5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

With stirring to a solution of 3-butyl-3-ethyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepine-1,1-dioxide (intermediate 128, 1.0 g, 1.85 mmol) in THF (10 mL) was added DABCO (0.02 g, 0.18 mmol) and tert-butyl propiolate (0.28 g , 2.22 mmol) at 0°C and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (20 ml). The organic layer was washed with water (2×15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 15% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 49% (0.6 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.66 (d, J =12.3 Hz, 1H), 7.43 (s, 1H), 7.37 (t, J= 7.6 Hz, 2H), 7.26 (d, J =8.6 Hz, 2H), 7.20-7.14 (m, 3H), 6.87 (d, J =8.6 Hz, 2H), 6.48-6.25 (m, 1H), 5.27 (d, J =12.1 Hz, 1H), 4.51 (s, 2H), 4.30-4.10 (m, 2H) , 3.73 (s, 3H), 2.08 (s, 3H), 1.50-1.36 (m, 13H), 1.18-0.84 (m, 4H), 0.72-0 .48 (m, 6H). LCMS: (Method A) 611.2 (M ⁺ - ^t Bu+H), Rt. 3.94 min, 98.16% (maximum).

Intermediate 130

Methyl-(Z)-3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro -1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate

To a suspension of NaH (60%, 29 mg, 1.21 mmol) in DMF (1 ml) was added N- (tert-butyl)-4-(3,3-dibutyl-8-hydroxy-7-( methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepin-5(2H)-yl)benzamide (intermediate 105; 550 mg, 1.0 mmol) in DMF (2 ml) and the mixture was stirred for 30 minutes at room temperature. Then methyl 3-bromo-2,2-difluoropropionate (164 mg, 0.8 mmol) was added at 0°C and the reaction mixture was heated for 8 h at 65°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, the reaction was stopped with diluted HCl (1.5 N., 2 ml) and the reaction mixture was concentrated in vacuo. The resulting crude material was subjected to distribution between ice-cold water (15 ml) and EtOAc (15 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with ice-cold water (10 ml), brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic part was concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 40% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 14% (90 mg, pale brown solid).

LCMS: (Method E) 649.3 (M ⁺ +H), Rt. 2.76 min, 73.56% (maximum).

Intermediate 131

3-Butyl-7-(dimethylamino)-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (3 g, 6.43 mmol) in toluene (10 ml) was added Cs ₂ CO ₃ (5.2 g, 16.1 mmol) and the reaction mixture was degassed with N ₂ for 10 min. Dimethylamine (2M in THF; 6.4 ml, 12.8 mmol), Pd(OAc) ₂ (0.04 g, 0.16 mmol) was then added followed by X-Phos (0.08 g, 0.16 mmol ) and the reaction mixture was heated for 16 h at 90°C. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and washed with EtOAc (100 ml). The combined organics were concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 13-15% EtOAc/PE, silica gel: 230-400 mesh) to give the title compound. Yield: 26% (0.7 g, yellow gum).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.28 (s, 1H), 7.22-7.18 (m, 2H), 6.97 (d, J =8.0 Hz, 2H ), 6.83 (t, J = 7.2 Hz, 1H), 6.34 (s, 1H), 3.85 (s, 3H), 3.70 (bs, 2H), 3.29 (s , 2H), 2.66 (s, 6H), 1.54-1.41 (m, 2H), 1.35-1.24 (m, 2H), 1.20-1.12 (m, 4H ), 0.85-0.75 (m, 6H). LCMS: (Method A) 431.2 (M ⁺ +H), Rt. 3.19 min, 83.34% (maximum).

Intermediate 132

3-Butyl-7-(dimethylamino)-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3-butyl-7-(dimethylamino)-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 131; 1.9 g, 4.41 mmol) in DMF (15 ml) at room temperature was added sodium thiomethoxide (1.54 g, 22.06 mmol) and the reaction mixture was stirred for 12 h at 80°C. After completion of the reaction (monitored by TLC), the reaction mass was cooled to room temperature and the reaction was stopped with water (15 ml). The aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (20 ml) and brine (20 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give a crude compound which was passed on to the next step without any further purification. Yield: 1.8 g (crude brown gum).

LCMS: (Method E) 417.2 (M ⁺ +H), Rt. 2.11 min, 55.04% (maximum).

Intermediate 133

tert-Butyl-(E)-3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring, to a solution of 3,3-butyl-7-(dimethylamino)-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide ( intermediate 132; 0.1 g, 0.24 mmol) in dry THF (3 ml) was added tert-butyl propiolate (0.046 g, 0.36 mmol) and DABCO (2.7 mg, 0.024 mmol) and then the reaction mixture was stirred for 30 min at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organics were washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 85% (0.11 g, white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.56 (d, J =12.2 Hz, 1H), 7.38 (s, 1H), 7.29 (t, J =7, 9 Hz, 2H), 7.15 (d, J =7.3 Hz, 2H), 6.97 (t, J =7.4 Hz, 1H), 6.26 (s, 1H), 5.38 (d, J =12.2 Hz, 1H), 3.72 (bs, 2H), 3.29 (s, 2H), 2.67 (s, 6H), 1.44 (s, 9H), 1 .37-1.30 (m, 4H), 1.17-0.95 (m, 4H), 0.75-0.70 (m, 6H). LCMS: (Method A) 543.3 (M ⁺ +H), Rt. 3.5 min, 97.69% (maximum).

Intermediate 134

5-Fluoro-6-methoxybenzo[d]thiazol-2-amine

Ammonium thiocyanate (2.96 g, 0.04 mmol) was added with stirring to a solution of 3-fluoro-4-methoxyaniline (5 g, 0.04 mmol) in acetic acid (50 ml) and the reaction mixture was stirred for 45 min at room temperature. Then, bromine (5.7 g, 0.04 mmol) dissolved in acetic acid (10 ml) was added dropwise to the reaction mixture at 15° C., and the resulting reaction mixture was stirred for 3 hours at room temperature. After completion of the reaction, the resulting solid was filtered off and the solid was washed with acetic acid (10 ml) and then dried in vacuo. The resulting solid was suspended in water (20 ml), basified with 10% NaOH solution (pH~10) and filtered. The resulting solid was washed with water (3 x 20 ml) and dried in vacuo to give the title compound. Yield: 84% (5.9 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.52 (d, J =8.6 Hz, 1H), 7.39 (s, 2H), 7.18 (d, J =12.4 Hz, 1H), 3.81(s, 3H). LCMS: (Method A) 199.04 (M ⁺ +H), Rt. 1.08 min, 98.24% (maximum).

Intermediate 135

2-(((2-Amino-4-fluoro-5-methoxyphenyl)thio)methyl)-2-butylhexanoic acid

KOH (27 g, 0.47 mmol) was added to a solution of 5-fluoro-6-methoxybenzo[d]thiazol-2-amine (intermediate 134; 5.9 g, 0.03 mmol) in water (60 ml) with stirring. ) and the reaction mixture was stirred for 16 h at 120°C. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to room temperature. Then 2-(bromomethyl)-2-butylhexanoic acid (4.5 g, 0.04 mmol) was added dropwise (dissolved in 20 ml THF) and the reaction mixture was stirred for 16 h at room temperature. After consumption of the starting material (monitored by LCMS), the reaction mixture was cooled to 0°C and acidified with concentrated HCl (pH ~2). The aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with water (20 ml) and brine (20 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was passed on to the next step without any further purification. Yield: 12.5 g (crude brown gum).

LCMS: (Method A) 358.2 (M ⁺ +H), Rt. 2.67 min, 61.03% (maximum).

Intermediate 136

3,3-Dibutyl-7-fluoro-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring, to a solution of 2-(((2-amino-4-fluoro-5-methoxyphenyl)thio)methyl)-2-butylhexanoic acid (intermediate 135; 12.5 g, 0.04 mmol) in EtOAc (80 ml ) at 0°C, triethylamine (9.04 g, 0.07 mmol) and a solution of 1-propane phosphoric anhydride (50% in EtOAc; 16.7 g, 0.05 mmol) were added dropwise and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by UPLC), the reaction was stopped with water (100 ml) and the aqueous layer was extracted with EtOAc (2×50 ml). The combined organic layer was washed with brine (25 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. 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: 48% (5.7 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.60 (s, 1H), 7.18 (d, J =9.2 Hz, 1H), 6.99 (d, J =12.7 Hz, 1H), 3.82 (s, 3H), 2.98 (s, 2H), 1.64-1.50 (m, 2H), 1.49-1.45 (m, 2H), 1 .22-1.17 (m, 8H), 0.83 (t, J = 6.7 Hz, 6H). LCMS: (Method A) 340.2 (M ⁺ +H), Rt. 2.96 min, 99.47% (maximum).

Intermediate 137

3,3-Dibutyl-7-fluoro-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring to a solution of 3,3-dibutyl-7-fluoro-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 136; 5.7 g, 0.02 mol ) in iodobenzene (30 ml) copper(I) iodide (0.67 g, 0.003 mol) and K ₂ CO ₃ (4.84 g, 0.035 mol) were added and the solution was purged with nitrogen for 20 min to degas. Tris[2-(2-methoxyethoxy)ethyl]amine (0.56 ml, 0.017 mol) was then added under nitrogen atmosphere and the resulting reaction mixture was heated for 40 h at 135°C. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through celite and the celite layer was washed with EtOAc (100 ml). The filtrate was washed with water (50 ml) and brine (50 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 3-5% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 63% (4.6 g, pale brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.44-7.38 (m, 3H), 7.29-7.26 (t, J =7.6 Hz, 1H), 7.07 (d, J =7.6 Hz, 2H), 6.79 (d, J =11.96 Hz, 1H), 3.89 (s, 3H), 3.46 (s, 2H), 1.37 -1.38 (m, 4H), 1.18-1.37 (m, 8H), 0.79-0.81 (m, 6H). LCMS: (Method A) 416.3 (M ⁺ +H), Rt. 3.32 min, 99.63% (maximum).

Intermediate 138

3,3-Dibutyl-7-fluoro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine

With stirring to a solution of 3,3-dibutyl-7-fluoro-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 137; 4.6 g, 11.07 mmol) in THF (45 ml) at 0°C was added dropwise borane dimethyl sulfide (2M in THF; 17.2 ml, 33.2 mmol) and the reaction mixture was refluxed for 40 h at 75°C . After completion of the reaction (monitored by UPLC), the reaction mixture was cooled to 0°C, the reaction was stopped with methanol (10 ml) and heated for 2 h at 65°C. The resulting reaction mixture was then cooled to room temperature and concentrated in vacuo to give a crude material which was passed on to the next step without any further purification. Yield: 5 g (crude colorless liquid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.20 (t, J =7.5 Hz, 2H), 7.10 (d, J =9.5 Hz, 1H), 6.93 ( d, J =6.0 Hz, 2H), 6.81 (t, J =7.1 Hz, 1H), 6.62 (d, J =12.6 Hz, 1H), 3.81 (s, 3H), 3.33(s, 2H), 2.73(s, 2H), 1.18-1.11(m, 12H), 0.79-0.78(m, 6H). LCMS: (Method D) 402.4 (M ⁺ +H), Rt. 3.9 min, 99.4% (maximum).

Intermediate 139

3,3-Dibutyl-7-fluoro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 3,3-dibutyl-7-fluoro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine (intermediate 138; 5 g, 0.01 mmol) in THF (75 ml) and water (7.5 ml) was added oxone (38.3 g, 0.13 mmol) at room temperature and the reaction mixture was stirred for 24 h at this temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a Buchner funnel and the filtrate was extracted with EtOAc (2×25 ml). The combined organic layer was washed with water (25 ml) and brine (25 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 10-12% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 59% (3.2 g, off-white solid).

LCMS: (Method D) 434.2 (M ⁺ +H), Rt. 3.21 min, 92.6% (maximum).

Intermediate 140

3,3-Dibutyl-7-fluoro-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 3,3-dibutyl-7-fluoro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 139; 1, 0 g, 2.3 mmol) in DCM (10 ml) was added BBr ₃ (1M in DCM; 7 ml, 6.92 mmol) at 0°C and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), methanol (10 ml) was added dropwise at 0° C. until gas evolution ceased. Then the reaction mixture was diluted with DCM (20 ml). The DCM layer was washed with water (2×20 ml) and brine (10 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 25-30% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 93% (0.9 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.45 (s, 1H), 7.48 (d, J =9.4 Hz, 1H), 7.24 (t, J =8.3 Hz, 2H), 7.02 (d, J =7.6 Hz, 2H), 6.90 (t, J =7.3 Hz, 1H), 6.74 (d, J =12.1 Hz, 1H), 3.68 (s, 2H), 3.27 (s, 2H), 1.40-1.32 (m, 4H), 1.18-1.01 (m, 8H), 0.75 (t, J =6.80 Hz, 6H). LCMS: (Method A) 420.3 (M ⁺ +H), Rt. 2.99 min, 95.69% (maximum).

Intermediate 141

tert-butyl-(E)-3-((3,3-dibutyl-7-fluoro-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylate

With stirring to a solution of 3,3-dibutyl-7-fluoro-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 140; 0. 15 g, 0.35 mmol) in dry THF (5 ml) was added ethyl propiolate (0.067 g, 0.53 mmol) and DABCO (4.0 mg, 0.035 mmol) and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between water (10 ml) and EtOAc (15 ml). The aqueous layer was extracted with EtOAc (2×15 ml). The combined organics were washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 82% (0.16 g, white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.73 (s, 1H), 7.71 (d, J =3.6 Hz, 1H), 7.37 (t, J =8, 0 Hz, 2H), 7.31 (d, J =7.6 Hz, 2H), 7.11 (t, J =7.2 Hz, 1H), 6.68 (d, J =11.2 Hz , 1H), 5.36 (d, J =12.0 Hz, 1H), 3.83 (bs, 2H), 3.47 (s, 2H), 1.42 (m, 11H), 1.35 -1.28 (m, 2H), 1.27-1.08 (m, 4H), 1.05-1.01 (m, 4H), 0.73 (t, J =8.00 Hz, 6H ). LCMS: (Method A) 490.2 (M- ^t Bu+H), Rt. 3.78 min, 95.03% (maximum).

Intermediate 142

3,3-Dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-7-carbonitrile-1,1-dioxide

To a degassed solution of 7-bromo-3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (2 g, 4.03 mmol ) in DMA (10 ml) was added sodium carbonate (0.42 g, 4.03 mmol) and K ₄ [Fe(CN)] ₆ (1.7 g, 4.03 mmol) at room temperature and the reaction mixture was degassed with using N ₂ for 15 min. Then Pd(OAc) ₂ (90 mg, 0.4 mmol) was added and the reaction mixture was heated for 24 h at 120°C. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo. The resulting mass was subjected to distribution between water (10 ml) and EtOAc (10 ml) and the aqueous layer was extracted with EtOAc (2×25 ml). The combined organic layer was washed with ice-cold water (50 ml) and brine (50 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 90-100% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 44% (1.2 g, light yellow solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.56 (s, 1H), 7.36 (s, 1H), 7.28 (t, J =7.6 Hz, 2H), 7, 09 (d, J =7.6 Hz, 2H), 6.96 (t, J =7.0 Hz, 1H), 3.99 (s, 3H), 3.43 (s, 2H), 3, 34 (s, 2H), 1.40-1.30 (m, 4H), 1.20-1.00 (m, 8H), 0.75 (t, J =6.1 Hz, 6H). LCMS: (Method A) 441.3 (M ⁺ +H), Rt. 3.15 min, 87.84% (maximum).

Intermediate 143

3,3-Dibutyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-7-carbonitrile-1,1-dioxide

To a solution of 3,3-dibutyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-7-carbonitrile-1,1-dioxide (intermediate 142; 0.76 g , 1.72 mmol) in DCM (10 ml) at -10°C was added BBr ₃ (1M in DCM, 3.4 ml, 3.45 mmol) and the reaction mixture was stirred for 12 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C and the reaction was stopped with ice-cold water (5 ml). The aqueous layer was extracted with DCM (2×10 ml). The combined organic layer was washed with ice-cold water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo 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: 28% (0.21 g, yellow solid).

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.68 (s, 1H), 7.34 (t, J= 8.0 Hz, 2H), 7.11-7.05 (m, 4H), 3.75 (s, 2H), 3.26 (s, 2H), 1.42-1.27 (m, 4H), 1.18-1.04 (m, 8H), 0.80 (t, J =6.8Hz, 6H). LCMS: (Method E) 427.2 (M ⁺ +H), Rt. 2.76 min, 68.44% (maximum).

Intermediate 144

tert-Butyl-(E)-3-((3,3-dibutyl-7-cyano-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylate

With stirring to a solution of 3,3-dibutyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-7-carbonitrile-1,1-dioxide (intermediate 143; 0. 21 g, 0.49 mmol) in THF (2 ml) was added DABCO (5.50 mg, 0.05 mmol) and tert-butyl propiolate (81 mg, 0.69 mmol) and the reaction mixture was stirred for 30 min at room temperature. temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organic layer was washed with ice-cold water (5 ml) and brine (5 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 81% (220 mg, yellow solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.80 (d, J= 12.0 Hz, 1H), 7.75 (s, 1H), 7.37 (t, J =7.6 Hz, 3H), 7.29 (d, J =6.8 Hz, 2H), 7.11 (t, J =7.6 Hz, 1H), 5.57 (d, J =12.0 Hz, 1H), 3.82 (s, 2H), 3.53 (s, 2H), 1.46-1.27 (m, 13H), 1.21-0.85 (m, 8H), 0.74 (t, J =6.4 Hz, 6H). LCMS: (Method E) 497.2 (M ⁺ - ^t Bu+H), Rt, 3.09 min, 80.74% (maximum).

Intermediate 145

Methyl-(E)-3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)but-2-enoate

With stirring to a solution of 3,3-dibutyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 1,1-dioxide (0.5 g, 0.88 mmol) in THF (10 mL) was added DABCO (0.09 g, 0.88 mmol) and methyl 2-butinoate (0.13 g, 1.32 mmol) at room temperature and the reaction mixture was stirred for 16 h at 50°C. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (30 ml) and the organic layer was washed with water (2×15 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 7% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 64% (0.38 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.41-7.34 (m, 3H), 7.23 (d, J =8.0 Hz, 2H), 7.16-7.13 (m, 3H), 6.84 (d, J =8.0 Hz, 2H), 6.34 (s, 1H), 4.76 (s, 1H), 4.58 (s, 2H), 4 .28 (s, 2H), 3.73 (s, 3H), 3.56 (d, J = 8.0 Hz, 3H), 2.42 (s, 3H), 1.99 (s, 3H) , 2.00-1.75 (m, 2H), 1.50-1.32 (m, 2H), 1.20-0.75 (m, 8H), 0.75-0.50 (m, 6H). LCMS: (Method A) 667.3 (M ⁺ +H), Rt. 3.84 min, 99.29% (maximum).

Intermediate 146

(E) -3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1, 2,5-Benzothiadiazepin-8-yl)oxy)but-2-enoic acid

With stirring, to a solution of methyl-(E)-3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4, 5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)but-2-enoate (intermediate 145; 0.38 g, 0.56 mmol) in 1,4-dioxane/water (10 mL) , 4:1) lithium hydroxide (0.047 g, 1.13 mmol) was added and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1 ml, 1.5 N). The aqueous layer was extracted with EtOAc (2×20 ml) and the combined organic layer was dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 20% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 40% (150 mg, white solid). LCMS: (Method E) 653.2 (M ⁺ +H), Rt. 3.01 min, 98.64% (maximum)

Intermediate 147

Ethyl-(Z)-3-((3-butyl-3-ethyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5- tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

With stirring to a solution of 3-butyl-3-ethyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepine-1,1-dioxide (intermediate 128; 1 g, 1.85 mmol) in DMF (10 ml) NaH (60%, 0.037 g, 9.25 mmol) was added in portions at 0°C and the reaction mixture was stirred in for 15 min at 0°C. Then methyl 3-bromo-2,2-difluoropropanoate (1.1 g, 5.55 mmol) was added and the reaction mixture was heated for 16 h at 80°C. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to 0°C, the reaction was stopped by diluting with HCl (1.5 N, pH ~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with brine (5 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 30% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 33% (0.04 g, brown gum).

LCMS: (Method A) 643.2 (M ⁺ +H), Rt. 3.35 min, 90.58% (maximum).

Intermediate 148

(Z) -3-((3-butyl-3-ethyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring to a solution of ethyl-(Z)-3-((3-butyl-3-ethyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3, 4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 147; 0.04 g, 0.62 mmol) in a mixture of 1,4-dioxane and water (10 ml, 4:1) lithium hydroxide (0.13 g, 3.11 mmol) was added and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by LCMS), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and then diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (8 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was evaporated in vacuo and the resulting crude material was passed on to the next step without any further purification. Yield: 89.9% (350 mg, pale brown gum).

LCMS: (Method A) 629.1 (M ⁺ +H), Rt. 3.08 min, 87.52% (maximum).

Intermediate 149

7-Bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (intermediate product 126; 1.0 g, 2.13 mmol) in DCM (10 ml) was added BBr ₃ (0.32 ml, 3.37 mmol) at -78°C and then the reaction mixture was stirred for 2 h at room temperature. After completion of the reaction (monitored by TLC), the reaction was stopped with methanol (10 ml) and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 24% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 51% (0.5 g, colorless gum).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.58 (s, 1H), 7.37-7.35 (m, 2H), 7.27 (t, J =7.6 Hz, 2H ), 7.06-7.04 (m, 2H), 6.97-6.94 (m, 2H), 3.83 (bs, 2H), 1.61-1.58 (m, 2H), 1.50-1.41 (m, 2H), 1.37-1.33 (m, 4H), 0.73 (t, J =6.80 Hz, 6H). LCMS: (Method A) 455.1 (M ⁺ +2H), Rt 2.81 min, 94.49% (maximum).

Intermediate 150

tert-Butyl-(E)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepin-8-yl)oxy)acrylate

With stirring to a solution of 7-bromo-3-butyl-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (intermediate product 149; 0.5 g, 1.10 mmol) in THF (5 ml) was added DABCO (12.3 mg, 0.1 mmol) and tert-butyl propiolate (0.22 ml, 1.60 mmol) and the reaction mixture stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (25 ml). The organic layer was washed with water (2×15 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 8% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 73% (0.46 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.75 (bs, 1H), 7.65 (d, J =16.4 Hz, 1H), 7.54 (s, 1H), 7, 45-7.42(m, 2H), 7.33-7.31(m, 2H), 7.19-7.15(m, 1H), 6.84(s, 1H), 5.19( d, J = 16.4 Hz, 1H), 4.02 (bs, 2H), 1.60-1.51 (m, 2H), 1.42 (s, 9H), 1.28-0.75 (m, 6H), 0.72-0.47 (m, 6H). LCMS: (Method A) 577.2 (M ⁺ -2H), Rt. 3.41 min, 94.96%.

Intermediate 151

7-Bromo-3-butyl-3-ethyl-8-methoxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (intermediate product 126; 2 g, 4.27 mmol) in N-methyl-2-pyrrolidone (10 ml) was added Cs ₂ CO ₃ (2.78 g, 8.53 mmol) and methyl iodide (1.33 ml, 21.36 mmol ) at room temperature and then the reaction mixture was stirred for 8 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (25 ml) and the organic layer was washed with water (2×15 ml). The combined organics were dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 8% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 65% (1.3 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.37-7.32 (m, 3H), 7.22-7.11 (m, 2H), 7.05-7.01 (m, 2H), 4.02-3.93(m, 2H), 3.89(s, 3H), 2.82(s, 3H), 1.85-1.73(m, 2H), 1.55 -1.46 (m, 2H), 1.20-0.91 (m, 4H), 0.78-0.72 (m, 6H). LCMS: (method E) 483.2 (M ⁺ +2H), Rt. 2.95 min, 96.64% (maximum).

Intermediate 152

3-Butyl-3-ethyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide

With stirring to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1- dioxide (intermediate 151; 1.3 g, 2.7 mmol) in DMF (10 ml) was added sodium thiomethoxide (0.72 g, 13.50 mmol) at room temperature and the reaction mixture was stirred for 16 h at 60° C. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (25 ml) and the organic layer was washed with water (2×15 ml). The organic was dried over anhydrous Na ₂ SO ₄ , concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 7% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 72% (0.87 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.35 (s, 1H), 7.25-7.18 (m, 3H), 7.01-6.99 (m, 2H), 6 .87 (t, J = 7.2 Hz, 1H), 6.61 (s, 1H), 3.82 (bs, 2H), 2.68 (s, 3H), 2.13 (s, 3H) , 1.84-1.71 (m, 2H), 1.54-1.47 (m, 2H), 1.24-0.95 (m, 4H), 0.86-0.68 (m, 6H). LCMS: (Method E) 435.2 (M ⁺ +H), Rt. 2.69 min, 97.42% (maximum).

Intermediate 153

tert-Butyl-(E)-3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)acrylate

With stirring to a solution of 3-butyl-3-ethyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1, 1-dioxide (intermediate 152; 200 mg, 0.46 mmol) in dry THF (3 ml) was added DABCO (5.10 mg, 0.05 mmol) and tert-butyl propiolate (0.1 ml, 0.7 mmol ) at room temperature and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by LCMS), the reaction mixture was diluted with ice-cold water (10 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and evaporated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 6% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 75% (190 mg, white solid).

H NMR (400 MHz, DMSO- d ₆ ): δ 7.61 (d, J =12.4 Hz, 1H), 7.38 (t, J =8.0 Hz, 3H), 7.30 (d , J= 7.6 Hz, 2H), 7.12 (t, J =7.2 Hz, 1H), 6.46 (s, 1H), 5.31 (d, J =12.4 Hz, 1H ), 4.12 (s, 2H), 2.88 (s, 3H), 2.08 (s, 3H), 1.92-1.86 (m, 1H), 1.82-1.76 ( m, 1H), 1.55-1.43 (m, 10H), 1.19-1.08 (m, 5H), 0.83-0.62 (m, 6H). LCMS: (Method A) 505.2 (M ⁺ - ^t Bu+H), Rt. 3.65 min, 97.40%.

Intermediate 154

3-Butyl-3-ethyl-8-methoxy-7-(methylamino)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of a mixture of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide and 3-butyl -3-ethyl-7-iodine-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 91; 1.0 g, 2, 1 mmol) in toluene (10 ml) was added Cs ₂ CO ₃ (1.74 g, 5.3 mmol) and the reaction mixture was degassed for 10 min with N ₂ . Then Pd(OAc) ₂ (0.048 g, 0.2 mmol), then X-Phos (0.102 g, 0.2 mmol) and methylamine (2M THF solution; 2.14 ml, 4.3 mmol) were added and the reaction mixture heated for 16 h at 90°C. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and washed with EtOAc (15 ml). The combined organics were concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 13-15% EtOAc/PE, silica gel: 230-400 mesh) to give the title compound. Yield: 62% (0.55 g, colorless gum).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.19 (d, J =11.2 Hz, 1H), 7.15 (s, 2H), 6.93 (s, 2H), 6, 79 (t, J =9.6 Hz, 1H), 5.97 (s, 2H), 3.85 (s, 3H), 3.62 (bs, 2H), 3.11 (s, 2H), 2.57 (d, J =2.8 Hz, 3H), 1.36-1.29 (m, 4H), 1.18-1.08 (m, 4H), 0.75 (t, J = 9.20Hz, 6H). LCMS: (Method A) 417.1 (M ⁺ +H), Rt. 2.78 min, 98.76% (maximum).

Intermediate 155

3-Butyl-3-ethyl-8-hydroxy-7-(methylamino)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3-butyl-3-ethyl-8-methoxy-7-(methylamino)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 154; 1.0 g, 2.4 mmol) in DMF (10 ml) at room temperature was added sodium thiomethoxide (0.84 g, 12.01 mmol) and the reaction mixture was stirred for 12 h at 80°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and the reaction was stopped with water (15 ml). The aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (20 ml) and brine (20 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give a crude material which was passed on to the next step without any further purification. Yield: 0.6 g (crude brown gum).

LCMS: (Method A) 403.1 (M+H), Rt. 2.76 min, 78.51% (maximum).

Intermediate 156

tert-Butyl-(E)-3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate

With stirring to a solution of 3-butyl-3-ethyl-8-hydroxy-7-(methylamino)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 155; 0.15 g, 0.37 mmol) in dry THF (5 ml) was added tert-butyl propiolate (0.07 g, 5.59 mmol) and DABCO (4 mg, 0.037 mmol) and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between water (10 ml) and EtOAc (15 ml). The aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 89% (0.18 g, white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.64 (d, J =12.4 Hz, 1H), 7.59 (s, 1H), 7.32 (t, J =8, 0 Hz, 2H), 7.27 (s, 1H), 7.23-7.20 (m, 2H), 7.02 (t, J = 7.2 Hz, 1H), 6.65 (s, 1H), 5.44 (d, J =12.0 Hz, 1H), 3.81 (bs, 2H), 3.41 (s, 2H), 2.97 (s, 3H), 1.48- 1.25 (m, 13H), 1.09-1.05 (m, 4H), 0.74 (t, J =6.80 Hz, 6H). LCMS: (Method A) 527.0 (M ⁺ +H), Rt. 3.65 min, 97.28% (maximum).

Intermediate 157

3,3-Dibutyl-8-hydroxy-5-(4-methoxyphenyl)-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 5-(4-bromophenyl)-3,3-dibutyl-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide ( intermediate 20; 5 g, 4.74 mmol) in DMF (30 ml) at room temperature was added NaOMe (30%, 2 ml, 9.50 mmol) and the reaction mixture was degassed for 15 min with N ₂ . Then Cu(I)Br (68 mg, 0.47 mmol) was added and the reaction mixture was heated for 48 h at 120°C. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (5 ml) and the aqueous layer was extracted with EtOAc (2×100 ml). The combined organic layer was washed with ice-cold water (100 ml) and brine (100 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and 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: 69% (3.1 g, brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.29 (s, 1H), 7.25 (s, 1H), 7.04 (d, J =8.4 Hz, 2H), 6, 85 (d, J = 8.8 Hz, 2H), 6.47 (s, 1H), 3.71 (s, 3H), 3.66 (bs, 2H), 3.23 (s, 2H), 2.11 (s, 3H), 1.46-1.26 (m, 4H), 1.11-1.02 (m, 8H), 0.76 (t, J =6.4 Hz, 6H) . LCMS: (Method E) 478.1 (M ⁺ +H), Rt. 2.80 min, 80.65% (maximum).

Intermediate 158

Ethyl-(Z)-3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate

To a suspension of NaH (60%, 0.85 g, 21.2 mmol) in DMF (15 mL) at 0°C was added 3,3-dibutyl-8-hydroxy-5-(4-methoxyphenyl)-7- (methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 157; 3.1 g, 6.5 mmol) in DMF (7.5 ml) and reaction the mixture was stirred for 30 minutes at room temperature. Ethyl 3-bromo-2,2-difluoropropionate (3.5 g, 16.2 mmol) in DMF (7.5 ml) was then added at 0°C and the reaction mixture was heated for 8 h at 65°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, the reaction was stopped with dilute HCl (1.5 N., 20 ml) and concentrated in vacuo. The resulting residue was dissolved in ice-cold water (100 ml) and the aqueous layer was extracted with EtOAc (2×100 ml). The combined organic layer was washed with ice-cold water (100 ml) and brine (100 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 40% (1.5 g, yellow gum).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.56-7.52 (m, 2H), 7.25 (d, J= 8.4 Hz, 2H), 6.95 (d, J =8.0 Hz, 2H), 6.42 (s, 1H), 4.27-4.24 (m, 2H), 3.78-3.73 (m, 5H), 3.40 (s, 2H), 2.12 (s, 3H), 1.42-1.34 (m, 7H), 1.27-0.99 (m, 8H), 0.75-0.73 (m, 6H) . LCMS: (Method A) 594.2 (M ⁺ +H), Rt. 3.36 min, 93.27% (maximum).

Intermediate 159

Ethyl-(Z)-3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate

With stirring, to a solution of ethyl-(Z)-3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (1.5 g, 2.52 mmol) in DCM (20 mL) BBr ₃ (1M in DCM, 5.1 mL) was added at -78°C , 5.0 mmol) and the reaction mixture was stirred for 6 h at -10°C. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (5 ml) and the aqueous layer was extracted with DCM (2×50 ml). The combined organic layer was washed with ice-cold water (50 ml) and brine (50 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 31% EtOAc PE; silica gel: 230-400 mesh) to give the title compound. Yield: 36% (0.52 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.39 (s, 1H), 7.53-7.49 (m, 2H), 7.16 (d, J =8.4 Hz, 2H ), 6.78 (d, J = 8.8 Hz, 2H), 6.34 (s, 1H), 4.24 (q, J = 7.2 Hz, 2H), 3.73 (s, 2H ), 3.40 (s, 2H), 2.10 (s, 3H), 1.43-1.24 (m, 4H), 1.16-1.11 (m, 3H), 1.09- 0.98 (m, 8H), 0.75 (t, J =7.2 Hz, 6H). LCMS: (Method A) 580.2 (M ⁺ +H), Rt. 2.99 min, 98.42% (maximum).

Intermediate 160

5-fluoro-6-methoxybenzo[d]thiazol-2-amine

While stirring, to a solution of 3-fluoro-4-methoxyaniline (50 g, 0.354 mol) in acetic acid (300 ml) was added ammonium thiocyanate (29.69 g, 0.39 mol) at room temperature, and then the reaction mixture was stirred for 45 min at room temperature. Bromine (57 g, 0.354 mol) dissolved in acetic acid (100 ml) at 15° C. was then added dropwise to the reaction mixture, and the resulting reaction mixture was stirred for 3 hours at room temperature. After completion of the reaction, the resulting solid was filtered off, washed with acetic acid (50 ml) and then dried in vacuo. The resulting solid was suspended in water (200 ml) and basified with 10% NaOH solution (pH~10). The resulting solid was filtered off, washed with water (3 x 200 ml) and dried in vacuo to give the title compound. Yield: 86% (60 g, almost white solid).

LCMS: (Method A) 199.0 (M ⁺ +H), Rt. 1.12 min, 90.09% (maximum).

Intermediate 161

2-(((2-amino-4-fluoro-5-methoxyphenyl)thio)methyl)-2-ethylhexanoic acid

KOH (135 g, 2.42 mol) was added to a stirred solution of 5-fluoro-6-methoxybenzo[d]thiazol-2-amine (intermediate 160; 30 g, 0.151 mol) in water (300 ml) and the reaction mixture stirred for 16 h at 120°C. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to room temperature. Then 2-(bromomethyl)-2-ethylhexanoic acid (43.05 g, 0.18 mol) was added dropwise (dissolved in 100 ml THF) and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to 0°C and acidified with concentrated HCl (pH ~2). The aqueous part was extracted with EtOAc (2×25 ml). The combined organic layer was washed with water (30 ml) and brine (30 ml). The organic portion was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give a crude material which was passed on to the next step without any further purification. Yield: 60 g (crude brown gum).

Intermediate 162

3-Butyl-3-ethyl-7-fluoro-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring to a solution of 2-(((2-amino-4-fluro-5-methoxyphenyl)thio)methyl)-2-ethylhexanoic acid (intermediate 161; 60 g, 0.18 mol) in EtOAc (600 mL) at 0°C, triethylamine (36.7 g, 0.3642 mol) and a solution of 1-propane phosphoric anhydride (50% in EtOAc; 69.5 g, 0.2185 mol) were added dropwise and the reaction mixture was stirred for 16 h at room temperature. temperature. After completion of the reaction (monitored by UPLC), water (500 ml) was added to the reaction mixture and the aqueous layer was extracted with EtOAc (2×500 ml). The combined organic layer was washed with brine (250 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. 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: 30% (17 g, almost white solid).

LCMS: (Method A) 312.3 (M ⁺ +H), Rt. 2.64 min, 99.63% (maximum).

Intermediate 163

3-Butyl-3-ethyl-7-fluoro-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring to a solution of 3-butyl-3-ethyl-7-fluoro-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 162; 17 g, 0.05 mol ) in iodobenzene (170 ml) was added copper(I) iodide (1.03 g, 0.01 mol) and K ₂ CO ₃ (15.08 g, 0.11 mol) and the reaction mixture was purged with nitrogen for 20 min to degassing. Tris[2-(2-methoxyethoxy)ethyl]amine (3.52 g, 0.01 mol) was then added under nitrogen atmosphere and the resulting reaction mixture was heated for 40 h at 135°C. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through celite and washed with EtOAc (250 ml). The filtrate was concentrated in vacuo to give the crude material, which was purified by Isolera column chromatography (eluent: 3-5% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 86% (16 g, pale brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.37-7.39 (m, 2H), 7.29 (d, J =6.80 Hz, 1H), 7.08 (d, J =6.80 Hz, 2H), 6.82 (d, J =12.00 Hz, 2H), 3.89 (s, 3H), 3.46 (s, 2H), 1.37-1.38 (m, 4H), 1.18-1.37 (m, 4H), 0.79-0.81 (m, 6H). LCMS: (method A) 387.9 (M ⁺ ), Rt. 3.09 min, 99.25% (maximum).

Intermediate 164

3-Butyl-3-ethyl-7-fluoro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine

With stirring to a solution of 3-butyl-3-ethyl-7-fluoro-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 163; 16 g, 0.04 mol) in THF (160 ml) at 0°C was added dropwise BH ₃ .DMS (2M in THF, 62 ml, 0.12 mol) and the reaction mixture was refluxed for 40 h at 75°C . After completion of the reaction (monitored by UPLC), the reaction mixture was cooled to 0°C and the reaction was stopped with methanol (100 ml). The resulting solution was heated for 2 h at 65°C and then cooled to room temperature and concentrated in vacuo. The resulting crude mixture was sent to the next stage without any further purification. Yield: 100% (15 g, colorless liquid).

LCMS: (Method A) 374.3 (M ⁺ +H), Rt. 2.72 min, 92.66% (maximum).

Intermediate 165

3-Butyl-3-ethyl-7-fluoro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 3-butyl-3-ethyl-7-fluoro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine (intermediate 164; 15 g, 0.04 mol) in THF (100 ml) was added water (45 ml) and oxone (125 g, 0.40 mol) at room temperature and then the reaction mixture was stirred for 24 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a Buchner funnel and the filtrate was extracted with EtOAc (2×250 ml). The combined organic layer was washed with water (250 ml) and brine (250 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. 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: 92% (15 g, yellowish solid).

¹ H NMR (300 MHz, DMSO- d ₆ ): δ 7.52 (d, J =8.70 Hz, 1H), 7.23-7.25 (m, 2H), 7.04-7.07 (m, 2H), 6.93-6.95 (m, 1H), 6.80 (d, J =12.60 Hz, 1H), 3.90 (s, 3H), 3.28 (s, 2H), 3.31 (m, 2H), 1.17-1.24 (m, 4H), 0.93-0.95 (m, 4H), 0.73-0.83 (m, 6H) . LCMS: (Method A) 406.2 (M ⁺ +H), Rt. 3.04 min, 95.49% (maximum).

Intermediate 166

3-Butyl-3-ethyl-7-fluoro-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 3-butyl-3-ethyl-7-fluoro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 165; 15 g, 0.036 mol) in DCM (200 ml) was added BBr ₃ (1M in DCM; 74 ml, 0.074 mmol) at 0°C and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), methanol (100 ml) was added dropwise at 0° C. until gas evolution ceased. The reaction mixture was diluted with DCM (100 ml) and the DCM layer was washed with water (2×200 ml) and brine (200 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 30-32% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 94% (7 g, almost white solid).

¹ H NMR (300 MHz, DMSO- d ₆ ): δ 7.48 (d, J =9.60 Hz, 2H), 7.22 (t, J =7.50 Hz, 2H), 6.99 ( d, J = 7.80 Hz, 2H), 6.74-6.79 (m, 2H), 3.66 (s, 2H), 3.18 (s, 2H), 1.36-1.47 (m, 4H), 1.01-1.10 (m, 4H), 0.73-0.75 (m, 6H). LCMS: (Method A) 392.2 (M ⁺ +H), Rt. 2.08 min, 96.59% (maximum).

Intermediate 167

Methyl-(Z)-3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)-2-fluoroacrylate

With stirring, to a solution of 3-butyl-3-ethyl-7-fluoro-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 166; 0.5 g, 0.2 mol) in DMF (5 mL) at 0°C NaH (60%, 0.10 g, 0.02 mmol) was added in portions and the reaction mixture was stirred for 15 min. Then methyl 3-bromo-2,2-difluoropropanoate (0.29 g, 0.02 mol) was added and the reaction mixture was heated for 16 h at 85°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, the reaction was stopped with dilute HCl (1.5 N, pH ~4) and then diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organic layer was washed with brine (5 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 15-18% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 25% (0.12 g, off-white solid).

LCMS: (Method A) 494.2 (M ⁺ +H), Rt. 3.04 min, 85% (maximum).

Intermediate 168

3,3-Dibutyl-7-(dimethylamino)-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1-oxide

With stirring to a solution of 7-bromo-3,3-dibutyl-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1, 1-dioxide (1.0 g, 1.81 mmol) in toluene (10 ml) was added dimethylamine (2M in THF; 2.7 ml, 5.44 mmol) and Cs ₂ CO ₃ (1.31 g, 4, 0 mmol) and the reaction mixture was degassed with N ₂ for 10 min. Then Pd(OAc) ₂ (0.036 g, 0.16 mmol) and X-Phos (0.077 g, 0.16 mmol) were added and the reaction mixture was heated for 16 h at 90°C. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and washed with EtOAc (15 ml). The combined organics were concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 13-15% EtOAc/PE, silica gel: 230-400 mesh) to give the title compound. Yield: 50% (0.53 g, yellow gum).

LCMS: (Method A) 580.3 (M ⁺ +H), Rt. 2.63 min, 85.42% (maximum).

Intermediate 169

3,3-Dibutyl-7-(dimethylamino)-8-hydroxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1- dioxide

With stirring, to a solution of 3,3-dibutyl-7-(dimethylamino)-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 1-oxide (intermediate 168; 0.53 g, 0.91 mmol) in dry DMF (5 ml) was added sodium thiomethoxide (320 mg, 4.57 mmol) at room temperature and the reaction mixture was stirred for 16 h at 60 °C. After completion of the reaction (monitored by LCMS), the reaction was stopped with ice-cold water (2 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (15 ml) and brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 10-15% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 87% (450 mg, almost white solid).

LCMS: (Method A) 566.3 (M ⁺ +H), Rt. 3.31 min, 70.45% (maximum).

Intermediate 170

Ethyl-(Z)-3-((3,3-dibutyl-7-(dimethylamino)-2-(4-methoxybenzyl)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

With stirring, to a solution of 3,3-dibutyl-7-(dimethylamino)-8-hydroxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 1,1-dioxide (intermediate 169; 0.25 g, 0.44 mmol) in DMF (5 mL) NaH (60%, 0.058 g, 1.43 mmol) was added in portions at 0°C and the reaction mixture was stirred in within 15 min. Then ethyl 3-bromo-2,2-difluoropropanoate (0.24 g, 1.1 mmol) was added and the reaction mixture was heated for 16 h at 65°C. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C, the reaction was stopped with dilute HCl (1.5 N, pH ~4) and then diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic part was concentrated in vacuo. The resulting crude material was then triturated with Et ₂ O and dried in vacuo. The resulting compound was repurified by Isolera column chromatography (eluent: 20% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 64% (0.02 g, off-white solid).

LCMS: (Method E) 682.3 (M ⁺ +H), Rt. 3.18 min, 80.56% (maximum).

Intermediate 171

(Z) -3-((3,3-dibutyl-7-(dimethylamino)-2-(4-methoxybenzyl)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1, 2,5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring to a solution of ethyl-(Z)-3-((3,3-dibutyl-7-(dimethylamino)-2-(4-methoxybenzyl)-1,1-dioxido-5-phenyl-2,3,4, 5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 170; 0.02 g, 0.28 mmol) in a mixture of 1,4-dioxane and water (5 ml, 4:1) lithium hydroxide (0.04 g, 0.84 mmol) was added and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with water (5 ml). Then the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (8 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 2-3% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 98% (0.18 g, almost white solid).

LCMS: (Method E) 654.3 (M ⁺ +H), Rt. 3.00 min, 78.1% (maximum).

Intermediate 172

Ethyl-(Z)-3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 2,5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

To a suspension of NaH (60%, 118 mg, 4.91 mmol) in DMA (3 ml) at 0°C was added a solution of 3-butyl-3-ethyl-8-hydroxy-2-methyl-7-(methylthio)-5 -phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (intermediate 152; 0.4 g, 0.92 mmol) in DMA (3 mL) and reaction mixture stirred for 30 min at room temperature. Ethyl 3-bromo-2,2-difluoropropionate (0.26 ml, 2.00 mmol) in DMA (3 ml) was then added dropwise at 0°C and the reaction mixture was heated for 3 h at 60°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, the reaction was stopped with dilute HCl (1.5 N, 3 ml, pH~4) and concentrated in vacuo. The resulting residue was subjected to distribution between ice-cold water (10 ml) and EtOAc (10 ml) and the aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with ice-cold water (15 ml) and brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and 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: 78% (390 mg, colorless gum).

Intermediate 173

tert-Butyl-(E)-3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylate

With stirring, to a solution of 3-butyl-3-ethyl-7-fluoro-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate 166; 500 mg, 1.28 mmol) in THF (5 ml) at room temperature were added DABCO (14 mg, 0.13 mmol) and tert-butyl propiolate (161 mg, 1.28 mmol) and the reaction mixture was stirred for 2 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with ice-cold water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and 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: 76% (500 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.70-7.72 (m, 2H), 7.27-7.29 (m, 4H), 7.10 (t, J =7, 16 Hz, 1H), 6.69 (d, J =12.16 Hz, 1H), 5.37 (d, J =12.20 Hz, 1H), 3.83 (s, 2H), 3.45 (s, 2H), 1.53-1.54 (m, 2H), 1.43 (s, 9H), 1.27-1.30 (m, 2H), 0.96-0.98 (m , 4H), 0.69-0.70 (m, 6H). LCMS: (Method E) 462.1 (M ⁺ - ^t Bu+H), Rt. 3.04 min, 98.65% (maximum).

Intermediate 174

4-(3,3-Dibutyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-dihydro-1,5-benzothiazepin-5(2H)-yl)-N , N-dimethylbenzamide

With stirring, to a solution of 4-(3-butyl-3-ethyl-8-hydroxy-7-(methylthio)-1,1-dioxido-2,3,4,5-dihydro-1,5-benzothiazepine-5(2H )-yl)-benzoic acid (intermediate 104; 500 mg, 1.01 mmol) in DMF (5 ml) was added triethylamine (0.4 g, 4.04 mmol), N,N -dimethylamine (165 mg, 2.04 mmol). 03 mmol) and HATU (772 mg, 2.03 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with ice-cold water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 7% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 99% (0.6 g, yellow solid).

LCMS: (Method A) 519.3 (M ⁺ +H), Rt. 2.53 min, 91.00% (maximum).

Intermediate 175

Ethyl-(Z)-3-((3,3-dibutyl-5-(4-(dimethylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate

A solution of 4-(3,3-dibutyl-8-hydroxy-7-(methylthio)-1, 1-dioxido-3,4-dihydro-1,5-benzothiazepin-5(2H)-yl)-N,N-dimethylbenzamide (intermediate 175; 600 mg, 1.15 mmol) in DMF (3 ml) and reaction the mixture was stirred for 30 minutes. Ethyl 3-bromo-2,2-difluoropropionate (627 mg, 2.89 mmol) in DMF (3 ml) was then added dropwise at 0°C and the reaction mixture was heated for 8 h at 65°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, the reaction was stopped with dilute HCl (1.5 N., 3 ml) and concentrated in vacuo. The resulting residue was subjected to distribution between ice-cold water (10 ml) and EtOAc (10 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with ice-cold water (15 ml) and brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 50% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 61% (450 mg, yellow gum).

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.68 (s, 1H), 7.40-7.30 (m, 3H), 7.05 (d, J =8.2 Hz, 2H), 6.79 (s, 1H), 4.57-4.51 (m, 2H), 3.19 (s, 2H), 3.16 (s, 6H), 3.07 (s, 2H), 2 .26 (s, 3H), 1.62-1.33 (m, 7H), 1.32-1.15 (m, 8H), 0.91-0.69 (m, 6H). LCMS: (Method E) 635.3 (M ⁺ +H), Rt. 2.69 min, 94.79% (maximum).

Intermediate 176

Ethyl-(Z)-3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

To a suspension of NaH (60%, 183 mg, 4.5 mmol) in dry DMA (3 ml) was added a solution of 3,3-dibutyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio )-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (0.8 g, 1.40 mmol) in DMA (3 ml) and the reaction mixture was stirred for 10 min at room temperature. Ethyl 3-bromo-2,2-difluoropropionate (0.41 ml, 3.10 mmol) in DMA (3 ml) was then added dropwise at 0°C and the reaction mixture was heated for 3 h at 60°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, the reaction was stopped with dilute HCl (1.5 N, 3 ml, pH~4) and concentrated in vacuo. The resulting residue was subjected to distribution between ice-cold water (10 ml) and EtOAc (10 ml) and the aqueous layer was extracted with EtOAc (2×30 ml). The combined organic layer was washed with ice-cold water (15 ml) and brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and 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: 83% (800 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.63 (s, 2H), 7.37 (s, 2H), 7.28-7.14 (m, 6H), 6.87 (d , J = 7.6 Hz, 2H), 4.49 (bs, 2H), 4.33-4.22 (m, 3H), 3.73 (s, 3H), 2.11 (s, 3H) , 1.40 (bs, 2H), 1.27-1.24 (m, 6H), 1.01-0.98 (m, 8H), 0.86-0.65 (m, 6H). LCMS: (Method E) 685.2 (M ⁺ +H), Rt. 3.18 min, 92.22% (maximum). HPLC: (method B) Rt. 7.46 min, 96.07% (maximum).

Intermediate 177

Ethyl-(Z)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 8-yl)oxy)-2-fluoroacrylate

With stirring, to a solution of ethyl-(Z)-3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4, 5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 176; 0.8 g, 1.10 mmol) in dry DCM (3 ml) TFA (3 ml) was added and triethylsilane at 0°C and the reaction mixture was stirred for 3 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and 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: 76% (500 mg, almost white solid).

LCMS: (Method A) 565.2 (M ⁺ +H), Rt. 3.28 min, 97.21% (maximum).

Intermediate 178

Ethyl-(Z)-3-((3,3-dibutyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

With stirring to a solution of ethyl-(Z)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 177; 0.5 g, 0.88 mmol) in N-methyl-2-pyrrolidone (5 ml) was added Cs ₂ CO ₃ (0.57 g, 1.75 mmol) and methyl iodide (0.28 ml, 4.49 mmol) at room temperature and the reaction mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (25 ml) and the organic layer was washed with water (2×15 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The obtained crude material was sent to the next stage without any further purification. Yield: 98% (0.5 g, brown gum).

Intermediate 179

7-Bromo-3-butyl-3-ethyl-8-methoxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (intermediate product 126; 1 g, 2.10 mmol) in N-methyl-2-pyrrolidone (10 ml) were added Cs ₂ CO ₃ (1.39 g, 4.20 mmol) and methyl iodide (0.66 ml, 10.6 mmol ) at room temperature and the reaction mixture was stirred for 3 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (25 ml) and the organic layer was washed with water (2×15 ml). The combined organics were dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The obtained crude material was sent to the next stage without any further purification. Yield: 88% (900 mg, almost white solid).

LCMS: (method E) 481.1 (M ⁺ ), Rt. 2.97 min, 61.59% (maximum).

Intermediate 180

7-Bromo-3-butyl-3-ethyl-8-hydroxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide

With stirring to a solution of 7-bromo-3-butyl-3-ethyl-8-methoxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1- dioxide (intermediate 179; 0.75 g, 1.50 mmol) in DCM (10 ml) was added BBr ₃ (1M in DCM, 2.33 ml, 2.30 mmol) at -78°C and then the reaction mixture was stirred for 3 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction was stopped with methanol (10 ml) and concentrated in vacuo. The resulting residue was dissolved in DCM (20 ml) and the organics washed with brine (20 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 23% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 27% (0.27 g, off-white solid).

LCMS: (method E) 469.0 (M ⁺ +2), Rt. 2.74 min, 86.35% (maximum).

Intermediate 181

Ethyl-(Z)-3-((7-bromo-3-butyl-3-ethyl-2-methyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

To a suspension of NaH (0.05 g, 1.39 mmol) in dry DMA (2 ml) at 0°C was added a solution of 7-bromo-3-butyl-3-ethyl-8-hydroxy-2-methyl-5-phenyl -2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-1,1-dioxide (intermediate 180; 0.2 g, 0.43 mmol) in DMA (2 ml) and the reaction mixture was stirred in for 10 min at room temperature. Ethyl 3-bromo-2,2-difluoropropionate (0.12 ml, 0.96 mmol) in DMA (3 ml) was then added dropwise at 0°C and the reaction mixture was heated for 3 h at 60°C. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0°C, the reaction was stopped with dilute HCl (1.5 N, 2 ml, pH~4) and concentrated in vacuo. The resulting residue was subjected to distribution between ice-cold water (10 ml) and EtOAc (20 ml) and the aqueous layer was extracted with EtOAc (2×30 ml). The combined organic layer was washed with ice-cold water (15 ml) and brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 8% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 60% (150 mg, almost white solid).

LCMS: (Method E) 585.1 (M ⁺ +2) Rt. 3.01 min, 88.73%

Intermediate 182

7-Bromo-3,3-dibutyl-5-(3,4-difluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

With stirring, to a solution of 7-bromo-3,3-dibutyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (3.0 g, 0.007 mol) in 3,4- difluorobromobenzene (5 ml) was added copper(I) iodide (0.286 mg, 0.0015 mol) and K ₂ CO ₃ (2.07 g, 0.015 mol) and the reaction mixture was purged with nitrogen for 20 min to degas. Tris[2-(2-methoxyethoxy)ethyl]amine (0.242 ml, 0.00075 mol) was then added under nitrogen atmosphere and the resulting reaction mixture was heated for 16 h at 130°C. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through celite and the celite layer was washed with EtOAc (50 ml). The filtrate was washed with water (50 ml) and brine (50 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 8-10% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 74% (2.8 g, pale brown solid).

LCMS: (Method E) 512.1 (M ⁺ ), Rt. 3.07 min, 85% (maximum).

Intermediate 183

7-Bromo-3,3-dibutyl-5-(3,4-difluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine

With stirring, to a solution of 7-bromo-3,3-dibutyl-5-(3,4-difluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (intermediate 182 ; 2.8 g, 0.0054 mmol) in THF (15 ml) at 0°C was added dropwise borane dimethyl sulfide (2M in THF, 8 ml, 0.016 mmol) and the reaction mixture was refluxed for 3 h at 60 °C After completion of the reaction (monitored by UPLC), the reaction mixture was cooled to 0°C, the reaction was stopped with methanol (10 ml) and then heated for 2 h at 60°C. The resulting reaction mixture was cooled to room temperature and concentrated in vacuo to give a crude product which was passed to the next step without any further purification. Yield: 1.49 g (crude colorless liquid).

Intermediate 184

7-Bromo-3,3-dibutyl-5-(3,4-difluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-5-(3,4-difluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine (intermediate 183; 1, 49 g, 0.01 mmol) in THF (75 ml) and water (7.5 ml) at room temperature was added oxone (38.3 g, 0.13 mmol) and the reaction mixture was stirred for 24 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a Buchner funnel and the filtrate was extracted with EtOAc (2×25 ml). The combined organic layer was washed with water (25 ml) and brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo 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: 62% (1 g, almost white solid).

LCMS: (Method A) 532.0 (M ⁺ +2), Rt. 3.31 min, 93.84% (maximum).

Intermediate 185

3,3-Dibutyl-5-(3,4-difluorophenyl)-8-methoxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring, to a solution of 7-bromo-3,3-dibutyl-5-(3,4-difluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide ( intermediate 184; 500 mg, 0.9757 mmol) in DMF (5 ml) was added sodium thiomethoxide (102 mg, 1.46 mmol) and the resulting reaction mixture was heated for 2 h at 60°C. After completion of the reaction (monitored by TLC), water (10 ml) was added to the reaction mixture and the aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with brine (500 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 18-20% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 82% (0.45 g, off-white solid).

LCMS: (Method E) 498.2 (M ⁺ +H), Rt. 3.57 min, 90.72% (maximum).

Intermediate 186

3,3-dibutyl-5-(3,4-difluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide

With stirring to a solution of 3,3-dibutyl-5-(3,4-difluorophenyl)-8-methoxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1- dioxide (intermediate 185; 450 mg, 0.9042 mmol) in DCM (5 ml) was added BBr ₃ (1M in DCM; 3 ml, 2.712 mmol) at 0°C and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), methanol (10 ml) was added dropwise at 0° C. until gas evolution ceased. Then the reaction mixture was diluted with DCM (10 ml) and the organic layer was washed with water (2×10 ml) and brine (10 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 100% EtOAc; silica gel: 230-400 mesh) to give the title compound. Yield: 91% (0.4 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.63 (s, 1H), 7.29 (s, 1H), 7.28-7.19 (m, 1H), 7.03-6 .98 (m, 1H), 6.72 (s, 1H), 6.64-6.59 (m, 1H), 3.62 (bs, 2H), 3.22 (s, 2H), 2, 22 (s, 3H), 1.40-1.34 (m, 4H), 1.15-1.05 (m, 8H), 0.79 (t, J =6.80 Hz, 6H). LCMS: (Method A) 484.2 (M ⁺ +H), Rt. 2.97 min, 94% (maximum).

Intermediate 187

Ethyl-(Z)-3-((3,3-dibutyl-5-(3,4-difluorophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate

With stirring to a solution of 3,3-dibutyl-5-(3,4-difluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1- dioxide (intermediate 186; 0.1 g, 0.021 mmol) in DMA (5 mL) NaH (60%, 0.03 g, 0.67 mmol) was added in portions at 0°C and the reaction mixture was stirred for 15 min at 0°C. Ethyl 3-bromo-2,2-difluoropropanoate (0.112 g, 0.052 mmol) was then added and the reaction mixture was heated for 5 h at 60°C. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C, the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was triturated with Et ₂ O. The resulting compound was dried in vacuo and passed on to the next step without any further purification. Yield: 56% (0.07 g, colorless gum).

Intermediate 188

Ethyl-(Z)-3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylate

With stirring to a solution of 3-butyl-7-(dimethylamino)-3-ethyl-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-1,1-dioxide (intermediate product 132; 0.2 g, 0.48 mmol) in DMA (5 ml) at 0°C NaH (60%, 0.04 g, 1.56 mmol) was added in portions and the reaction mixture was stirred for 15 min at 0°C. Ethyl 3-bromo-2,2-difluoropropanoate (0.26 g, 1.20 mmol) was then added and the reaction mixture was heated for 3 h at 70°C. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0°C, the reaction was stopped with 1.5 N. HCl (pH ~4) and diluted with water (15 ml). The aqueous layer was extracted with EtOAc (2×15 ml), the combined organic layer was washed with brine (10 ml) and then dried over anhydrous Na₂SO₄. The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 15-20% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 27% (0.07 g, colorless gum).

LCMS: (Method A) 533.2 (M ⁺ +H), Rt. 3.37 min, 80.27% (maximum).

Intermediate 189

Ethyl-(S)- (Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate and ethyl-(R)-(Z)-3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5 -phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate

The two enantiomers of racemic ( Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine -8-yl)oxy)-2-fluoroacrylate (0.340 g, 0.64 mmol) was separated by chiral preparative SFC (method G); wavelength: 280 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1 : Yield: 38% (130 mg, almost white solid). LCMS: (Method E) 533.1 (M ⁺ +H), Rt. 3.35 min, 95.24% (maximum). HPLC: (method B) Rt. 6.63 min, 96.05% (maximum). SFC: (method I) Rt. 2.35 min, 100% (maximum).

Enantiomer 2: Yield: 44% (150 mg, almost white solid). LCMS: (Method E) 533.1 (M ⁺ +H), Rt. 3.35 min, 93.83% (maximum). HPLC: (method B) Rt. 6.63 min, 97.12% (maximum). SFC: (method I) Rt. 2.97 min, 99.54% (maximum).

Example 1

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 1; 0.32 g, 0.58 mmol) in a mixture of 1,4-dioxane and water (6 ml, 2:1) was added lithium hydroxide (0.037 g, 0.58 mmol). 88 mmol) and the resulting mixture was stirred at room temperature for 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo. The resulting residue was acidified with dilute HCl (1.5 N., 2 ml) and the aqueous part was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (15 ml), brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the crude material was purified by preparative HPLC (method B) to give the title compound. Yield: 86% (300 mg, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.28 (bs, 1H), 7.66 (d, J =12.0 Hz, 1H), 7.47 (d, J =4.8 Hz, 1H), 7.33-7.29 (m, 2H), 7.18 (bs, 2H), 7.01 (t, J=6.8 Hz, 1H), 6.66 (d, J =4.0 Hz, 1H), 5.42 (dd, J =12.0, 4.4 Hz, 1H), 3.76 (bs, 2H), 3.38 (d, J =4.8 Hz , 2H), 2.16 (d, J =4.40 Hz, 3H), 1.41-1.33 (m, 4H), 1.08-1.01 (m, 8H), 0.75 ( t, J = 5.60 Hz, 6H). LCMS: (Method D) 518.3 (M+H), Rt. 3.04 min, 98.5% (maximum). HPLC: (method A) Rt. 5.75 min, 99.10% (maximum).

Example 2

(E) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring to a solution of (E) -3-((3,3-dibutyl-7-(methylthio)-5-(4-nitrophenyl)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-benzothiazepin-8-yl)oxy)acrylic acid (intermediate 2; 0.9 g, 1.60 mmol) in THF (20 ml) was added concentrated HCl (1 ml) and SnCl ₂ (1.2 g, 6 .40 mmol) at room temperature and the reaction mixture was stirred for 12 h at 70°C. After completion of the reaction (monitored by TLC), the reaction was stopped with a saturated solution of NaHCO ₃ , filtered through a pad of celite and then washed with EtOAc. The organic portion was washed with water (25 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the title compound. Yield: 94% (800 mg, crude brown solid).

LCMS: (Method A) 533.9 (M+H), Rt. 2.32 min, 81.07% (maximum).

Example 3

(E) -3-((5-(4-((tert-butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

With stirring to a solution of ( E )-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-benzothiazepin-8-yl)oxy)acrylic acid (Example 2; 0.2 g, 0.37 mol) in 1,4-dioxane (5 ml) at 0°C was added a solution of NaOH (0.03 g, 0 .56 mmol) in water (1 ml) followed by Boc anhydride (0.16 g, 0.75 mmol). Stirring was continued for 12 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with citric acid (pH ~3-4) and the aqueous layer was extracted with EtOAc (2×25 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method C) to give the title compound. Yield: 30% (70 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.24 (s, 1H), 9.32 (s, 1H), 7.64 (d, J =12.2 Hz, 1H), 7, 44 (s, 1H), 7.41 (d, J =9.2 Hz, 2H), 7.18 (d, J =8.4 Hz, 2H), 6.50 (s, 1H), 5, 38 (d, J =12.2 Hz, 1H), 3.74 (s, 2H), 3.40 (s, 2H), 2.13 (s, 3H), 1.47 (s, 9H), 1.43-1.40(m, 2H), 1.35-1.29(m, 2H), 1.13-1.02(m, 8H), 0.77-0.75(m, 6H ); LCMS: (Method B) 577.3 (M- ^t Bu), Rt. 3.14 min, 97.7% (maximum). HPLC: (method A) Rt. 5.95 min, 99.6% (maximum).

Example 4

(E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 3; 0.28 g, 0.48 mmol) in a mixture of 1,4-dioxane and water (3 ml; 5:1) was added lithium hydroxide (0.04 g, 0.96 mmol) and the resulting mixture was stirred for 12 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo. The resulting residue was acidified with dilute HCl (1.5 N., 2 ml) and the aqueous part was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (15 ml), brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the crude material was purified by Isolera column chromatography (eluent: 2-3% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 60% (150 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.29 (s, 1H), 7.73 (d, J= 12.0 Hz, 1H), 7.65-7.64 (m, 1H ), 7.37-7.34 (m, 2H), 7.28-7.25 (m, 2H), 7.09-7.05 (m, 2H), 5.39 (d, J= 12 .0 Hz, 1H), 3.88-3.73 (m, 2H), 3.46 (s, 2H), 1.52-1.34 (m, 4H), 1.08-0.98 ( m, 4H), 0.72-0.69 (m, 6H). LCMS: (Method A) 296.0 (M+H), Rt. 2.86 min, 95.59% (maximum). HPLC: (method B) Rt. 6.03 min, 97.02% (maximum).

Examples 5 and 6

(R)- (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylic acid and (S)- (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

The two enantiomers of racemic (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8 -yl)oxy)acrylic acid ((Example 4; 150 mg, 0.29 mmol) was separated by chiral preparative SFC (method A), mobile phase: CO ₂ : IPA (70:30), wavelength: 220 nm; cycle time: 5 min back pressure: 100 bar The material was concentrated in vacuo at 40° C. The first eluting fraction was enantiomer 1 and the second eluting fraction was enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 17% (25 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.29 (s, 1H), 7.73 (d, J= 11.6 Hz, 1H), 7.64 (s, 1H), 7, 38-7.34 (m, 2H), 7.27-7.26 (m, 2H), 7.10-7.05 (m, 2H), 5.39 (d, J= 12.4 Hz, 1H), 3.80-3.78 (m, 2H), 3.46 (s, 2H), 1.52-1.34 (m, 4H), 1.08-0.98 (m, 4H) , 0.72-0.69 (m, 6H). LCMS: (method A) 521.8 (M ⁺ ), Rt. 2.86 min, 99.14% (maximum). HPLC: (method B) Rt. 5.99 min, 96.82% (maximum). SFC: (method A) Rt. 4.88 min, 99.31%.

Enantiomer 2: Yield: 20% (30 mg, almost white solid). ¹ H NMR (400 MHz, DMSO): δ 12.28 (s, 1H), 7.73 (d, J= 12.4 Hz, 1H), 7.64 (s, 1H), 7.38-7 .34 (m, 2H), 7.26-7.24 (m, 2H), 7.10-7.05 (m, 2H), 5.39 (d, J= 12.0 Hz, 1H), 3.80-3.75 (m, 2H), 3.46 (s, 2H), 1.51-1.30 (m, 4H), 1.17-0.98 (m, 4H), 0. 76-0.68 (m, 6H). LCMS: (Method A) 524.0 (M+ 2), Rt. 2.96 min, 98.77% (maximum). HPLC: (method B) Rt. 5.99 min, 97.71% (maximum). SFC: (method A) Rt. 5.95 min, 98.99%.

Example 7

(E) -3-((3,3-dibutyl-7-cyclopropyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-cyclopropyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylate (intermediate 6; 0.150 g, 0.28 mmol) in a mixture of 1,4-dioxane and water (6 ml, 5:1) was added lithium hydroxide (24 mg, 0.06 mmol) and the resulting mixture was stirred at room temperature for 12 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo to give a crude residue which was acidified with dilute HCl (1.5 N., 2 ml) and the aqueous layer was extracted with EtOAc (2×15 ml). The organic part was washed with water (15 ml), brine (15 ml), dried over anhydrous Na₂SO₄ and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 50% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 34% (48 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.19 (bs, 1H), 7.76 (d, J=12.0 Hz, 1H), 7.47 (s, 1H), 7, 28 (t, J=8.0 Hz, 2H), 7.11 (d, J=7.6 Hz, 2H), 6.98 (t, J=7.2 Hz, 1H), 6.41 ( s, 1H), 5.37 (d, J=12.4 Hz, 1H), 3.73 (bs, 2H), 3.36 (s, 2H), 1.97-1.90 (m, 1H ), 1.42-1.34 (m, 2H), 1.31-1.24 (m, 2H), 1.10-1.03 (m, 8H), 1.00-0.98 (m , 2H), 0.9 (t, J=4.4 Hz, 6H), 0.42-0.41 (m, 2H). LCMS: (Method A) 512.2 (M+H), Rt. 3.13 min, 99.21% (maximum). HPLC: (method A) Rt. 6.05 min, 98.83% (maximum).

Example 8

(E) -3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 9; 0.08 g, 0.15 mmol) in a mixture of 1,4-dioxane and water (6 ml, 5:1) was added lithium hydroxide (0.012 g, 0.15 mmol). 29 mmol) and the resulting mixture was stirred at room temperature overnight. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo to give a crude residue which was acidified with dilute HCl (1.5 N, 2 ml) and the aqueous layer was extracted with EtOAc (2×15 ml). The organic portion was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 4-5% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. The resulting material was further purified by preparative HPLC (method A) to give the title compound. Yield: 9% (7 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.16 (s, 1H), 7.58 (d, J =12.4 Hz, 1H), 7.38 (s, 1H), 7, 31-7.29 (m, 2H), 7.17-7.15 (m, 2H), 6.99-6.96 (m, 2H), 6.26 (s, 1H), 5.40 ( d, J =12.0 Hz, 2H), 3.73(s, 2H), 3.29(s, 2H), 2.68(s, 6H), 1.41-1.32(m, 4H ), 1.15-1.00 (m, 8H), 0.77-0.73 (m, 6H). LCMS: (Method C) 515.2 (M+ 2H), Rt. 3.13 min, 97.01% (maximum). HPLC: (method B) Rt. 5.80 min, 94.38% (maximum).

Example 9

(E) -3-((3,3-dibutyl-5-(4-(cyclopropanesulfonamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of ( E )-3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-benzothiazepin-8-yl)oxy)acrylic acid (Example 2; 0.12 g, 0.22 mmol) in pyridine (3 ml) was added DMAP (0.03 mg, 0.02 mmol) and cyclopropanesulfonyl chloride (0.063 mg 0.45 mmol) at 0°C and the reaction mixture was stirred for 12 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (25 ml). The organic layer was then washed with saturated NaHCO ₃ solution (15 ml) and water (15 ml). The organic was dried over anhydrous Na ₂ SO ₄ , concentrated in vacuo and the resulting crude material was purified by preparative HPLC (method C) to give the title compound. Yield: 14% (20 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.45-7.40 (m, 2H), 7.17 (m, 4H), 6.60 (s, 1H), 5.41 (d , J =12.0 Hz, 1H), 3.73(m, 2H), 3.54(m, 2H), 2.16(s, 3H), 1.89(s, 1H), 1.42 -1.30 (m, 4H), 1.12-1.08 (m, 8H), 1.04-1.03 (m, 4H), 0.88-0.85 (m, 6H). LCMS: (Method B) 635.2 (M ⁺ -H), Rt. 3.14 min, 94.6% (maximum). HPLC: (method A) Rt. 5.28 min, 99.49% (maximum).

Example 10

(E) -3-((3,3-dibutyl-5-(4-(methylsulfonamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of (E) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-benzothiazepin-8-yl)oxy)acrylic acid (Example 2; 0.11 g, 0.18 mmol) in pyridine (3 ml) was added DMAP (0.03 mg, 0.02 mmol) and methanesulfonyl chloride (0. 03 ml, 0.28 mmol) at 0°C and the reaction mixture was stirred for 12 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (25 ml). The organic layer was then washed with saturated NaHCO ₃ solution (15 ml) and water (15 ml). The organic was dried over anhydrous Na ₂ SO ₄ , concentrated in vacuo and the resulting crude material was purified by preparative HPLC (method B) to give the title compound. Yield: 5% (7 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d6 ): 7.79 (d, J =1.2 Hz, 1H), 7.68 (s, 1H), 7.28 (s _, 1H), 7.23 (d, J =8.8 Hz, 1H), 7.13 (d, J =8.8 Hz, 2H), 6.66 (s, 1H), 6.62 (s, 1H), 5.56 (d, J =12.0 Hz, 1H), 3.75 (bs, 2H), 3.22 (s, 2H), 3.02 (d, J =5.60 Hz, 3H), 2.22 (s, 3H), 1.57-1.52 (m, 2H), 1.48-1.41 (m, 6H), 1.28-1.21 (m, 2H), 1.19-1 .08 (m, 2H), 0.8 (t, J = 6.8 Hz, 6H). LCMS: (Method D) 609.2 (M ⁺ -H), Rt. 2.42 min, 91.68% (maximum). HPLC: (Method C) Rt. 9.87 min, 96.13% (maximum).

Example 11

(Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 10; 8.2 g, 15.0 mmol) in a mixture of 1,4-dioxane and water (82 ml, 4:1) was added LiOH.H ₂ O (1.25 g, 30.0 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with 1.5 N sodium hydroxide. HCl (pH~4) and diluted with water (100 ml). The aqueous portion was extracted with EtOAc (150 ml) and then the organic layer was washed with brine (100 ml) and dried over anhydrous Na ₂ SO _{4 .} -4% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 52% (6.73 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.59 (s, 1H), 7.56 (d, J =18.4 Hz, 1H), 7.56 (s, 1H), 7, 30 (t, J =7.6 Hz, 2H), 7.16 (d, J =7.6 Hz, 2H), 6.99 (t, J =7.2 Hz, 1H), 6.67 ( s, 1H), 3.75 (s, 2H), 3.36 (s, 2H), 2.18 (s, 3H), 1.43-1.41 (m, 2H), 1.36-1 .30 (m, 2H), 1.18-1.01 (m, 8H), 0.77-0.72 (m, 6H). LCMS: (Method A) 536.1 (M ⁺ +H), Rt. 3.19 min, 97.92% (maximum). HPLC: (method B) Rt. 6.45 min, 99.04% (maximum).

Example 12

(E) -3-((3,3-dibutyl-7-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylate (intermediate 14; 0.055 g, 0.1 mmol) in a mixture of 1,4-dioxane and water (6 ml, 5:1) was added lithium hydroxide (8 mg, 0.2 mmol) and the resulting mixture was stirred at room temperature for 4 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo to give a crude residue, which was acidified with dilute HCl (1.5 N, 2 ml) and the aqueous layer was extracted with EtOAc (2×15 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 80% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 14.4% (7.5 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.74 (d, J= 12.1 Hz, 1H), 7.48 (s, 1H), 7.29 (t, J =7.5 Hz, 2H), 7.17-7.15 (m, 2H), 6.98 (t, J = 7.3 Hz, 1H), 6.84 (s, 1H), 5.40 (d, J = 12.1 Hz, 1H), 3.88-3.65 (m, 2H), 3.37 (s, 2H), 2.49-2.48 (m, 2H), 1.41-1, 24 (m, 6H), 1.09-1.04 (m, 6H), 1.01-0.97 (m, 4H), 0.80-0.75 (m, 6H), LCMS: (method A) 500.3 (M+H), Rt. 3.16 min, 98.8% (maximum). HPLC: (method B) Rt. 6.58 min, 97.03% (maximum).

Example 13

(E) -3-((3-butyl-3-ethyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3-butyl-3-ethyl-7-methoxy-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 16; 0.125 g, 0.24 mmol) in a mixture of 1,4-dioxane and water (3.5 ml; 6:1) was added lithium hydroxide (32 mg, 0. 74 mmol) and the resulting mixture was stirred at room temperature for 4 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo to give a crude residue which was acidified with dilute HCl (1.5 N, 2 mL). Then the aqueous layer was extracted with EtOAc (2×15 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 80% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 21% (23 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.12 (s, 1H), 7.65 (d, J =12.0 Hz, 1H), 7.53 (s, 1H), 7, 30 (t, J= 7.6 Hz, 2H), 7.17-7.15 (m, 2H), 6.98 (t, J =6.8 Hz, 1H), 6.56 (s, 1H ), 5.31 (d, J =12.0 Hz, 1H), 3.85-3.65 (m, 2H), 3.61 (s, 3H), 3.31 (s, 2H), 1 .53-1.31 (m, 2H), 1.28-1.25 (m, 2H), 1.24-1.09 (m, 4H), 0.75-0.71 (m, 6H) . LCMS: (Method D) 474.2 (M+H), Rt. 2.55 min, 99.31% (maximum). HPLC: (method B) Rt. 5.59 min, 98.87% (maximum).

Example 14

(E) -3-((3,3-dibutyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-methoxy-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylate (intermediate 19; 0.23 g, 0.43 mmol) in a mixture of 1,4-dioxane and water (7 ml, 6:1) was added lithium hydroxide (0.13 g, 3. 03 mmol) and the resulting mixture was stirred at room temperature for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo to give a crude residue which was acidified with dilute HCl (1.5 N, 2 mL). Then the aqueous layer was extracted with EtOAc (2×15 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by Isolera column chromatography (eluent: 80% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 75% (160 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.64 (d, J =12.4 Hz, 1H), 7.52 (s, 1H), 7.30 (t, J= 8.0 Hz, 2H), 7.19 (d, J=7.2 Hz, 2H), 7.00 (t, J =7.2 Hz, 1H), 6.52 (s, 1H), 5.30 ( d, J =12.4 Hz, 1H), 3.76(bs, 2H), 3.61(s, 3H), 3.31(s, 2H), 1.42-1.33(m, 2H ), 1.29-1.20 (m, 2H), 1.10-0.98 (m, 8H), 0.86-0.84 (m, 6H). LCMS: (Method C) 502.1 (M+H), Rt. 2.99 min, 96.68% (maximum). HPLC: (method B) Rt. 6.12 min, 96.05% (maximum).

Example 15

(E) -3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- Lithium hydroxide (33 mg, 0.80 mmol) and the resulting mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated in vacuo to give a crude residue which was acidified with dilute HCl (1.5 N, 2 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 89% (35 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.26 (bs, 1H), 7.71 (d, J =12.4 Hz, 1H), 7.50-7.40 (m, 3H ), 7.12-6.91 (m, 2H), 6.84 (s, 1H), 5.48-5.44 (m, 1H), 3.85-3.62 (m, 2H), 3.33 (s, 2H), 2.26 (s, 3H), 1.44-1.05 (m, 12H), 0.85-0.75 (m, 6H). LCMS: (Method D) 596.1 (M+H), Rt. 3.23 min, 97.39% (maximum). HPLC: (method B) Rt. 6.63 min, 97.05% (maximum).

Example 16

(E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-benzothiazepin-8-yl)oxy)acrylate (intermediate 24; 0.3 g, 0.58 mmol) in a mixture of 1,4-dioxane (10 ml) and water (3 ml) was added lithium hydroxide (0. 12 g, 2.89 mmol) and the resulting mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated in vacuo to give a crude residue which was acidified with dilute HCl (1.5 N., 2 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 42% (120 mg, pale brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.28 (bs, 1H), 7.67 (d, J =12.4 Hz, 1H), 7.49 (s, 1H), 7, 30 (t, J =8.0 Hz, 2H), 7.16 (d, J =7.6 Hz, 2H), 6.99 (t, J =7.2 Hz, 1H), 6.69 ( s, 1H), 5.43 (d, J =12.0 Hz, 1H), 3.82-3.62 (m, 2H), 3.38 (s, 2H), 2.18 (s, 3H ), 1.56-1.32 (m, 4H), 1.20-1.02 (m, 4H), 0.75-0.69 (m, 6H). LCMS: (Method C) 490.0 (M+H), Rt. 2.89 min, 97.98% (maximum). HPLC: (method B) Rt. 5.84 min, 96.52% (maximum).

Examples 17 and 18

(R)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid and (S)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3 ,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

Racemic enantiomers (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 8-yl)hydroxy)acrylic acid (Example 16; 100 mg, 0.20 mmol) was separated by chiral preparative SFC (method B); mobile phase: CO ₂ : IPA (70:30); wavelength: 220 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction gave enantiomer 1 of 98.01% chiral purity and the second fraction gave the second enantiomer as a mixture. This mixture was separated by chiral preparative SFC (method C); mobile phase: CO ₂ : ammonia in methanol (70:30); wavelength: 220 nm; cycle time: 5 min; back pressure: 100 bar. The first eluting fraction was concentrated in vacuo at 40°C to give the second enantiomer with a chiral purity of 100%. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 20% (20 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.58-7.57 (m, 1H), 7.46 (s, 1H), 7.29 (t, J =8.0 Hz, 2H ), 7.14 (d, J =7.6 Hz, 2H), 6.98 (t, J =7.2 Hz, 1H), 6.69 (s, 1H), 5.43 (d, J =12.4 Hz, 1H), 3.82-3.62 (m, 2H), 3.37 (s, 2H), 2.18 (s, 3H), 1.54-0.98 (m, 8H), 0.76-0.72 (m, 6H). LCMS: (Method C) 490.1 (M+H), Rt. 2.87 min, 95.62% (maximum). HPLC: (method B) Rt. 5.86 min, 97.74% (maximum). Chiral purity: (method B, chiral SFC) Rt. 4.69 min, 98.01% (maximum).

Enantiomer 2: Yield: 13% (15 mg, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.66-7.63 (m, 1H), 7.47 (s, 1H), 7.29 (t, J =7.6 Hz, 2H ), 7.15 (d, J = 8.0 Hz, 2H), 6.98 (t, J = 7.2 Hz, 1H), 6.68 (s, 1H), 5.42 (d, J =12.4 Hz, 1H), 3.82-3.62 (m, 2H), 3.36 (s, 2H), 2.17 (s, 3H), 1.53-0.98 (m, 8H), 0.76-0.72 (m, 6H). LCMS: (Method C) 490.1 (M+H), Rt. 2.87 min, 98.68% (maximum). HPLC: (method B) Rt. 5.86 min, 98.65% (maximum). Chiral purity: (method B, chiral SFC) Rt. 5.28 min, 100% (maximum).

Example 19

(E) -3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- Lithium hydroxide (0 04 g, 0.85 mmol) and the resulting mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated in vacuo to give a crude residue, which was acidified with dilute HCl (1.5 N. , 2 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 89% (35 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.35-7.33 (m, 2H), 7.23 (d, J =8.0 Hz, 2H), 6.94-6.91 (m, 2H), 6.42 (s, 1H), 5.35 (d, J = 11.6 Hz, 1H), 3.82-3.68 (m, 5H), 3.39 (s, 2H), 2.09 (s, 3H), 1.41-0.95 (m, 12H), 0.75-0.72 (m, 6H). LCMS: (Method B) 548.2 (M+H), Rt. 2.97 min, 98.03% (maximum). HPLC: (method B) Rt. 6.28 min, 99.26% (maximum).

Example 20

(E) -3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (Intermediate 27; 0.09 g, 0.17 mmol) in a mixture of 1,4-dioxane (10 mL) and water (3 mL) was added lithium hydroxide ( 0.04 g, 0.85 mmol) and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated in vacuo to give a crude residue which was acidified with dilute HCl (1.5 N ., 2 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 21% (20 mg, brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.61 (d, J =12.4 Hz, 1H), 7.41 (s, 1H), 7.18 (d, J =8.4 Hz, 2H), 6.78 (d, J = 8.4 Hz, 2H), 6.35 (s, 1H), 5.33 (d, J = 12.4 Hz, 1H), 3.78- 3.63 (m, 2H), 3.45-3.39 (m, 2H), 2.09 (s, 3H), 1.43-0.98 (m, 12H), 0.77-0, 73(m, 6H). LCMS: (Method B) 534.2 (M+H), Rt. 2.71 min, 95.25% (maximum). HPLC: (method B) Rt. 5.51 min, 95.27% (maximum).

Example 21

(E) -3-((3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)acrylate (intermediate 28; 0.156 g, 0.33 mmol) in a mixture of 1,4-dioxane (10 ml) and water (3 ml) was added lithium hydroxide (0.03 g, 0.85 mmol) and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated in vacuo to give a crude residue which was acidified with dilute HCl (1.5N, 2 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 2-3% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 21% (25 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d6 ): δ 7.64 (d, J =12.2 Hz, 1H) _, 7.52 (d, J =2.7 Hz, 1H), 7.35 -7.26 (m, 3H), 7.13 (d, J =7.4 Hz, 2H), 6.96 (t, J= 7.7 Hz, 2H), 5.49 (d, J = 12.1 Hz, 1H), 3.77-3.74 (bs, 2H), 3.38-3.33 (bs, 2H), 1.52-1.35 (m, 4H), 1.33 -1.15 (m, 4H), 1.01-0.96 (m, 6H). LCMS: (Method A) 441.9 (MH), Rt. 2.69 min, 96.87% (maximum). HPLC: (method A) Rt. 5.25 min, 99.10% (maximum).

Example 22

( E )-3-((5-(4-(benzylamino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of (E) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylic acid (Example 2; 0.1 g, 0.18 mol) in MeOH (6 ml) at 0°C was added dropwise benzaldehyde (0.04 g, 0.37 mmol ) and acetic acid (0.05 ml). Stirring was continued for 1 hour at room temperature. Then NaCNBH ₃ (0.03 g, 0.37 mmol) was added and the reaction mixture was stirred for 12 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was made basic with saturated NaHCO ₃ solution (15 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (15 ml), brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method C) to give the title compound. Yield: 16% (17.79 mg, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.45 (d, J =12.0 Hz, 1H), 7.35-7.28 (m, 5H), 7.23-7.19 (m, 1H), 7.07 (d, J =8.4 Hz, 2H), 6.61 (d, J =8.8 Hz, 2H), 6.31 (t, J =6.0 Hz , 1H), 6.26 (s, 1H), 5.30 (d, J =12.0 Hz, 1H), 4.30 (d, J =6.0 Hz, 2H), 3.71 (bs , 2H), 3.40 (s, 2H), 2.08 (s, 3H), 1.45-1.37 (m, 4H), 1.11-1.07 (m, 8H), 1, 04 (t, J =7.20 Hz, 6H); LCMS: (Method C) 623.2 (M ⁺ +H), Rt. 3.18 min, 97.02% (maximum). HPLC: (method A) Rt. 6.08 min, 97.17% (maximum).

Example 23

( E )-3-((7-bromo-5-(4-((tert-butoxycarbonyl)amino)phenyl)-3-butyl-3-ethyl-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

With stirring to a solution of ( E )-3-((5-(4-aminophenyl)-7-bromo-3-butyl-3-ethyl-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylic acid (intermediate 30; 0.12 g, 0.22 mol) in 1,4-dioxane (5 ml) was added at 0°C a solution of NaOH (0.013 g, 0. 33 mmol) in water (1 ml) followed by Boc anhydride (0.097 g, 0.44 mmol). Stirring was continued for 12 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with citric acid (~pH 3-4) and the aqueous layer was extracted with EtOAc (2×25 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method C) to give the title compound. Yield: 10% (16 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.15 (s, 1H), 9.39 (s, 1H), 7.66 (d, J =12.0 Hz, 1H), 7, 59 (s, 1H), 7.46 (d, J =8.8 Hz, 2H), 7.24 (d, J =8.4 Hz, 2H), 6.85 (s, 1H), 5, 33 (d, J =12.0 Hz, 1H), 3.76 (bs, 2H), 3.48 (s, 2H), 1.48 (s, 9H), 1.33-1.31 (m , 4H), 1.11-1.07 (m, 2H), 1.01-0.97 (m, 2H), 0.73 (t, J =6.80 Hz, 6H); LCMS: (method B) 635.1 (M ⁺ -H), Rt. 2.96 min, 97.27% (maximum). HPLC: (method A) Rt. 5.19 min, 97.97% (maximum).

Example 24

(E) -3-((3,3-dibutyl-5-(4-((methoxycarbonyl)amino)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylic acid

With stirring to a solution of methyl-(E)-3-((3,3-dibutyl-5-(4-((methoxycarbonyl)amino)phenyl)-7-(methylthio)-1,1-dioxido-2,3, 4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (Intermediate 34; 0.06 g, 0.09 mol) in 1,4-dioxane/water (5 ml, 4:1) ) was added lithium hydroxide (6 mg, 0.14 mmol) and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1 ml, 1.5 N.) and the aqueous layer was extracted with EtOAc (2×10 ml). The organic portion was washed with water (8 ml) and brine (8 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method C) to give the title compound. Yield: 29% (17 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.62 (s, 1H), 7.65 (d, J =12.0 Hz, 1H), 7.44 (d, J =3.6 Hz, 2H), 7.42 (s, 1H), 7.20 (d, J = 8.4 Hz, 2H), 7.13 (s, 1H), 6.52 (s, 1H), 5, 38 (d, J =12.4 Hz, 1H), 3.74 (bs, 2H), 3.66 (s, 3H), 3.40 (s, 2H), 2.13 (s, 3H), 1.24-1.13 (m, 2H), 1.11-1.10 (m, 2H), 1.02-1.00 (m, 8H), 0.7 (t, J =6.4 Hz, 6H). LCMS: (Method B) 591.2 (M ⁺ +H), Rt. 2.78 min, 98.32% (maximum). HPLC: (method A) Rt. 5.18 min, 98.08% (maximum).

Example 25

(E) -3-((3,3-dibutyl-5-(4-(dimethylamino)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring, to a solution of methyl-(E)-3-((3,3-dibutyl-5-(4-(dimethylamino)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 36; 46 mg, 0.08 mmol) in a mixture of 1,4-dioxane and water (5 ml, 4:1) was added lithium hydroxide ( 6.7 mg, 0.16 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 1 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The organic portion was washed with water (10 ml), brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was triturated with cold methanol to give the title compound. Yield: 29% (13 mg, pale yellow solid).

¹ H-NMR (400 MHz, DMSO- d6 ): 12.19 (s, 1H) _, 7.62 (d, J =9.6 Hz, 1H), 7.41 (s, 1H), 7, 20 (d, J =6.4 Hz, 2H), 6.76 (d, J =6.0 Hz, 2H), 6.38 (s, 1H), 5.33 (d, J= 11.2 Hz, 1H), 3.76 (s, 2H), 3.43 (s, 2H), 2.89 (s, 6H), 2.09 (s, 3H), 1.44-1.39 (m , 4H), 1.34-0.99 (m, 8H), 0.76-0.73 (m, 6H). LCMS: (Method C) 561.2 (M+H), Rt. 2.85 min, 96.89% (maximum), HPLC: (method B) Rt. 4.45 min, 97.85% (maximum).

Example 26

(Z) -3-((5-(4-((tert-butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (Intermediate 38; 0.24 g, 0.43 mol) in 1,4-dioxane (2.4 mL) NaOH (0.026 g, 0.65 mmol) in water (0.48 ml) and then Boc anhydride (0.19 g, 0.87 mmol) and stirring was continued for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with citric acid (~pH 3-4) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 5% (15 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.57 (s, 1H), 9.30 (s, 1H), 7.50-7.47 (m, 2H), 7.42 (d , J= 8.4 Hz, 2H), 7.14 (d, J = 8.4 Hz, 2H), 6.52 (s, 1H), 3.72 (s, 2H), 3.37 (s , 2H), 2.15 (s, 3H), 1.48 (s, 9H), 1.40-1.35 (m, 4H), 1.16-1.03 (m, 8H), 0, 77-0.74 (m, 6H); LCMS: (Method A) 651.3 (M+H), Rt. 3.01 min, 95.75% (maximum). HPLC: (method B) Rt. 6.46 min, 95.23% (maximum).

Example 27

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro- Lithium hydroxide (0.013 g 0.31 mmol) and the reaction mixture was stirred at room temperature overnight. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1 ml, 1.5 N.) and the aqueous layer was extracted with EtOAc (2×10 ml). The organic portion was washed with water (8 ml) and brine (8 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method C) to give the title compound. Yield: 48% (30 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.24 (s, 1H), 9.16 (s, 1H), 7.66 (d, J =12.0 Hz, 2H), 7 .60 (d, J = 8.4 Hz, 1H), 7.46 (s, 1H), 7.16 (d, J = 8.4 Hz, 2H), 6.61 (s, 1H), 5 .41 (d, J = 12.4 Hz, 1H), 3.74 (bs, 2H), 3.38 (s, 2H), 2.17 (s, 3H), 1.22-1.14 ( m, 2H), 1.13-1.11 (m, 2H), 1.09 (s, 9H), 1.06-1.05 (m, 8H), 0.8 (t, J =6, 8Hz, 6H). LCMS: (Method B) 617.2 (M ⁺ +H), Rt. 2.92 min, 97.89% (maximum). HPLC: (Method C) Rt. 6.04 min, 95.46% (maximum).

Example 28

(Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, triethylamine (0.61 ml, 4.40 mmol). Then pivaloyl chloride (0.22 ml, 1.76 mmol) was added and stirring was continued for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml) and the organic layer was washed with cold water (15 ml) and brine (15 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give the crude material. The resulting crude material was purified by preparative HPLC (method B) to give the title compound. Yield: 21% (0.2 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ) B362441: δ 13.58 (s, 1H), 9.15 (s, 1H), 7.59-7.49 (m, 4H), 7.12 ( d, J = 8.4 Hz, 2H), 6.62 (s, 1H), 3.71 (s, 2H), 3.36 (s, 2H), 2.19 (s, 3H), 1, 46-1.43 (m, 2H), 1.36-1.30 (m, 2H), 1.22 (s, 9H), 1.13-1.07 (m, 8H), 0.78- 0.75 (m, 6H); LCMS (B361751): (Method A) 635.2 (M ⁺ +H), Rt. 2.88 min, 96.98% (maximum). HPLC: (method B) Rt. 6.12 min, 97.61% (maximum).

Example 29

(E) -3-((5-(4-((butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((5-(4-((butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3, 4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 40; 0.041 g, 0.06 mmol) in 1,4-dioxane and water (3 mL, 4:1) was added lithium hydroxide (0.008 g, 0.19 mmol) and the reaction mixture was stirred at room temperature overnight. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1 ml, 1.5 N.) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organics were washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method C) to give the title compound. Yield: 20% (6.1 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.23 (bs, 1H), 9.58 (s, 1H), 7.65 (d, J =12.0 Hz, 1H), 7, 44-7.42 (m, 3H), 7.19 (d, J =8.8 Hz, 2H), 6.52 (s, 1H), 5.38 (d, J =12.0 Hz, 1H ), 4.07 (t, J =6.4 Hz, 2H), 3.75 (bs, 2H), 3.40 (s, 2H), 2.13 (s, 3H), 1.62-1 .58 (m, 2H), 1.43-1.39 (m, 5H), 1.37-1.33 (m, 5H), 1.24-1.10 (m, 4H), 0.9 (t, J =7.6 Hz, 3H), 0.7 (t, J =6.4 Hz, 6H). LCMS: (method C) 633.2 (M ⁺ +1), Rt. 3.06 min, 98.93% (maximum). HPLC: (method B) Rt. 6.41 min, 97.46% (maximum).

Example 30

(E) -3-((3,3-dibutyl-5-(4-(3,3-dimethylbutanamido)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro -1,5-Benzothiazepin-8-yl)oxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-(3,3-dimethylbutanamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3 ,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 41; 0.15 g, 0.22 mmol) in 1,4-dioxane and water (3 ml, 4: 1) lithium hydroxide (0.038 g, 0.91 mmol) was added at 0° C. and then the reaction mixture was stirred at room temperature overnight. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1 ml, 1.5 N.) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organics were washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude compound was purified by fractional crystallization using a combination of DCM/methanol and hexane. Yield: 59% (84.5 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.22 (s, 1H), 9.75 (s, 1H), 7.66 (d, J =12.0 Hz, 1H), 7 .56 (d, J =8.8 Hz, 2H), 7.46 (s, 1H), 7.18 (d, J =8.0 Hz, 2H), 6.59 (s, 1H), 5 .39 (d, J =12.4 Hz, 1H), 3.75 (bs, 2H), 3.39 (s, 2H), 2.17 (s, 3H), 2.16 (s, 2H) , 1.43-1.32 (m, 4H), 1.13-1.07 (m, 8H), 1.02 (s, 9H), 0.8 (t, J =6.8 Hz, 6H ). LCMS: (Method C) 631.2 (M ⁺ +H), Rt. 2.97 min, 96.38% (maximum). HPLC: (method B) Rt. 6.14 min, 97.17% (maximum).

Example 31

(Z) -3-((3,3-dibutyl-5-(4-isobutyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (intermediate 38; 0.2 g, 0.36 mmol) in DMF (2 ml) was added isobutyric acid (0.041 g, 0.47 mmol), HATU (0.27 g, 0.73 mmol) and N-methylmorpholine (0.12 ml, 1.10 mmol) and the resulting mixture was stirred overnight at room temperature. After completion of the reaction (monitored by TLC), water (5 ml) was added and the reaction mixture was extracted with EtOAc (2×10 ml). The combined organic layer was washed with brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method B) to give the title compound. Yield: 10% (10 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.77 (s, 1H), 7.57-7.51 (m, 4H), 7.13 (d, J =8.0 Hz, 2H ), 6.60 (s, 1H), 3.71 (s, 2H), 3.36 (s, 2H), 2.59-2.57 (m, 1H), 2.18 (s, 3H) , 1.44-1.42 (m, 2H), 1.36-1.30 (m, 2H), 1.11-1.09 (m, 14H), 0.78-0.76 (m, 6H). LCMS: (Method A) 621.2 (M+H), Rt. 2.71 min, 93.97% (maximum). HPLC: (method B) Rt. 5.75 min, 93.01% (maximum).

Example 32

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-(trifluoromethyl)phenyl)-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 46; 0.09 g, 0.14 mmol) in a mixture of 1,4-dioxane and water (5 ml, 4:1) hydroxide was added lithium (0.012 g, 0.29 mmol) and the reaction mixture was stirred for 3 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N., 3 ml) and the aqueous layer was extracted with EtOAc (2×8 ml). The combined organic layer was washed with water (8 ml) and brine (8 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 5% MeOH DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 16% (0.014 g, white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.31 (s, 1H), 7.75 (d, J =12.1 Hz, 1H), 7.54-7.51 (m, 3H), 7.07-7.05 (m, 3H), 5.53 (d, J =12.2 Hz, 1H), 3.80 (s, 2H), 3.37 (s, 2H), 2.33 (s, 3H), 1.51-1.48 (m, 2H), 1.33-1.11 (m, 10H), 0.82-0.75 (m, 6H). LCMS: (method C) 585.8 (M ⁺ +H), Rt. 3.07 min, 99.24% (maximum). HPLC: (method A) Rt. 6.50 min, 93.77% (maximum).

Example 33

(E) -3-((3,3-dibutyl-5-(4-isobutyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-isobutyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- Lithium hydroxide (0.035 g, 0.82 mmol) at 0°C and the reaction mixture was stirred for 3 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N., 4 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The organic portion was washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method C). Yield: 29% (36 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.23 (s, 1H), 9.79 (s, 1H), 7.66 (d, J =12.4 Hz, 1H), 7 .57 (d, J = 8.8 Hz, 2H), 7.46 (s, 1H), 7.17 (d, J = 8.4 Hz, 2H), 6.58 (s, 1H), 5 .40 (d, J = 12.0 Hz, 1H), 3.73 (bs, 2H), 3.39 (s, 2H), 3.35-3.34 (m, 1H), 2.16 ( s, 3H), 1.44-1.33 (m, 4H), 1.25-1.20 (m, 6H), 1.11-1.01 (m, 8H), 0.8 (t, J =5.6Hz, 6H). LCMS: (Method C) 603.2 (M ⁺ +H), Rt. 2.81 min, 95.01% (maximum). HPLC: (method B) Rt. 5.7 min, 95.73% (maximum).

Example 34

(E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 51; 0.135 g, 0.21 mmol) in a mixture of 1,4-dioxane and water (5 ml, 4:1) at 0°C was added lithium hydroxide (0.027 g, 0.65 mmol) and then the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N., 4 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The organic portion was washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method C). Yield: 46% (58 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.22 (bs, 1H), 9.15 (s, 1H), 7.65 (d, J =12.4 Hz, 1H), 7, 58 (d, J =8.8 Hz, 2H), 7.46 (s, 1H), 7.14 (d, J =8.0 Hz, 2H), 6.61 (s, 1H), 5, 40 (d, J =12.4 Hz, 1H), 3.73 (bs, 2H), 3.37 (s, 2H), 2.16 (s, 3H), 1.51-1.41 (m , 4H), 1.21 (s, 9H), 1.14-1.07 (m, 4H), 0.77 (t, J =6.80 Hz, 6H). LCMS: (Method B) 587.2 (M ⁺ -H), Rt. 2.69 min, 98.5% (maximum). HPLC: (method B) Rt. 5.57 min, 99.14% (maximum).

Example 35

(Z) -3-((3,3-dibutyl-5-(4-(cyclopentanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (intermediate 38; 0.05 g, 0.091 mmol) in DMF (0.5 ml) was added cyclopentanecarboxylic acid (0.016 g, 0.13 mmol), HATU (0.069 g, 0.18 mmol) and N-methylmorpholine (0.03 ml, 0.27 mmol). The reaction mixture was stirred overnight at room temperature. After completion of the reaction (monitored by TLC), water (5 ml) was added to the reaction mixture and the resulting solid was filtered off and dried in vacuo. The crude material was purified by preparative HPLC (method A) to give the title compound. Yield: 16% (10 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.56 (s, 1H), 9.82 (s, 1H), 7.57-7.52 (m, 4H), 7.14 (d , J = 8.4 Hz, 2H), 6.58 (s, 1H), 3.73 (s, 2H), 3.36 (s, 2H), 2.77-2.73 (m, 1H) , 2.17 (s, 3H), 1.85-1.83 (m, 2H), 1.73-1.69 (m, 4H), 1.56-1.54 (m, 2H), 1 .44-1.42 (m, 2H), 1.36-1.30 (m, 2H), 1.12-1.06 (m, 8H), 0.77-0.76 (m, 6H) . LCMS: (Method C) 647.2 (M+H), Rt. 3.07 min, 94.09% (maximum). HPLC: (method B) Rt. 6.08 min, 95.76% (maximum).

Example 36

(Z) -3-((3,3-dibutyl-5-(4-(cyclopropanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (intermediate 38; 0.2 g, 0.36 mmol) in DMF (2 ml) was added cyclopropanecarboxylic acid (0.041 g, 0.47 mmol), HATU (0.27 g, 0.73 mmol) and N-methylmorpholine (0.12 ml, 1.10 mmol). The reaction mixture was stirred overnight at room temperature. After completion of the reaction (monitored by TLC), water (5 ml) was added to the reaction mixture and the resulting solid was filtered off and dried in vacuo. The crude material was purified by preparative HPLC (method B) to give the title compound. Yield: 20% (45 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 10.14 (s, 1H), 7.55-7.48 (m, 3H), 7.37-7.33 (m, 1H), 7 .14 (d, J = 8.0 Hz, 2H), 6.57 (s, 1H), 3.72 (s, 2H), 3.36 (s, 2H), 2.17 (s, 3H) , 1.77-1.75 (m, 1H), 1.43-1.40 (m, 2H), 1.37-1.31 (m, 2H), 1.12-1.08 (m, 8H), 0.78-0.74 (m, 10H). LCMS: (Method C) 619.0 (M ⁺ +H), Rt. 2.80 min, 99.01% (maximum). HPLC: (method B) Rt. 5.71 min, 97.39% (maximum).

Example 37

(E) -3-((3,3-dibutyl-5-(4-(cyclopentanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-(cyclopentanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 52; 0.14 g, 0.21 mmol) in a mixture of 1,4-dioxane and water (3 ml, 4:1) at 0 °C lithium hydroxide (0.036 g, 0.85 mmol) was added and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N., 4 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The organic portion was washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method C). Yield: 11% (14.8 mg, off-white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.35 (s, 1H), 9.83 (s, 1H), 7.63 (d, J =12.1 Hz, 1H), 7 .57 (d, J =7.8 Hz, 2H), 7.45 (s, 1H), 7.17 (d, J =7.6 Hz, 2H), 6.58 (s, 1H), 5 .40 (d, J = 11.3 Hz, 1H), 3.74 (bs, 2H), 3.38 (s, 2H), 2.77-2.74 (m, 1H), 2.16 ( s, 3H), 1.85-1.84 (m, 2H), 1.71-1.69 (m, 4H), 1.56-1.44 (m, 2H), 1.41-1, 33 (m, 4H), 1.12-1.04 (m, 8H), 0.8 (t, J =6.0 Hz, 6H). LCMS: (Method A) 629.2 (M ⁺ +H), Rt. 2.89 min, 97.32% (maximum). HPLC: (method B) Rt. 6.08 min, 96.14% (maximum).

Example 38

(E) -3-((3-butyl-5-(4-(cyclopentanecarboxamido)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3-butyl-5-(4-(cyclopentanecarboxamido)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4 ,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 53; 0.14 g, 0.22 mmol) in 1,4-dioxane/water (3 mL, 4:1) lithium hydroxide (0.037 g, 0.89 mmol) was added at 0° C. and the reaction mixture was stirred for 12 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1 ml, 1.5 N.) and the aqueous layer was extracted with EtOAc (2×15 ml). The organic layer was dried over Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method C) to give the title compound. Yield: 25% (33 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 9.83 (s, 1H), 7.62 (d, J =12.2 Hz, 1H), 7.57 (d, J =8, 9 Hz, 2H), 7.45 (s, 1H), 7.16 (d, J = 8.4 Hz, 2H), 6.59 (s, 1H), 5.41 (d, J = 12, 2 Hz, 1H), 3.38 (bs, 2H), 3.33 (s, 2H), 2.75-2.74 (m, 1H), 2.16 (s, 3H), 1.85- 1.83 (m, 2H), 1.74-1.70 (m, 4H), 1.70-1.56 (m, 3H), 1.55-1.54 (m, 3H), 1, 38-1.11 (m, 4H), 0.70 (t, J = 7.2 Hz, 6H). LCMS: (Method A) 601.2 (M ⁺ +H), Rt. 2.67 min, 99.09% (maximum). HPLC: (method B) Rt. 5.61 min, 98.96% (maximum).

Example 39

(E) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 61; 0.17 g, 0.34 mmol) in a mixture of 1,4-dioxane and water (4 ml, 4:1) was added lithium hydroxide (0.03 g, 0.70 mmol) and the reaction mixture was stirred at room temperature overnight. 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 organics were washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method A) to give the title compound. Yield: 42% (68 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.25 (s, 1H), 7.70 (dd, J =12.2, 2.0 Hz, 1H), 7.50 (d, J = 2.4 Hz, 1H), 7.29 (t, J = 7.2 Hz, 2H), 7.13 (d, J = 6.4 Hz, 2H), 6.97 (t, J = 6.8 Hz, 1H), 6.72 (s, 1H), 5.44 (dd, J =12.0, 2.4 Hz, 1H), 3.76 (s, 2H), 3.37 ( s, 2H), 2.19 (s, 3H), 1.55-1.53 (m, 2H), 1.39-1.34 (m, 2H), 0.73 (t, J =6, 4Hz, 6H). LCMS: (Method A) 462.1 (M+H), Rt. 2.33 min, 97.56% (maximum), HPLC: (method B) Rt. 5.29 min, 97.10% (maximum).

Example 40

(E) -3-((3,3-dibutyl-5-(4-butyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-butyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- Lithium hydroxide (0.038 g 0.038 g .91 mmol) and the reaction mixture was stirred at room temperature overnight. 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 organics were washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude compound was purified by preparative HPLC (method C). Yield: 28.2% (39.3 mg, off-white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.22 (s, 1H), 9.83 (s, 1H), 7.66 (d, J =12.4 Hz, 1H), 7 .56 (d, J =8.4 Hz, 2H), 7.46 (s, 1H), 7.18 (d, J =8.4 Hz, 2H), 6.57 (s, 1H), 5 .39 (d, J =12.4 Hz, 1H), 3.75 (bs, 2H), 3.39 (s, 2H), 2.27 (t, J =7.20 Hz, 2H), 2 .15 (s, 3H), 1.64-1.60 (m, 2H), 1.58-1.36 (m, 4H), 1.33-1.08 (m, 4H), 1.04 -1.02 (m, 4H), 0.9 (t, J =4.0 Hz, 3H), 0.7 (t, J =8.0 Hz, 6H). LCMS: (Method A) 603.3 (M ⁺ +H), Rt. 2.49 min, 97.34% (maximum). HPLC: (method B) Rt. 5.71 min, 98.16% (maximum).

Example 41

(Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of methyl-(Z)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 63; 0.11 g, 0.21 mmol) in a mixture of 1,4-dioxane and water (3 mL, 4:1) was added lithium hydroxide ( 0.018 g, 0.42 mmol) and the reaction mixture was stirred overnight at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml) and water (5 ml) was added. Then the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (8 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 42-44% EtOAc/PE, silica gel: 230-400 mesh) to give the title compound. Yield: 62% (68 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.57 (s, 1H), 7.59-7.56 (m, 2H), 7.31-7.29 (m, 2H), 7 .14-7.12(m, 1H), 6.70(s, 1H), 3.76-3.75(m, 2H), 3.36(s, 2H), 2.19(s, 3H ), 1.54-1.43 (m, 4H), 1.18-1.11 (m, 4H), 0.74 (t, J =5.60 Hz, 6H). LCMS: (Method A) 506.2 (M ⁺ -H), Rt. 2.85 min, 97.26% (maximum). HPLC: (method B) Rt. 5.96 min, 96.72% (maximum).

Examples 42 and 43

(R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid and (S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl- 2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

The two enantiomers of racemic (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine -8-yl)hydroxy)-2-fluoroacrylic acid (Example 41; 50 mg, 0.098 mmol) was separated by chiral preparative SFC (method A); mobile phase: CO ₂ : IPA (70:30); wavelength: 280 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 40% (20 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.57-7.56 (m, 2H), 7.31-7.29 (m, 2H), 7.14-7.12 (m, 2H), 6.99-6.97(m, 1H), 6.70(s, 1H), 3.74(s, 2H), 3.35(s, 2H), 2.20(s, 3H ), 1 1.42-1.35 (m, 4H), 1.11-1.03 (m, 4H), 0.75-0.74 (m, 6H). LCMS: (Method E) 508.2 (M+H), Rt. 2.68 min, 96.09% (maximum). HPLC: (method B) Rt. 5.96 min, 97.80% (maximum). SFC: (method A) Rt. 6.23 min, 100% (maximum).

Enantiomer 2: Yield: 16% (8 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.54-7.46 (m, 2H), 7.30-7.27 (m, 2H), 7.13-7.12 (m, 2H), 6.98-6.96(m, 1H), 6.70(s, 1H), 3.73(s, 2H), 3.34(s, 2H), 2.19(s, 3H ), 1.54 (s, 1H), 1.43-1.32 (m, 3H), 1.11-1.01 (m, 4H), 0.75-0.74 (m, 6H). LCMS: (Method A) 508.2 (M ⁺ +H), Rt. 2.85 min, 99.63% (maximum). HPLC: (method A) Rt. 5.99 min, 98.89% (maximum). SFC: (method A) Rt. 7.38 min, 97.68% (maximum).

Example 44

(Z) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 64; 0.06 g, 0.12 mmol) in a mixture of 1,4-dioxane and water (3 ml, 4:1) was added lithium hydroxide (0. 01 g, 0.24 mmol) and the reaction mixture was stirred overnight at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml) and water (5 ml) was added. Then the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (8 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method A) to give the title compound. Yield: 24% (37 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.55 (s, 1H), 7.57-7.56 (m, 2H), 7.29-7.27 (m, 2H), 7 .10-7.08(m, 2H), 6.96-6.94(m, 1H), 6.72(s, 1H), 3.73(s, 2H), 3.35(s, 2H ), 2.20 (s, 3H), 1.57-1.55 (m, 2H), 1.38-1.36 (m, 2H), 0.72 (t, J = 7.60 Hz, 6H). LCMS: (Method A) 480.1 (M+H), Rt. 2.63 min, 99.01% (maximum). HPLC: (method B) Rt. 5.42 min, 99.46% (maximum).

Example 45

(Z) -3-((3,3-dibutyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring to a solution of methyl-(Z)-3-((3,3-dibutyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylate (intermediate 67; 0.08 g, 0.15 mmol) in a mixture of 1,4-dioxane and water (3 ml, 4:1) was added lithium hydroxide (0.013 g, 0.31 mmol) and the reaction mixture was stirred at room temperature overnight. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1 ml, 1.5 N.) and the aqueous layer was extracted with EtOAc (2×10 ml). The combined organics were washed with water (8 ml) and brine (8 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by preparative HPLC (method A) to give the title compound. Yield: 45% (35 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.44 (d, J =2.8 Hz, 1H), 7.30-7.24 (m, 3H), 7.10-7, 08 (m, 2H), 6.98-6.91 (m, 3H), 3.73 (s, 2H), 3.37 (s, 2H), 1.37-1.31 (m, 4H) , 1.14-1.00 (m, 8H), 0.85-0.69 (m, 6H). LCMS: (Method C) 490.1 (M+H), Rt. 3.22 min, 98.94% (maximum), HPLC: (method B) Rt. 6.27 min, 99.58% (maximum).

Example 46

(Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 75; 0.74 g, 1.45 mmol) in a mixture of 1,4-dioxane and water (8 ml, 4:1) was added lithium hydroxide (0. 12 g, 2.90 mmol) and the reaction mixture was stirred overnight at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml) and water (10 ml) was added. Then the aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 3-4% MeOH/DCM, silica gel: 230-400 mesh) to give the title compound. Yield: 72% (0.52 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.57 (s, 1H), 7.59-7.56 (m, 2H), 7.31-7.29 (m, 2H), 7 .14-7.12(m, 1H), 6.70(s, 1H), 3.76-3.75(m, 2H), 3.36(s, 2H), 2.19(s, 3H ), 1.54-1.43 (m, 4H), 1.18-1.11 (m, 4H), 0.74 (t, J =5.60 Hz, 6H). LCMS: (Method A) 497.2 (M ⁺ +H), Rt. 2.86 min, 97.24% (maximum). HPLC: (method B) Rt. 6.06 min, 95.11% (maximum).

Examples 47 and 48

(R) -(Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid and (S)- (Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3, 4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

The two enantiomers of racemic (Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8 -yl)oxy)-2-fluoroacrylic acid (Example 46; 0.52 g, 1.05 mmol) was separated by chiral preparative SFC (method A); mobile phase: CO ₂ : IPA (70:30); wavelength: 280 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 26% (140 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.62 (s, 1H), 7.74 (s, 1H), 7.60 (d, J =18.4 Hz, 1H), 7, 34 (t, J =7.2 Hz, 2H), 7.22 (d, J =6.8 Hz, 2H), 7.06 (t, J =7.2 Hz, 1H), 6.94 ( s, 1H), 3.78 (s, 2H), 3.44 (s, 2H), 1.53-1.32 (m, 4H), 1.10-1.05 (m, 4H), 0 .72-0.72 (m, 6H). LCMS: (Method C) 494.0 (M ⁺ -H), Rt. 3.03 min, 98.39% (maximum). HPLC: (method B) Rt. 6.05 min, 97.65% (maximum). SFC: (method A) Rt. 2.82 min, 99.06% (maximum).

Enantiomer 2: Yield: 30% (160 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.61 (s, 1H), 7.74 (s, 1H), 7.59 (d, J =18.4 Hz, 1H), 7, 34 (t, J =7.2 Hz, 2H), 7.22 (d, J =7.6 Hz, 2H), 7.06 (t, J =7.2 Hz, 1H), 6.94 ( s, 1H), 3.78 (s, 2H), 3.44 (s, 2H), 1.53-1.32 (m, 4H), 1.12-1.07 (m, 4H), 0 .73-0.70 (m, 6H). LCMS: (method C) 494.1 (M ⁺ -H), Rt. 3.03 min, 97.42% (maximum). HPLC: (method B) Rt. 6.04 min, 96.90% (maximum). SFC: (method A) Rt. 3.35 min, 98.76% (maximum).

Example 49

(E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 76; 1.0 g, 1.97 mmol) in a mixture of 1,4-dioxane and water (10 ml; 5:1) was added lithium hydroxide (0.25 g, 5.92 mmol) and the resulting mixture was stirred for 12 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo. The resulting residue was acidified with dilute HCl (1.5 N., 2 ml) and the aqueous part was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (15 ml) and brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the crude material was purified by Isolera column chromatography (eluent: 2-3% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 64% (600 mg, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.29 (s, 1H), 7.75 (d, J =12.0 Hz, 1H), 7.69 (s, 1H), 7, 38-7.36 (m, 1H), 7.34 (s, 1H), 7.27-7.25 (m, 2H), 7.09 (t, J =7.2 Hz, 1H), 6 .91 (s, 1H), 5.40 (d, J =12.4 Hz, 1H), 3.81 (bs, 2H), 3.47 (s, 2H), 1.54-1.51 ( m, 1H), 1.45-1.34 (m, 3H), 1.11-0.98 (m, 4H), 0.74 (t, J =4.40 Hz, 6H). LCMS: (Method A) 522.0 (M ⁺ ), Rt. 2.78 min, 98.84% (maximum). HPLC: (method B) Rt. 5.95 min, 98.16% (maximum).

Examples 50 and 51

(R)- (E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylic acid and (S)- (E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

The two enantiomers of racemic (E)-3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8 -yl)oxy)acrylic acid (Example 49; 600 mg, 1.25 mmol) was separated by chiral preparative SFC (instrument: Pic SFC 10-150); mobile phase: CO ₂ : methanol; column: Lux A1; Flow rate: 3 ml/min; wavelength: 220 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 37% (222 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.32 (bs, 1H), 7.74 (d, J= 12.0 Hz, 1H), 7.69 (s, 1H), 7, 38-7.34 (m, 2H), 7.27-7.25 (m, 2H), 7.08 (t, J =7.2 Hz, 1H), 6.91 (s, 1H), 5 .40 (d, J = 12.0 Hz, 1H), 3.81 (bs, 2H), 3.47 (s, 2H), 1.54-1.36 (m, 4H), 1.34- 1.04 (m, 4H), 0.73 (t, J =6.80 Hz, 6H). LCMS: (method C) 480.0 (M ⁺ +2), Rt. 2.91 min, 99.26% (maximum). HPLC: (method B) Rt. 5.95 min, 98.81% (maximum). SFC: (method D) Rt. 3.2 min, 100% (maximum).

Enantiomer 2: Yield: 32% (195 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.37 (bs, 1H), 7.75 (d, J =12.4 Hz, 1H), 7.68 (d, J =4.8 Hz, 1H), 7.38-7.35 (m, 2H), 7.27 (d, J =6.4 Hz, 2H), 7.11-7.09 (m, 1H), 6.91 (s, 1H), 5.41 (d, J =12.4 Hz, 1H), 3.80 (bs, 2H), 3.40 (s, 2H), 1.53-1.38 (m, 2H), 1.36-1.33 (m, 2H), 1.09-1.07 (m, 2H), 1.04-1.01 (m, 2H), 0.72 (t, J = 4.8Hz, 6H). LCMS: (Method C) 479.1.0 (M ⁺ +H), Rt. 2.91 min, 97.56% (maximum). HPLC: (method B) Rt. 5.95 min, 95.56% (maximum). SFC: (method D) Rt. 4.51 min, 99.16% (maximum).

Example 52

(Z) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 79; 0.18 g, 0.3 mmol) in a mixture of 1,4-dioxane and water (10 ml; 5:1) hydroxide was added lithium (0.038 g, 0.91 mmol) and the resulting mixture was stirred for 12 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo. The resulting residue was acidified with dilute HCl (1.5 N., 2 ml) and the aqueous part was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (15 ml) and brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo to give the crude material, which was purified by preparative HPLC (method A). Yield: 25% (42 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.14 (s, 1H), 7.57 (d, J =8.8 Hz, 2H), 7.52 (s, 1H), 7, 48-7.43 (m, 1H), 7.10 (d, J =8.0 Hz, 2H), 6.65 (s, 1H), 3.72 (bs, 2H), 3.35 (s , 2H), 2.20 (s, 3H), 1.55-1.50 (m, 2H), 1.42-1.34 (m, 2H), 1.22 (s, 9H), 0, 71 (t, J =7.20 Hz, 6H). LCMS: (Method A) 579.2 (M ⁺ +H), Rt. 2.42 min, 99.24% (maximum). HPLC: (method B) Rt. 5.17 min, 99.04% (maximum).

Example 53

(Z) -3-((3,3-dibutyl-7-chloro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) -2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((3,3-dibutyl-7-chloro-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylate (intermediate 80; 617 mg, 1.14 mmol) in a mixture of 1,4-dioxane and water (10 ml, 5:1) was added lithium hydroxide (241 mg, 41.96 mmol) and the reaction mixture was stirred overnight at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml) and water (5 ml) was added. Then the aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (8 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method C). Yield: 20% (70 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.61 (s, 1H), 7.33 (t, J=7.6 Hz, 2H), 7.26-7.20 (m, 3H ), 7.04 (t, J=7.2 Hz, 1H), 6.93 (s, 1H), 3.75 (s, 2H), 3.38 (d, J=35.8 Hz, 3H ), 1.42-1.32 (m, 4H), 1.14-1.08 (m, 8H), 0.75 (s, 6H). LCMS: (Method B) 522.2 (M ⁺ -2H), Rt. 2.46 min, 99.75% (maximum). HPLC: (method A) Rt. 6.12 min, 99.28% (maximum).

Example 54

(E) -3-((3,3-diethyl-7-iodo-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid

With stirring, to a solution of tert-butyl-(E)-3-((3,3-diethyl-7-iodine-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 82; 130 mg, 0.2175 mmol) in DCM (2 ml) at 0°C was added TFA (2 ml) and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (15 ml) and the aqueous layer was extracted with DCM (3×10 ml). The combined organic layer was washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method B) to give the title compound. Yield: 21% (25 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.28 (s, 1H), 7.70 (d, J =12.0 Hz, 1H), 7.53 (s, 1H), 7, 35-7.17 (m, 5H), 7.03 (t, J =15.2 Hz, 1H), 5.39 (d, J =12.4 Hz, 1H), 3.74 (s, 2H ), 3.41 (s, 2H), 1.53-1.48 (m, 2H), 1.37-1.32 (m, 2H), 0.69 (t, J = 14.4 Hz, 6H). LCMS: (Method B) 542.0 (M ⁺ +H), Rt. 2.09 min, 95.08% (maximum). HPLC: (method B) Rt. 5.47 min, 93.13% (maximum).

Example 55

(E) -3-((7-bromo-3,3-diethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid

With stirring, to a solution of ethyl-(E)-3-((7-bromo-3,3-diethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylate (intermediate 84; 250 mg, 0.478 mmol) in a mixture of 1,4-dioxane and water (5 ml, 4:1) was added lithium hydroxide (100 mg, 2.3925 mmol) and the reaction mixture stirred overnight at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml) and water (10 ml) was added. The aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with water (15 ml) and brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method C), and the resulting fraction was concentrated under reduced pressure to give the title compound. Yield: 10% (15 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.28 (s, 1H), 7.72 (d, J =12.4 Hz, 1H), 7.64 (s, 1H), 7, 34 (t, J =15.6 Hz, 2H), 7.21 (d, J =7.6 Hz, 2H), 7.05 (t, J =15.6 Hz, 2H), 5.40 ( d, J= 12.0 Hz, 1H), 3.77 (s, 2H), 3.43 (d, J =8.0 Hz, 2H), 1.52-1.49 (m, 2H), 1.38-1.34 (m, 2H), 0.69 (t, J =14.4 Hz, 6H). LCMS: (Method E) 494.0 (M ⁺ +H), Rt. 2.48 min, 98.56% (maximum). HPLC: (method B) Rt. 5.43 min, 98.18% (maximum).

Example 56

(Z) -3-((7-bromo-3,3-diethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) -2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((7-bromo-3,3-diethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylate (intermediate 85; 250 mg, 0.475 mmol) in a mixture of 1,4-dioxane and water (5 ml, 4:1) was added lithium hydroxide (100 mg, 2.376 mmol) and the reaction the mixture was stirred overnight at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml) and water (10 ml) was added. The aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (10 ml) and brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method C) and the resulting fraction was concentrated in vacuo to give the title compound. Yield: 10% (15 mg, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.69 (s, 1H), 7.58 (d, J =18.4 Hz, 1H), 7.32 (t, J =15.6 Hz, 2H), 7.17 (d, J =7.6 Hz, 2H), 7.10 (s, 1H), 7.03 (t, J =14.4 Hz, 1H), 3.73 ( d, J = 11.2 Hz, 2H), 3.41 (s, 2H), 1.54-1.49 (m, 2H), 1.37-1.32 (m, 2H), 0.69 (t, J =14.8 Hz, 6H). LCMS: (Method A) 512.0 (M ⁺ +H), Rt. 2.63 min, 99.15% (maximum). HPLC: (method B) Rt. 5.56 min, 95.79% (maximum).

Examples 57 and 58

(S)- (Z) -3-((3-butyl-3-ethyl-7-iodine-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylic acid and (R)- (Z) -3-((3-butyl-3-ethyl-7-iodo-1,1-dioxydo-5-phenyl-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

Enantiomers of racemic (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)oxy)acrylic acid and (E) -3-((3-butyl-3-ethyl-7-iodo-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid (intermediate 94; 3.38 g, 5.93 mmol) was separated by chiral preparative SFC (instrument: Pic SFC 10-150); mobile phase: CO ₂ : 0.5% isopropylamine in IPA (70:30); column: Lux A1; flow rate: 3 ml/min; wavelength: 220 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction was enantiomer 1 of Examples 5 and 6; the second eluting fraction corresponded to a mixture of enantiomer 2 of Examples 5 and 6 and enantiomer 1 of the title compound; and the third fraction corresponded to enantiomer 2 of the desired compound.

A mixture of enantiomer 2 of Examples 5 and 6 and enantiomer 1 of the title compound was re-separated with a chiral preparative SFC (instrument: Pic SFC 10-150); mobile phase: CO ₂ : 0.5% isopropylamine in IPA (70:30); column: YMC Cellulose-SB; Flow rate: 3 ml/min; wavelength: 220 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction was enantiomer 2 of Examples 5 and 6 and the second eluting fraction was enantiomer 1 of the title compound. The absolute configuration of these two enantiomers of the target compound is unknown.

Enantiomer 1: Yield: 9% (0.305 g, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.27 (s, 1H), 7.71 (d, J =12.4 Hz, 1H), 7.53 (s, 1H), 7, 37-7.33 (m, 2H), 7.27-7.21 (m, 3H), 7.09-7.07 (m, 1H), 5.38 (d, J =12.4 Hz, 1H), 3.66 (s, 2H), 3.43 (s, 2H), 1.53-1.51 (m, 1H), 1.35-1.32 (m, 3H), 1.14 -0.99 (m, 4H), 0.74-0.73 (m, 6H). LCMS: (Method A) 568.1 (M ⁺ -H), Rt. 2.92 min, 97.41% (maximum). HPLC: (method B) Rt. 6.03 min, 95.00% (maximum). SFC: (method A) Rt. 3.55 min, 99.53% (maximum).

Enantiomer 2: Yield: 3.5% (0.12 g, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.27 (s, 1H), 7.71 (d, J =12.0 Hz, 1H), 7.53 (s, 1H), 7, 37-7.33 (m, 2H), 7.27-7.21 (m, 3H), 7.09-7.07 (m, 1H), 5.38 (d, J =12.4 Hz, 1H), 3.68 (s, 2H), 3.43 (s, 2H), 1.51-1.50 (m, 1H), 1.37-1.30 (m, 3H), 1.14 -0.98 (m, 4H), 0.75-0.74 (m, 6H. LCMS: (Method A) 568.1 (M ⁺ -H), Rt. 2.92 min, 96.49% ( HPLC: (Method B) Rt 6.03 min, 96.77% (max) SFC: (Method A) Rt 6.78 min, 94.72% (max).

Example 59

(Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(Z)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, Lithium hydroxide (0.12 g, 2.89 mmol) and the resulting mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated in vacuo. The crude residue was acidified with dilute HCl (1.5 N, 2 ml) and the aqueous layer was extracted with EtOAc (2×10 ml). The organic layer was washed with water (15 ml) and brine (15 ml), dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 56% (160 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.13 (bs, 1H), 7.44 (s, 1H), 7.29 (t, J =8.0 Hz, 2H), 7, 13 (d, J =7.6 Hz, 2H), 7.09 (d, J =7.2 Hz, 1H), 6.97 (t, J =7.2 Hz, 1H), 6.69 ( s, 1H), 5.20 (d, J =6.8 Hz, 1H), 3.74-3.71 (m, 2H), 3.38 (s, 2H), 2.19 (s, 3H ), 1.55-1.31 (m, 4H), 1.10-0.98 (m, 4H), 0.75-0.72 (m, 6H). LCMS: (Method A) 490.2 (M+H), Rt. 2.70 min, 99.89% (maximum). HPLC: (method B) Rt. 5.69 min, 99% (maximum).

Examples 60 and 61

(S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid and (R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3 ,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

The two enantiomers of racemic (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine -8-yl)hydroxy)acrylic acid (Example 59; 97 mg, 0.20 mmol) was separated by chiral preparative SFC (method B); mobile phase: CO ₂ : IPA (70:30); wavelength: 220 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 20% (20 mg, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.07 (bs, 1H), 7.43 (s, 1H), 7.29 (t, J =8.4 Hz, 2H), 7, 12 (d, J =8.4 Hz, 2H), 6.99 (d, J =7.2 Hz, 1H), 6.97 (t, J =7.2 Hz, 1H), 6.69 ( s, 1H), 5.21 (d, J = 6.8 Hz, 1H), 3.74-3.71 (m, 2H), 3.38 (s, 2H), 2.19 (s, 3H ), 1.54-1.31 (m, 4H), 1.16-0.89 (m, 4H), 0.75-0.72 (m, 6H). LCMS: (Method C) 490.1 (M ⁺ +H), Rt. 2.78 min, 98.11% (maximum). HPLC: (method B) Rt. 5.69 min, 97.25% (maximum). Chiral SFC: (method B) Rt. 8.86 min, 100% (maximum).

Enantiomer 2: Yield: 18% (18 mg, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 11.92 (bs, 1H), 7.44 (s, 1H), 7.29 (t, J =7.6 Hz, 2H), 7, 12 (d, J =7.6 Hz, 2H), 7.05 (d, J =7.2 Hz, 1H), 6.97 (t, J =7.2 Hz, 1H), 6.69 ( s, 1H), 5.21 (d, J = 6.8 Hz, 1H), 3.74-3.71 (m, 2H), 3.38 (s, 2H), 2.19 (s, 3H ), 1.54-1.31 (m, 4H), 1.16-0.89 (m, 4H), 0.75-0.72 (m, 6H). LCMS: (Method C) 490.0 (M ⁺ +H), Rt. 2.78 min, 98.21% (maximum). HPLC: (method B) Rt. 5.69 min, 96.69% (maximum). Chiral purity: (method B) Rt. 9.84 min, 98.07% (maximum).

Example 62

(E) -3-((3-butyl-5-(4-(tert-butylcarbamoyl)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro -1,5-Benzothiazepin-8-yl)oxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3-butyl-5-(4-(tert-butylcarbamoyl)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3 ,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (Intermediate 99; 50 mg, 0.08 mmol) in 1,4-dioxane/water (4:1, 5 mL) lithium hydroxide (6.8 mg, 0.16 mmol) was added and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was dissolved in ice-cold water (2 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organic layer was washed with ice-cold water (5 ml), brine (5 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 10% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 12% (5.9 mg, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.72-7.67 (m, 3H), 7.53 (s, 2H), 6.98-6.94 (m, 3H), 5 .51 (d, J = 12.0 Hz, 1H), 3.85-3.65 (m, 2H), 3.41 (s, 2H), 2.29 (s, 3H), 1.59- 1.49 (m, 2H), 1.36 (s, 9H), 1.33-1.29 (m, 2H), 1.24-1.13 (m, 4H), 0.83-0, 79(m, 6H). LCMS: (Method B) 589.3 (M ⁺ +H), Rt. 2.11 min, 95.20% (maximum). HPLC: (method A) Rt. 5.21 min, 96.87% (maximum).

Example 63:

(Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 100; 0.55 g, 0.99 mmol) in a mixture of 1,4-dioxane and water (4:1.5 ml) was added lithium hydroxide (0. 08 g, 1.98 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and then diluted with ice-cold water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was then washed with water (8 ml), brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 40-45% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 49% (0.26 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.62 (s, 1H), 7.71 (s, 1H), 7.59 (s, 1H), 7.37-7.33 (m , 2H), 7.31-7.29 (m, 2H), 7.22-7.08 (m, 2H), 3.78 (bs, 2H), 3.43 (s, 2H), 1, 38-1.30 (m, 4H), 1.11-1.03 (m, 4H), 0.73 (t, J =6.40 Hz, 6H). LCMS: (Method A) 538.0 (M ⁺ - 2H), Rt. 2.84 min, 98.53% (maximum). HPLC: (method B) Rt. 6.09 min, 99.67% (maximum).

Examples 64 and 65

(R)- (Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid and (S)- (Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3, 4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

The two enantiomers of racemic (Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8 -yl)oxy)-2-fluoroacrylic acid (Example 63; 0.24 g, 4.44 mmol) was separated by chiral preparative SFC (method F); mobile phase: CO ₂ : 0.5% isopropylamine in IPA (70:30); wavelength: 280 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 14% (34 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.68 (bs, 1H), 7.69 (s, 1H), 7.56 (d, J =18.8 Hz, 1H), 7, 34 (t, J =8.0 Hz, 2H), 7.21 (d, J =7.2 Hz, 2H), 7.08-7.06 (m, 2H), 3.75 (bs, 2H ), 3.43 (s, 2H), 1.54-1.45 (m, 1H), 1.41-1.28 (m, 3H), 1.17-0.80 (m, 4H), 0.76-0.64 (m, 6H). LCMS: (Method A) 540.1 (M ⁺ ), Rt. 2.93 min, 98.05% (maximum). HPLC: (method B) Rt. 6.09 min, 95.88% (maximum). SFC: (method F) Rt. 6.98 min, 99.57% (maximum).

Enantiomer 2: Yield: 14% (35 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.59 (bs, 1H), 7.70 (s, 1H), 7.60 (d, J =18.4 Hz, 1H), 7, 37-7.35 (m, 2H), 7.21 (d, J =7.2 Hz, 2H), 7.08-7.04 (m, 2H), 3.76 (bs, 2H), 3 .43 (s, 2H), 1.55-1.53 (m, 1H), 1.42-1.30 (m, 3H), 1.18-0.90 (m, 4H), 0.68 -0.78 (m, 6H). LCMS: (Method A) 540.2 (M ⁺ ), Rt. 2.92 min, 97.84% (maximum). HPLC: (method B) Rt. 6.09 min, 93.15% (maximum). SFC: (method F) Rt. 8.38 min, 97.28% (maximum).

Example 66

(Z) -3-((5-(4-(benzylamino)phenyl)-3,3-diethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((5-(4-aminophenyl)-3,3-diethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 78; 0.02 g, 0.03 mmol) in 1,4-dioxane/water (4:1, 5 mL) at room temperature temperature, lithium hydroxide (0.01 g, 0.07 mmol) was added and the mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with 1.5N HCl. HCl solution (3 ml) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with water (10 ml), brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by preparative HPLC (method A) to give the title compound. Yield: 17% (3.5 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.43 (s, 1H), 7.37-7.30 (m, 5H), 7.24 (d, J =6.8 Hz, 1H ), 7.05 (d, J =8.4 Hz, 2H), 6.63 (d, J =8.4 Hz, 2H), 6.24 (s, 2H), 4.28 (s, 2H ), 3.69 (s, 2H), 3.39 (s, 2H), 2.05 (s, 3H), 1.52 (q, J = 7.20 Hz, 2H), 1.40 (q , J = 7.20 Hz, 2H), 0.64 (t, J = 7.20 Hz, 6H). LCMS: (Method A) 585.1 (M ⁺ +H), Rt. 2.63 min, 90.29% (maximum). HPLC: (method B) Rt. 4.33 min, 93.44% (maximum).

Example 67

(E) -3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid

With stirring, to a solution of ethyl-(E)-3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 102; 0.23 g, 0.41 mmol) in a mixture of 1,4-dioxane and water (4:1.5 ml) was added lithium hydroxide (0.07 g, 1.64 mmol) and the reaction mixture was stirred for 16 hours at room temperature. 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 layer was washed with water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by preparative HPLC (method A) to give the title compound. Yield: 42% (68 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.24 (s, 1H), 7.68 (d, J =12.2 Hz, 1H), 7.49 (s, 1H), 7 .32 (t, J =7.7 Hz, 2H), 7.21 (d, J =7.4 Hz, 2H), 7.03 (t, J =7.2 Hz, 1H), 6.67 (s, 1H), 5.40 (d, J =12.2 Hz, 1H), 3.78 (bs, 2H), 3.40 (s, 2H), 2.69 (q, J =6, 92 Hz, 2H), 1.43-1.39 (m, 3H), 1.36-1.13 (m, 12H), 0.86-0.67 (m, 6H). LCMS: (Method A) 532.2 (M ⁺ +H), Rt. 3.13 min, 97.39% (maximum). HPLC: (method A) Rt. 6.07 min, 96.57% (maximum).

Example 68

(E) -3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4 ,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 106; 55 mg, 0.08 mmol) in 1,4-dioxane/water (4:1, 5 ml) at room temperature temperature, lithium hydroxide (7.17 mg, 0.17 mmol) was added and the reaction mixture was stirred for 6 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organic layer was washed with ice-cold water (5 ml) and brine (5 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 55% EtOAc/PE; silica gel: 230-400 mesh). The resulting compound was repurified by preparative HPLC (method A) to give the pure title compound. Yield: 4% (2.1 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.75-7.68 (m, 3H), 7.52 (s, 2H), 6.99-6.92 (m, 3H), 5 .51 (d, J= 12.4 Hz, 1H), 3.75 (bs, 2H), 3.42 (s, 2H), 2.39 (s, 3H), 1.46-1.57 ( m, 2H), 1.36 (s, 9H), 1.22-1.11 (m, 10H), 0.80 (t, J =6.8 Hz, 6H). LCMS: (Method A) 617.3 (M ⁺ +H), Rt. 2.74 min, 96.23% (maximum). HPLC: (method B) Rt. 5.91 min, 93.46% (maximum).

Example 69

(E) -3-((3,3-dibutyl-5-(4-(isopropylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3,3-dibutyl-5-(4-(isopropylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (intermediate 108; 50 mg, 0.08 mmol) in a mixture of 1,4-dioxane and water (4:1, 5 ml) was added lithium hydroxide ( 6.6 mg, 0.16 mmol) and the reaction mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (2 ml) and EtOAc (2 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organic layer was washed with ice-cold water (5 ml) and brine (5 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 9% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 10% (5 mg, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.28 (bs, 1H), 8.00 (d, J =8.0 Hz, 1H), 7.75-7.72 (m, 3H ), 7.53 (s, 1H), 7.03-7.01 (m, 2H), 6.92 (s, 1H), 5.51 (d, J =12.0 Hz, 1H), 4 .11-4.06(m, 1H), 3.78(bs, 2H), 3.29(s, 2H), 2.28(s, 3H), 1.58-1.45(m, 2H ), 1.38-1.25 (m, 2H), 1.18-1.10 (m, 14H), 0.80 (t, J =6.8 Hz, 6H). LCMS: (Method A) 603.2 (M ⁺ +H), Rt. 2.52 min, 91.99% (maximum). HPLC: (method B) Rt. 5.51 min, 85.91% (maximum).

Example 70

(Z) -3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 109; 0.36 g, 0.64 mmol) in a mixture of 1,4-dioxane and water (5 ml, 4:1) was added lithium hydroxide (0. 05 g, 1.3 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml) to pH~4 and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (8 ml) and brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 40-45% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 18% (70 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.55 (s, 1H), 7.56 (d, J =4.8 Hz, 1H), 7.52 (s, 1H), 7, 31 (t, J= 8.0 Hz, 2H), 7.18 (d, J =7.6 Hz, 2H), 7.02 (d, J =6.8 Hz, 1H), 6.75 ( s, 1H), 3.76 (bs, 2H), 3.37 (s, 2H), 2.71 (q, J = 7.2 Hz, 2H), 1.44-1.42 (m, 2H ), 1.36-1.30 (m, 2H), 1.18-1.10 (m, 3H), 1.07-1.04 (m, 8H), 0.75 (t, J =6 .80Hz, 6H). LCMS: (Method A) 550.2 (M ⁺ +1), Rt. 3.19 min, 97.86% (maximum). HPLC: (method A) Rt. 6.2 min, 98.44% (maximum).

Example 71

(Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-propionamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (intermediate 38; 0.15 g, 0.27 mol) in DCM (5 ml) at 0°C was added triethylamine (0.06 g, 0. 54 mmol), then propionyl chloride (0.03 g, 0.32 mmol) and stirring continued for 1 hour at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with DCM (15 ml) and washed with saturated NaHCO ₃ solution (10 ml) and brine (10 ml). The organic layer was then dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 5% MeOH in DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 37% (60 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.81 (s, 1H), 7.54 (d, J =8.4 Hz, 2H), 7.42 (s, 1H), 7, 30 (d, J =12.0 Hz, 1H), 7.13 (d, J =7.6 Hz, 2H), 6.58 (s, 1H), 3.72 (bs, 2H), 3, 18 (s, 2H), 2.30 (q, J =7.6 Hz, 2H), 2.16 (s, 3H), 1.43-1.36 (m, 4H), 1.24-1 .06 (m, 11H), 0.88-0.65 (m, 6H). LCMS: (Method C) 604.6 (M ⁺ -2H), Rt. 2.90 min, 91.45% (maximum). HPLC: (method B) Rt. 5.57 min, 93.38% (maximum).

Example 72

(Z) -3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of methyl-(Z)-3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 111; 0.23 g, 0.42 mmol) in a mixture of 1,4-dioxane and water (4:1.5 ml) was added lithium hydroxide ( 0.04 g, 0.85 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with water (8 ml) and brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 35-40% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 30% (0.07 g, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.58 (s, 1H), 7.57 (d, J =8.4 Hz, 1H), 7.52 (s, 1H), 7, 30 (t, J =8.0 Hz, 2H), 7.15 (d, J =7.6 Hz, 2H), 6.99 (t, J =7.2 Hz, 1H), 6.71 ( s, 1H), 3.74 (bs, 2H), 3.37 (s, 2H), 2.75-2.68 (m, 2H), 1.53-1.32 (m, 4H), 1 .08-1.01 (m, 7H), 0.74 (t, J =4.80 Hz, 6H). LCMS: (Method A) 522.1 (M ⁺ +H), Rt. 2.94 min, 99.01% (maximum). HPLC: (method A) Rt. 5.74 min, 95.24% (maximum).

Example 73

(E) -3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid

With stirring to a solution of ethyl-(E)-3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, Lithium hydroxide (0.6 g, 1.35 mmol) and the reaction mixture was stirred for 16 hours at room temperature. 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 layer was washed with water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by preparative HPLC (method B) to give the title compound. Yield: 54% (180 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.25 (s, 1H), 7.68 (d, J =12.0 Hz, 1H), 7.49 (s, 1H), 7, 31 (t, J =7.6 Hz, 2H), 7.18 (d, J =7.6 Hz, 2H), 7.01 (t, J =7.2 Hz, 1H), 6.70 ( s, 1H), 5.41 (d, J =12.0 Hz, 1H), 3.77 (bs, 2H), 3.39 (s, 2H), 2.70 (q, J =7.20 Hz, 2H), 1.56-1.32 (m, 4H), 1.18-1.03 (m, 7H), 0.73 (t, J =5.20 Hz, 6H). LCMS: (Method A) 504.1 (M ⁺ +H), Rt. 2.88 min, 99.31% (maximum). HPLC: (method B) Rt. 6.02 min, 96.02% (maximum).

Example 74

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)-2-methacrylic acid

With stirring, to a solution of (E) -3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5- tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methacrylic acid (intermediate 114; 0.2 g, 0.30 mmol) in dry DCM (2 ml) TFA (0.6 ml) was added ) and triethylsilane (0.6 ml) at 0°C and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with ice-cold water (5 ml). The aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and evaporated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 20% EtOAc/PE; silica gel: 230-400 mesh) and the resulting product was further triturated with diethyl ether to give the title compound. Yield: 46% (75 mg, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.31 (s, 1H), 7.50 (s, 2H), 7.38-7.32 (m, 3H), 7.29-7 .23 (m, 2H), 7.16-7.02 (m, 1H), 6.52 (s, 1H), 4.18-3.71 (bs, 2H), 2.11 (s, 3H ), 1.80 (s, 3H), 1.58-1.46 (m, 2H), 1.41-1.35 (m, 2H), 1.29-1.18 (m, 3H), 1.17-1.04 (m, 2H), 1.03-0.92 (m, 3H), 0.74-0.72 (m, 6H). LCMS: (Method E) 533.2 (M ⁺ +H), Rt. 2.81 min, 95.34% (maximum). HPLC: (method B) Rt. 6.39 min, 95.95% (maximum).

Example 75

(E) -3-((7-bromo-3,3-dibutyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl) hydroxy)acrylic acid

With stirring, to a solution of tert-butyl-(E)-3-((7-bromo-3,3-dibutyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-Benzothiadiazepin-8-yl)oxy)acrylate (intermediate 116; 0.11 g, 0.18 mmol) in DCM (5 ml) TFA (2 ml) was added at 0°C and the reaction mixture was stirred for 2 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (15 ml) and the aqueous layer was extracted with DCM (2×10 ml). The combined organic layer was washed with water (10 ml) and brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 25% EtOAc in hexanes; silica gel: 230-400 mesh) to give the title compound. Yield: 25% (28 mg, light brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.23 (s, 1H), 7.76 (s, 1H), 7.68 (d, J =12.0 Hz, 1H), 7, 51 (s, 1H), 7.42 (t, J =8.0 Hz, 2H), 7.38-7.29 (m, 2H), 7.17 (t, J =8.0 Hz, 1H ), 6.86 (s, 1H), 5.27 (d, J =12.0 Hz, 1H), 4.01 (bs, 2H), 1.60-1.42 (m, 2H), 1 .42-1.35 (m, 2H), 1.30-1.11 (m, 2H), 1.00-1.12 (m, 2H), 1.00-0.80 (m, 4H) , 0.80-0.60 (m, 6H). LCMS: (Method A) 553.1 (M ⁺ +2H), Rt. 2.98 min, 96.92% (maximum). HPLC: (method B) Rt. 6.28 min, 98.09% (maximum).

Example 76

(E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 8-yl)hydroxy)acrylic acid

With stirring to a solution of tert-butyl-(E)-3-((3-butyl-3-ethyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxydo-5-phenyl-2, 3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (intermediate 129; 0.6 g, 0.9 mmol) in dry DCM (5 mL) TFA (2. 5 ml) and triethylsilane (2.5 ml) at 0°C and the reaction mixture was stirred for 3 hours at room temperature. After completion of the reaction (monitored by LCMS), the reaction mixture was diluted with ice-cold water (5 ml) and the aqueous layer was extracted with DCM (2×10 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and evaporated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 60% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 68% (0.3 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.20 (bs, 1H), 7.64 (d, J =12.2 Hz, 1H), 7.54-7.51 (m, 1H ), 7.39-7.32 (m, 3H), 7.27-7.21 (m, 2H), 7.11-7.08 (m, 1H), 6.51 (s, 1H), 5.32 (d, J =12.2 Hz, 1H), 4.11-3.97 (m, 2H), 2.10 (s, 3H), 1.63-1.62 (m, 1H) , 1.50-1.36 (m, 3H), 1.24-0.93 (m, 4H), 0.86-0.70 (m, 6H). LCMS: (Method A) 491.1 (M ⁺ +H), Rt. 2.73 min, 95.09% (maximum). HPLC: (method B) Rt. 5.66 min, 94.36% (maximum).

Examples 77 and 78

(S)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)acrylic acid and (R)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2 ,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

The two enantiomers of racemic (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5 -benzothiadiazepin-8-yl)oxy)acrylic acid (Example 77; 0.3 g, 0.61 mmol) was separated by chiral preparative SFC (Method E); mobile phase: CO ₂ : 0.5% isopropylamine in methanol (50:50); wavelength: 210 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 33.3% (100 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.21 (bs, 1H), 7.64 (d, J =12.0 Hz, 1H), 7.54-7.51 (m, 1H ), 7.36-7.10 (m, 6H), 6.51 (s, 1H), 5.32 (d, J = 12.4 Hz, 1H), 4.02-3.87 (m, 2H), 2.10 (s, 3H), 1.63-1.39 (m, 4H), 1.24-0.93 (m, 4H), 0.86-0.70 (m, 6H) . LCMS: (Method A) 491.1 (M ⁺ +H), Rt. 2.71 min, 96.91% (maximum). HPLC: (method B) Rt. 5.70 min, 95.47% (maximum). SFC: (Method E) Rt. 2.03 min, 96.17% (maximum).

Enantiomer 2: Yield: 34% (110 mg, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.21 (bs, 1H), 7.64 (d, J =12.4 Hz, 1H), 7.55-7.51 (m, 1H ), 7.39-7.36 (m, 3H), 7.27-7.21 (m, 2H), 7.11-7.08 (m, 1H), 6.51 (s, 1H), 5.32 (d, J =12.4 Hz, 1H), 4.02-3.87 (m, 2H), 2.10 (s, 3H), 1.62-1.38 (m, 4H) , 1.24-0.91 (m, 4H), 0.89-0.71 (m, 6H). LCMS: (Method A) 491.2 (M ⁺ +H), Rt. 2.71 min, 97.88% (maximum). HPLC: (method B) Rt. 5.70 min, 98.57% (maximum). SFC: (Method E) Rt. 2.31 min, 97.27% (maximum).

Example 79

(Z) -3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro-1 ,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4 ,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 130; 90 mg, 0.14 mmol) in 1,4-dioxane/water (4:1, 5 mL) ) lithium hydroxide (11.7 mg, 0.27 mmol) was added and the mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the residue was subjected to distribution between ice-cold water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organic layer was washed with ice-cold water (5 ml) and brine (5 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by preparative HPLC (method A) to give the title compound. Yield: 13% (11.3 mg, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.67 (d, J= 8.8 Hz, 2H), 7.51-7.47(m, 2H), 7.27-7.23 (m, 1H), 6.94-6.90 (m, 3H), 3.71 (bs, 2H), 3.42 (s, 2H), 2.28 (s, 3H), 1.61- 1.58 (m, 2H), 1.36 (s, 9H), 1.31-1.07 (m, 10H), 0.80 (t, J =6.8 Hz, 6H). LCMS: (Method A) 635.2 (M ⁺ +H), Rt. 2.83 min, 97.38% (maximum). HPLC: (method B) Rt. 5.96 min, 96.71% (maximum).

Example 80

(E) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid

With stirring, to a solution of tert-butyl-(E)-3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (Intermediate 133; 0.11 g, 0.20 mmol) in DCM (3 mL) TFA (0.02 g, 2.02 mmol) was added at 0° C and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between water (5 ml) and EtOAc (5 ml). The organic layer was washed with water (5 ml) and brine (5 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 2-3% MeOH in DCM, silica gel: 230-400 mesh) to give the title compound. Yield: 51% (50 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.59 (d, J =12.0 Hz, 1H), 7.39 (s, 1H), 7.28 (t, J =8, 0 Hz, 2H), 7.14 (d, J =8.0 Hz, 2H), 6.96 (t, J =7.2 Hz, 1H), 6.29 (s, 1H), 5.41 (d, J = 12.4 Hz, 1H), 3.76 (bs, 2H), 3.28 (s, 2H), 2.72 (s, 6H), 1.37-1.30 (m, 4H), 1.24-1.07 (m, 4H), 0.74 (t, J =8.00 Hz, 6H). LCMS: (Method A) 487.2 (M ⁺ +H), Rt. 2.81 min, 95.8% (maximum), HPLC: (method B) Rt. 5.75 min, 95.33% (maximum).

Example 81

(E) -3-((3,3-dibutyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid

With stirring, to a solution of tert-butyl-(E)-3-((3,3-dibutyl-7-fluoro-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 141; 0.16 g, 0.29 mmol) in DCM (5 ml) was added TFA (0.2 ml, 2.93 mmol) at 0°C and the reaction mixture stirred for 3 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between EtOAc (10 ml) and water (10 ml). The organic layer was washed with water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified with Isolera (eluent: 20% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 55% (78 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.26 (s, 1H), 7.77 (s, 1H), 7.73-7.70 (m, 1H), 7.36 ( t, J =8.4 Hz, 2H), 7.29 (d, J =7.6 Hz, 2H), 7.10 (t, J =7.2 Hz, 1H), 6.69 (d, J =12.0 Hz, 1H), 5.41(d, J =12.0 Hz, 1H), 3.82(bs, 2H), 3.45(s, 2H), 1.43-1, 28 (m, 4H), 1.13-0.99 (m, 8H), 0.73 (t, J =8.00 Hz, 6H). LCMS: (Method A) 490.2 (M ⁺ +H), Rt. 3.0 min, 97.39% (maximum), HPLC: (method B) Rt. 6.28 min, 97.94% (maximum).

Example 82

(E) -3-((3,3-dibutyl-7-cyano-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid

With stirring, to a solution of tert-butyl-(E)-3-((3,3-dibutyl-7-cyano-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylate (intermediate 144; 220 mg, 0.39 mmol) in DCM (2 ml) at 0° C. TFA (0.15 ml, 2.0 mmol) was added dropwise followed by the reaction the mixture was stirred for 6 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the residue was subjected to distribution between ice-cold water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with ice-cold water (5 ml) and brine (5 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and 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: 24% (49 mg, yellow solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.85 (d, J =12.0 Hz, 1H), 7.75 (s, 1H), 7.36 (t, J =7.2 Hz, 2H), 7.28 (t, J =7.6 Hz, 3H), 7.10 (t, J =7.6 Hz, 1H), 5.62 (d, J =12.4 Hz, 1H), 3.81 (s, 2H), 3.53 (s, 2H), 1.42-1.24 (m, 4H), 1.17-0.99 (m, 8H), 0.74 (t, J =6.4 Hz, 6H). LCMS: (Method A) 497.2 (M ⁺ +H), Rt. 2.91 min, 98.47% (maximum). HPLC: (method B) Rt. 6.17 min, 98.03% (maximum).

Example 83

(E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)but-2-enoic acid

With stirring, to a solution of (E) -3-((3,3-dibutyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5- tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)but-2-enoic acid (intermediate 146; 0.15 g, 0.22 mmol) in dry DCM (10 mL) TFA was added at 0°C (0.4 ml) and triethylsilane (0.4 ml). The reaction mixture was then stirred for 30 minutes at room temperature. After completion of the reaction (monitored by LCMS), the reaction mixture was diluted with ice-cold water (5 ml) and the aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 14% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 40% (50 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 11.88 (s, 1H), 7.61 (s, 1H), 7.41-7.32 (m, 5H), 7.15-7 .13 (m, 1H), 6.48 (s, 1H), 4.65 (s, 1H), 4.01 (s, 2H), 2.38 (s, 3H), 2.05 (s, 3H), 1.60-1.30 (m, 4H), 1.30-1.15 (m, 2H), 1.15-1.05 (m, 2H), 1.05-0.85 ( m, 4H), 0.72 (s, 6H). LCMS: (Method A) 533.3 (M ⁺ +H), Rt. 2.99 min, 98.60% (maximum). HPLC: (method B) Rt. 6.36 min, 98.35% (maximum).

Example 84

(Z)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of (Z)-3-((3-butyl-3-ethyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4, 5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (intermediate 148; 0.35 g, 0.55 mmol) in dry DCM (5 ml) at 0°C was added TFA (2.5 ml) and triethylsilane (2.5 ml). The reaction mixture was then stirred for 3 hours at room temperature. After completion of the reaction (monitored by LCMS), the reaction mixture was diluted with ice-cold water (10 ml) and the aqueous layer was extracted with DCM (2×10 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and evaporated in vacuo. The resulting crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 35% (100 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.49 (bs, 1H), 7.52-7.34 (m, 5H), 7.32-7.22 (m, 2H), 7 .09-7.08(m, 1H), 6.52(s, 1H), 4.15-3.62(m, 2H), 2.12(s, 3H), 1.63-1.36 (m, 4H), 1.24-1.06 (m, 4H), 0.93-0.71 (m, 6H). LCMS: (Method A) 509.2 (M ⁺ +H), Rt. 2.72 min, 98.96% (maximum). HPLC: (method B) Rt. 5.77 min, 99.77% (maximum).

Examples 85 and 86

(S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5- phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

The two enantiomers of racemic (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5 -benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 84; 0.08 g, 0.16 mmol) was separated by chiral preparative SFC; mobile phase: CO ₂ : 0.5% isopropylamine in methanol (70:30); column: YMC Cellulose-SC; Flow rate: 3 ml/min; wavelength: 210 nm; cycle time: 7 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 25% (20 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.58 (bs, 1H), 7.57-7.31 (m, 5H), 7.32-7.22 (m, 2H), 7 .15-7.14(m, 1H), 6.58(s, 1H), 4.04-3.82(m, 2H), 2.18(s, 3H), 1.69-1.42 (m, 4H), 1.30-1.13 (m, 4H), 0.94-0.71 (m, 6H). LCMS: (Method A) 509.1 (M ⁺ +H), Rt. 2.74 min, 98.86% (maximum). HPLC: (method B) Rt. 5.77 min, 98.84% (maximum). SFC: (method F) Rt. 5.25 min, 100% (maximum).

Enantiomer 2: Yield: 25% (20 mg, pale brown solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.50 (bs, 1H), 7.57-7.43 (m, 3H), 7.36 (t, J =7.2 Hz, 2H ), 7.39-7.18 (m, 2H), 7.09-7.07 (m, 1H), 6.52 (s, 1H), 4.04-3.82 (m, 2H), 2.11 (s, 3H), 1.62-1.36 (m, 4H), 1.24-1.06 (m, 4H), 0.94-0.71 (m, 6H). LCMS: (Method A) 509.2 (M ⁺ +H), Rt. 2.72 min, 98.97% (maximum). HPLC: (method B) Rt. 5.77 min, 99.50% (maximum). SFC: (method F) Rt. 5.93 min, 95.00% (maximum).

Example 87

(E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)acrylic acid

With stirring to a solution of tert-butyl-(E)-3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 2,5-Benzothiazepin-8-yl)oxy)acrylate (Intermediate 150; 0.2 g, 0.34 mmol) in dry DCM (5 ml) TFA (2 ml) was added at 0°C and the reaction mixture was stirred at for 2 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 28% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 81% (148 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.23 (s, 1H), 7.74 (bs, 1H), 7.68 (d, J =6.80 Hz, 1H), 7, 51 (s, 1H), 7.41 (t, J =7.6 Hz, 2H), 7.31 (d, J =7.2 Hz, 2H), 7.16 (t, J =7.2 Hz, 1H), 6.85 (bs, 1H), 5.26 (d, J = 12.4 Hz, 1H), 4.03 (bs, 2H), 1.60-1.55 (m, 1H ), 1.54-1.35(m, 3H), 1.29-0.98(m, 2H), 0.95-0.81(m, 2H), 0.75-0.61(m , 6H). LCMS: (Method A) 525.2 (M ⁺ +2H), Rt. 2.76 min, 90.66% (maximum). HPLC: (method B) Rt. 5.77 min, 90.61% (maximum).

Example 88

(E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-Benzothiadiazepin-8-yl)hydroxy)acrylic acid

With stirring to a solution of tert-butyl-(E)-3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4, 5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (intermediate 153; 190 mg, 0.33 mmol) in dry DCM (3 ml) TFA (2 ml) was added at 0°C and the reaction mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and diluted with ice-cold water (10 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was dried over anhydrous Na ₂ SO ₄ . The organic portion was evaporated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 20% EtOAc in hexane; silica gel: 230-400 mesh) to give the title compound. Yield: 80% (148 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.22 (s, 1H), 7.66 (d, J =12.2 Hz, 1H), 7.40-7.36 (m, 3H ), 7.30 (d, J = 7.4 Hz, 2H), 7.12 (t, J = 7.2 Hz, 1H), 6.49 (s, 1H), 5.39 (d, J =12.2 Hz, 1H), 4.12 (bs, 2H), 2.88 (s, 3H), 2.10 (s, 3H), 1.90-1.87 (m, 2H), 1 .52-1.49 (m, 2H), 1.16-0.90 (m, 4H), 0.82-0.61 (m, 6H). LCMS: (Method A) 504.9 (M ⁺ ), Rt. 2.91 min, 97.98% (maximum). HPLC: (method B) Rt. 6.01 min, 97.03% (maximum).

Examples 89 and 90

(S)- (E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)acrylic acid and (R)- (E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1 -dioxido-5-phenyl-2,3,4,5-tetrahydro1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

The two enantiomers of racemic (E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1 ,2,5-Benzothiadiazepin-8-yl)oxy)acrylic acid (Example 88; 0.14 g, 0.27 mmol) was separated by chiral preparative SFC (method F); mobile phase: CO ₂ : 0.5% isopropylamine in methanol; wavelength: 280 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 28% (40 mg, white solid). ¹ H NMR (400 MHz, DMSO- d6 ): δ 12.20 (bs, 1H) _, 7.66 (d, J =12.4 Hz, 1H), 7.40-7.36 (m, 5H ), 7.12 (t, J = 7.2 Hz, 1H), 6.49 (s, 1H), 5.39 (d, J = 12.0 Hz, 1H), 4.12 (bs, 2H ), 2.88 (s, 3H), 2.09 (s, 3H), 1.91-1.87 (m, 1H), 1.79-1.72 (m, 1H), 1.56- 1.50 (m, 2H), 1.24-1.05 (m, 4H), 0.76-0.70 (m, 6H). LCMS: (Method A) 505.2 (M ⁺ +H), Rt. 2.89 min, 97.38% (maximum). HPLC: (method B) Rt. 5.99 min, 98.97% (maximum). SFC: (Method E) Rt. 2.77 min, 100% (maximum).

Enantiomer 2: Yield: 28% (40 mg, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.19 (bs, 1H), 7.65 (d, J =12.0 Hz, 1H), 7.38 (t, J =8.0 Hz, 3H), 7.29 (d, J =7.6 Hz, 2H), 7.11 (t, J =7.2 Hz, 1H), 6.48 (s, 1H), 5.38 ( d, J =12.4 Hz, 1H), 4.06(s, 2H), 2.87(s, 3H), 2.09(s, 3H), 1.91-1.86(m, 1H ), 1.78-1.76 (m, 1H), 1.55-1.52 (m, 2H), 1.46-0.90 (m, 4H), 0.81-0.62 (m , 6H), LCMS: (method E) 505.1 (M ⁺ +H), Rt. 2.69 min, 97.32% (maximum). HPLC: (method B) Rt. 5.99 min, 97.57% (maximum). SFC: (Method E) Rt. 3.06 min, 93.78% (maximum).

Example 91

(E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid

With stirring, to a solution of tert-butyl-(E)-3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)acrylate (intermediate 156; 0.18 g, 0.33 mmol) in DCM (5 mL) TFA (0.04 g, 3.31 mmol) was added at 0° C and the reaction mixture was stirred for 1 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the residue was subjected to distribution between EtOAc (5 ml) and water (5 ml). The organic layer was washed with water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 2-3% MeOH in DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 58% (90 mg, almost white solid).

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 12.15 (s, 1H), 7.64 (d, J =12.0 Hz, 1H), 7.32 (s, 1H), 7 .25 (t, J =8.0 Hz, 2H), 7.08 (d, J =7.6 Hz, 2H), 6.90 (t, J =7.2 Hz, 1H), 6.30 (d, J =4.8 Hz, 1H), 6.01 (s, 1H), 5.34 (d, J =12.4 Hz, 1H), 3.69 (bs, 2H), 3.18 (s, 2H), 2.67 (s, 3H), 1.54-1.41 (m, 2H), 1.37-1.30 (m, 2H), 1.11-1.00 (m , 4H), 0.75 (t, J = 8.00 Hz, 6H). LCMS: (Method A) 473.3 (M ⁺ +H), Rt. 2.67 min, 98.1% (maximum). HPLC: (method B) Rt. 5.59 min, 97.1% (maximum).

Examples 92 and 93

(S)- (E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid and (R)- (E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxydo-5-phenyl-2,3 ,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

The two enantiomers of racemic (E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine -8-yl)hydroxy)acrylic acid (Example 91; 80 mg, 0.16 mmol) was separated by chiral preparative SFC (Method E); wavelength: 220 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 11% (10 mg, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.38 (d, J =12.0 Hz, 1H), 7.26 (s, 1H), 7.23 (t, J =8.4 Hz, 2H), 7.05 (d, J =7.2 Hz, 2H), 6.88 (t, J =7.2 Hz, 1H), 6.23 (d, J =4.8 Hz, 1H), 6.01 (s, 1H), 5.37 (d, J =12.0 Hz, 1H), 3.67 (bs, 2H), 3.19 (s, 2H), 2.51 ( s, 3H), 1.52-1.32 (m, 2H), 1.30-1.01 (m, 6H), 0.73 (t, J =8.00 Hz, 6H). LCMS: (Method A) 473.3 (M ⁺ +H), Rt. 2.67 min, 95.41% (maximum). HPLC: (method B) Rt. 5.60 min, 97.83% (maximum). Chiral SFC: (method D) Rt. 2.95 min, 95.67% (maximum).

Enantiomer 2: Yield: 18% (15 mg, white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.39 (d, J =12.0 Hz, 1H), 7.26 (s, 1H), 7.23 (t, J =8.4 Hz, 2H), 7.05 (d, J =7.6 Hz, 2H), 6.88 (t, J =7.2 Hz, 1H), 6.23 (d, J =4.8 Hz, 1H), 6.01 (s, 1H), 5.37 (d, J =12.0 Hz, 1H), 3.71 (bs, 2H), 3.18 (s, 2H), 2.67 ( s, 3H), 1.52-1.35 (m, 2H), 1.30-1.08 (m, 6H), 0.73 (t, J =8.00 Hz, 6H). LCMS: (Method A) 473.2 (M ⁺ +H), Rt. 2.69 min, 91.43% (maximum). HPLC: (method B) Rt. 5.6 min, 96.86% (maximum). Chiral SFC: (method D) Rt. 3.77 min, 95.82% (maximum).

Example 94

(Z) -3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid (Example 11; 100 mg, 0.18 mmol) in DMF (5 ml) at 0°C was added N-bromosuccinimide (35.5 mg, 0.2 mmol) and the reaction the mixture was stirred for 1 h at 0°C. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (5 ml) and EtOAc (5 ml). The aqueous layer was extracted with EtOAc (2×5 ml). The combined organic layer was washed with ice-cold water (5 ml) and brine (5 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 60% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 52% (60 mg, brown solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.47 (bs, 1H), 7.62 (s, 1H), 7.57 (s, 1H), 7.40 (d, J =9 .2 Hz, 2H), 7.01-6.99 (m, 2H), 6.84 (s, 1H), 3.48 (bs, 2H), 3.32 (s, 2H), 2.28 (s, 3H), 1.55-1.39 (m, 2H), 1.47-1.21 (m, 2H), 1.20-1.04 (m, 8H), 0.79 (t , J = 6.8 Hz, 6H). LCMS: (Method E) 616.0 (M ⁺ +2H), Rt, 2.92 min, 96.99% (maximum). HPLC: (method B) Rt. 6.61 min, 97.49% (maximum).

Example 95

(Z) -3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)-2-fluoroacrylic acid

With stirring, to a solution of ethyl-(Z)-3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 159; 0.51 g, 0.88 mmol) in 1,4-dioxane/water (4:1, 10 mL) at room temperature temperature, lithium hydroxide (74 mg, 1.75 mmol) was added and the reaction mixture was stirred for 30 minutes at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (10 ml) and EtOAc (10 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with ice-cold water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 8% MeOH/DCM; silica gel: 230-400 mesh) to give the title compound. Yield: 42% (235 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 9.37 (s, 1H), 7.47 (s, 1H), 7.42 (d, J =18.8 Hz, 1H), 7, 14 (d, J =8.0 Hz, 2H), 6.77 (d, J =8.8 Hz, 2H), 6.34 (s, 1H), 3.71 (s, 2H), 3, 39 (s, 2H), 2.09 (s, 3H), 1.42-1.35 (m, 4H), 1.11-1.07 (m, 4H), 1.04-0.97 ( m, 4H), 0.76-0.74 (m, 6H). LCMS: (Method A) 552.1 (M ⁺ +H), Rt. 2.58 min, 97.48% (maximum). HPLC: (method B) Rt. 5.52 min, 97.70% (maximum).

Example 96

(Z) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid

With stirring, to a solution of methyl-(Z)-3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 167; 0.12 g, 0.24 mmol) in a mixture of 1,4-dioxane and water (4:1.5 ml) was added lithium hydroxide (0. 05 g, 1.22 mmol) and the reaction mixture was stirred for 4 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with ice-cold water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml). The combined organic layer was washed with water (5 ml) and brine (5 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 18-20% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 43% (50 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.78 (d, J =8.80 Hz, 1H), 7.57 (d, J =18.00 Hz, 1H), 7.32- 7.30 (m, 2H), 7.18-7.16 (m, 2H), 7.07 (t, J =7.20 Hz, 1H), 6.75 (d, J =12.00 Hz , 1H), 3.78 (d, J = 6.40 Hz, 2H), 3.38 (m, 2H), 1.24-1.20 (m, 4H), 1.06-1.05 ( m, 4H), 0.84-0.60 (m, 6H). LCMS: (Method E) 480.1 (M ⁺ +H), Rt. 2.61 min, 99.74% (maximum). HPLC: (method B) Rt. 5.84 min, 98.24% (maximum).

Example 97

(Z) -3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of (Z) -3-((3,3-dibutyl-7-(dimethylamino)-2-(4-methoxybenzyl)-1,1-dioxydo-5-phenyl-2,3,4,5- tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (intermediate 171; 0.02 g, 0.27 mmol) in dry DCM (2 ml) trifluoroacetic acid (1 ml) was added and triethylsilane (1 ml) at 0°C and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between EtOAc (5 ml) and water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was dried over anhydrous Na ₂ SO ₄ and evaporated in vacuum. The resulting crude material was purified by preparative HPLC (method A) to give the title compound. Yield: 11% (17 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.42 (s, 1H), 7.37-7.35 (m, 3H), 7.33-7.30 (m, 2H), 7 .26 (d, J = 9.6 Hz, 2H), 7.07 (s, 1H), 6.15 (s, 1H), 4.05 (bs, 2H), 2.59 (s, 6H) , 1.51-1.36 (m, 4H), 1.33-1.19 (m, 4H), 1.09-0.95 (m, 4H), 0.72 (t, J =8, 00Hz, 6H). LCMS: (Method E) 534.2 (M ⁺ +H), Rt. 2.75 min, 99.62% (maximum). HPLC: (method B) Rt. 6.12 min, 98.21% (maximum).

Example 98

(Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring, to a solution of ethyl-(Z)-3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5- tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 172; 0.39 g, 0.70 mmol) in a mixture of THF and water (10 ml, 4:1) hydroxide was added lithium (89 mg, 2.12 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with water (10 ml). The aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layer was washed with water (10 ml) and brine (10 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 35% (130 mg, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.50 (d, J =18.4 Hz, 1H), 7.43 (s, 1H), 7.36 (t, J =7.6 Hz, 2H), 7.25 (d, J =6.8 Hz, 2H), 7.09 (s, 1H), 6.51 (s, 1H), 4.08 (bs, 2H), 2, 85 (s, 3H), 2.12 (s, 3H), 1.91-1.76 (m, 2H), 1.55-1.47 (m, 2H), 1.24-0.90 ( m, 4H), 0.77-0.71 (m, 6H). LCMS: (Method A) 523.2 (M ⁺ +H), Rt. 2.90 min, 99.26% (maximum). HPLC: (method B) Rt. 6.07 min, 99.61% (maximum)

Examples 99 and 100

(S)- (Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)- (Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)- 1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

The two enantiomers of racemic (Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1 ,2,5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 98; 0.13 g, 0.25 mmol) was separated by chiral preparative SFC (method F); wavelength: 280 nm; cycle time: 5 min; back pressure: 100 bar. The material was concentrated in vacuo at 40°C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 38% (50 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.5 (bs, 1H), 7.53 (d, J =18.4 Hz, 1H), 7.49 (s, 1H), 7, 36 (t, J =7.6 Hz, 2H), 7.25 (d, J =7.6 Hz, 2H), 7.09 (t, J =7.2 Hz, 1H), 6.51 ( s, 1H), 4.08 (bs, 2H), 2.85 (s, 3H), 2.15 (s, 3H), 1.90-1.78 (m, 2H), 1.56-1 .47 (m, 2H), 1.25-0.90 (m, 4H), 0.77-0.71 (m, 6H). LCMS: (Method E) 523.1 (M ⁺ +H), Rt. 2.72 min, 98.30% (maximum). HPLC: (method B) Rt. 6.07 min, 97.89% (maximum). SFC: (method H) Rt. 5.71 min, 99.46% (maximum).

Enantiomer 2: Yield: 40% (52 mg, almost white solid). ¹ HNMR (400 MHz, DMSO- d ₆ ): δ 7.50-7.35 (m, 4H), 7.25 (d, J =6.8 Hz, 2H), 7.09 (t, J = 6.8 Hz, 1H), 6.51 (s, 1H), 4.08 (s, 2H), 2.85 (s, 3H), 2.12 (s, 3H), 1.91-1, 76 (m, 2H), 1.55-1.49 (m, 2H), 1.26-1.23 (m, 1H), 1.18-1.07 (m, 2H), 0.96- 0.81 (m, 1H), 0.77-0.73 (m, 6H). LCMS: (Method E) 523.1 (M ⁺ +H), Rt. 2.72 min, 98.06% (maximum). HPLC: (method B) Rt. 6.07 min, 97.19% (maximum). SFC: (method H) Rt. 6.7 min, 98.20% (maximum).

Example 101

(E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid

With stirring to a solution of tert-butyl-(E)-3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)acrylate (Intermediate 173; 0.5 g, 0.97 mmol) in DCM (3 mL) was added dropwise with TFA (3 mL) at 0°C and stirred for 5 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 14-15% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 90% (400 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 12.26 (s, 1H), 7.74 (t, J =9.20 Hz, 2H), 7.36 (t, J =7.20 Hz, 2H), 7.27 (d, J =6.80 Hz, 2H), 7.09 (t, J =7.20 Hz, 1H), 5.43 (d, J =12.40 Hz, 1H), 6.72 (d, J =12.80 Hz, 1H), 3.80 (d, J =14.80 Hz, 2H), 3.45 (s, 2H), 1.29-1, 32 (m, 4H), 0.95-0.97 (m, 4H), 0.69-0.71 (m, 6H). LCMS: (Method E) 462.1 (M ⁺ +H), Rt. 2.59 min, 96.24% (maximum). HPLC: (method B) Rt. 5.73 min, 97.13% (maximum).

Examples 102 and 103

(R)- (E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)acrylic acid and (S)- (E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxido-5-phenyl-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

The two enantiomers of racemic (E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8 -yl)hydroxy)acrylic acid (Example 101; 0.40 g, 0.87 mmol separated by chiral preparative SFC (method E), wavelength: 280 nm, cycle time: 5 min, backpressure: 100 bar. in vacuo at 40° C. The first eluting fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configuration of these two enantiomers is unknown.

Enantiomer 1: Yield: 26% (85 mg, almost white solid). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.69 (d, J =8.80 Hz, 1H), 7.63 (d, J =12.40 Hz, 1H), 7.35 ( t, J =7.20 Hz, 2H), 7.25-7.27 (m, 2H), 7.08 (t, J =7.20 Hz, 1H), 6.73 (d, J =12 .80 Hz, 1H), 5.43 (d, J = 12.40 Hz, 1H), 3.44 (s, 2H), 3.35 (s, 2H), 1.36-1.38 (m , 1H), 1.31-1.32(m, 3H), 1.10-1.11(m, 4H), 0.69-0.71(m, 6H). LCMS: (Method E) 462.1 (M ⁺ +H), Rt. 2.61 min, 95.03% (maximum). HPLC: (method B) Rt. 5.73 min, 97.89% (maximum). SFC: (method D) Rt. 3.47 min, 99.44% (maximum).

Enantiomer 2: Yield: 30% (160 mg, almost white solid). ¹ HNMR (400 MHz, DMSO- d ₆ ): δ 7.64 (d, J =8.80 Hz, 1H), 7.46 (d, J =12.40 Hz, 1H), 7.34 (t , J = 7.60 Hz, 2H), 7.24 (d, J = 7.60 Hz, 2H), 7.06 (t, J = 7.20 Hz, 1H), 6.74 (d, J =12.00 Hz, 1H), 5.41 (d, J =12.00 Hz, 1H), 3.79 (s, 2H), 3.34 (s, 2H), 1.41-1.53 (m, 1H), 1.31-1.32 (m, 3H), 0.99-1.02 (m, 4H), 0.71-0.73 (m, 6H). LCMS: (Method E) 462.1 (M ⁺ +H), Rt. 2.61 min, 95.27% (maximum). HPLC: (method B) Rt. 5.73 min, 96.74% (maximum). SFC: (method D) Rt. 4.04 min, 95.44% (maximum).

Example 104

(Z) -3-((3,3-dibutyl-5-(4-(dimethylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring to a solution of ethyl-(Z)-3-((3,3-dibutyl-5-(4-(dimethylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5 -tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 175; 450 mg, 0.7 mmol) in a mixture of 1,4-dioxane and water (4:1, 5 ml) was added lithium hydroxide (59.5 mg, 0.16 mmol) and the reaction mixture was stirred for 3 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated in vacuo and the resulting residue was subjected to distribution between ice-cold water (10 ml) and EtOAc (10 ml). The aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with ice-cold water (15 ml) and brine (15 ml) and dried over anhydrous Na ₂ SO ₄ . The organic portion was concentrated in vacuo and the resulting crude material was purified by Isolera column chromatography (eluent: 18% MeOH/DCM; silica gel: 230-400 mesh). The resulting compound was repurified by preparative HPLC (method A) to give the title compound. Yield: 3% (7 mg, almost white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.50 (s, 1H), 7.43-7.33 (m, 1H), 7.30 (d, J =8.4 Hz, 2H ), 7.03-7.01 (m, 2H), 6.87 (s, 1H), 3.92 (bs, 2H), 3.35 (s, 2H), 2.94 (s, 6H) , 2.27 (s, 3H), 1.44-1.29 (m, 4H), 1.23-1.03 (m, 8H), 0.77 (t, J =6.4 Hz, 6H ). LCMS: (Method A) 607.3 (M ⁺ +H), Rt. 2.50 min, 95.26% (maximum). HPLC: (Method E) Rt. 5.38 min, 97.77% (maximum).

Example 105

(Z) -3-((3,3-dibutyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring, to a solution of ethyl-(Z)-3-((3,3-dibutyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 178; 0.5 g, 0.86 mmol) in a mixture of THF and water (10 ml, 4:1) was added lithium hydroxide ( 108 mg, 2.57 mmol) and the reaction mixture was stirred for 16 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with water (10 ml). The aqueous layer was extracted with EtOAc (2×20 ml) and the combined organic layer was washed with water (10 ml) and brine (10 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 55% EtOAc/PE; silica gel: 230-400 mesh) to give the title compound. Yield: 36% (0.17 g, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.46 (bs, 1H), 7.53 (d, J =18.8 Hz, 1H), 7.44 (s, 1H), 7, 39-7.35 (m, 2H), 7.28 (d, J =7.6 Hz, 2H), 7.11 (t, J =7.2 Hz, 1H), 6.49 (s, 1H ), 4.12 (bs, 2H), 2.87 (s, 3H), 2.11 (s, 3H), 1.82-1.77 (m, 2H), 1.51-1.45 ( m, 2H), 1.24-0.95 (m, 8H), 0.76-0.74 (m, 6H). LCMS: (Method E) 551.2 (M ⁺ +H), Rt. 2.88 min, 97.46% (maximum). HPLC: (method B) Rt. 6.55 min, 97.54% (maximum).

Example 106

(Z) -3-((7-bromo-3-butyl-3-ethyl-2-methyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring, to a solution of ethyl-(Z)-3-((7-bromo-3-butyl-3-ethyl-2-methyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-Benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 181; 0.15 g, 0.25 mmol) in 1,4-dioxane/water (5 mL, 4:1) lithium hydroxide (32 mg, 0.77 mmol) was added and the reaction mixture was stirred for 16 h at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 1.5 ml, pH~4) and diluted with water (15 ml). The aqueous layer was extracted with EtOAc (2×15 ml) and the combined organic layer was washed with water (10 ml) and brine (10 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method D) to give the title compound. Yield: 42% (60 mg, white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.57 (s, 1H), 7.46-7.29 (m, 5H), 7.16 (d, J =6.4 Hz, 1H ), 6.88 (s, 1H), 4.12 (bs, 2H), 2.89 (s, 3H), 1.88-1.75 (m, 2H), 1.54-1.48 ( m, 2H), 1.17-1.13 (m, 2H), 1.07-0.96 (m, 2H), 0.87-0.85 (m, 6H). LCMS: (method E) 555.0 (M ⁺ ), Rt. 2.73 min, 98.87% (maximum). HPLC: (method B) Rt. 6.67 min, 99.69% (maximum)

Example 107

(Z) -3-((3,3-dibutyl-5-(3,4-difluorophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

With stirring, to a solution of ethyl-(Z)-3-((3,3-dibutyl-5-(3,4-difluorophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 187; 0.05 g, 0.083 mmol) in a mixture of 1,4-dioxane and water (4:1, 5 ml) hydroxide was added lithium (0.01 g, 0.25 mmol) and the reaction mixture was stirred for 4 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with ice-cold water (5 ml). The aqueous layer was extracted with EtOAc (2×5 ml) and the combined organic layer was washed with water (5 ml) and brine (5 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC to give the pure title compound. Yield: 32% (0.015 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 13.63 (bs, 1H), 7.59-7.54 (m, 2H), 7.34-7.24 (m, 2H), 6 .81-6.78(m, 2H), 3.76(s, 2H), 3.35(s, 2H), 2.28(s, 3H), 1.35-1.33(m, 4H ), 1.23-1.13 (m, 8H), 0.79-0.76 (m, 6H). LCMS: (Method A) 572.1 (M ⁺ +H), Rt. 2.98 min, 95.80% (maximum). HPLC: (method B) Rt. 6.36 min, 98.43% (maximum).

Example 108

(Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid

With stirring to a solution of ethyl-(Z)-3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1, 5-Benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 188; 0.07 g, 0.13 mmol) in a mixture of 1,4-dioxane and water (4:1.5 ml) was added lithium hydroxide ( 0.02 g, 0.39 mmol) and the reaction mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with ice-cold water (5 ml). The aqueous layer was extracted with EtOAc (2×10 ml) and the combined organic layer was washed with water (8 ml) and brine (10 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (method A). Yield: 39% (0.026 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.39 (s, 1H), 7.26 (t, J= 7.2 Hz, 2H), 7.09 (d, J =7.6 Hz, 2H), 7.03 (s, 1H), 6.92 (t, J = 7.2 Hz, 1H), 6.31 (s, 1H), 3.70 (bs, 2H), 3, 26 (s, 2H), 2.70 (s, 6H), 1.54-1.51 (m, 1H), 1.42-1.30 (m, 3H), 1.24-1.01 ( m, 4H), 0.77-0.75 (m, 6H). LCMS: (Method A) 505.1 (M ⁺ + H), Rt. 2.88 min, 99.93% (maximum). HPLC: (method B) Rt. 5.88 min, 98.74% (maximum).

Examples 109 and 110

(S)- (Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)- (Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxydo-5-phenyl- 2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

Lithium hydroxide (0.021 g, 0.48 mmol) was added with stirring to a solution of enantiomer 1 of intermediate 189 (0.13 g, 0.24 mmol) in a mixture of 1,4-dioxane and water (4:1.5 ml) and the reaction mixture was stirred for 1 hour at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with ice-cold water (10 ml). The aqueous layer was extracted with EtOAc (2×15 ml) and the combined organic layer was washed with water (8 ml) and brine (10 ml). The organic part was dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. The resulting material was triturated with Et ₂ O and concentrated in vacuo to give the title compound. The absolute configuration of the enantiomer is unknown. Yield: 39% (0.026 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.39 (s, 1H), 7.25 (t, J =7.6 Hz, 2H), 7.09 (d, J =7.6 Hz, 3H), 6.91 (t, J =6.8 Hz, 1H), 6.31 (s, 1H), 3.69 (bs, 2H), 3.26 (s, 2H), 2, 70 (s, 6H), 1.52-1.41 (m, 1H), 1.40-1.30 (m, 3H), 1.24-1.02 (m, 4H), 0.75 ( t, J = 6.40 Hz, 6H). LCMS: (Method A) 505.2 (M ⁺ + H), Rt. 2.84 min, 99.51% (maximum). HPLC: (method B) Rt. 5.87 min, 98.75% (maximum). SFC: (method F) Rt. 8.93 min, 100% (maximum).

Lithium hydroxide (0.024 g, 0.56 mmol) was added to a solution of enantiomer 2 of intermediate 189 (0.15 g, 0.28 mmol) in a mixture of 1,4-dioxane and water (4:1, 5 ml) with stirring, and the reaction mixture was stirred for 2 hours at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5N, 3 ml, pH~4) and diluted with ice-cold water (10 ml). The aqueous layer was extracted with EtOAc (2×15 ml). The combined organic layer was washed with water (8 ml) and brine (10 ml) and then dried over anhydrous Na ₂ SO ₄ . The organic part was concentrated in vacuo and the resulting material was triturated with Et ₂ O and received the desired compound. The absolute configuration of the enantiomer is unknown. Yield: 39% (0.026 g, off-white solid).

¹ H NMR (400 MHz, DMSO- d ₆ ): δ 7.40 (s, 1H), 7.26 (t, J =7.6 Hz, 2H), 7.15 (s, 1H), 7, 09 (d, J =7.6 Hz, 2H), 6.92 (t, J =6.8 Hz, 1H), 6.31 (s, 1H), 3.69 (bs, 2H), 3, 26 (s, 2H), 2.69 (s, 6H), 1.51-1.49 (m, 1H), 1.35-1.32 (m, 3H), 1.21-1.11 ( m, 4H), 0.73 (t, J = 7.20 Hz, 6H). LCMS: (Method A) 505.2 (M ⁺ + H), Rt. 2.85 min, 96.20% (maximum). HPLC: (method B) Rt. 5.87 min, 99.29% (maximum). SFC: (method F) Rt. 9.62 min, 98.93% (maximum).

BIOLOGICAL RESEARCH

ASBT analysis protocol (h/m)

10,000 cells (human or mouse cells overexpressing ASBT) were seeded in a 96-well plate (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 ₂ for 48 hours. After incubation, the medium was decanted from the wells and the cells were washed twice with 300 µl of MEM-alpha basic medium (containing no FBS). Each time the MEM-alpha base medium was decanted, the plates were tapped on paper towel to ensure maximum removal of remaining media.

Dilutions of test inhibitors (in the study, the highest concentration was 10 μM, 3-fold serial dilution, 10 samples) prepared in DMSO (Sigma D2650) were added to the incubation mixture (maintaining a 0.2% final concentration of DMSO) containing 0. 25 μM 3H-taurocholic acid (ARC ART-1368) and 5 μM cold taurocholinic acid (Sigma T4009). Then, 50 μl of the incubation mix containing test inhibitors was added to the wells (repeated twice) and the plates were incubated for 20 min in a CO ₂ incubator at 37°C. After the incubation, the reaction was stopped by placing the plates in ice water for 2-3 minutes, and then the incubation mixture was completely aspirated from the wells. The wells were washed twice with 250 μl of cold unlabeled 1 mM taurocholinic acid dissolved in HEPES buffered (Gibco 15630080) (10 mM) HBSS (Gibco 14175079) (pH 7.4). The plates were tapped on paper towel after each wash to ensure maximum removal of the blocking buffer.

100 µl of MicroScint-20 (PerkinElmer 6013621) was added to the wells and kept overnight at room temperature and then read on a PerkinElmer microplate scintillation and luminescence counter using the 3H assay protocol (set to read for 120 seconds per well).

LBAT analysis protocol (h/m)

20,000 cells (human or mouse cells overexpressing LBAT) were seeded in a 96-well plate (Corning CLS3809) in 100 µl of MEM-alpha medium (Gibco 12571-063) supplemented with 10% FBS (Gibco 10438026) containing geneticin (Gibco 10131- 027) (1 mg/ml) and incubated at 37° C. in 5% CO ₂ for 24 h. After incubation, the medium was decanted from the wells and the cells were washed twice with 300 μl of MEM-alpha basic medium (without FBS). Each time the MEM-alpha base medium was decanted, the plates were tapped on paper towel to ensure maximum removal of remaining media.

For human LBAT, the incubation mixture was prepared by adding dilutions of the inhibitors under study (3-fold serial dilution in DMSO (Sigma D2650), 10 samples) in MEM-alpha (without FBS) containing 0.3 μM 3H-taurocholinic acid (ARC ART- 1368) and 7.5 μM cold taurocholinic acid (Sigma T4009) (maintaining a 0.2% final concentration of DMSO). For mouse LBAT, an incubation mix was prepared by adding dilutions of the inhibitors under study (3-fold serial dilution in DMSO, 10 samples) in MEM-alpha (without FBS) containing 0.3 μM 3H-taurocholinic acid and 25 μM cold taurocholinic acid (maintaining equal to 0.2% final concentration of DMSO).

Then, 50 μl of the incubation mix containing test inhibitors was added to the wells (repeated twice) and the plates were incubated for 20 min in a CO ₂ incubator at 37°C. After the incubation, the reaction was stopped by placing the plates in ice water for 2-3 minutes, and then the incubation mixture was completely aspirated from the wells. The wells were washed twice with 250 μl of cold unlabeled 1 mM taurocholinic acid dissolved in HEPES buffered (Gibco 15630080) (10 mM) HBSS (Gibco 14175079) (pH 7.4). The plates were tapped on paper towel after each wash to ensure maximum removal of the blocking buffer.

100 µl of MicroScint-20 (PerkinElmer 6013621) was added to the wells and kept overnight at room temperature and then read on a PerkinElmer microplate scintillation and luminescence counter according to the 3H assay protocol (when set to read for 120 s per well in normal orientation tablet).

HepaRG-based assay protocol

A frozen vessel with differentiated HepaRG cells (Biopredic International HPR116080) was thawed in HepaRG Thawing/Plating/General Purpose (Biopredic International ADD670C) supplemented with 200 mM Glutamax (Gibco 35050061) according to the protocol provided by Biopredic International. 70,000 cells per well were seeded in a 96-well plate (Corning CLS3809) in 100 µl of HepaRG Thawing/Plating/General Purpose medium supplemented with 200 mM Glutamax and incubated at 37°C in 5% CO ₂ for 24 hours. the platings were changed to HepaRG Maintenance/Metabolism medium (Biopredic International ADD620C) and incubated for 6 days, replenishing with fresh HepaRG Maintenance/Metabolism medium every 48 h. µl of basic Williams medium E (Gibco 12551032). After each decantation of the basic Williams medium E, the plates were tapped on a paper towel to ensure maximum removal of the remaining media.

The incubation mixture was prepared by adding dilutions of the inhibitors under study (3-fold serial dilution in DMSO (Sigma D2650)) in William's E (basic) medium containing 0.3 μM 3H-taurocholinic acid (ARC ART-1368) and 7, 5 μM cold taurocholinic acid (Sigma T4009) (maintaining a 0.2% final concentration of DMSO). Then 50 μl of the incubation mixture containing the test inhibitors was added to the wells (repeated twice) and the plates were incubated for 30 min in a 5% CO ₂ incubator at 37°C. After the incubation, the reaction was stopped by placing the plates in ice water for 2-3 minutes, and then the incubation mixture was completely aspirated from the wells. The wells were washed twice with 250 μl of cold unlabeled 1 mM taurocholinic acid dissolved in HEPES buffered (Gibco 15630080) (10 mM) HBSS (Gibco 14175079) (pH 7.4). The plates were tapped on paper towel after each wash to ensure maximum removal of the blocking buffer.

100 µl of MicroScint-20 (PerkinElmer 6013621) was added to the wells and kept overnight at room temperature and then read on a PerkinElmer microplate scintillation and luminescence counter according to the 3H assay protocol (when set to read for 120 s per well in normal orientation tablet).

Preparation of dilutions of the test compound

All test compounds were provided in powder form at room temperature. Stock solutions of test compounds were prepared in 10 mM DMSO, aliquots were taken and stored at -20°C. From stock solutions of compounds in 10 mM DMSO, a 3-fold serial dilution in DMSO was prepared and a total of 10 dilutions of test compounds were obtained. To prepare the incubation mixture, 0.5 μl of this dilution in DMSO was added to 250 μl of FBS-free base medium containing 3H-taurocholinic acid and cold taurocholinic acid.

Bioavailability studies

Used mice C57BL/6 at the age of 8-9 weeks. For each test compound, two groups of 3 animals were used. Animals of one group received a single dose of 1 mg/kg (diluent 100% DMSO) intravenously via the tail vein, and animals of the second group received a single dose of 10 mg/kg orally with a feeding needle. Animals of the group dosed orally were fasted overnight. Blood samples were taken 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 h after oral administration. Blood samples were taken from the saphenous vein of the father. 0.2% EDTA was used as the anticoagulant. Samples were analyzed according to bioanalytical research methods developed for the determination of test compound in plasma using the LC-MS/MS system.

results

Biological data for the compounds of examples are shown below in table 7.

Table 7

Example hLBAT IC ₅₀ (nM) hASBT IC ₅₀ (nM) IC ₅₀ HepaRG cells (nM) Bioavailability (%) 1 145 70 2 1405 354 >6666 3 177 >10000 885 4 218 167 5 290 1231 6 156 57 809 100 7 749 217 3188 8 1475 40 9 1093 1195 3085 10 3977 1409 4704 eleven 45 48 1087 27 12 393 482 2610 13 991 144 4458 14 1426 304 5156 15 532 253 2251 16 82 33 1029 17 116 18 80 19 1264 35 19 783 128 2893 20 1298 20 21 4153 929 5231 22 343 2143 23 100 1204 24 225 4665 2282 25 351 370 3311 26 53 3244 477 27 46 4799 350 28 13 4447 89 29 205 666 thirty 107 336 31 17 310 32 506 209 672 33 214 691 34 76 469 35 66 590 36 39 877 37 45 2004 592 38 122 454 39 75 796 399 40 108 4546 1507 41 8.3 26 336 42 10 220 378 43 8 21 405 44 44 9.1 257 205 100 45 201 1458 3158 46 24 97 590 47 25 339 362 48 8.1 42 260 18 49 94 220 570 50 89 77 339 51 96 52 84 191 53 62 280 800 54 100 186 260 55 100 487 328 56 37 380 175 57 137 361 1161 58 84 24 1502 59 85 60 4220 39 61 1621 62 235 >20000 772 63 10 35 358 64 28 524 455 65 20 27 265 66 thirty 2758 518 67 461 192 68 130 6667 344 69 364 >3000 630 70 81 125 716 71 47 >3000 296 72 13 76 319 73 120 155 766 74 649 57 3380 75 349 148 76 217 41 77 297 24 5331 78 96 156 79 8 >1000 246 80 828 7.5 81 210 771 82 270 873 83 4919 18 84 31 28 559 85 71 7 1107 86 31 88 527 87 220 157 1075 88 70 2246 1153 89 290 2006 90 49 10000 1398 91 1299 41 92 1466 748 93 3163 52 94 96 42 95 77 17 1223 96 231 15 785 97 341 9 98 22 286 671 99 4.4 325 20 100 49 631 1447 101 102 298 3333 103 187 320 104 290 3333 105 68 106 31 673 246 107 50 59 365 108 5.9 8.5 447 109 68 6.5 >2222 110 33 138

Model PD: Investigation of the effect of test compound on total bile acids in male C57BL6 mice.

C57BL/6N Tac mice aged 8-9 weeks were used to study the effect of bile acid modulators on bile acid levels. After completion of the quarantine and acclimatization period, animals were randomized into x treatment groups based on body weight: (i) diluent control and (ii) oral administration of test compound at y mg/kg once daily. Animals were treated with test compound for 7 days. On day 5 of the study, animals were placed individually in clean cages. On day 7, faeces were collected from each cage, then blood was taken from each animal by the retrobulbar route. Animals were sacrificed to collect liver and terminal ileum from each animal for analysis. Body weight and food intake were determined twice a week. Serum lipid profiles were analyzed in day 7 serum samples. Serum total bile acids were determined in day 7 serum samples. Fecal bile excretion was determined in day 7 fecal samples. 7. The content of triglycerides and total cholesterol was determined in liver samples on day 7.

Urinary bile acid model: Investigation of the effect of test compounds on urinary bile acids in male C57BL/6N mice.

C57BL/6N Tac mice aged 8-9 weeks were used to study the effect of bile acid modulators on bile acid levels. After completion of the quarantine and acclimatization period, animals were randomized into x treatment groups based on body weight: (i) diluent control and (ii) oral administration of test compound at y mg/kg once daily. Animals were treated with test compound for 7 days. On day 6 of the study, animals were transferred to a metabolic cage. On day 7, faeces and urine were collected from each cell in the metabolic study, then blood was taken from each animal by the retrobulbar route. Animals were sacrificed to collect kidneys from each animal for analysis. Body weight was determined twice a week. Serum total bile acids were determined in day 7 serum samples. Fecal bile excretion was determined in day 7 faecal samples. Urinary bile acid excretion was determined in day 7 samples. in samples on day 7.

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SEQUENCE LIST

<110> ALBIREO AB

<120> BENZOTIA(DI)AZEPINES AND THEIR USE AS BILE ACID MODULATORS

<130> NP0376WO

<150> IN201811021016

<151> 2018-06-05

<150> SE 1850915-8

<151> 2018-07-18

<150> IN201911000892

<151> 2019-01-08

<160> 4

<170>PatentIn version 3.5

<210> 1

<211> 1251

<212> PRT

<213> Homo sapiens

<400> 1

Met Ser Thr Glu Arg Asp Ser Glu Thr Thr Phe Asp Glu Asp Ser Gln

1 5 10 15

Pro Asn Asp Glu Val Val Pro Tyr Ser Asp Asp Glu Thr Glu Asp Glu

20 25 30

Leu Asp Asp Gln Gly Ser Ala Val Glu Pro Glu Gln Asn Arg Val Asn

35 40 45

Arg Glu Ala Glu Glu Asn Arg Glu Pro Phe Arg Lys Glu Cys Thr Trp

50 55 60

Gln Val Lys Ala Asn Asp Arg Lys Tyr His Glu Gln Pro His Phe Met

65 70 75 80

Asn Thr Lys Phe Leu Cys Ile Lys Glu Ser Lys Tyr Ala Asn Asn Ala

85 90 95

Ile Lys Thr Tyr Lys Tyr Asn Ala Phe Thr Phe Ile Pro Met Asn Leu

100 105 110

Phe Glu Gln Phe Lys Arg Ala Ala Asn Leu Tyr Phe Leu Ala Leu Leu

115 120 125

Ile Leu Gln Ala Val Pro Gln Ile Ser Thr Leu Ala Trp Tyr Thr Thr

130 135 140

Leu Val Pro Leu Leu Val Val Leu Gly Val Thr Ala Ile Lys Asp Leu

145 150 155 160

Val Asp Asp Val Ala Arg His Lys Met Asp Lys Glu Ile Asn Asn Arg

165 170 175

Thr Cys Glu Val Ile Lys Asp Gly Arg Phe Lys Val Ala Lys Trp Lys

180 185 190

Glu Ile Gln Val Gly Asp Val Ile Arg Leu Lys Lys Asn Asp Phe Val

195 200 205

Pro Ala Asp Ile Leu Leu Leu Ser Ser Ser Glu Pro Asn Ser Leu Cys

210 215 220

Tyr Val Glu Thr Ala Glu Leu Asp Gly Glu Thr Asn Leu Lys Phe Lys

225 230 235 240

Met Ser Leu Glu Ile Thr Asp Gln Tyr Leu Gln Arg Glu Asp Thr Leu

245 250 255

Ala Thr Phe Asp Gly Phe Ile Glu Cys Glu Glu Pro Asn Asn Arg Leu

260 265 270

Asp Lys Phe Thr Gly Thr Leu Phe Trp Arg Asn Thr Ser Phe Pro Leu

275 280 285

Asp Ala Asp Lys Ile Leu Leu Arg Gly Cys Val Ile Arg Asn Thr Asp

290 295 300

Phe Cys His Gly Leu Val Ile Phe Ala Gly Ala Asp Thr Lys Ile Met

305 310 315 320

Lys Asn Ser Gly Lys Thr Arg Phe Lys Arg Thr Lys Ile Asp Tyr Leu

325 330 335

Met Asn Tyr Met Val Tyr Thr Ile Phe Val Val Leu Ile Leu Leu Ser

340 345 350

Ala Gly Leu Ala Ile Gly His Ala Tyr Trp Glu Ala Gln Val Gly Asn

355 360 365

Ser Ser Trp Tyr Leu Tyr Asp Gly Glu Asp Asp Thr Pro Ser Tyr Arg

370 375 380

Gly Phe Leu Ile Phe Trp Gly Tyr Ile Ile Val Leu Asn Thr Met Val

385 390 395 400

Pro Ile Ser Leu Tyr Val Ser Val Glu Val Ile Arg Leu Gly Gln Ser

405 410 415

His Phe Ile Asn Trp Asp Leu Gln Met Tyr Tyr Ala Glu Lys Asp Thr

420 425 430

Pro Ala Lys Ala Arg Thr Thr Thr Leu Asn Glu Gln Leu Gly Gln Ile

435 440 445

His Tyr Ile Phe Ser Asp Lys Thr Gly Thr Leu Thr Gln Asn Ile Met

450 455 460

Thr Phe Lys Lys Cys Cys Ile Asn Gly Gln Ile Tyr Gly Asp His Arg

465 470 475 480

Asp Ala Ser Gln His Asn His Asn Lys Ile Glu Gln Val Asp Phe Ser

485 490 495

Trp Asn Thr Tyr Ala Asp Gly Lys Leu Ala Phe Tyr Asp His Tyr Leu

500 505 510

Ile Glu Gln Ile Gln Ser Gly Lys Glu Pro Glu Val Arg Gln Phe Phe

515 520 525

Phe Leu Leu Ala Val Cys His Thr Val Met Val Asp Arg Thr Asp Gly

530 535 540

Gln Leu Asn Tyr Gln Ala Ala Ser Pro Asp Glu Gly Ala Leu Val Asn

545 550 555 560

Ala Ala Arg Asn Phe Gly Phe Ala Phe Leu Ala Arg Thr Gln Asn Thr

565 570 575

Ile Thr Ile Ser Glu Leu Gly Thr Glu Arg Thr Tyr Asn Val Leu Ala

580 585 590

Ile Leu Asp Phe Asn Ser Asp Arg Lys Arg Met Ser Ile Ile Val Arg

595 600 605

Thr Pro Glu Gly Asn Ile Lys Leu Tyr Cys Lys Gly Ala Asp Thr Val

610 615 620

Ile Tyr Glu Arg Leu His Arg Met Asn Pro Thr Lys Gln Glu Thr Gln

625 630 635 640

Asp Ala Leu Asp Ile Phe Ala Asn Glu Thr Leu Arg Thr Leu Cys Leu

645 650 655

Cys Tyr Lys Glu Ile Glu Glu Lys Glu Phe Thr Glu Trp Asn Lys Lys

660 665 670

Phe Met Ala Ala Ser Val Ala Ser Thr Asn Arg Asp Glu Ala Leu Asp

675 680 685

Lys Val Tyr Glu Glu Ile Glu Lys Asp Leu Ile Leu Leu Gly Ala Thr

690 695 700

Ala Ile Glu Asp Lys Leu Gln Asp Gly Val Pro Glu Thr Ile Ser Lys

705 710 715 720

Leu Ala Lys Ala Asp Ile Lys Ile Trp Val Leu Thr Gly Asp Lys Lys

725 730 735

Glu Thr Ala Glu Asn Ile Gly Phe Ala Cys Glu Leu Leu Thr Glu Asp

740 745 750

Thr Thr Ile Cys Tyr Gly Glu Asp Ile Asn Ser Leu Leu His Ala Arg

755 760 765

Met Glu Asn Gln Arg Asn Arg Gly Gly Val Tyr Ala Lys Phe Ala Pro

770 775 780

Pro Val Gln Glu Ser Phe Phe Pro Pro Gly Gly Asn Arg Ala Leu Ile

785 790 795 800

Ile Thr Gly Ser Trp Leu Asn Glu Ile Leu Leu Glu Lys Lys Thr Lys

805 810 815

Arg Asn Lys Ile Leu Lys Leu Lys Phe Pro Arg Thr Glu Glu Glu Arg

820 825 830

Arg Met Arg Thr Gln Ser Lys Arg Arg Leu Glu Ala Lys Lys Glu Gln

835 840 845

Arg Gln Lys Asn Phe Val Asp Leu Ala Cys Glu Cys Ser Ala Val Ile

850 855 860

Cys Cys Arg Val Thr Pro Lys Gln Lys Ala Met Val Val Asp Leu Val

865 870 875 880

Lys Arg Tyr Lys Lys Ala Ile Thr Leu Ala Ile Gly Asp Gly Ala Asn

885 890 895

Asp Val Asn Met Ile Lys Thr Ala His Ile Gly Val Gly Ile Ser Gly

900 905 910

Gln Glu Gly Met Gln Ala Val Met Ser Ser Asp Tyr Ser Phe Ala Gln

915 920 925

Phe Arg Tyr Leu Gln Arg Leu Leu Leu Val His Gly Arg Trp Ser Tyr

930 935 940

Ile Arg Met Cys Lys Phe Leu Arg Tyr Phe Phe Tyr Lys Asn Phe Ala

945 950 955 960

Phe Thr Leu Val His Phe Trp Tyr Ser Phe Phe Asn Gly Tyr Ser Ala

965 970 975

Gln Thr Ala Tyr Glu Asp Trp Phe Ile Thr Leu Tyr Asn Val Leu Tyr

980 985 990

Thr Ser Leu Pro Val Leu Leu Met Gly Leu Leu Asp Gln Asp Val Ser

995 1000 1005

Asp Lys Leu Ser Leu Arg Phe Pro Gly Leu Tyr Ile Val Gly Gln

1010 1015 1020

Arg Asp Leu Leu Phe Asn Tyr Lys Arg Phe Phe Val Ser Leu Leu

1025 1030 1035

His Gly Val Leu Thr Ser Met Ile Leu Phe Phe Ile Pro Leu Gly

1040 1045 1050

Ala Tyr Leu Gln Thr Val Gly Gln Asp Gly Glu Ala Pro Ser Asp

1055 1060 1065

Tyr Gln Ser Phe Ala Val Thr Ile Ala Ser Ala Leu Val Ile Thr

1070 1075 1080

Val Asn Phe Gln Ile Gly Leu Asp Thr Ser Tyr Trp Thr Phe Val

1085 1090 1095

Asn Ala Phe Ser Ile Phe Gly Ser Ile Ala Leu Tyr Phe Gly Ile

1100 1105 1110

Met Phe Asp Phe His Ser Ala Gly Ile His Val Leu Phe Pro Ser

1115 1120 1125

Ala Phe Gln Phe Thr Gly Thr Ala Ser Asn Ala Leu Arg Gln Pro

1130 1135 1140

Tyr Ile Trp Leu Thr Ile Ile Leu Ala Val Ala Val Cys Leu Leu

1145 1150 1155

Pro Val Val Ala Ile Arg Phe Leu Ser Met Thr Ile Trp Pro Ser

1160 1165 1170

Glu Ser Asp Lys Ile Gln Lys His Arg Lys Arg Leu Lys Ala Glu

1175 1180 1185

Glu Gln Trp Gln Arg Arg Gln Gln Val Phe Arg Arg Gly Val Ser

1190 1195 1200

Thr Arg Arg Ser Ala Tyr Ala Phe Ser His Gln Arg Gly Tyr Ala

1205 1210 1215

Asp Leu Ile Ser Ser Gly Arg Ser Ile Arg Lys Lys Arg Ser Pro

1220 1225 1230

Leu Asp Ala Ile Val Ala Asp Gly Thr Ala Glu Tyr Arg Arg Thr

1235 1240 1245

Gly Asp Ser

1250

<210> 2

<211> 3756

<212> DNA

<213> Homo sapiens

<400> 2

atgagtacag aaagagactc agaaacgaca tttgacgagg attctcagcc taatgacgaa 60

gtggttccct acagtgatga tgaaacagaa gatgaacttg atgaccaggg gtctgctgtt 120

gaaccagaac aaaaccgagt caacagggaa gcagaggaga accgggagcc attcagaaaa 180

gaatgtacat ggcaagtcaa agcaaacgat cgcaagtacc acgaacaacc tcactttatg 240

aacacaaaat tcttgtgtat taaggagagt aaatatgcga ataatgcaat taaaacatac 300

aagtacaacg catttacctt tataccaatg aatctgtttg agcagtttaa gagagcagcc 360

aatttatatt tcctggctct tcttatctta caggcagttc ctcaaatctc taccctggct 420

tggtacacca cactagtgcc cctgcttgtg gtgctgggcg tcactgcaat caaagacctg 480

gtggacgatg tggctcgcca taaaatggat aaggaaatca acaataggac gtgtgaagtc 540

attaaggatg gcaggttcaa agttgctaag tggaaagaaa ttcaagttgg agacgtcatt 600

cgtctgaaaa aaaatgattt tgttccagct gacattctcc tgctgtctag ctctgagcct 660

aacagcctct gctatgtgga aacagcagaa ctggatggag aaaccaattt aaaatttaag 720

atgtcacttg aaatcacaga ccagtacctc caaagagaag atacattggc tacatttgat 780

ggttttattg aatgtgaaga acccaataac agactagata agtttacagg aacactattt 840

tggagaaaca caagttttcc tttggatgct gataaaattt tgttacgtgg ctgtgtaatt 900

aggaacaccg atttctgcca cggcttagtc atttttgcag gtgctgacac taaaataatg 960

aagaatagtg ggaaaaccag atttaaaaga actaaaattg attacttgat gaactacatg 1020

gtttacacga tctttgttgt tcttattctg ctttctgctg gtcttgccat cggccatgct 1080

tattgggaag cacaggtggg caattcctct tggtacctct atgatggaga agacgataca 1140

ccctcctacc gtggattcct cattttctgg ggctatatca ttgttctcaa caccatggta 1200

cccatctctc tctatgtcag cgtggaagtg attcgtcttg gacagagtca cttcatcaac 1260

tgggacctgc aaatgtacta tgctgagaag gacacacccg caaaagctag aaccaccaca 1320

ctcaatgaac agctcgggca gatccattat atcttctctg ataagacgggg gacactcaca 1380

caaaatatca tgacctttaa aaagtgctgt atcaacgggc agatatatgg ggaccatcgg 1440

gatgcctctc aacacaacca caacaaaata gagcaagttg attttagctg gaatacatat 1500

gctgatggga agcttgcatt ttatgaccac tatcttattg agcaaatcca gtcagggaaa 1560

gagccagaag tacgacagtt cttcttcttg ctcgcagttt gccacacagt catggtggat 1620

aggactgatg gtcagctcaa ctaccaggca gcctctcccg atgaaggtgc cctggtaaac 1680

gctgccagga actttggctt tgccttcctc gccaggaccc agaacaccat caccatcagt 1740

gaactgggca ctgaaaggac ttacaatgtt cttgccattt tggacttcaa cagtgaccgg 1800

aagcgaatgt ctatcattgt aagaacccca gaaggcaata tcaagcttta ctgtaaaggt 1860

gctgacactg ttatttatga acggttacat cgaatgaatc ctactaagca agaaacacag 1920

gatgccctgg atatctttgc aaatgaaact cttagaaccc tatgcctttg ctacaaggaa 1980

attgaagaaa aagaatttac agaatggaat aaaaagttta tggctgccag tgtggcctcc 2040

accaaccggg acgaagctct ggataaagta tatgaggaga ttgaaaaaga cttaattctc 2100

ctgggagcta cagctattga agacaagcta caggatggag ttccagaaac catttcaaaa 2160

cttgcaaaag ctgacattaa gatctgggtg cttactggag acaaaaagga aactgctgaa 2220

aatataggat ttgcttgtga acttctgact gaagacacca ccatctgcta tggggaggat 2280

attaattctc ttcttcatgc aaggatggaa aaccagagga atagaggtgg cgtctacgca 2340

aagtttgcac ctcctgtgca ggaatctttt tttccacccg gtggaaaccg tgccttaatc 2400

atcactggtt cttggttgaa tgaaattctt ctcgagaaaa agaccaagag aaataagatt 2460

ctgaagctga agttcccaag aacagaagaa gaaagacgga tgcggaccca aagtaaaagg 2520

aggctagaag ctaagaaaga gcagcggcag aaaaactttg tggacctggc ctgcgagtgc 2580

agcgcagtca tctgctgccg cgtcaccccc aagcagaagg ccatggtggt ggacctggtg 2640

aagaggtaca agaaagccat cacgctggcc atcggagatg gggccaatga cgtgaacatg 2700

atcaaaactg cccacattgg cgttggaata agtggacaag aaggaatgca agctgtcatg 2760

tcgagtgact attcctttgc tcagttccga tatctgcaga ggctactgct ggtgcatggc 2820

cgatggtctt acataaggat gtgcaagttc ctacgatact tcttttacaa aaactttgcc 2880

tttactttgg ttcatttctg gtactccttc ttcaatggct actctgcgca gactgcatac 2940

gaggattggt tcatcaccct ctacaacgtg ctgtacacca gcctgcccgt gctcctcatg 3000

gggctgctcg accaggatgt gagtgacaaa ctgagcctcc gattccctgg gttatacata 3060

gtgggacaaa gagacttact attcaactat aagagattct ttgtaagctt gttgcatggg 3120

gtcctaacat cgatgatcct cttcttcata cctcttggag cttatctgca aaccgtaggg 3180

caggatggag aggcaccttc cgactaccag tcttttgccg tcaccattgc ctctgctctt 3240

gtaataacag tcaatttcca gattggcttg gatacttctt attggacttt tgtgaatgct 3300

3360

ggaatacatg ttctctttcc atctgcattt caatttacag gcacagcttc aaacgctctg 3420

agacagccat acatttggtt aactatcatc ctggctgttg ctgtgtgctt actacccgtc 3480

gttgccattc gattcctgtc aatgaccatc tggccatcag aaagtgataa gatccagaag 3540

catcgcaagc ggttgaaggc ggaggagcag tggcagcgac ggcagcaggt gttccgccgg 3600

ggcgtgtcaa cgcggcgctc ggcctacgcc ttctcgcacc agcggggcta cgcggacctc 3660

atctcctccg ggcgcagcat ccgcaagaag cgctcgccgc ttgatgccat cgtggcggat 3720

ggcaccgcgg agtacaggcg caccggggac agctga 3756

<210> 3

<211> 1321

<212> PRT

<213> Homo sapiens

<400> 3

Met Ser Asp Ser Val Ile Leu Arg Ser Ile Lys Lys Phe Gly Glu Glu

1 5 10 15

Asn Asp Gly Phe Glu Ser Asp Lys Ser Tyr Asn Asn Asp Lys Lys Ser

20 25 30

Arg Leu Gln Asp Glu Lys Lys Gly Asp Gly Val Arg Val Gly Phe Phe

35 40 45

Gln Leu Phe Arg Phe Ser Ser Ser Thr Asp Ile Trp Leu Met Phe Val

50 55 60

Gly Ser Leu Cys Ala Phe Leu His Gly Ile Ala Gln Pro Gly Val Leu

65 70 75 80

Leu Ile Phe Gly Thr Met Thr Asp Val Phe Ile Asp Tyr Asp Val Glu

85 90 95

Leu Gln Glu Leu Gln Ile Pro Gly Lys Ala Cys Val Asn Asn Thr Ile

100 105 110

Val Trp Thr Asn Ser Ser Leu Asn Gln Asn Met Thr Asn Gly Thr Arg

115 120 125

Cys Gly Leu Leu Asn Ile Glu Ser Glu Met Ile Lys Phe Ala Ser Tyr

130 135 140

Tyr Ala Gly Ile Ala Val Ala Val Leu Ile Thr Gly Tyr Ile Gln Ile

145 150 155 160

Cys Phe Trp Val Ile Ala Ala Ala Arg Gln Ile Gln Lys Met Arg Lys

165 170 175

Phe Tyr Phe Arg Arg Ile Met Arg Met Glu Ile Gly Trp Phe Asp Cys

180 185 190

Asn Ser Val Gly Glu Leu Asn Thr Arg Phe Ser Asp Asp Ile Asn Lys

195 200 205

Ile Asn Asp Ala Ile Ala Asp Gln Met Ala Leu Phe Ile Gln Arg Met

210 215 220

Thr Ser Thr Ile Cys Gly Phe Leu Leu Gly Phe Phe Arg Gly Trp Lys

225 230 235 240

Leu Thr Leu Val Ile Ile Ser Val Ser Pro Leu Ile Gly Ile Gly Ala

245 250 255

Ala Thr Ile Gly Leu Ser Val Ser Lys Phe Thr Asp Tyr Glu Leu Lys

260 265 270

Ala Tyr Ala Lys Ala Gly Val Val Ala Asp Glu Val Ile Ser Ser Met

275 280 285

Arg Thr Val Ala Ala Phe Gly Gly Glu Lys Arg Glu Val Glu Arg Tyr

290 295 300

Glu Lys Asn Leu Val Phe Ala Gln Arg Trp Gly Ile Arg Lys Gly Ile

305 310 315 320

Val Met Gly Phe Phe Thr Gly Phe Val Trp Cys Leu Ile Phe Leu Cys

325 330 335

Tyr Ala Leu Ala Phe Trp Tyr Gly Ser Thr Leu Val Leu Asp Glu Gly

340 345 350

Glu Tyr Thr Pro Gly Thr Leu Val Gln Ile Phe Leu Ser Val Ile Val

355 360 365

Gly Ala Leu Asn Leu Gly Asn Ala Ser Pro Cys Leu Glu Ala Phe Ala

370 375 380

Thr Gly Arg Ala Ala Ala Thr Ser Ile Phe Glu Thr Ile Asp Arg Lys

385 390 395 400

Pro Ile Ile Asp Cys Met Ser Glu Asp Gly Tyr Lys Leu Asp Arg Ile

405 410 415

Lys Gly Glu Ile Glu Phe His Asn Val Thr Phe His Tyr Pro Ser Arg

420 425 430

Pro Glu Val Lys Ile Leu Asn Asp Leu Asn Met Val Ile Lys Pro Gly

435 440 445

Glu Met Thr Ala Leu Val Gly Pro Ser Gly Ala Gly Lys Ser Thr Ala

450 455 460

Leu Gln Leu Ile Gln Arg Phe Tyr Asp Pro Cys Glu Gly Met Val Thr

465 470 475 480

Val Asp Gly His Asp Ile Arg Ser Leu Asn Ile Gln Trp Leu Arg Asp

485 490 495

Gln Ile Gly Ile Val Glu Gln Glu Pro Val Leu Phe Ser Thr Thr Ile

500 505 510

Ala Glu Asn Ile Arg Tyr Gly Arg Glu Asp Ala Thr Met Glu Asp Ile

515 520 525

Val Gln Ala Ala Lys Glu Ala Asn Ala Tyr Asn Phe Ile Met Asp Leu

530 535 540

Pro Gln Gln Phe Asp Thr Leu Val Gly Glu Gly Gly Gly Gln Met Ser

545 550 555 560

Gly Gly Gln Lys Gln Arg Val Ala Ile Ala Arg Ala Leu Ile Arg Asn

565 570 575

Pro Lys Ile Leu Leu Leu Asp Met Ala Thr Ser Ala Leu Asp Asn Glu

580 585 590

Ser Glu Ala Met Val Gln Glu Val Leu Ser Lys Ile Gln His Gly His

595 600 605

Thr Ile Ile Ser Val Ala His Arg Leu Ser Thr Val Arg Ala Ala Asp

610 615 620

Thr Ile Ile Gly Phe Glu His Gly Thr Ala Val Glu Arg Gly Thr His

625 630 635 640

Glu Glu Leu Leu Glu Arg Lys Gly Val Tyr Phe Thr Leu Val Thr Leu

645 650 655

Gln Ser Gln Gly Asn Gln Ala Leu Asn Glu Glu Asp Ile Lys Asp Ala

660 665 670

Thr Glu Asp Asp Met Leu Ala Arg Thr Phe Ser Arg Gly Ser Tyr Gln

675 680 685

Asp Ser Leu Arg Ala Ser Ile Arg Gln Arg Ser Lys Ser Gln Leu Ser

690 695 700

Tyr Leu Val His Glu Pro Pro Leu Ala Val Val Asp His Lys Ser Thr

705 710 715 720

Tyr Glu Glu Asp Arg Lys Asp Lys Asp Ile Pro Val Gln Glu Glu Val

725 730 735

Glu Pro Ala Pro Val Arg Arg Ile Leu Lys Phe Ser Ala Pro Glu Trp

740 745 750

Pro Tyr Met Leu Val Gly Ser Val Gly Ala Ala Val Asn Gly Thr Val

755 760 765

Thr Pro Leu Tyr Ala Phe Leu Phe Ser Gln Ile Leu Gly Thr Phe Ser

770 775 780

Ile Pro Asp Lys Glu Glu Gln Arg Ser Gln Ile Asn Gly Val Cys Leu

785 790 795 800

Leu Phe Val Ala Met Gly Cys Val Ser Leu Phe Thr Gln Phe Leu Gln

805 810 815

Gly Tyr Ala Phe Ala Lys Ser Gly Glu Leu Leu Thr Lys Arg Leu Arg

820 825 830

Lys Phe Gly Phe Arg Ala Met Leu Gly Gln Asp Ile Ala Trp Phe Asp

835 840 845

Asp Leu Arg Asn Ser Pro Gly Ala Leu Thr Thr Arg Leu Ala Thr Asp

850 855 860

Ala Ser Gln Val Gln Gly Ala Ala Gly Ser Gln Ile Gly Met Ile Val

865 870 875 880

Asn Ser Phe Thr Asn Val Thr Val Ala Met Ile Ile Ala Phe Ser Phe

885 890 895

Ser Trp Lys Leu Ser Leu Val Ile Leu Cys Phe Phe Pro Phe Leu Ala

900 905 910

Leu Ser Gly Ala Thr Gln Thr Arg Met Leu Thr Gly Phe Ala Ser Arg

915 920 925

Asp Lys Gln Ala Leu Glu Met Val Gly Gln Ile Thr Asn Glu Ala Leu

930 935 940

Ser Asn Ile Arg Thr Val Ala Gly Ile Gly Lys Glu Arg Arg Phe Ile

945 950 955 960

Glu Ala Leu Glu Thr Glu Leu Glu Lys Pro Phe Lys Thr Ala Ile Gln

965 970 975

Lys Ala Asn Ile Tyr Gly Phe Cys Phe Ala Phe Ala Gln Cys Ile Met

980 985 990

Phe Ile Ala Asn Ser Ala Ser Tyr Arg Tyr Gly Gly Tyr Leu Ile Ser

995 1000 1005

Asn Glu Gly Leu His Phe Ser Tyr Val Phe Arg Val Ile Ser Ala

1010 1015 1020

Val Val Leu Ser Ala Thr Ala Leu Gly Arg Ala Phe Ser Tyr Thr

1025 1030 1035

Pro Ser Tyr Ala Lys Ala Lys Ile Ser Ala Ala Arg Phe Phe Gln

1040 1045 1050

Leu Leu Asp Arg Gln Pro Pro Ile Ser Val Tyr Asn Thr Ala Gly

1055 1060 1065

Glu Lys Trp Asp Asn Phe Gln Gly Lys Ile Asp Phe Val Asp Cys

1070 1075 1080

Lys Phe Thr Tyr Pro Ser Arg Pro Asp Ser Gln Val Leu Asn Gly

1085 1090 1095

Leu Ser Val Ser Ile Ser Pro Gly Gln Thr Leu Ala Phe Val Gly

1100 1105 1110

Ser Ser Gly Cys Gly Lys Ser Thr Ser Ile Gln Leu Leu Glu Arg

1115 1120 1125

Phe Tyr Asp Pro Asp Gln Gly Lys Val Met Ile Asp Gly His Asp

1130 1135 1140

Ser Lys Lys Val Asn Val Gln Phe Leu Arg Ser Asn Ile Gly Ile

1145 1150 1155

Val Ser Gln Glu Pro Val Leu Phe Ala Cys Ser Ile Met Asp Asn

1160 1165 1170

Ile Lys Tyr Gly Asp Asn Thr Lys Glu Ile Pro Met Glu Arg Val

1175 1180 1185

Ile Ala Ala Ala Lys Gln Ala Gln Leu His Asp Phe Val Met Ser

1190 1195 1200

Leu Pro Glu Lys Tyr Glu Thr Asn Val Gly Ser Gln Gly Ser Gln

1205 1210 1215

Leu Ser Arg Gly Glu Lys Gln Arg Ile Ala Ile Ala Arg Ala Ile

1220 1225 1230

Val Arg Asp Pro Lys Ile Leu Leu Leu Asp Glu Ala Thr Ser Ala

1235 1240 1245

Leu Asp Thr Glu Ser Glu Lys Thr Val Gln Val Ala Leu Asp Lys

1250 1255 1260

Ala Arg Glu Gly Arg Thr Cys Ile Val Ile Ala His Arg Leu Ser

1265 1270 1275

Thr Ile Gln Asn Ala Asp Ile Ile Ala Val Met Ala Gln Gly Val

1280 1285 1290

Val Ile Glu Lys Gly Thr His Glu Glu Leu Met Ala Gln Lys Gly

1295 1300 1305

Ala Tyr Tyr Lys Leu Val Thr Thr Gly Ser Pro Ile Ser

1310 1315 1320

<210> 4

<211> 3966

<212> DNA

<213> Homo sapiens

<400> 4

atgtctgact cagtaattct tcgaagtata aagaaatttg gagaggagaa tgatggtttt 60

gagtcagata aatcatataa taatgataag aaatcaaggt tacaagatga gaagaaaggt 120

gatggcgtta gagttggctt ctttcaattg tttcggtttt cttcatcaac tgacatttgg 180

ctgatgtttg tgggaagttt gtgtgcattt ctccatggaa tagcccagcc aggcgtgcta 240

ctcatttttg gcacaatgac agatgttttt attgactacg acgttgagtt acaagaactc 300

cagattccag gaaaagcatg tgtgaataac accattgtat ggactaacag ttccctcaac 360

cagaacatga caaatggaac acgttgtggg ttgctgaaca tcgagagcga aatgatcaaa 420

tttgccagtt actatgctgg aattgctgtc gcagtactta tcacaggata tattcaaata 480

tgcttttggg tcattgccgc agctcgtcag atacagaaaa tgagaaaatt ttactttagg 540

agaataatga gaatggaaat agggtggttt gactgcaatt cagtggggga gctgaataca 600

agattctctg atgatattaa taaaatcaat gatgccatag ctgaccaaat ggcccttttc 660

attcagcgca tgacctcgac catctgtggt ttcctgttgg gatttttcag gggttggaaa 720

ctgaccttgg ttattatttc tgtcagccct ctcattggga ttggagcagc caccattggt 780

ctgagtgtgt ccaagtttac ggactatgag ctgaaggcct atgccaaagc aggggtggtg 840

gctgatgaag tcatttcatc aatgagaaca gtggctgctt ttggtggtga gaaaagagag 900

gttgaaaggt atgagaaaaa tcttgtgttc gcccagcgtt ggggaattag aaaaggaata 960

gtgatgggat tctttactgg attcgtgtgg tgtctcatct ttttgtgtta tgcactggcc 1020

ttctggtacg gctccacact tgtcctggat gaaggagaat atacaccagg aacccttgtc 1080

cagattttcc tcagtgtcat agtaggagct ttaaatcttg gcaatgcctc tccttgtttg 1140

gaagcctttg caactggacg tgcagcagcc accagcattt ttgagacaat agacaggaaa 1200

cccatcattg actgcatgtc agaagatggt tacaagttgg atcgaatcaa gggtgaaatt 1260

gaattccata atgtgacctt ccattatcct tccagaccag aggtgaagat tctaaatgac 1320

ctcaacatgg tcattaaacc aggggaaatg acagctctgg taggacccag tggagctgga 1380

aaaagtacag cactgcaact cattcagcga ttctatgacc cctgtgaagg aatggtgacc 1440

gtggatggcc atgacattcg ctctcttaac attcagtggc ttagagatca gattgggata 1500

gtggagcaag agccagttct gttctctacc accattgcag aaaatattcg ctatggcaga 1560

gaagatgcaa caatggaaga catagtccaa gctgccaagg aggccaatgc ctacaacttc 1620

1680

ggtggccaga aacaaagggt agctatcgcc agagccctca tccgaaatcc caagattctg 1740

cttttggaca tggccacctc agctctggac aatgagagtg aagccatggt gcaagaagtg 1800

ctgagtaaga ttcagcatgg gcacacaatc atttcagttg ctcatcgctt gtctacggtc 1860

agagctgcag ataccatcat tggttttgaa catggcactg cagtggaaag agggacccat 1920

gaagaattac tggaaaggaa aggtgtttac ttcactctag tgactttgca aagccaggga 1980

aatcaagctc ttaatgaaga ggacataaag gatgcaactg aagatgacat gcttgcgagg 2040

acctttagca gagggagcta cgggatagt ttaagggctt ccatccggca acgctccaag 2100

tctcagcttt cttacctggt gcacgaacct ccattagctg ttgtagatca taagtctacc 2160

tatgaagaag atagaaagga caaggacatt cctgtgcagg aagaagttga acctgcccca 2220

gttaggagga ttctgaaatt cagtgctcca gaatggccct acatgctggt agggtctgtg 2280

ggtgcagctg tgaacgggac agtcacaccc ttgtatgcct tttttattcag ccagattctt 2340

gggacttttt caattcctga taaagggaa caaaggtcac agatcaatgg tgtgtgccta 2400

ctttttgtag caatgggctg tgtatctctt ttcacccaat ttctacaggg atatgccttt 2460

gctaaatctg gggagctcct aacaaaaagg ctacgtaaat ttggtttcag ggcaatgctg 2520

gggcaagata ttgcctggtt tgatgacctc agaaaatagcc ctggagcatt gacaacaaga 2580

cttgctacag atgcttccca agttcaaggg gctgccggct ctcagatcgg gatgatagtc 2640

aattccttca ctaacgtcac tgtggccatg atcattgcct tctcctttag ctggaagctg 2700

agcctggtca tcttgtgctt cttccccttc ttggctttat caggagccac acagaccagg 2760

atgttgacag gatttgcctc tcgagataag caggccctgg agatggtggg acagattaca 2820

aatgaagccc tcagtaacat ccgcactgtt gctggaattg gaaagggag gcggttcatt 2880

2940

tacggattct gctttgcctt tgcccagtgc atcatgttta ttgcgaattc tgcttcctac 3000

agatatggag gttacttaat ctccaatgag gggctccatt tcagctatgt gttcagggtg 3060

atctctgcag ttgtactgag tgcaacagct cttggaagag ccttctctta caccccaagt 3120

tatgcaaaag ctaaaatatc agctgcacgc ttttttcaac tgctggaccg acaaccccca 3180

atcagtgtat acaatactgc aggtgaaaaa tgggacaact tccaggggaa gattgatttt 3240

gttgattgta aatttacata tccttctcga cctgactcgc aagttctgaa tggtctctca 3300

gtgtcgatta gtccagggca gacactggcg tttgttggga gcagtggatg tggcaaaagc 3360

actagcattc agctgttgga acgtttctat gatcctgatc aagggaaggt gatgatagat 3420

ggtcatgaca gcaaaaaagt aaatgtccag ttcctccgct caaacattgg aattgtttcc 3480

caggaaccag tgttgtttgc ctgtagcata atggacaata tcaagtatgg agacaacacc 3540

3600

gtcatgtcac tcccagagaa atatgaaact aacgttggggt cccaggggtc tcaactctct 3660

agaggggaga aacaacgcat tgctattgct cgggccattg tacgagatcc taaaatcttg 3720

ctactagatg aagccacttc tgccttagac acagaaagtg aaaagacggt gcaggttgct 3780

ctagacaaag ccagagaggg tcggacctgc attgtcattg cccatcgctt gtccaccatc 3840

cagaacgcgg atatcattgc tgtcatggca cagggggtgg tgattgaaaa ggggacccat 3900

gaagaactga tggcccaaaa aggagcctac tacaaactag tcaccactgg atcccccatc 3960

agttga 3966

<---

Claims (137)

1. Compound of formula (I)

wherein M is selected from the group consisting of -CH ₂ - and -NR ⁷ -; R ¹ and R ² are all independently C _1-4 alkyl; R ³ is selected from the group consisting of hydrogen, halogen, hydroxy, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, cyano, nitro, amino, N- (C _1-4 alkyl)amino, N, N -di(C _1-4 alkyl)amino, N- (aryl-C _1-4 alkyl)amino, C _1-6 alkylcarbonylamino, C _3-6 cycloalkylcarbonylamino, N- (C _1-4 alkyl)aminocarbonyl, N, N -di(C _1-4 alkyl)aminocarbonyl, C _1-4 alkyloxycarbonylamino group, C _3-6 cycloalkyloxycarbonylamino group, C _1-4 alkylsulfonamide group and C _3-6 cycloalkylsulfonamide group; n is an integer equal to 1, 2 or 3; R ⁴ is selected from the group consisting of hydrogen, halogen, cyano, C _1-4 alkyl, C _3-6 cycloalkyl, C _1-4 alkoxy, C _3-6 cycloalkyloxy, C _1-4 alkylthio, C _3-6 cycloalkylthio, amino , N -(C _1-4 alkyl)amino and N,N -di(C _1-4 alkyl)amino; one of R ⁵ and R ⁶ is carboxy and the other of R ⁵ and R ⁶ is selected from the group consisting of hydrogen, fluorine, C _1-4 alkyl and C _1-4 haloalkyl; And R ⁷ is selected from the group consisting of hydrogen and C _1-4 alkyl; R ⁸ is selected from the group consisting of hydrogen and C _1-4 alkyl; or a pharmaceutically acceptable salt thereof. 2. A compound according to claim 1 wherein all R ¹ and R ² are n-butyl. 3. A compound according to claim 1 wherein all R ¹ and R ² are ethyl. 4. A compound according to claim 1 wherein R ¹ is n-butyl and R ² is ethyl. 5. Connection according to any one of paragraphs. 1-4, wherein R ³ is selected from the group consisting of hydrogen, bromine, hydroxy, methoxy, amino, t-butoxycarbonylamino, methylsulfonamide, and cyclopropylsulfonamide. 6. The connection according to any one of paragraphs. 1-5, wherein R ⁴ is selected from the group consisting of hydrogen, bromine, ethyl, cyclopropyl, methoxy, methylthio and dimethylamino. 7. Connection according to any one of paragraphs. 1-6, in which R ⁵ is selected from the group consisting of hydrogen and fluorine. 8. Connection according to any one of paragraphs. 1-7, in which R ⁶ represents a carboxy group. 9. The compound according to claim 1, selected from the group consisting of: (E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid; (E) -3-((5-(4-aminophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (E) -3-((5-(4-((tert-butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid; (R)- (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (S)- (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (E) -3-((3,3-dibutyl-7-cyclopropyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid; (E) -3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-(cyclopropanesulfonamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-(methylsulfonamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid; (Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid; (E) -3-((3,3-dibutyl-7-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid; (E) -3-((3-butyl-3-ethyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid; (E) -3-((3,3-dibutyl-7-methoxy-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid; (E) -3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid; (R)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid; (S) -(E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-methoxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (E) -3-((3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid ; ( E )-3-((5-(4-(benzylamino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid; ( E )-3-((7-bromo-5-(4-((tert-butoxycarbonyl)amino)phenyl)-3-butyl-3-ethyl-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-((methoxycarbonyl)amino)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro- 1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-(dimethylamino)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid; (Z) -3-((5-(4-((tert-butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5- tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((5-(4-((butoxycarbonyl)amino)phenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro- 1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-(3,3-dimethylbutanamido)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro -1,5-benzothiazepin-8-yl)oxy)acrylic acid; (Z) -3-((3,3-dibutyl-5-(4-isobutyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-isobutyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid; (Z) -3-((3,3-dibutyl-5-(4-(cyclopentanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3,3-dibutyl-5-(4-(cyclopropanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((3,3-dibutyl-5-(4-(cyclopentanecarboxamido)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3-butyl-5-(4-(cyclopentanecarboxamido)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1 ,5-benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-butyramidophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid; (R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid; (Z) -3-((3,3-dibutyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid; (R) -(Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (S)- (Z) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid; (R)- (E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (S)- (E) -3-((3-butyl-7-chloro-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (Z) -3-((3,3-diethyl-7-(methylthio)-1,1-dioxido-5-(4-pivalamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3,3-dibutyl-7-chloro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) -2-fluoroacrylic acid; (E) -3-((3,3-diethyl-7-iodo-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid; (E) -3-((7-bromo-3,3-diethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid; (Z) -3-((7-bromo-3,3-diethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) -2-fluoroacrylic acid; (S)- (Z) -3-((3-butyl-3-ethyl-7-iodine-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (R)- (Z) -3-((3-butyl-3-ethyl-7-iodine-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid; (S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid; (R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3-butyl-5-(4-(tert-butylcarbamoyl)phenyl)-3-ethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro -1,5-benzothiazepin-8-yl)oxy)acrylic acid; (Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid; (R)- (Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (S)- (Z) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((5-(4-(benzylamino)phenyl)-3,3-diethyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro-1 ,5-benzothiazepin-8-yl)oxy)acrylic acid; (E) -3-((3,3-dibutyl-5-(4-(isopropylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)acrylic acid; (Z) -3-((3,3-dibutyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid; (Z) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxido-5-(4-propionamidophenyl)-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid; (E) -3-((3-butyl-3-ethyl-7-(ethylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid; (E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)-2-methacrylic acid; (E) -3-((7-bromo-3,3-dibutyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl) hydroxy)acrylic acid; (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 8-yl)hydroxy)acrylic acid; (S)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)- (E) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)acrylic acid; (Z) -3-((3,3-dibutyl-5-(4-(tert-butylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxydo-2,3,4,5-tetrahydro-1 ,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid; (E) -3-((3,3-dibutyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid; (E) -3-((3,3-dibutyl-7-cyano-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy) acrylic acid; (E) -3-((3,3-dibutyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)oxy)but-2-enoic acid; (Z)-3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (S)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (R)- (Z) -3-((3-butyl-3-ethyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((7-bromo-3-butyl-3-ethyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)acrylic acid; (E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)- (E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)- (E) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)acrylic acid; (S)- (E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid; (R)- (E) -3-((3-butyl-3-ethyl-7-(methylamino)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)acrylic acid; (Z) -3-((5-(4-bromophenyl)-3,3-dibutyl-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3,3-dibutyl-5-(4-hydroxyphenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)-2-fluoroacrylic acid; (Z) -3-((3,3-dibutyl-7-(dimethylamino)-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine-8- yl)hydroxy)-2-fluoroacrylic acid; (Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2, 5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (S)- (Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (R)- (Z) -3-((3-butyl-3-ethyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro- 1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl) hydroxy)acrylic acid; (R)- (E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (S)- (E) -3-((3-butyl-3-ethyl-7-fluoro-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine- 8-yl)hydroxy)acrylic acid; (Z) -3-((3,3-dibutyl-5-(4-(dimethylcarbamoyl)phenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5 -benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3,3-dibutyl-2-methyl-7-(methylthio)-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((7-bromo-3-butyl-3-ethyl-2-methyl-1,1-dioxydo-5-phenyl-2,3,4,5-tetrahydro-1,2,5- benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3,3-dibutyl-5-(3,4-difluorophenyl)-7-(methylthio)-1,1-dioxido-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine-8- yl)hydroxy)-2-fluoroacrylic acid; (S)- (Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; And (R)- (Z) -3-((3-butyl-7-(dimethylamino)-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,5- benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; or a pharmaceutically acceptable salt thereof. 10. Pharmaceutical composition for inhibiting the apical sodium-dependent bile acid transporter (ASBT) and/or liver bile acid transporter (LBAT), containing a therapeutically effective amount of a compound according to any one of paragraphs. 1-9 and one or more pharmaceutically acceptable excipients. 11. The connection according to any one of paragraphs. 1-9 for use as a medicine. 12. Connection according to any one of paragraphs. 1-9 for use in the treatment or prevention of cardiovascular disease, or fatty acid metabolism disorders, or glucose utilization disorders such as hypercholesterolemia; metabolic disorders of fatty acids; diabetes mellitus type 1 and type 2; complications of diabetes, including cataracts, micro- and macrovascular diseases, retinopathy, neuropathy, nephropathy and slow 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 rate disorders and vascular restenosis; diabetes-related diseases such as insulin resistance (impaired glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood fatty acids or glycerol, obesity, dyslipidemia, hyperlipidemia, including hypertriglyceridemia, metabolic syndrome (syndrome X), atherosclerosis, and hypertension; and to increase high-density lipoprotein content. 13. Connection according to any one of paragraphs. 1-9 for use in the treatment or prevention of a gastrointestinal disease or disorder such as constipation (including chronic constipation, functional constipation, chronic idiopathic constipation (CIC), intermittent/sporadic constipation, constipation secondary to diabetes mellitus, constipation secondary to constipation secondary to chronic kidney disease constipation secondary to multiple sclerosis constipation secondary to Parkinson's disease constipation secondary to systemic sclerosis constipation secondary to drug-induced constipation, irritable bowel syndrome with constipation (IBS) -C), mixed irritable bowel syndrome (IBS-M), functional constipation in children, and opioid-induced constipation); Crohn's disease; primary malabsorption of bile acids; irritable bowel syndrome (IBS); inflammatory bowel disease (IBD); inflammation of the ileum; and reflux disease and its complications such as Barrett's esophagus, biliary reflux esophagitis and biliary reflux gastritis. 14. Connection according to any one of paragraphs. 1-9 for use in the treatment or prevention of a disease or disorder of the liver, such as an inherited metabolic disorder in the liver; congenital defects in the synthesis of bile acids; congenital anomalies of the bile ducts; atresia of the bile ducts; atresia of the bile ducts after the Kasai operation; bile duct atresia after liver transplantation; congenital giant cell hepatitis; cholestasis of newborns; hereditary forms of cholestasis; cerebrotendon xanthomatosis; secondary defect in BA synthesis; Zellweger's syndrome; cystic fibrosis-associated liver disease; alpha-1 antitrypsin deficiency; Alagille syndrome (ALGS); Byler's syndrome; primary defect in bile acid synthesis (BA); progressive familial intrahepatic cholestasis (PFIC), including PFIC-1, PFIC-2, PFIC-3 and non-specific PFIC, PFIC after bile diversion and PFIC after liver transplantation; benign recurrent intrahepatic cholestasis (BRIC), including BRIC1, BRIC2 and non-specific BRIC, BRIC after bile diversion and BRIC after liver transplantation; autoimmune hepatitis; primary biliary cirrhosis (PBC); liver fibrosis; non-alcoholic fatty liver disease (NAFLD); non-alcoholic steatohepatitis (NASH); portal hypertension; cholestasis; cholestasis in Down syndrome; drug-induced cholestasis; intrahepatic cholestasis of pregnancy (jaundice during pregnancy); intrahepatic cholestasis; extrahepatic cholestasis; associated with parenteral nutrition cholestasis (PNAC); associated with a low content of phospholipids cholestasis; Aagenes syndrome 1 (LSC1); primary sclerosing cholangitis (PSC); associated with immunoglobulin G4 cholangitis; primary biliary cholangitis; cholelithiasis (gallstones); biliary lithiasis; choledocholithiasis; gallstone pancreatitis; Caroli disease; malignant tumor of the bile ducts; a malignant tumor that causes obstruction of the biliary tree; strictures of the bile ducts; AIDS-associated cholangiopathy; ischemic cholangiopathy; pruritis due to cholestasis or jaundice; pancreatitis; chronic autoimmune liver disease leading to progressive cholestasis; fatty hepatosis; alcoholic hepatitis; acute fatty infiltration of the liver; fatty infiltration of the liver of pregnant women; medicinal hepatitis; iron overload disorders; congenital defect in bile acid synthesis type 1 (BAS type 1); drug-induced liver injury (DILI); liver fibrosis; congenital fibrosis of the liver; cirrhosis of the liver; histiocytosis of Langerhans cells (LCH); ichthyosis of newborns - sclerosing cholangitis (NISCH); erythropoietic protoporphyria (EPP); adult idiopathic ductopenia (IAD); idiopathic congenital giant cell hepatitis (INH); non-syndromic interlobular bile duct insufficiency (NS PILBD); North American Indian Infantile Cirrhosis (NAIC); sarcoidosis of the liver; amyloidosis; necrotizing enterocolitis; toxicities caused by serum bile acids, including cardiac arrhythmias (eg, atrial fibrillation) in cases of abnormal serum bile acid profile, cardiomyopathy associated with cirrhosis of the liver ("cholecardia"), and skeletal muscle weakness associated with cholestatic liver disease; viral hepatitis (including hepatitis A, hepatitis B, hepatitis C, hepatitis D and hepatitis E); hepatocellular carcinoma (hepatoma); cholangiocarcinoma; bile acid-related gastrointestinal cancers and cholestasis caused by tumors and neoplasms in the liver, biliary tract and pancreas. 15. Connection according to any one of paragraphs. 1-9 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 and pruritis in renal failure.