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

Abstract

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

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to Indian Patent Application No. 201911049982, filed on December 4, 2019 (the disclosure of which is incorporated herein by reference in its entirety).

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

Background Art

[0003] Bile acids are physiological surfactants that play an important role in the intestinal absorption and transport of lipids, nutrients, and vitamins. It is also a signaling molecule that activates nuclear receptors and a cellular signaling pathway that regulates lipid, glucose, and energy metabolism. Bile acids are steroid acids synthesized from cholesterol in the liver and stored in the gallbladder as mixed micelles. During digestion, the duodenum induces the release of hormones that cause the gallbladder to contract, thereby releasing bile acids into the small intestine, where they enable the absorption of fat-soluble vitamins and cholesterol. When bile acids reach the ileum, they are reabsorbed from the intestine, secreted into the portal blood, and returned to the liver via the portal circulation. In this way, more than 90% of bile acids are recycled and returned to the liver. These bile acids are then transported across the sinusoidal cell membrane of hepatocytes and re-secreted into bile across the canalicular membrane. In this first pass, 75-90% of bile acids are taken up by hepatocytes, completing one enterohepatic circulation. A portion of the bile acids that were not removed by the liver enters the systemic circulation, where free bile acids are filtered by the renal glomeruli, efficiently reabsorbed in the proximal renal tubules, and returned to the systemic circulation. Interestingly, most of the bile acids secreted into bile across the canalicular membrane are derived from the recycling pool, and less than 10% arise from de novo hepatic synthesis. The small amount of bile acids that were not reabsorbed in the ileum reaches the colon. In the intestinal lumen, primary bile acids are converted to secondary bile acids mainly by mono- or di-dehydroxylation reactions of the steroid nucleus under the action of intestinal bacteria. Bile acids that were not absorbed by the intestine are then excreted in feces.

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

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

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

Prior Art Documents

Patent Documents

[0007]

Patent Document 1

Patent Document 2

[0008] [Non-Patent Document 1] Kosters and Karpen (Xenobiotica 2008, Vol. 38, pp. 1043 - 1071) [Non-Patent Document 2] Chiang (J. Lipid Res. 2009, Vol. 50, pp. 1955 - 1966) [Non-Patent Document 3] Dawson (Handb. Exp. Pharmacol. 2011, Vol. 201, pp. 169 - 203)

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Summary of the Invention

[0009] Despite several previously reported ASBT inhibitor compounds, there is a need for further bile acid modulating compounds having an optimized profile with respect to potency, selectivity, and bioavailability.

Brief Description of the Drawings

[0010]

Figure 1

Figure 2

Figure 3

Figure 4

[0011] Certain 1,5-benzothiazepine and 1,2,5-benzothiadiazepine derivatives have been discovered to be potent inhibitors of the apical sodium-dependent bile acid transporter (ASBT) and / or the liver bile acid transporter (LBAT) and may be useful in treating diseases where inhibition of bile acid circulation is desirable.

Modes for Carrying Out the Invention

[0012] In a first aspect, the present invention relates to a compound of formula (I)

[0013] [Chem.]

[0014] (wherein, M is selected from -CH2- and -NR 7 -; R 1 is C 1~4 alkyl; R 2 is independently 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, C 3~6 cycloalkyloxycarbonylamino, C 1~4 alkylsulfonamide, and C 3~6 cycloalkylsulfonamide; n is an integer of 1, 2, or 3; R 3 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; R 4 and R5 One of them is carboxyl, and R 4 and R 5 The other is selected from the group consisting of hydrogen, fluoro, C 1~4 alkyl, and C 1~4 haloalkyl; R 6 is selected from the group consisting of hydrogen and C 1~4 alkyl; R 7 is selected from the group consisting of hydrogen and C 1~4 alkyl) or a pharmaceutically acceptable salt thereof.

[0015] In some embodiments, R 1 is C 2~4 alkyl. In a preferred embodiment, R 1 is n-propyl. In another preferred embodiment, R 1 is n-butyl.

[0016] In some embodiments, R 2 is selected from the group consisting of hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, amino, methylamino, dimethylamino, isopropylcarbonylamino, tert-butylcarbonylamino, tert-butylaminocarbonyl, tert-butoxycarbonylamino, methylsulfonamide, and cyclopropylsulfonamide. In a preferred embodiment, n is 1, that is, the phenyl ring is substituted with only one substituent R 2 In another preferred embodiment, R 2 is in the para position.

[0017] In some embodiments, R 3 is selected from the group consisting of hydrogen, fluoro, chloro, bromo, methyl, cyclopropyl, methoxy, ethoxy, methylthio, ethylthio, amino, methylamino, and dimethylamino.

[0018] In some embodiments, R 4 is hydrogen or fluoro.

[0019] In some embodiments, R 5 is carboxyl.

[0020] In some embodiments, R 6 is hydrogen.

[0021] In some embodiments, R 7 is hydrogen or methyl.

[0022] In a preferred embodiment, the compound of formula (I) is a compound of formula (I-a):

[0023]

Chemical formula

[0024] (wherein, M is selected from the group consisting of -CH2-, -NH-, and -NCH3-; R 1 is C 2~4 alkyl; R 2 is independently hydrogen, halogen, hydroxy, C 1~4 alkyl, C 1~4 haloalkyl, C 1~4 alkoxy, amino, N-(C 1~4 alkyl)amino, N,N-di(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, C 1~4 alkylsulfonamide, and C 3~6 cycloalkylsulfonamide; n is an integer of 1 or 2; R 3 is hydrogen, halogen, C 1~4 alkyl, C 3~6Cycloalkyl, C 1~4 Alkoxy, C 1~4 Alkylthio, amino, N-(C 1~4 Alkyl)amino, and N,N-di(C 1~4 Alkyl)amino selected from the group consisting of; R 4 is hydrogen or fluoro) or a pharmaceutically acceptable salt thereof.

[0025] In a preferred embodiment, the compound of formula (I) is a compound of formula (I-b):

[0026]

Chemical formula

[0027] (wherein, M is selected from the group consisting of -CH2-, -NH-, and -NCH3-; R 1 is C 2~4 alkyl, more preferably n-propyl or n-butyl; R 2 is independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, amino, methylamino, dimethylamino, isopropylcarbonylamino, tert-butylcarbonylamino, tert-butylaminocarbonyl, tert-butoxycarbonylamino, methylsulfonamide, and cyclopropylsulfonamide; R 3 is selected from the group consisting of fluoro, chloro, bromo, methyl, cyclopropyl, methoxy, ethoxy, methylthio, ethylthio, amino, methylamino, and dimethylamino; R 4 is hydrogen or fluoro) or a pharmaceutically acceptable salt thereof.

[0028] A preferred compound of the present invention is a compound of formula (I-b) as defined above (wherein, M and R 1 from R4 is as shown in Table 1 below) or a pharmaceutically acceptable salt thereof:

[0029] [Table 1A]

[0030] [Table 1B]

[0031] [Table 1C]

[0032] [Table 1D]

[0033] [Table 1E]

[0034] [Table 1F]

[0035] [Table 1G]

[0036] [Table 1H]

[0037] In certain embodiments, the compound of formula (I) is (E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (S)-(Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (R)-(Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E)-3-((3-ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (S)-(Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (R)-(Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-Butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-Butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-Butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; and (E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; or a salt thereof acceptable as a medicament, selected from the group consisting of.

[0038] As used herein, the term "halo" refers to fluoro, chloro, bromo, and iodo.

[0039] As used herein, "C 1~6 alkyl" refers to a straight-chain or branched alkyl group having 1 to 6 carbon atoms, and "C 1~4 alkyl" refers to a straight-chain or branched alkyl group having 1 to 4 carbon atoms. C 1~4Examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0040] As used herein, "C" 1~4 The term "haloalkyl" refers to a straight or branched C 1~4 alkyl group as defined herein, in which one or more hydrogen atoms are replaced by halogen. C 1~4 Examples of haloalkyl include chloromethyl, fluoroethyl, and trifluoromethyl.

[0041] As used herein, "C" 1~4 "alkoxy" and "C" 1~4 "alkylthio" refer to a straight or branched C 1~4 alkyl group bonded through an oxygen or sulfur atom, respectively, to the remainder of the molecule.

[0042] As used herein, "C" 3~6 The term "cycloalkyl" refers to a monocyclic saturated hydrocarbon ring having 3 to 6 carbon atoms. C 3~6 Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0043] The term "aryl" refers to an aromatic monocyclic ring composed of 6 carbon atoms or an aromatic bicyclic ring system composed of 10 carbon atoms. Examples of aryl include phenyl, naphthyl, and azulenyl.

[0044] The term "amino" refers to the -NH2 group. As used herein, "N-(C" 1~4 "alkyl)amino" and "N,N-di(C" 1~4 "alkyl)amino" refer to an amino group in which one or both hydrogen atoms are each replaced by a straight or branched C 1~4 alkyl group. N-(C 1~4Examples of (alkyl)amino include methylamino, ethylamino, and tert-butylamino, and N,N-di-(C 1~4 Examples of (alkyl)amino include dimethylamino and diethylamino.

[0045] As used herein, the term "N-(aryl-C 1~4 alkyl)amino" refers to an amino group in which a hydrogen atom is replaced with an aryl-C 1~4 alkyl group. Examples of N-(aryl-C 1~4 alkyl)amino include benzylamino and phenylethylamino. "C 1~6 alkylcarbonylamino" refers to an amino group in which a hydrogen atom is replaced with a C 1~6 alkylcarbonyl group. Examples of C 1~6 alkanoylamino include acetylamino and tert-butylcarbonylamino. "C 1~4 alkyloxycarbonylamino" refers to an amino group in which a hydrogen atom is replaced with a C 1~4 alkyloxycarbonyl group. An example of C 1~4 alkyloxycarbonylamino is tert-butoxycarbonylamino. "C 1~4 alkylsulfonamide" and "C 3~6 cycloalkylsulfonamide" refer to amino groups in which a hydrogen atom is replaced with a C 1~4 alkylsulfonyl or C 3~6 cycloalkylsulfonyl group, respectively.

[0046] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are suitable for human pharmaceutical use, are generally safe, non-toxic, and neither biologically nor otherwise undesirable.

[0047] As used herein, the term "about" refers to a value or parameter that, in this specification, includes (and describes) embodiments directed to that value or parameter itself. For example, a recitation of "about 20" includes a recitation of "20". Numerical ranges include the numbers defining the range. Generally speaking, the term "about" refers to either the value indicated by the variable, all values within the experimental error of the value indicated by the variable (e.g., within the 95% confidence interval of the mean), or the greater of the two, within 10 percent of the value indicated by the variable.

[0048] The 1,5-benzothiazepine and 1,2,5-benzothiadiazepine compounds of formula (I), or a pharmaceutically acceptable salt thereof, are inhibitors of the apical sodium-dependent bile acid transporter, the hepatic bile acid transporter, or both the apical sodium-dependent bile acid transporter and the hepatic bile acid transporter (an ASBT inhibitor, an LBAT inhibitor, and a dual ASBT / LBAT inhibitor, respectively). Thus, they are useful for the treatment or prevention of 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.

[0049] Examples of cardiovascular diseases and disorders of fatty acid metabolism and glucose utilization include, but are not limited to, hypercholesterolemia; disorders of fatty acid metabolism; type 1 and type 2 diabetes mellitus; complications of diabetes, such as cataracts, micro- and macrovascular diseases, retinopathy, neuropathy, nephropathy, and delayed wound healing, tissue ischemia, diabetic foot lesions, atherosclerosis, myocardial infarction, acute coronary syndrome, unstable angina, stable angina, stroke, peripheral arterial occlusive disease, cardiomyopathy, heart failure, cardiac arrhythmias, and vascular restenosis; diabetes-related diseases, such as insulin resistance (disorders of glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, obesity, dyslipidemia, hyperlipidemia, such as hypertriglyceridemia, metabolic syndrome (syndrome X), atherosclerotic cardiovascular disease, and hypertension; and increased high-density lipoprotein levels.

[0050] Gastrointestinal diseases and disorders include constipation (chronic constipation, functional constipation, chronic idiopathic constipation (CIC), intermittent / sporadic constipation, constipation secondary to type 2 diabetes, 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, constipation-predominant irritable bowel syndrome (IBS-C), mixed irritable bowel syndrome (IBS-M), pediatric functional constipation, and opioid-induced constipation); Crohn's disease; primary bile acid malabsorption; irritable bowel syndrome (IBS); inflammatory bowel disease (IBD); ileal inflammation; and reflux disease and its complications, such as Barrett's esophagus, bile reflux esophagitis, and bile reflux gastritis.

[0051] A liver disease, as defined herein, is any disease in the liver and related organs such as the pancreas, portal vein, liver parenchyma, intrahepatic biliary system, extrahepatic biliary system, and gallbladder. In some cases, the liver disease is a bile acid-dependent liver disease. Liver diseases and disorders include: hereditary metabolic disorders of the liver; congenital abnormalities in bile acid synthesis; congenital biliary tract malformations; biliary atresia; biliary atresia after Kasai operation; biliary atresia after liver transplantation; neonatal hepatitis; neonatal cholestasis; genetic forms of cholestasis; cerebrotendinous xanthomatosis; secondary defects in BA synthesis; Zellweger syndrome; liver diseases associated with cystic fibrosis; alpha1-antitrypsin deficiency; Alagille syndrome (ALGS); Byler syndrome; primary defects in bile acid (BA) synthesis; progressive familial intrahepatic cholestasis (PFIC), such as PFIC-1, PFIC-2, PFIC-3, and unspecified PFIC, PFIC after bile shunt, and PFIC after liver transplantation; benign recurrent intrahepatic cholestasis (BRIC), such as BRIC1, BRIC2, and unspecified BRIC, BRIC after bile shunt, 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; parenteral nutrition-associated cholestasis (PNAC); cholestasis associated with low phospholipids; lymphedema cholestasis syndrome 1 (LSC1); primary sclerosing cholangitis (PSC); cholangitis associated with immunoglobulin G4; primary biliary cholangitis; cholelithiasis (gallstones); biliary lithiasis; common bile duct stones; gallstone pancreatitis; Caroli disease; malignant tumors of the bile duct; malignant tumors causing obstruction of the biliary tree; bile duct stricture; AIDS cholangiopathy; ischemic cholangiopathy; pruritus due to cholestasis or jaundice; pancreatitis; chronic autoimmune liver disease leading to progressive cholestasis; hepatic steatosis; alcoholic hepatitis; acute fatty liver; fatty liver of pregnancy; drug-induced hepatitis; hemosiderosis; congenital bile acid metabolic disorder type 1 (BAS type 1); drug-induced liver injury (DILI); liver fibrosis; congenital hepatic fibrosis; cirrhosis; Langerhans cell histiocytosis (LCH); neonatal ichthyosis sclerosing cholangitis (NISCH); erythropoietic protoporphyria (EPP); idiopathic adult ductopenia (IAD); idiopathic neonatal hepatitis (INH);Non-symptomatic intrahepatic bile ductopenia (NS PILBD); autosomal recessive hereditary intrahepatic cholestasis (North American Indian childhood cirrhosis) (NAIC); hepatic sarcoidosis; amyloidosis; necrotizing enteritis; toxicities caused by serum bile acids, e.g., cardiac arrhythmias (e.g., atrial fibrillation), cardiomyopathy associated with cirrhosis ("cholecardia"), and skeletal muscle wasting associated with cholestatic liver disease in the setting of an abnormal serum bile acid profile; polycystic liver disease; viral hepatitis (including hepatitis A, B, C, D, and E); hepatocellular carcinoma (hepatocellular tumor); cholangiocarcinoma; gastrointestinal cancers related to bile acids; and cholestasis caused by tumors and neoplasms of the liver, biliary tract, and pancreas, including but not limited to these. The compounds of formula (I) or pharmaceutically acceptable salts thereof are also useful for enhancing corticosteroid therapy in liver diseases.;

[0052] Other diseases that can be treated or prevented by the compounds of formula (I) or pharmaceutically acceptable salts thereof include malabsorption syndromes (including abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicron retention disease (CRD), and sitosterolemia); vitamin excess and marble bone disease; hypertension; glomerular hyperfiltration; polycystic kidney disease (PKD), e.g., autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD); and uremic pruritus. The compounds are also useful for protecting against kidney injury associated with liver or metabolic diseases.

[0053] Bile acid transport in the human body is by the action of members of the SLC10 family of solute transporter proteins, particularly Na expressed on the sinusoidal cell membrane of hepatocytes +- Taurocholic acid cotransporting polypeptide (NTCP, also known as liver bile acid transporter (LBAT); gene symbol SLC10A1), and apical sodium-dependent bile acid transporter (ASBT, also known as ileal bile acid transporter (IBAT), ISBT, ABAT, or NTCP2; gene symbol SLC10A2) expressed on the apical membranes of ileal enterocytes, proximal tubular cells, bile duct epithelium, large bile duct cells, and gallbladder epithelial cells. In the liver, bile acids are efficiently extracted from portal blood by the liver bile acid transporter (LBAT) and resecreted across the canalicular membrane by the bile salt export pump (BSEP; gene symbol ABCB11). Ileal bile acid reabsorption is handled by the apical sodium-dependent bile acid transporter (ASBT), generally called the ileal bile acid transporter (IBAT) in the ileum. Both LBAT and ASBT function as electrogenic sodium-solute cotransporters that move more than two Na + ions per molecule of solute.

[0054] Endogenous substances, including xenobiotics and bile acids, are taken up from portal blood by the liver and secreted into bile by distinct transport proteins with individualized substrate specificities. Glycine- and taurine-conjugated bile acids exist in anionic form and cannot cross membranes by diffusion, and thus are completely dependent on membrane transport proteins to enter and exit hepatocytes (Kosters and Karpen, Xenobiotica 2008, Vol. 38, pp. 1043-1071). ASBT and LBAT prefer glycine- and taurine-conjugated bile salts over their unconjugated counterparts and show higher affinity for dihydroxy bile salts than trihydroxy bile salts. The non-bile acid substrates of ASBT have not yet been identified, but LBAT has been found to transport various steroid sulfates, hormones, and xenobiotics as well.

[0055] LBAT has not been as fully characterized as ASBT with respect to drug inhibition requirements. Dong et al. identified FDA-approved drugs that inhibit human LBAT and compared the inhibition requirements of LBAT and ASBT. A series of LBAT inhibition tests were conducted using FDA-approved drugs in conjunction with the development of an iterative computational model. From the screening tests, 27 drugs, including irbesartan (Ki = 11.9 μM) and ezetimibe (Ki = 25.0 μM), were identified as novel LBAT inhibitors. Since there are common features in the pharmacophore, it was shown that two hydrophobic substances and one hydrogen bond acceptor are important for the inhibition of LBAT. From 72 drugs screened in vitro, a total of 31 drugs inhibited LBAT, while 51 drugs (i.e., more than half) inhibited ASBT. Thus, although there is overlap in inhibitors, ASBT is unexpectedly more permissive than LBAT with respect to drug inhibition, which may be related to the fact that LBAT has fewer pharmacophore features (Dong et al., Mol. Pharm. 2013, Vol. 10, pp. 1008 - 1019).

[0056] Vaz et al. described the identification of LBAT deficiency as a new congenital disorder with a relatively mild clinical phenotype. The identification of LBAT deficiency confirmed that this transporter is the major uptake system for conjugated bile salts into the liver, but also showed that auxiliary transporters can sustain the enterohepatic circulation even in its absence (Vaz et al., Hepatology 2015, Vol. 61, pp. 260 - 267). These findings support the hypothesis that LBAT inhibition is a safe mechanism of action, since hepatocytes still have the potential to take up the necessary amount of bile acids.

[0057] Liu et al. described the identification of a new type of hypercholanemia associated with homozygosity for the p.Ser267Phe mutation in SLC10A1 (LBAT). The allele frequency of this mutation in the SLC10A1 gene varies by ethnic group, with the highest incidence rates occurring in South China (8% and 12% in Han Chinese and Dai ethnic groups in China, respectively) and Vietnam (11%). This "hidden" hypercholanemia was thought to affect 0.64% of the South Han Chinese ethnic group, 1.44% of the Dai ethnic group, and 1.21% of the ethnic group in Vietnam. An increase in conjugated and unconjugated serum BA levels was also observed in homozygous individuals. Liu et al. suggested that this finding is most likely due to a reduction in BA transport from the portal circulation to hepatocytes. This supports the hypothesis that the physiological function of the enterohepatic circulation is not only the recirculation of bile acids, but also the removal of bile acids from this circulation to achieve homeostasis (Karpen and Dawson, Hepatology 2015, Vol. 61, pp. 24 - 27). Alternatively, in homozygous carriers, the liver may synthesize increased levels of bile acids to compensate for the reduced enterohepatic recirculation. Since LBAT also transports unconjugated bile acids, the increase in unconjugated bile acids in this study was not surprising (Liu et al., Scientific Reports 2017, 7: 9214, pp. 1 - 7).

[0058] LBAT has been found to be downregulated in several forms of cholestatic liver injury and cholestasis, while ASBT is downregulated in various gastrointestinal disorders such as Crohn's disease, primary bile acid malabsorption, inflammatory bowel disease, and ileal inflammation, but is upregulated in cholestasis. LBAT also functions as a cellular receptor for viral entry of hepatitis B virus (HBV) and hepatitis D virus (HDV), which ultimately leads to the main causes of liver disease and hepatocellular carcinoma.

[0059] ASBT inhibition has been investigated with respect to decreasing plasma cholesterol levels, improving insulin resistance, and reducing the burden of bile acids in the liver in cholestatic liver diseases. In addition, ASBT inhibition has been found to restore insulin levels and euglycemia, whereby ASBT inhibition has been established as a promising treatment for type 2 diabetes mellitus. ASBT inhibitors are also used in the treatment of functional constipation.

[0060] Since ASBT is mainly expressed in the ileum (often called IBAT in the ileum), ASBT inhibitors do not need to be systemically absorbed. On the other hand, ASBT is also expressed in proximal tubular cells of the kidney. Therefore, systemically absorbed ASBT inhibitors may also inhibit the reuptake of bile acids in the kidney. This is thought to increase the level of bile acids in urine and enhance the removal of bile acids from the body via urine. Therefore, it is predicted that systemically absorbed ASBT inhibitors that act on both the ileum and the kidney will lead to a greater reduction in bile acid levels than non-systemically absorbed ASBT inhibitors that act only on the ileum.

[0061] Compounds with high ASBT inhibitory potency are particularly suitable for the treatment of liver diseases that cause cholestasis, such as progressive familial intrahepatic cholestasis (PFIC), Alagille syndrome, biliary atresia, and non-alcoholic steatohepatitis (NASH).

[0062] Biliary atresia is a rare pediatric liver disease that is accompanied by (or even the absence of) partial or total obstruction of the large bile ducts. This obstruction or absence causes cholestasis leading to the accumulation of bile acids that damage the liver. In some embodiments, the accumulation of bile acids occurs in the extrahepatic biliary system. In some embodiments, the accumulation of bile acids occurs in the intrahepatic biliary system. The current standard treatment is the Kasai procedure, a surgical operation that removes the obstructed bile duct and connects a part of the small intestine directly to the liver. Currently, there is no approved drug therapy for this disorder.

[0063] A method for treating biliary atresia in a subject in need of treatment for biliary atresia is provided herein, the method comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is administered a compound of formula (I) or a pharmaceutically acceptable salt thereof after undergoing the Kasai procedure. In some embodiments, the subject is administered a compound of formula (I) or a pharmaceutically acceptable salt thereof before undergoing the Kasai procedure. In some embodiments, treatment of biliary atresia results in a decrease in the level of serum bile acids in the subject. In some embodiments, the level of serum bile acids is determined by, for example, an ELISA enzyme assay or a total bile acid measurement assay as described in Danese et al., PLoS One. 2017, Vol. 12(6): e0179200, which is incorporated herein by reference in its entirety. In some embodiments, the level of serum bile acids can be reduced by, for example, 10% to 40%, 20% to 50%, 30% to 60%, 40% to 70%, 50% to 80%, or more than 90% of the level of serum bile acids before administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, treatment of biliary atresia includes treatment of pruritus.

[0064] PFIC is a rare genetic disorder that is estimated to affect one in 50,000 to 100,000 births worldwide and causes a progressive, life-threatening liver disease.

[0065] One symptom of PFIC is pruritus, which often significantly reduces quality of life. In some cases, PFIC progresses to cirrhosis and liver failure. Current therapies include partial external biliary drainage (PEFD) and liver transplantation, but these options carry a substantial risk of postoperative complications as well as potential psychological and social problems.

[0066] Three alternative genetic defects have been identified that correlate with three separate PFIC subtypes known as type 1, type 2, and type 3. · Type 1 PFIC, sometimes called "Byler's disease," is caused by a disorder of bile secretion resulting from mutations in the ATP8B1 gene, which encodes a protein that helps maintain the proper balance of fats known as phospholipids in the cell membranes within the bile ducts. These phospholipid imbalances are associated with bile stasis and elevated bile acids in the liver. Subjects affected by type 1 PFIC typically develop bile stasis at 1 month of age and, without surgical treatment, progress to cirrhosis and end-stage liver disease before 10 years of age. · Type 2 PFIC, sometimes called "Byler's syndrome," is caused by a disorder of bile salt secretion resulting from mutations in the ABCB11 gene, which encodes a protein known as the bile salt export pump that moves bile acids out of the liver. Subjects with type 2 PFIC often develop liver failure within several years of birth and have a high risk of developing a certain type of liver cancer known as hepatocellular carcinoma. · Type 3 PFIC typically presents with progressive bile stasis in the first few years of childhood and is caused by mutations in the ABCB4 gene, which encodes a transporter that moves phospholipids across cell membranes.

[0067] In addition, mutations in the TJP2 gene, NR1H4 gene, or Myo5b gene have been proposed as causes of PFIC. In addition, there are subjects with PFIC who do not have mutations in any of the ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b genes. In these cases, the cause of this condition is unknown.

[0068] Exemplary mutations in the ATP8B1 gene or the resulting protein are numbered based on the human wild-type ATP8B1 protein (e.g., SEQ ID NO: 1) or gene (e.g., SEQ ID NO: 2) and listed in Tables 2 and 3. Exemplary mutations in the ABCB11 gene or the resulting protein are numbered based on the human wild-type ABCB11 protein (e.g., SEQ ID NO: 3) or gene (e.g., SEQ ID NO: 4) and listed in Tables 4 and 5.

[0069] As will be understood by those skilled in the art, the amino acid positions in the reference protein sequence corresponding to specific amino acid positions in SEQ ID NO: 1 or 3 can be determined by aligning the reference protein sequence with SEQ ID NO: 1 or 3 (for example, using a software program such as ClustalW2). Changes to these residues (referred to herein as "mutations") can include single or multiple amino acid substitutions, insertions, and deletions within or adjacent to the sequence. As will be understood by those skilled in the art, the nucleotide positions in the reference gene sequence corresponding to specific nucleotide positions in SEQ ID NO: 2 or 4 can be determined by aligning the reference gene sequence with SEQ ID NO: 2 or 4 (for example, using a software program such as ClustalW2). Changes to these residues (referred to herein as "mutations") can include single or multiple nucleotide substitutions, insertions, and deletions within or adjacent to the sequence. See also Kooistra et al., "KLIFS: A structural kinase-ligand interaction database", Nucleic Acids Res. 2016, Vol. 44, No. D1, pp. D365-D371 (which is hereby incorporated by reference in its entirety).

[0070] Canonical protein sequence of ATP8B1 (SEQ ID NO: 1) - Uniprot ID O43520

[0071]

Chem.

[0072] Canonical DNA sequence of ATP8B1 (SEQ ID NO: 2)

[0073]

Chem.

[0074]

Chem.

[0075]

Table 2A

[0076]

Table 2B

[0077]

Table 2C

[0078]

Table 2D

[0079]

Table 2E

[0080]

Table 2F

[0081]

Table 2G

[0082]

Table 2H

[0083]

Table 2I

[0084]

Table 3A

[0085]

Table 3B

[0086]

Table 3C

[0087]

Table 3D

[0088] References related to Table 2 and Table 3 1 Folmer et al., Hepatology. 2009, vol. 50(5), p. 1597-1605. 2 Hsu et al., Hepatol Res. 2009, vol. 39(6), p. 625-631. 3 Alvarez et al., Hum Mol Genet. 2004, vol. 13(20), p. 2451-2460. 4 Davit-Spraul et al., Hepatology 2010, vol. 51(5), p. 1645-1655. 5 Vitale et al., J Gastroenterol. 2018, vol. 53(8), p. 945-958. 6 Klomp et al., Hepatology 2004, vol. 40(1), p. 27-38. 7 Zarenezhad et al., Hepatitis Monthly: 2017, vol. 17(2); e43500. 8 Dixon et al., Scientific Reports 2017, vol. 7, 11823. 9 Painter et al., Eur J Hum Genet. 2005, vol. 13(4), p. 435-439. 10 Deng et al., World J Gastroenterol. 2012, vol. 18(44), p. 6504-6509. 11 Giovannoni et al., PLoS One. 2015, vol. 10(12): e0145021. 12 Li et al., Hepatology International 2017, vol. 11, No. 1, Supp. Supplement 1, pp. S180. Abstract Number: OP284. 13 Togawa et al., Journal of Pediatric Gastroenterology and Nutrition 2018, vol. 67, Supp. Supplement 1, pp. S363. Abstract Number: 615. 14 Miloh et al., Gastroenterology 2006, vol. 130, No. 4, Suppl. 2, pp. A759-A760. Meeting Info.: Digestive Disease Week Meeting / 107th Annual Meeting of the American-Gastroenterological-Association. Los Angeles, CA, USA. May 19. 15Droege et al., Zeitschrift fur Gastroenterologie 2015, vol. 53, No. 12. Abstract Number: A3-27. Meeting Info: 32. Jahrestagung der Deutschen Arbeitsgemeinschaft zum Studium der Leber. Dusseldorf, Germany. 22 Jan 2016-23 Jan 2016 16 Mizuochi et al., Clin Chim Acta. 2012, vol. 413(15-16), p. 1301-1304. 17 Liu et al., Hepatology International 2009, vol. 3, No. 1, p. 184-185. Abstract Number: PE405. Meeting Info: 19th Conference of the Asian Pacific Association for the Study of the Liver. Hong Kong, China. 13 Feb 2009-16 Feb 2009 18 McKay et al., Version 2. F1000Res. 2013; 2: 32. DOI: 10.12688 / f1000research.2-32.v2 19 Hasegawa et al., Orphanet J Rare Dis. 2014, vol. 9:89. 20 Stone et al., J Biol Chem. 2012, vol. 287(49), p. 41139-51. 21 Kang et al., J Pathol Transl Med. 2019 May 16. doi: 10.4132 / jptm.2019.05.03. [Epub ahead of print] 22Sharma et al., BMC Gastroenterol. 2018, vol. 18(1), p. 107. 23 Uegaki et al., Intern Med. 2008, vol. 47(7), p. 599-602. 24 Goldschmidt et al., Hepatol Res. 2016, vol. 46(4), p. 306-311. 25 Liu et al., J Pediatr Gastroenterol Nutr. 2010, vol. 50(2), p. 179-183. 26 Jung et al., J Pediatr Gastroenterol Nutr. 2007, vol. 44(4), p. 453-458. 27 Bounford. University of Birmingham. Dissertation Abstracts International, (2016) Vol. 75, No. 1C. Order No.: AAI10588329. ProQuest Dissertations & Theses. 28 Stolz et al., Aliment Pharmacol Ther. 2019, vol. 49(9), p. 1195-1204. 29 Ivashkin et al., Hepatology International 2016, vol. 10, No. 1, Supp. SUPPL. 1, pp. S461. Abstract Number: LBO-38. Meeting Info: 25th Annual Conference of the Asian Pacific Association for the Study of the Liver, APASL 2016. Tokyo, Japan. 20 Feb 2016-24 Feb 2016 30 Blackmore et al., J Clin Exp Hepatol. 2013, vol. 3(2), p. 159-161. 31 Matte et al., J Pediatr Gastroenterol Nutr. 2010, vol. 51(4), p. 488-493. 32 Squires et al., J Pediatr Gastroenterol Nutr. 2017, vol. 64(3), p. 425-430. 33 Hayshi et al., EBioMedicine. 2018, vol. 27, p. 187-199. 34 Nagasaka et al., J Pediatr Gastroenterol Nutr. 2007, vol. 45(1), p. 96-105. 35 Wang et al., PLoS One. 2016; vol. 11(4): e0153114. 36 Narchi et al., Saudi J Gastroenterol. 2017, vol. 23(5), p. 303-305. 37 Alashkar et al., Blood 2015, vol. 126, No. 23. Meeting Info.: 57th Annual Meeting of the American-Society-of-Hematology. Orlando, FL, USA. December 05 -08, 2015. Amer Soc Hematol. 38Ferreira et al., Pediatric Transplantation 2013, vol. 17, Supp. SUPPL. 1, pp. 99. Abstract Number: 239. Meeting Info: IPTA 7th Congress on Pediatric Transplantation. Warsaw, Poland. 13 Jul 2013-16 Jul 2013. 39 Pauli-Magnus et al., J Hepatol. 2005, vol. 43(2), p. 342-357. 40 Jericho et al., Journal of Pediatric Gastroenterology and Nutrition 2015, vol. 60(3), p. 368-374. 41 van der Woerd et al., PLoS One. 2013, vol. 8(11): e80553. 42 Copeland et al., J Gastroenterol Hepatol. 2013, vol. 28(3), p. 560-564. 43 Droege et al., J Hepatol. 2017, vol. 67(6), p. 1253-1264. 44 Chen et al., Journal of Pediatrics 2002, vol. 140(1), p. 119-124. 45 Jirsa et al., Hepatol Res. 2004, vol. 30(1), p. 1-3. 46 van der Woerd et al., Hepatology 2015, vol. 61(4), p. 1382-1391.

[0089] In some embodiments, the mutations in ATP8B1 are selected from L127P, G308V, T456M, D554N, F529del, I661T, E665X, R930X, R952X, R1014X, and G1040R.

[0090] Canonical protein sequence of ABCB11 (SEQ ID NO: 3) - Uniprot ID O95342

[0091]

Chemical formula

[0092] Canonical DNA sequence of ABCB11 (SEQ ID NO: 4)

[0093]

Chemical formula

[0094]

Chemical formula

[0095]

Table 4A

[0096]

Table 4B

[0097]

Table 4C

[0098]

Table 4D

[0099]

Table 4E

[0100]

Table 4F

[0101]

Table 4G

[0102]

Table 4H

[0103]

Table 4I

[0104]

Table 4J

[0105]

Table 4K

[0106]

Table 4L

[0107]

Table 4M

[0108]

Table 4N

[0109]

Table 5A

[0110]

Table 5B

[0111]

Table 5C

[0112]

Table 5D

[0113]

Table 5E

[0114]

Table 5F

[0115]

Table 5G

[0116]

Table 5H

[0117] References related to Table 4 and Table 5 1 Noe et al., J Hepatol. 2005, vol. 43(3), p. 536-543. 2 Lam et al., Am J Physiol Cell Physiol. 2007, vol. 293(5), p. C1709-16. 3 Stindt et al., Liver Int. 2013, vol. 33(10), p. 1527-1735. 4Gao et al., Shandong Yiyao 2012, vol. 52(10), p. 14-16. 5 Strautnieks et al., Gastroenterology. 2008, vol. 134(4), p. 1203-1214. 6 Kagawa et al., Am J Physiol Gastrointest Liver Physiol. 2008, vol. 294(1), p. G58-67. 7 Byrne et al., Hepatology. 2009, vol. 49(2), p. 553-567. 8 Chen et al., J Pediatr. 2008, vol. 153(6), p. 825-832. 9 Davit-Spraul et al., Hepatology 2010, vol. 51(5), p. 1645-1655. 10 Droege et al., Sci Rep. 2016, vol. 6: 24827. 11 Lang et al., Pharmacogenet Genomics. 2007, vol. 17(1), p. 47-60. 12 Ellinger et al., World J Gastroenterol. 2017, vol. 23(29), p. :5295-5303. 13 Vitale et al., J Gastroenterol. 2018, vol. 53(8), p. 945-958. 14 Knisely et al., Hepatology. 2006, vol. 44(2), p. 478-86. 15Ellis et al., Hepatology. 2018, vol. 67(4), p. 1531-1545. 16 Lam et al., J Hepatol. 2006, vol. 44(1), p. 240-242. 17 Varma et al., Hepatology 2015, vol. 62(1), p. 198-206. 18 Treepongkaruna et al., World J Gastroenterol. 2009, vol. 15(34), p. 4339-4342. 19 Zarenezhad et al., Hepatitis Monthly: 2017, vol. 17(2); e43500. 20 Hayashi et al., Hepatol Res. 2016, vol. 46(2), p. 192-200. 21 Guorui et al., Linchuang Erke Zazhi 2013, vol. 31(10), 905-909. 22 van Mil et al., Gastroenterology. 2004, vol. 127(2), p. 379-384. 23 Anzivino et al., Dig Liver Dis. 2013, vol. 45(3), p. 226-232. 24 Park et al., World J Gastroenterol. 2016, vol. 22(20), p. 4901-4907. 25 Imagawa et al., J Hum Genet. 2018, vol. 63(5), p. 569-577. 26Giovannoni et al., PLoS One. 2015, vol. 10(12): e0145021. 27 Hu et al., Mol Med Rep. 2014, vol. 10(3), p. 1264-1274. 28 Lang et al,. Drug Metab Dispos. 2006, vol. 34(9), p. 1582-1599. 29 Masahata et al., Transplant Proc. 2016, vol. 48(9), p. 3156-3162. 30 Holz et al., Hepatol Commun. 2018, vol. 2(2), p. 152-154. 31 Li et al., Hepatology International 2017, vol. 11, No. 1, Supp. Supplement 1, pp. S180. Abstract Number: OP284. 32 Francalanci et al., Laboratory Investigation 2011, vol. 91, Supp. SUPPL. 1, pp. 360A. Abstract Number: 1526. 33 Francalanci et al., Digestive and Liver Disease 2010, vol. 42, Supp. SUPPL. 1, pp. S16. Abstract Number: T.N.5. 34 Shah et al., J Pediatr Genet. 2017, vol. 6(2), p. 126-127. 35 Gao et al., Hepatitis Monthly 2017, vol. 17(10), e55087 / 1-e55087 / 6. 36 Evason et al., Am J Surg Pathol. 2011, vol. 35(5), p. 687-696. 37 Davit-Spraul et al., Mol Genet Metab. 2014, vol. 113(3), p. 225-229. 38 Maggiore et al., J Hepatol. 2010, vol. 53(5), p. 981-6. 39 McKay et al., Version 2. F1000Res. 2013; 2: 32. DOI: 10.12688 / f1000research.2-32.v2 40 Liu et al., Pediatr Int. 2013, vol. 55(2), p. 138-144. 41 Waisbourd-Zinman et al., Ann Hepatol. 2017, vol. 16(3), p. 465-468. 42 Griffin, et al., Canadian Journal of Gastroenterology and Hepatology 2016, vol. 2016. Abstract Number: A200. Meeting Info: 2016 Canadian Digestive Diseases Week, CDDW 2016. Montreal, QC, United States. 26 Feb 2016-29 Feb 2016 43 Qiu et al., Hepatology 2017, vol. 65(5), p. 1655-1669. 44 Imagawa et al., Sci Rep. 2017, 7:41806. 45Kang et al., J Pathol Transl Med. 2019 May 16. doi: 10.4132 / jptm.2019.05.03. [Epub ahead of print] 46 Takahashi et al., Eur J Gastroenterol Hepatol. 2007, vol. 19(11), p. 942-6. 47 Shimizu et al., Am J Transplant. 2011, vol. 11(2), p. 394-398. 48 Krawczyk et al., Ann Hepatol. 2012, vol. 11(5), p. 710-744. 49 Sharma et al., BMC Gastroenterol. 2018, vol. 18(1), p. 107. 50 Sattler et al., Journal of Hepatology 2017, vol. 66, No. 1, Suppl. S, pp. S177. Meeting Info.: International Liver Congress / 52nd Annual Meeting of the European-Association-for-the-Study-of-the-Liver. Amsterdam, NETHERLANDS. April 19 -23, 2017. European Assoc Study Liver. 51 Jung et al., J Pediatr Gastroenterol Nutr. 2007, vol. 44(4), p. 453-458. 52Sciveres. Digestive and Liver Disease 2010, vol. 42, Supp. SUPPL. 5, pp. S329. Abstract Number: CO18. Meeting Info: 17th National Congress SIGENP. Pescara, Italy. 07 Oct 2010-09 Oct 2010 53 Sohn et al., Pediatr Gastroenterol Hepatol Nutr. 2019, vol. 22(2), p. 201-206. 54 Ho et al., Pharmacogenet Genomics. 2010, vol. 20(1), p. 45-57. 55 Wang et al., Hepatol Res. 2018, vol. 48(7), p. 574-584. 56 Shaprio et al., J Hum Genet. 2010, vol. 55(5), p. 308-313. 57 Bounford. University of Birmingham. Dissertation Abstracts International, (2016) Vol. 75, No. 1C. Order No.: AAI10588329. 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[0118] In some embodiments, the mutation in ABCB11 is selected from A167T, G238V, V284L, E297G, R470Q, R470X, D482G, R487H, A570T, N591S, A865V, G982R, R1153C, and R1268Q.

[0119] A method for treating PFIC (e.g., PFIC-1 and PFIC-2) in a subject is provided, the method comprising assaying a sample obtained from the subject to determine whether the subject has a mutation associated with PFIC (e.g., an ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b mutation), and administering to the subject determined to have a mutation associated with PFIC a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof (e.g., administering specifically or selectively). In some embodiments, the mutation is an ATP8B1 or ABCB11 mutation. For example, a mutation as shown in any one of Tables 1-4. In some embodiments, the mutation in ATP8B1 is selected from L127P, G308V, T456M, D554N, F529del, I661T, E665X, R930X, R952X, R1014X, and G1040R. In some embodiments, the mutation in ABCB11 is selected from A167T, G238V, V284L, E297G, R470Q, R470X, D482G, R487H, A570T, N591S, A865V, G982R, R1153C, and R1268Q.

[0120] Methods for treating PFIC (e.g., PFIC-1 and PFIC-2) in a subject in need of treatment for PFIC are also provided, the methods comprising: (a) detecting a mutation associated with PFIC in the subject (e.g., an ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b mutation); and (b) administering to the subject a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the method for treating PFIC may comprise administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a subject having a mutation associated with PFIC (e.g., an ATP8B1, ABCB11, ABCB4, TJP2, NR1H4, or Myo5b mutation). In some embodiments, the mutation is an ATP8B1 or ABCB11 mutation. For example, a mutation as shown in any one of Tables 1-4. In some embodiments, the mutation in ATP8B1 is selected from L127P, G308V, T456M, D554N, F529del, I661T, E665X, R930X, R952X, R1014X, and G1040R. In some embodiments, the mutation in ABCB11 is selected from A167T, G238V, V284L, E297G, R470Q, R470X, D482G, R487H, A570T, N591S, A865V, G982R, R1153C, and R1268Q.

[0121] In some embodiments, a subject is determined to have a mutation associated with PFIC in the subject or a biopsy specimen from the subject through the use of tests recognized by those of skill in the art, including next generation sequencing (NGS). In some embodiments, a subject is determined to have a mutation associated with PFIC using a test or assay approved by a regulatory agency, such as an assay approved by the FDA, or by performing any of the non-limiting examples of assays described herein, to identify a mutation associated with PFIC in the subject or a biopsy specimen from the subject. Further methods of diagnosing PFIC are described in Gunaydin, M. et al., Hepat Med. 2018, Vol. 10, pp. 95-104, which is incorporated herein by reference in its entirety.

[0122] In some embodiments, treatment of PFIC (e.g., PFIC-1 or PFIC-2) reduces the level of bile acids in the serum of a subject. In some embodiments, the level of bile acids in the serum is determined, for example, by an ELISA enzyme assay or by an assay for measuring total bile acids as described in Danese et al., PLoS One. 2017, Vol. 12(6): e0179200, which is incorporated herein by reference in its entirety. In some embodiments, the level of bile acids in the serum can be reduced by, for example, 10% - 40%, 20% - 50%, 30% - 60%, 40% - 70%, 50% - 80%, or more than 90% of the level of bile acids in the serum prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, treatment of PFIC includes treatment of pruritus.

[0123] Since LBAT is expressed in hepatocytes, LBAT and dual ASBT / LBAT inhibitor substances need to have at least a certain degree of bioavailability and free fraction in the blood. Since LBAT inhibitor compounds only need to remain from the intestine to the liver, relatively low systemic exposure of such compounds is sufficient, thereby potentially minimizing any risk of side effects at other sites of the body. Inhibition of LBAT and ASBT is expected to have at least an additive effect on the reduction of bile acid concentration in the liver. It is also expected that dual ASBT / LBAT inhibitors may be able to reduce bile acid levels without inducing diarrhea as sometimes observed with ASBT inhibitors.

[0124] Compounds having high LBAT inhibitory potency and sufficient bioavailability are expected to be particularly suitable for the treatment of hepatitis. Compounds having dual ASBT / LBAT inhibitory potency and sufficient bioavailability are expected to be particularly suitable for the treatment of non-alcoholic steatohepatitis (NASH).

[0125] NASH is a common and serious chronic liver disease similar to alcoholic liver disease, but occurs in people who rarely or never drink alcohol. In NASH patients, fat accumulation in the liver known as non-alcoholic fatty liver disease (NAFLD) or steatosis, and other factors such as high LDL cholesterol and insulin resistance induce chronic inflammation in the liver, leading to progressive scarring of tissue known as fibrosis, and cirrhosis, and ultimately potentially liver failure and death. Patients with NASH have been found to have significantly higher total serum bile acid concentrations at fasting and all postprandial time points compared to healthy subjects (a 2.2 - 2.4-fold increase in NASH at fasting and a 1.7 - 2.2-fold increase in NASH at all postprandial time points). These are driven by increases in taurine- and glycine-conjugated primary and secondary bile acids. Patients with NASH exhibited large variations in their fasting and postprandial bile acid profiles. From these results, it is shown that patients with NASH are more exposed to bile acids, including more hydrophobic and cytotoxic secondary species, at fasting and postprandially. The increased exposure to bile acids may be involved in liver injury and the development of NAFLD and NASH (Ferslew et al., Dig Dis Sci. 2015, Vol. 60, pp. 3318 - 3328). Therefore, ASBT and / or LBAT inhibition is likely to be beneficial for the treatment of NASH.

[0126] NAFLD is characterized by hepatic steatosis without secondary causes of hepatic steatosis, including excessive alcohol consumption, other known liver diseases, or long-term use of steatogenic pharmacotherapies (Chalasani et al., Hepatology 2018, Vol. 67(1), pp. 328-357). NAFLD can be classified into non-alcoholic fatty liver (NAFL) and non-alcoholic steatohepatitis (NASH). According to Chalasani et al., NAFL is defined as the presence of ≥5% hepatic steatosis without evidence of hepatocellular injury in the form of ballooning hepatocyte enlargement. NASH is defined as the presence of ≥5% hepatic steatosis and inflammation with hepatocyte injury (e.g., ballooning) regardless of the presence or absence of any hepatic fibrosis. NASH is generally also associated with liver inflammation and hepatic fibrosis, which can progress to cirrhosis, end-stage liver disease, and hepatocellular carcinoma. Hepatic fibrosis does not necessarily exist in NASH, but when present, the severity of fibrosis can be associated with long-term outcomes.

[0127] There are many techniques used to assess and grade the severity of the disease, including determining whether a subject has NAFLD and, if so, differentiating whether the NAFLD is NAFL or NASH. In some embodiments, the severity of NAFLD can be assessed using NAS. In some embodiments, the treatment of NAFLD can be evaluated 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 6 for a simplified NAS scheme based on Kleiner.

[0128]

Table 6

[0129] In some embodiments, NAS is determined non-invasively, for example, as described in U.S. Patent Application Publication No. 2018 / 0140219, which is hereby incorporated by reference in its entirety. In some embodiments, NAS is determined on a sample 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 the administration period of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a lower NAS score during or after the administration period of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as compared to prior to administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof, indicates treatment of NAFLD (e.g., NASH). For example, a decrease in NAS by 1, 2, 3, 4, 5, 6, or 7 indicates treatment of NAFLD (e.g., NASH). In some embodiments, NAS after administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof is 7 or less. In some embodiments, NAS during the administration period 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. In some embodiments, NAS during the administration period of a compound of formula (I) or a pharmaceutically acceptable salt thereof is 7 or less. In some embodiments, NAS during the administration period 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. In some embodiments, NAS after the administration period of a compound of formula (I) or a pharmaceutically acceptable salt thereof is 7 or less. In some embodiments, NAS after the administration period 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.

[0130] Additional methods for assessing or assaying NASH in a subject include determining one or more of hepatic steatosis (e.g., accumulation of fat in the liver); liver inflammation; and one or more biomarkers (e.g., serum biomarkers and panels) indicative of one or more of liver injury, liver inflammation, liver fibrosis, and / or cirrhosis. Further examples of physiological indicators of NASH can include the liver morphology, liver stiffness, and size or mass of the subject's liver. In some embodiments, NASH in a subject is evidenced by detection of accumulation of liver fat and a biomarker indicative of liver injury. For example, elevated serum ferritin and low titers of serum autoantibodies can be common features of NASH.

[0131] In some embodiments, methods for assessing NASH include magnetic resonance imaging (either by spectroscopy or proton density fat fraction measurement (MRI-PDFF)) for quantifying steatosis, transient elastography (FIBROSCAN®) for diagnosing significant liver fibrosis and / or cirrhosis, hepatic venous pressure gradient (HVPG), measurement of liver stiffness by MRE, and evaluation of histological features of a liver biopsy. In some embodiments, magnetic resonance imaging is used to detect one or more of non-alcoholic steatohepatitis (NASH-MRI), liver fibrosis (Fibro-MRI), and steatosis. See, for example, U.S. Patent Application Publication Nos. 2016 / 146715 and 2005 / 0215882, each of which is incorporated herein by reference in its entirety.

[0132] In some embodiments, treatment of NASH includes reduction of one or more symptoms associated with NASH in a subject after administration of one or more doses of a compound of formula (I) or a pharmaceutically acceptable salt thereof; reduction in the amount of hepatic steatosis; reduction in NAS; reduction in liver inflammation; reduction in the level of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis; and reduction of fibrosis and / or cirrhosis, absence of further progression of fibrosis and / or cirrhosis, or delay in the progression of fibrosis and / or cirrhosis.

[0133] In some embodiments, the treatment of NASH includes a reduction in one or more symptoms associated with NASH in a subject. Exemplary symptoms can include one or more of liver enlargement, fatigue, pain in the upper right abdomen, abdominal distension, enlargement of blood vessels just under the surface of the skin, breast enlargement in males, spleen enlargement, palmar erythema, jaundice, and pruritus. In some embodiments, the subject is asymptomatic. In some embodiments, the total weight of the subject does not increase. In some embodiments, the total weight of the subject decreases. 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 waist-to-hip ratio (WTH) of the subject does not increase. In some embodiments, the waist-to-hip ratio (WTH) of the subject decreases.

[0134] In some embodiments, the treatment of NASH can be evaluated by measuring hepatic steatosis. In some embodiments, the treatment of NASH comprises a reduction in hepatic steatosis after administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof as described herein. In some embodiments, hepatic steatosis is determined by one or more methods selected from the group consisting of ultrasonography, computed tomography (CT), magnetic resonance imaging, magnetic resonance spectroscopy (MRS), magnetic resonance elastography (MRE), transient elastography (TE) (e.g., FIBROSCAN®), measurement of liver size or mass, or by liver biopsy (see, e.g., Di Lascio et al., Ultrasound Med Biol. 2018, Vol. 44(8), pp. 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 Ledinghen V et al., J Gastroenterol Hepatol. 2016, Vol. 31(4), pp. 848-855, each of which is incorporated herein by reference in its entirety). Subjects diagnosed with NASH may have greater than about 5% hepatic steatosis, e.g., greater than about 5% to about 25%, about 25% to about 45%, about 45% to about 65%, or greater than about 65% hepatic steatosis. In some embodiments, subjects with greater than about 5% to about 33% hepatic steatosis have stage 1 hepatic steatosis, subjects with about 33% to about 66% hepatic steatosis have stage 2 hepatic steatosis, and subjects with greater than about 66% hepatic steatosis have stage 3 hepatic steatosis.

[0135] In some embodiments, the amount of hepatic steatosis is determined prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the amount of hepatic steatosis is determined during or after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a reduction in the amount of hepatic steatosis during or after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof as compared to prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof indicates treatment of NASH. For example, a reduction in the amount of hepatic steatosis of about 1% to about 50%, about 25% to about 75%, or about 50% to about 100% indicates treatment of NASH. In some embodiments, a reduction in the amount of hepatic steatosis of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% indicates treatment of NASH.

[0136] In some embodiments, the presence of liver inflammation is determined by one or more methods selected from the group consisting of biomarkers indicative of liver inflammation and liver biopsy samples from the subject. In some embodiments, the severity of liver inflammation is determined from a liver biopsy sample from the subject. For example, liver inflammation in a liver biopsy specimen can be evaluated as described in Kleiner et al., Hepatology 2005, Vol. 41(6), pp. 1313-1321, and Brunt et al., Am J Gastroenterol 1999, Vol. 94, pp. 2467-2474 (each of which is incorporated herein by reference in its entirety). In some embodiments, the severity of liver inflammation is determined prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the severity of liver inflammation is determined during or after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a decrease in the severity of liver inflammation during or after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof, compared to prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, indicates treatment of NASH. For example, a decrease in the severity of liver inflammation of about 1% to about 50%, about 25% to about 75%, or about 50% to about 100% indicates treatment of NASH. In some embodiments, a decrease in the severity of liver inflammation of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% indicates treatment of NASH.

[0137] In some embodiments, the treatment of NASH includes the treatment of fibrosis and / or cirrhosis, e.g., a decrease in the severity of fibrosis, the absence of further progression of fibrosis and / or cirrhosis, or a delay in the progression of fibrosis and / or cirrhosis. In some embodiments, the presence of fibrosis and / or cirrhosis is determined by one or more methods selected from the group consisting of transient elastography (e.g., FIBROSCAN®), non-invasive markers of fibrosis, and histological features of a liver biopsy. In some embodiments, the severity (e.g., stage) of fibrosis is determined by one or more methods selected from the group consisting of transient elastography (e.g., FIBROSCAN®), a fibrosis-scoring system, biomarkers of fibrosis (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), pp. 846-854), the fibrosis-scoring system of Brunt et al., Am. J. Gastroenterol. 1999, vol. 94, pp. 2467-2474, the fibrosis-scoring system of Kleiner et al., Hepatology 2005, vol. 41(6), pp. 1313-1321, and the Ishak fibrosis-scoring system (see Ishak et al., J. Hepatol. 1995, vol. 22, pp. 696-699) (the contents of each of which are hereby incorporated by reference in their entirety).

[0138] In some embodiments, the severity of fibrosis is determined before administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the severity of fibrosis is determined during or after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a decrease in the severity of fibrosis during or after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof compared to before administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof indicates treatment of NASH. In some embodiments, a decrease in the severity of fibrosis, absence of further progression of fibrosis and / or cirrhosis, or delay in the progression of fibrosis and / or cirrhosis indicates treatment of NASH. In some embodiments, the severity of fibrosis is determined using a scoring system such as any of the fibrosis scoring systems described herein, for example, the score can indicate the stage of fibrosis, for example, stage 0 (no fibrosis), stage 1, stage 2, stage 3, and stage 4 (cirrhosis) (see, e.g., Kleiner et al.). In some embodiments, a decrease in the stage of fibrosis is a decrease in the severity of fibrosis. For example, a decrease of 1, 2, 3, or 4 stages is a decrease in the severity of fibrosis. In some embodiments, a decrease in stage, for example, a decrease from stage 4 to stage 3, stage 4 to stage 2, stage 4 to stage 1, stage 4 to stage 0, stage 3 to stage 2, stage 3 to stage 1, stage 3 to stage 0, stage 2 to stage 1, stage 2 to stage 0, or stage 1 to stage 0 indicates treatment of NASH. In some embodiments, the stage of fibrosis decreases from stage 4 to stage 3, stage 4 to stage 2, stage 4 to stage 1, stage 4 to stage 0, stage 3 to stage 2, stage 3 to stage 1, stage 3 to stage 0, stage 2 to stage 1, stage 2 to stage 0, or stage 1 to stage 0 after administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof compared to before administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments. The stage of fibrosis is during the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable as the compound of formula (I) Compared with before administration of the salt thereof, it 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. In some embodiments, the stage of fibrosis, after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof, compared with before administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof, 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.

[0139] In some embodiments, the presence of NASH is determined by one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis, or a scoring system thereof. In some embodiments, the severity of NASH is determined by one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis, or a scoring system thereof. The level of a biomarker can be determined, for example, by measuring, quantifying, and monitoring the expression level of the gene or mRNA encoding the biomarker and / or the peptide or protein of the biomarker.Non-limiting examples of one or more biomarkers indicating liver impairment, inflammation, liver fibrosis, and / or cirrhosis, and / or their scoring systems include the ratio index of aspartate aminotransferase (AST) to platelets (APRI); the ratio of aspartate aminotransferase (AST) to alanine aminotransferase (ALT) (AAR); the FIB-4 score based on APRI, alanine aminotransferase (ALT) level, and the age of the subject (see, for example, McPherson et al., Gut 2010, Vol. 59(9), pp. 1265-1269, which is incorporated herein by reference in its entirety); hyaluronic acid; pro-inflammatory cytokines; a panel of biomarkers consisting of α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, γ-glutamyl transpeptidase (GGT) in combination with the age and sex of the subject to generate a measure of fibrotic and necroinflammatory activity in the liver (e.g., FIBROTEST®, FIBROSURE®); a panel of biomarkers consisting of bilirubin, γ-glutamyl transpeptidase, hyaluronic acid, α2-macroglobulin in combination with the age and sex of the subject (e.g., HEPASCORE®; see, for example, Adams et al., Clin. Chem. 2005, Vol. 51(10), pp. 1867-1873), and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1, hyaluronic acid, and α2-macroglobulin (e.g., FIBROSPECT®); a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1 (TIMP-1), amino-terminal propeptide of type III procollagen (PIIINP), and hyaluronic acid (HA) (e.g., Enhanced Liver Fibrosis (ELF) score, see, for example, Lichtinghagen R et al., J Hepatol. August 2013; 59(2):236-242, which is incorporated herein by reference in its entirety).In some embodiments, the presence of fibrosis is determined by one or more of a panel of biomarkers consisting of α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma-glutamyl transpeptidase (GGT) in combination with the age and sex of the subject to generate a measure of fibrosis and necroinflammatory activity in the liver (e.g., FIBROTEST®, FIBROSURE®), a panel of biomarkers consisting of bilirubin, gamma-glutamyl transferase, hyaluronic acid, α2-macroglobulin in combination with the age and sex of the subject (e.g., HEPASCORE®; see, e.g., Adams et al., Clin. Chem. 2005, Vol. 51(10), pp. 1867-1873), and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1, hyaluronic acid, and α2-macroglobulin (e.g., FIBROSPECT®); and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1 (TIMP-1), amino-terminal propeptide of type III procollagen (PIIINP), and hyaluronic acid (HA) (e.g., Enhanced Liver Fibrosis (ELF) score). In some embodiments, the level of aspartate aminotransferase (AST) does not increase. In some embodiments, the level of aspartate aminotransferase (AST) decreases. In some embodiments, the level of alanine aminotransferase (ALT) does not increase. In some embodiments, the level of alanine aminotransferase (ALT) decreases. In some embodiments, the "level" of an enzyme refers to the concentration of the enzyme, e.g., the concentration in blood. For example, the level of AST or ALT can be expressed as units / L.

[0140] In some embodiments, the severity of fibrosis is determined by one or more of the FIB-4 score, a panel of biomarkers consisting of α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma-glutamyl transpeptidase (GGT) in combination with the age and sex of the subject to generate a measure of fibrosis and necroinflammatory activity in the liver (e.g., FIBROTEST®, FIBROSURE®), a panel of biomarkers consisting of bilirubin, gamma-glutamyl transferase, hyaluronic acid, α2-macroglobulin in combination with the age and sex of the subject (e.g., HEPASCORE®; see, e.g., Adams et al., Clin. Chem. 2005, Vol. 51(10), pp. 1867-1873, which is incorporated herein by reference in its entirety), and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1, hyaluronic acid, and α2-macroglobulin (e.g., FIBROSPECT®); and a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1 (TIMP-1), amino-terminal propeptide of type III procollagen (PIIINP), and hyaluronic acid (HA) (e.g., Enhanced Liver Fibrosis (ELF) score).

[0141] In some embodiments, liver inflammation is determined by the level of biomarkers of liver inflammation, such as the level of 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 chemoattractant protein (MCP)-1, C-reactive protein (CRP), PAI-1, and collagen isoforms, such as Col1a1, Col1a2, and Col4a1 (see, e.g., Neuman et al., Can. J. Gastroenterol. Hepatol. 2014, Vol. 28(11), pp. 607-618, and U.S. Patent No. 9,872,844, each of which is incorporated herein by reference in its entirety). Liver inflammation can also be evaluated by changes in macrophage infiltration, for example, by measuring changes in CD68 expression levels. In some embodiments, liver inflammation can be determined by measuring or monitoring the serum or circulating levels of one or more of interleukin (IL)-6, interleukin (IL)-1β, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, monocyte chemoattractant protein (MCP)-1, and C-reactive protein (CRP).

[0142] In some embodiments, the level of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis is determined on a sample from a subject prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the level of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis is determined during or after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, a decrease in the level of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis during or after the administration period of the compound of formula (I) or a pharmaceutically acceptable salt thereof as compared to prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof indicates treatment of NASH. For example, a decrease of at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% in the level of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis indicates treatment of NASH. In some embodiments, the decrease in the level of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis after administration of the 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 decrease in the level of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis during the administration period of the 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 decrease in the level of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis after the administration period of the 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%.

[0143] In some embodiments, treatment of NASH results in a decrease in the level of serum bile acids in a subject. In some embodiments, the level of serum bile acids is determined by, for example, an ELISA enzyme assay or a total bile acid measurement assay such as that described in Danese et al., PLoS One. 2017, Vol. 12(6): e0179200, which is incorporated herein by reference in its entirety. In some embodiments, the level of serum bile acids can be reduced by, for example, 10% - 40%, 20% - 50%, 30% - 60%, 40% - 70%, 50% - 80%, or more than 90% of the level of serum bile acids prior to administration of the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, NASH is NASH with accompanying cholestasis. In cholestasis, the release of bile, including bile acids, from the liver is blocked. Bile acids can cause damage to hepatocytes (see, for example, Perez MJ, Briz O. World J. Gastroenterol. 2009, Vol. 15(14), pp. 1677 - 1689), thereby leading to or increasing the progression of fibrosis (e.g., cirrhosis) and increasing the risk of hepatocellular carcinoma (see, for example, Sorrentino P et al., Dig. Dis. Sci. 2005, Vol. 50(6), pp. 1130 - 1135, and Satapathy SK and Sanyal AJ. Semin. Liver Dis. 2015, Vol. 35(3), pp. 221 - 235, each of which is incorporated herein by reference in its entirety). In some embodiments, treatment of NASH includes treatment of pruritus. In some embodiments, treatment of NASH with accompanying cholestasis includes treatment of pruritus. In some embodiments, a subject having NASH with accompanying cholestasis has pruritus.

[0144] Exemplary biomarkers for NASH are shown in Table 7.

[0145]

Table 7

[0146] References related to Table 7 1 McPherson et al., Gut. 2010, vol. 59(9), p. 1265 - 1269. 2 Adams, et al. Clin Chem. 2005, vol. 51(10), p. 1867 - 1873. 3 Lichtinghagen, et al. J Hepatol. 2013, vol. 59(2), p. 236 - 242. 4 Neuman, et al. Can J Gastroenterol Hepatol. 2014, vol. 28(11), p. 607 - 618. 5 U.S. Patent No. 9,872,844

[0147] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may exhibit a higher free fraction in plasma. In some embodiments, the free fraction is greater than about 0.2%, such as greater than about 0.4%, such as greater than about 0.6%, such as greater than about 0.8%, such as greater than about 1.0%, such as greater than about 1.25%, such as greater than about 1.5%, such as greater than about 1.75%, such as greater than about 2.0%, such as greater than about 2.5%, such as greater than about 3%, such as greater than about 4%, such as greater than about 5%, such as greater than about 7.5%, such as greater than about 10%, or such as greater than about 20%.

[0148] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may be excreted in urine. In some embodiments, the fraction of the compound excreted in urine is greater than about 0.2%, such as greater than about 0.4%, such as greater than about 0.6%, such as greater than about 0.8%, such as greater than about 1.0%, such as greater than about 2%, such as greater than about 3%, such as greater than about 5%, such as greater than about 7.5%, such as greater than about 10%, such as greater than about 15%, such as greater than about 20%, such as greater than about 30%, or such as greater than about 50%.

[0149] After absorption from the intestine, some compounds of formula (I) or pharmaceutically acceptable salts thereof can be circulated via the enterohepatic circulation. In some embodiments, the fraction of the compound circulated via the enterohepatic circulation is greater than about 0.1%, such as greater than about 0.2%, such as greater than about 0.3%, such as greater than about 0.5%, such as greater than about 1.0%, such as greater than about 1.5%, such as greater than about 2%, such as greater than about 3%, such as greater than about 5%, such as greater than about 7%, such as greater than about 10%, such as greater than about 15%, such as greater than about 20%, such as greater than about 30%, or such as greater than about 50%.

[0150] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may cause renal excretion of bile salts. In some embodiments, the fraction of the circulating bile acids excreted via the renal pathway is greater than about 1%, such as greater than about 2%, such as greater than about 5%, such as greater than about 7%, such as greater than about 10%, such as greater than about 15%, such as greater than about 20%, or such as greater than about 25%.

[0151] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may exhibit improved or optimal permeability. Permeability can be measured in Caco2 cells and the value is expressed in cm / s as the Papp (apparent permeability) value. In some embodiments, the permeability is at least greater than about 0.1×10 -6 cm / s, such as greater than about 0.2×10 -6 cm / s, such as greater than about 0.4×10 -6 cm / s, such as greater than about 0.7×10 -6 cm / s, such as greater than about 1.0×10 -6 cm / s, such as greater than about 2×10 -6 cm / s, such as greater than about 3×10 -6 cm / s, such as greater than about 5×10 -6 cm / s, such as greater than about 7×10 -6 cm / s, such as greater than about 10×10 -6 cm / s, such as greater than about 15×10 -6 cm / s.

[0152] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may exhibit improved or optimal bioavailability. In some embodiments, the oral bioavailability is greater than about 5%, such as greater than about 7%, such as greater than about 10%, such as greater than about 15%, such as greater than about 20%, such as greater than about 30%, such as greater than about 40%, such as greater than about 50%, such as greater than about 60%, such as greater than about 70%, or such as greater than about 80%. In other embodiments, the oral bioavailability is between about 10% and about 90%, such as between about 20% and about 80%, such as between about 30% and about 70%, or such as between about 40% and about 60%.

[0153] Some compounds of formula (I) or pharmaceutically acceptable salts thereof can serve as substrates for relevant transporters in the kidney.

[0154] Some compounds of formula (I) or pharmaceutically acceptable salts thereof can give rise to concentrations of bile acids in the intestine, liver, and serum without causing adverse gastrointestinal effects.

[0155] Some compounds of formula (I) or pharmaceutically acceptable salts thereof can reduce the concentration of bile acids in the liver without causing gastrointestinal disorders such as diarrhea.

[0156] As used herein, the terms "treatment", "treating", and "treatment" refer to the regression, alleviation, delay in the onset, or inhibition of the progression of a disease, disorder, or one or more symptoms thereof as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual (e.g., taking into account a medical history of symptoms and / or genetic or other susceptibility factors) prior to the onset of symptoms. Treatment may also be continued after the symptoms have resolved, for example, to prevent or delay their recurrence.

[0157] Suitable pharmaceutically acceptable salts of the compounds of the present invention include, for example, base addition salts of the compounds of the present invention that are sufficiently acidic, such as alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., calcium or magnesium salts), ammonium salts, or salts with organic bases that provide physiologically acceptable cations, such as salts with methylamine, dimethylamine, trimethylamine, piperidine, morpholine, or tris-(2-hydroxyethyl)amine.

[0158] Some compounds of formula (I) or pharmaceutically acceptable salts thereof may have chiral centers and / or geometric isomer centers (E- and Z-isomers). It should be understood that the present invention encompasses all such optical isomers, diastereoisomers, and geometric isomers having ASBT and / or LBAT inhibitory activity. The present invention also encompasses all tautomers of the compounds of formula (I) or pharmaceutically acceptable salts thereof having ASBT and / or LBAT inhibitory activity. Some compounds of formula (I) or pharmaceutically acceptable salts thereof may exist not only in the non-solvated form but also, for example, in a solvated form such as a hydrated form. It should be understood that the present invention encompasses all such solvated forms having ASBT and / or LBAT inhibitory activity.

[0159] In another aspect, the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. Examples of excipients include fillers, binders, disintegrants, glidants, and lubricants. Generally, the pharmaceutical composition can be prepared in a conventional manner using conventional excipients.

[0160] Examples of suitable fillers include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose (such as lactose monohydrate), sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, dried starch, hydrolyzed starch, and pregelatinized starch. In certain embodiments, the filler is mannitol and / or microcrystalline cellulose.

[0161] 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 rubbers (such as gum arabic and tragacanth gum), sodium alginate, cellulose derivatives (such as hydroxypropylmethylcellulose (or hypromellose), hydroxypropylcellulose, and ethylcellulose), and synthetic polymers (such as copolymers of acrylic acid and methacrylic acid, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid / polymethacrylic acid copolymers, and polyvinylpyrrolidone (povidone)). In certain embodiments, the binder is hydroxypropylmethylcellulose (hypromellose).

[0162] Examples of suitable disintegrants include, but are not limited to, dried starch, modified starch ((partially) pregelatinized starch, sodium starch glycolate, and sodium carboxymethyl starch), alginic acid, cellulose derivatives (such as sodium carboxymethylcellulose, hydroxypropylcellulose, and low-substituted hydroxypropylcellulose (L-HPC)), and cross-linked polymers (such as carmellose, croscarmellose sodium, carmellose calcium, and cross-linked PVP (cross-povidone)). In certain embodiments, the disintegrant is croscarmellose sodium.

[0163] Examples of suitable flow promoters and lubricants include, but are not limited to, talc, magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, colloidal silica, aqueous silicon dioxide, synthetic magnesium silicate, micronized silicon dioxide, starch, sodium lauryl sulfate, boric acid, magnesium oxide, waxes (such as carnauba wax), hardened oils, polyethylene glycol, sodium benzoate, polyethylene glycol, and mineral oil. In certain embodiments, the flow promoter or lubricant is magnesium stearate or colloidal silica.

[0164] The pharmaceutical composition may be conventionally coated with one or more coating layers. Enteric coating layers or coating layers for delayed or targeted release of the compound of formula (I) or a pharmaceutically acceptable salt thereof are also contemplated. The coating layer may comprise one or more coating agents and optionally a plasticizer and / or a pigment (or colorant).

[0165] Examples of suitable coating agents include, but are not limited to, cellulose-based polymers (such as ethyl cellulose, hydroxypropylmethyl cellulose (or hypromellose), hydroxypropyl cellulose, cellulose acetate phthalate, cellulose acetate succinate, hydroxypropylmethyl cellulose acetate succinate, and hydroxypropylmethyl cellulose phthalate), vinyl-based polymers (such as polyvinyl alcohol), and polymers based on acrylic acid and its derivatives (such as copolymers of acrylic acid and methacrylic acid, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid / polymethacrylic acid copolymers). In certain embodiments, the coating agent is hydroxypropylmethyl cellulose. In other embodiments, the coating agent is polyvinyl alcohol.

[0166] Examples of suitable plasticizers include, but are not limited to, triethyl citrate, glyceryl triacetate, tributyl citrate, diethyl phthalate, tributyl acetylcitrate, dibutyl phthalate, dibutyl sebacate, and polyethylene glycol. In certain embodiments, the plasticizer is polyethylene glycol.

[0167] Examples of suitable pigments include, but are not limited to, titanium dioxide, iron oxide (such as yellow, brown, red, or black iron oxide), and barium sulfate.

[0168] The pharmaceutical composition may be in a form suitable for oral administration, injection (including intravenous, subcutaneous, intramuscular, and intracapsular injection), or topical administration for rectal administration. In a preferred embodiment, the pharmaceutical composition is in a form suitable for oral administration, such as tablets or capsules.

[0169] The dosage required for therapeutic or prophylactic treatment will depend on the route of administration, the severity of the disease, the age and weight of the patient, and other factors normally considered by the attending physician when determining a regimen and dosage level suitable for a particular patient.

[0170] The amount of the compound to be administered will vary depending on the patient to be treated and can range from about 1 μg / kg body weight to about 50 mg / kg body weight per day. Unit dosage forms such as tablets or capsules will usually contain 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, for example about 2.5 mg, or about 5 mg, or about 10 mg, or about 15 mg of the active ingredient. The daily dosage can be administered as a single dose or divided into one, two, three, or more unit doses. The daily dosage of the bile acid modulator administered orally is preferably within about 0.1 to about 250 mg, more preferably within about 1 to about 100 mg, for example within about 1 to about 5 mg, for example within about 1 to about 10 mg, for example within about 1 to about 15 mg, or for example within about 1 to about 20 mg.

[0171] In another aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use as a medicament. The present invention also relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as a medicament.

[0172] In another aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of any of the diseases described herein. The present invention also relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing any of the diseases described herein. The present invention also relates to a method of treating or preventing any of the diseases described herein in a subject such as a human, the method 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.

[0173] Combination therapy In one aspect of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with at least one other therapeutically active agent, for example, one, two, three, or more other therapeutically active agents. The compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one other therapeutically active agent may be administered simultaneously, sequentially, or separately. Suitable therapeutically active agents for combination with the compound of formula (I) include, but are not limited to, known active agents useful in the treatment of any of the above-mentioned conditions, disorders, and diseases.

[0174] In one embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with another ASBT inhibitor. Suitable ASBT inhibitors 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 / 38726, WO 00 / 38727, WO 00 / 38728, WO 00 / 38729, WO 01 / 66533, WO 01 / 68096, WO 02 / 32428, WO 02 / 50051, WO 03 / 020710, WO 03 / 022286, WO 03 / 022825, WO 03 / 022830, WO 03 / 061663, WO 03 / 091232, WO 03 / 106482, WO 2004 / 006899, WO 2004 / 076430, WO 2007 / 009655, WO 2007 / 009656, WO 2011 / 137135, WO 2019 / 234077, WO 2020 / 161216, WO 2020 / 161217, DE 19825804, EP 864582, EP 489423, EP 549967, EP 573848, EP 624593, EP 624594, EP 624595, EP 624596, EP 0864582, EP 1173205, EP 1535913 and EP 3210977 (all of which are hereby incorporated by reference in their entirety).

[0175] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a bile acid binder (also referred to as a bile acid sequestrant or resin), such as colesevelam, cholestyramine, or colestipol. In a preferred embodiment of such a combination, the bile acid binder is formulated for colonic release. Examples of such formulations are disclosed, for example, in WO 2017 / 138877, WO 2017 / 138878, WO 2019 / 032026, and WO 2019 / 032027 (all of which are incorporated herein by reference in their entirety).

[0176] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a DPP-IV inhibitor, such as gliptins, such as sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, treagliptin, omarigliptin, evogliptin, gosogliptin, and dutogliptin, or a pharmaceutically acceptable salt thereof.

[0177] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an HMG CoA reductase inhibitor, such as fluvastatin, lovastatin, pravastatin, simvastatin, atorvastatin, pitavastatin, cerivastatin, mevastatin, rosuvastatin, besivastatin, or dalvastatin, or a pharmaceutically acceptable salt thereof.

[0178] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a cholesterol absorption inhibitor, such as ezetimibe or a pharmaceutically acceptable salt thereof.

[0179] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a PPARα agonist, such as a fibrate, such as clofibrate, bezafibrate, ciprofibrate, clinofibrate, clofibride, fenofibrate, gemfibrozil, ronifibrate, and simfribrate, or a pharmaceutically acceptable salt thereof.

[0180] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a PPARγ agonist, such as a thiazolidinedione, such as pioglitazone, rosiglitazone, and lobeglitazone, or a pharmaceutically acceptable salt thereof.

[0181] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a dual PPARα / γ agonist, such as a glitazar, such as saroglitazar, aleglitazar, muraglitazar, or tesaglitazar, or a pharmaceutically acceptable salt thereof.

[0182] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a dual PPARα / δ agonist, such as elafibranor.

[0183] In yet another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a pan-PPAR agonist (i.e., a PPAR agonist having activity across all subtypes: α, γ, and δ), such as IVA337.

[0184] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a farnesoid X receptor (FXR) modulator, such as an FXR agonist, such as cafestol, chenodeoxycholic acid, 6α-ethyl-chenodeoxycholic acid (obeticholic acid; INT-747), fexaramine, tropifexor, cilofexor, and MET409.

[0185] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a TGR5 receptor modulator, such as a TGR5 agonist, such as 6α-ethyl-23(S)-methylcholic acid (INT-777).

[0186] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a dual FXR / TGR5 agonist, such as INT-767.

[0187] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with ursodeoxycholic acid (UDCA). In yet another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with norursodeoxycholic acid (norUDCA).

[0188] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an FGF19 modulator, such as NGM282.

[0189] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an FGF21 agonist, such as BMS-986036.

[0190] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an integrin inhibitor, such as PLN-74809 and PLN-1474.

[0191] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a CCR2 / CCR5 inhibitor, such as cenicriviroc.

[0192] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a caspase protease inhibitor, such as emricasan.

[0193] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a galectin-3 inhibitor, such as GR-MD-02.

[0194] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a stearoyl-CoA desaturase (SCD) inhibitor, such as arachidylamide cholanic acid.

[0195] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an apoptosis signal-regulating kinase 1 (ASK1) inhibitor, such as selonsertib.

[0196] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a LOXL2 inhibitor, such as simtuzumab.

[0197] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an ACC inhibitor, such as GS-0976.

[0198] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a thyroid hormone receptor beta agonist, such as MGL3196.

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

[0200] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a dual agonist of glucagon-like peptide and the glucagon receptor, such as SAR425899.

[0201] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a mitochondrial pyruvate transporter inhibitor, such as MSDC-0602K.

[0202] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an antioxidant, such as vitamin E.

[0203] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an SGLT1 inhibitor, an SGLT2 inhibitor, or a dual inhibitor of SGLT1 and SGLT2. Examples of such compounds are dapagliflozin, sotagliflozin, canagliflozin, empagliflozin, LIK066, and SGL5213.

[0204] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a diacylglycerol O-acyltransferase 2 (DGAT2) inhibitor, such as DGAT2RX and PF-06865571.

[0205] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a fatty acid synthase (FASN) inhibitor, such as TVB-2640.

[0206] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an AMP-activated protein kinase (AMPK) activator, such as PXL-770.

[0207] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a glucocorticoid receptor antagonist (GR), a mineralocorticoid receptor antagonist (MR), or a dual GR / MR antagonist. Examples of such compounds are MT-3995 and CORT-118335.

[0208] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a cannabinoid receptor 1 (CB1) antagonist, such as IM102.

[0209] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a fibroblast growth factor receptor (FGFR) and klotho-β (KLB) activator, such as MK-3655 (formerly known as NGM-313).

[0210] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a chemokine (C-C motif) ligand 24 (CCL24) inhibitor, such as CM101.

[0211] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an A3 antagonist, such as PBF-1650.

[0212] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a P2X7 receptor antagonist, such as SGM 1019.

[0213] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a P2Y13 receptor agonist, such as CER-209.

[0214] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a sulfated oxysterol, such as Dur-928.

[0215] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a leukotriene D4 (LTD4) receptor antagonist, such as MN-001.

[0216] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a type 1 natural killer T cell (NKT1) inhibitor, such as GRI-0621.

[0217] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an anti-lipopolysaccharide (LPS) compound, such as IMM-124E.

[0218] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a VAP1 inhibitor, such as BI1467335.

[0219] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an A3 adenosine receptor agonist, such as CF-102.

[0220] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a SIRT-1 activator, such as NS-20.

[0221] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a nicotinic acid receptor 1 agonist, such as ARI-3037MO.

[0222] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a TLR4 antagonist, such as JKB-121.

[0223] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a ketohexokinase inhibitor, such as PF-06835919.

[0224] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an adiponectin receptor agonist, such as ADP-335.

[0225] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an autotaxin inhibitor, such as PAT-505 and PF8380.

[0226] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a chemokine (C-C motif) receptor 3 (CCR3) antagonist, such as belrimumab.

[0227] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a chloride ion channel stimulant, such as cobiprostone and lubiprostone.

[0228] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a heat shock protein 47 (HSP47) inhibitor, such as ND-L02-s0201.

[0229] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a sterol regulatory element binding protein (SREBP) transcription factor inhibitor, such as CAT-2003 and MDV-4463.

[0230] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a biguanide, such as metformin.

[0231] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with insulin.

[0232] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a glycogen phosphorylase inhibitor and / or a glucose-6-phosphatase inhibitor.

[0233] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a sulfonylurea, such as glypidide, glibenclamide, and glimepiride.

[0234] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a meglitinide, such as repaglinide, nateglinide, and ormiglitinide.

[0235] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a glucosidase inhibitor, such as acarbose or miglitol.

[0236] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a squalene synthase inhibitor, such as TAK-475.

[0237] In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with a PTPB1 inhibitor, such as trodusquemine, ertiprotafib, JTT-551, and claramine.

[0238] Preparation of the Compounds The compounds of the present invention can be prepared by the processes described below as the free acid or a pharmaceutically acceptable salt thereof. Throughout the following description of such processes, where appropriate, suitable protecting groups will be added in a manner readily understood by those skilled in the art of organic synthesis and subsequently removed from the various reactants and intermediates. Conventional procedures for using such protecting groups, and examples of suitable protecting groups, are described, for example, in Greene's Protective Groups in Organic Synthesis, 4th Edition, by P.G.M Wutz and T.W. Greene, John Wiley & Sons, Hoboken, 2006.

[0239] General Methods All solvents used were of analytical grade. Commercially available anhydrous solvents were routinely used in the reactions. The starting materials were either available from commercial suppliers or prepared according to literature procedures. Room temperature refers to 20 - 25 °C. The composition of the solvent mixtures is expressed as volume percentages or volume ratios.

[0240] 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 NH4HCO3 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 NH4OAc 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. Method F: Mobile phase: A: 0.1% TFA in water, B: 0.1% TFA in ACN; Flow rate: 0.8 mL / min; Column: ZORBAX ECLIPSE PLUS C18 (50×2.1 mm), 1.8 μM. Method G: Mobile phase: A: 0.1% TFA in water, B: 0.1% TFA in ACN; Flow rate: 0.8 mL / min; Column: Acquity UPLC BEH C18 (2.1×50 mm), 1.7 μm. Method H: Mobile phase: A: 10 mM NH4OAc, B: 100% ACN; Flow rate: 0.8 mL / min; Column: Acquity UPLC BEH C18 (2.1×50) mm; 1.7 μm. Method I: Mobile phase: A: 0.1% HCOOH in water: ACN (95:5), B: ACN; Flow rate: 0.8 mL / min; Column: ZORBAX ECLIPSE PLUS C18 (2.1×50) mm, 1.8 μm. Method J: Mobile phase: A: 0.1% TFA in water, B: ACN; Flow rate: 1.0 mL / min; Column: Zorbax Extend C18 (50×4.6 mm), 5 μM.

[0241] UPLC: Instrument 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.

[0242] HPLC: Instrument name: Agilent 1260 Infinity II series instrument (as follows, using % in UV detection (max plot)). Method A: Mobile phase: A: 10 mM NH4HCO3 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 NH4OAc in Milli-Q water, B: ACN; Flow rate: 1.0 ml / min; Column: Phenomenex Gemini C18 (150×4.6 mm, 3.0 μm). Method D: Mobile phase: A: 0.1% TFA in water, B: ACN; Flow rate: 1.0 mL / min; Column: ATLANTIS dC18 (250×4.6 mm, 5.0 μm). Method E: Mobile phase: A: 0.1% TFA in water, B: ACN, Flow rate: 2.0 mL / min; Column: X-Bridge C8 (50×4.6 mm, 3.5 μm).

[0243] Chiral SFC: Machine name: PIC SFC 10 (for analysis) The ratio of CO2 to the co-solvent is within the range of 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. Method G: Mobile phase: 0.5% isopropylamine in methanol; Flow rate: 3 mL / min; Column: Lux A1. Method H: Mobile phase: 0.5% isopropylamine in IPA; Flow rate: 3 mL / min; Column: Lux A1 (250×4.6 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). Method J: Mobile phase: 0.5% isopropylamine in IPA; Flow rate: 5 mL / min; Column: YMC Cellulose-SC AD-H (250×4.6 mm, 5 μm). Method K: Mobile phase: 0.5% isopropylamine in methanol; Flow rate: 4 mL / min; Column: (R,R)-Whelk-01 (250×4.6 mm, 5 μm). Method L: Mobile phase: 0.5% isopropylamine in IPA; Flow rate: 3 mL / min; Column: Chiralcel OX-H (250×4.6 mm, 5 μm). Method M: Mobile phase: 0.5% isopropylamine in IPA; Flow rate: 5 mL / min; Column: YMC Cellulose-SC (250×4.6 mm, 5 μm). Method N: Mobile phase: Methanol, Flow rate: 5 mL / min; Column: Chiralcel OX-H (250×4.6 mm, 5 μm). Method O: Mobile phase: 0.1% isopropylamine in IPA:Methanol (1:1), Flow rate: 3 mL / min; Column: Chiralpak AS-H (250×4.6 mm, 5 μm). Method P: Mobile phase: 0.5% isopropylamine in methanol, Flow rate: 3 mL / min; Column: Chiralpak AS-H (250×4.6 mm, 5 μm). Method Q: Mobile phase: IPA, Flow rate: 3 mL / min; Column: Lux A1 (250×4.6 mm, 5 μm). Method R: Mobile phase: 0.1% isopropylamine in IPA:Methanol (1:1), Flow rate: 3 mL / min; Column: Lux A1 (250×4.6 mm, 5 μm). Method S: Mobile phase: 0.5% isopropylamine in methanol, Flow rate: 3 mL / min; Column: Chiralpak OX-H (250×4.6 mm, 5 μm). Method T: Mobile phase: 0.5% isopropylamine in IPA, Flow rate: 4 mL / min; Column: YMC Cellulose-SB (250×4.6 mm, 5 μm).

[0244] Preparative HPLC: Equipment name: Agilent 1290 Infinity II Method A: Mobile phase: A: 0.1% TFA in water; Mobile phase; B: 0.1% TFA in ACN; Flow rate: 2.0 mL / min; Column: X-Bridge C8 (50×4.6 mm, 3.5 μM). Method B: Mobile phase: A: 10 mM NH4OAc 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 NH4HCO3 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).

[0245] Chiral preparative SFC: Instrument name: PIC SFC 100 and PSC SFC 400 The ratio of CO2 to the co-solvent is in the range of 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). Method H: Mobile phase: 0.5% isopropylamine in methanol; Flow rate: 5 mL / min; Column: Chiralcel OX-H (250×30 mm, 5 μm).

[0246] Abbreviations ACN Acetonitrile BOC tert-Butoxycarbonyl DABCO 1,4-Diazabicyclo[2.2.2]octane DCM Dichloromethane DMA Dimethylacetamide DMF Dimethylformamide IPA Isopropyl alcohol LCMS Liquid chromatography mass spectrometry HPLC High performance liquid chromatography PE Petroleum ether SFC Supercritical fluid chromatography TFA Trifluoroacetic acid THF Tetrahydrofuran TLC Thin layer chromatography UPLC Ultra performance liquid chromatography

[0247] The present invention will now be described by the following examples, which do not limit the present invention in any way. All cited documents and references are incorporated by reference.

Example

[0248] Intermediate 1 2-(((2-Amino-5-methoxyphenyl)thio)methyl)hexanoic acid

[0249]

Chemical formula

[0250] To a stirred solution of 6-methoxybenzothiazol-2-amine (15.0 g, 0.08 mol) in water (300 mL) was added KOH (74.7 g, 1.33 mol), and the reaction mixture was stirred at 120 °C for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was cooled to room temperature. Then a solution of 2-(bromomethyl)hexanoic acid (20.0 g, 0.095 mol) in THF (100 mL) was added dropwise, and the reaction mixture was stirred at room temperature for 48 h. After consumption of the starting material (monitored by LCMS), the reaction mixture was concentrated to remove THF. The reaction mixture was then cooled to 0 °C and acidified with concentrated HCl (pH ~2). The aqueous layer was extracted with DCM (2 × 200 mL), and the combined organic layers were washed with water (100 mL) and brine (100 mL). The organic portion was dried over anhydrous Na2SO4 and then concentrated under vacuum. The crude material obtained was sent directly to the next step without further purification. Yield: 19.8 g (crude, brown gum). LCMS: (Method A) 284.2 (M + +H), Rt. 1.89 min, 86.69% (max).

[0251] Intermediate 2 3-Butyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0252]

Chemical Structure

[0253] To a stirred solution of 2-(((2-amino-5-methoxyphenyl)thio)methyl)hexanoic acid (Intermediate 1; 19.8 g, 0.069 mol) in EtOAc (200 mL) at 0 °C, triethylamine (19.5 mL, 0.139 mol) and 1-propanephosphonic anhydride solution (50% in EtOAc; 31.1 g, 0.09 mol) were added dropwise, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was quenched with water (100 mL), and the aqueous layer was extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. 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: 64% (12.0 g, brown liquid). 1 1H NMR (400 MHz, DMSO-d6): δ 7.09 (d, J = 2.8 Hz, 1H), 7.09 - 7.01 (m, 1H), 6.971 - 6.94 (m, 1H), 3.75 (s, 3H), 3.46 (q, J = 5.6 Hz, 1H), 2.88 (t, J = 12.0 Hz, 1H), 1.69 - 1.67 (m, 1H), 1.23 - 1.11 (m, 6H), 0.79 (t, J = 6.8 Hz, 3H). LCMS: (Method A) 266.2 (M + +H), Rt. 2.31 min, 98.96% (max).

[0254] Intermediate 3 7-Bromo-3-butyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0255]

Chemical Structure

[0256] A stirred solution of 3-butyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 2; 10 g, 0.03 mol) in ACN (50 mL) and DCM (50 mL) at 0 °C was added N-bromosuccinimide (8.04 g, 0.045 mol), and the resulting reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by UPLC), the reaction mixture was concentrated under vacuum. The obtained crude product was quenched with water (100 mL), and the aqueous layer was extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The obtained crude material was purified by Isolera column chromatography (eluent: 10% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 70% (9.16 g, brown gummy solid). 1 H NMR (400 MHz, DMSO-d6): δ 9.63 (s, 1H), 7.30 (s, 1H), 7.23 (s, 1H), 3.85 (s, 3H), 3.48 (q, J = 6.0 Hz, 1H), 2.93 (t, J = 12.4 Hz, 1H), 2.49-2.49 (m, 1H), 1.75-1.66 (m, 1H), 1.25-1.07 (m, 5H), 0.80 (t, J = 6.8 Hz, 3H). LCMS: (Method A) 344.1 (M + ), Rt. 2.53 min, 97.48% (max).

[0257] Intermediate 4 7-Bromo-3-butyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3-butyl-7-iodo-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0258]

Chemical Structure

[0259] To a stirred solution of 7-bromo-3-butyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 3; 5.5 g, 0.015 mol) in iodobenzene (55 mL) were added copper(I) iodide (0.304 g, 0.001 mol) and K2CO3 (4.41 g, 0.031 mol), and the solution was purged with nitrogen for 20 minutes for degassing. Then, tris[2-(2-methoxyethoxy)ethyl]amine (1.0 g, 0.003 mol) was added under a nitrogen atmosphere, and the resulting reaction mixture was heated at 135 °C for 16 hours. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through Celite, and the Celite pad was washed with EtOAc (100 mL). The filtrate was washed with water (50 mL) and brine (50 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude product was purified by Isolera column chromatography (eluent: 6% EtOAc / PE; silica gel: 230 - 400 mesh) to give a mixture of the title compounds. Yield: 66% (4.43 g, brown gum; overall yield). LCMS: (Method A) For the 7-bromo compound, 420.0 (M + ), Rt. 3.09 min, 49.30% (max) and for the 7-iodo compound, 468.0 (M + +H), Rt. 3.15 min, 47.54% (max).

[0260] Intermediate 5 7-Bromo-3-butyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3-Butyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine

[0261]

Chemical Structure

[0262] A stirred solution of a mixture of 7-bromo-3-butyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3-butyl-7-iodo-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 4; 7.0 g, 0.016 mol) in THF (70 mL) at 0 °C was added dropwise with borane dimethyl sulfide (2 M in THF; 25 mL, 0.049 mol), and the reaction mixture was refluxed at 65 °C for 16 h. After completion of the reaction (monitored by UPLC), the reaction mixture was cooled to 0 °C, quenched with methanol (15 mL), and heated at 65 °C for 2 h. Then, the resulting reaction mixture was cooled to room temperature and concentrated under vacuum. The obtained residue was diluted with water (100 mL), and the aqueous layer was extracted with DCM (2 × 200 mL). The combined organic layers were washed with water (100 mL) and brine (100 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the obtained crude material was sent directly to the next step without further purification. Yield: 7.2 g (crude, white gum). LCMS: (Method A) 406.1 (M + ) for the 7-bromo compound, Rt. 2.47 min, 57.90% (max), 454.1 (M + +H) for the 7-iodo compound, Rt. 2.66 min, 40.08% (max).

[0263] Intermediate 6 7-Bromo-3-butyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 3-butyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0264] [Chemical formula]

[0265] A stirred solution of a mixture of 7-bromo-3-butyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3-butyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine (Intermediate 5; 7.2 g, 0.02 mol) in THF (50 mL) and water (22 mL) was added Oxone (54.45 g, 0.177 mol) at 0 °C. The resulting reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through a Buchner funnel, and the filtrate was extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with water (100 mL) and brine (100 mL), dried over anhydrous Na2SO4, and then concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 7% EtOAc / PE; silica gel: 230 - 400 mesh) to give a mixture of the title compounds. Yield: 90% (7.0 g, off-white gum, overall yield). LCMS: (Method E) 438.0 (M + ) for the 7-bromo compound and 486.0 (M + +H) for the 7-iodo compound, Rt. 2.75 min, 92.14% (max).

[0266] Intermediate 7 3-Butyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetra-1,5-hydroxybenzothiazepine 1,1-dioxide

[0267]

Chemical Structure

[0268] A stirred solution of a mixture of 7-bromo-3-butyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 3-butyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 6; 3.0 g, 0.006 mol) in DMF (30 mL) was added sodium thiomethoxide (2.39 g, 0.034 mol) at room temperature, and then the reaction mixture was stirred at 65 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and quenched with water (100 mL). The aqueous layer was extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 18% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 93% (2.5 g, off-white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 10.75 (s, 1H), 7.37 (s, 1H), 7.15 - 7.11 (m, 2H), 6.99 (s, 1H), 6.65 (t, J = 7.2 Hz, 1H), 6.52 (d, J = 8.4 Hz, 2H), 4.19 (d, J = 14.8 Hz, 1H), 3.37 (s, 1H), 3.25 - 3.18 (m, 1H), 3.07 - 3.01 (m, 1H), 2.34 (s, 3H), 3.00 - 2.22 (m, 1H), 1.39 - 1.28 (m, 6H), 0.91 (t, J = 7.20 Hz, 3H). LCMS: (Method A) 392.2 (M + +H), Rt. 2.62 min, 96.86% (max).

[0269] Intermediate 8 (S)-3-Butyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and (R)-3-butyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0270] [Chemical formula]

[0271] The two enantiomers of racemic 3-butyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 7; 500 mg, 1.27 mmol) were separated by chiral preparative SFC (equipment: Pic SFC 100); mobile phase: CO2: methanol; flow rate: 4 mL / min; column: YMC Amylose-C; wavelength: 280 nm; cycle time: 5 min; back pressure: 100 bar. The substance was concentrated under vacuum at 40 °C. The first elution fraction corresponded to enantiomer 1 and the second elution fraction corresponded to enantiomer 2. The absolute configurations of the two enantiomers are unknown. Enantiomer 1: Yield: 45% (500 mg, light brown solid). LCMS: (Method E) 392.1 (M + +H), Rt. 3.048 min, 97.00% (max). SFC: (Method F) Rt. 1.77 min, 96.55% (max). Enantiomer 2: Yield: 45% (500 mg, light brown solid). LCMS: (Method E) 392.1 (M + +H), Rt. 2.907 min, 90.95% (max). SFC: (Method F) Rt. 3.65 min, 96.89% (max).

[0272] Intermediate 9 Ethyl (E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

[0273] [Chem.]

[0274] To a stirred solution of 3-butyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 7, 0.2 g, 0.51 mmol) in THF (5 mL) were added DABCO (0.06 g, 0.05 mmol) and ethyl propiolate (0.075 g, 0.76 mmol) at 0 °C, and the reaction mixture was stirred at room temperature for 1 h. 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 (3 × 15 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The resulting crude product was purified by Isolera column chromatography (eluent: 8 - 9% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound: Yield: 90% (0.23 g, colorless gum). LCMS: (Method E) 490.2 (M + +H), Rt. 2.75 min, 90.08% (max).

[0275] Intermediate 10 Ethyl (Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate

[0276] [Chem.]

[0277] To a stirred solution of 3-butyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 7; 300 mg, 0.76 mmol) in DMF (5 mL) at 0 °C was added portionwise 60% NaH (99 mg, 2.49 mmol), and the reaction mixture was stirred for 15 minutes. Then ethyl 3-bromo-2,2-difluoropropanoate (374 mg, 1.72 mmol) was added, and the reaction mixture was heated at 60 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mass was cooled to 0 °C, quenched with dilute HCl (1.5 N, pH ~4), and diluted with water (15 mL). The aqueous layer was extracted with EtOAc (2 × 50 mL), the combined organic layers were washed with brine (20 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum to afford the crude title compound, which was carried forward to the next step without further purification. Yield: 0.17 g (crude, brown gummy solid). LCMS: (Method A) 508.1 (M + +H), Rt. 2.95 min, 39.38% (max).

[0278] Intermediate 11 Ethyl (S)-(Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate and ethyl (R)-(Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate

[0279]

Chemical Structure

[0280] To a stirred solution of enantiomer 1 of 3-butyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (intermediate 8; 500 mg, 1.27 mmol) in DMF (8 mL) at 0 °C was added portionwise 60% NaH (166 mg, 4.1 mmol), and the reaction mixture was stirred for 15 minutes. Then ethyl 3-bromo-2,2-difluoropropanoate (0.41 mL, 3.19 mmol) was added, and the reaction mixture was heated at 70 °C for 3 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0 °C, quenched with 1.5 N HCl (pH ca. 4), and diluted with water (15 mL). The aqueous layer was extracted with EtOAc (2 × 25 mL). The combined organic layers were washed with water (20 mL) and brine (20 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 7–15% EtOAc / PE; silica gel: 230–400 mesh) to afford the title compound. Starting from 500 mg of enantiomer 2 of intermediate 8 and following the same procedure, enantiomer 2 of the title compound was obtained. The absolute configuration of the two enantiomers is unknown. Enantiomer 1: Yield: 650 mg (crude, white solid). LCMS: (method A) 508.2 (M + +H), Rt. 2.96 min, 92.04% (max). Enantiomer 2: Yield: 92% (600 mg, white solid). LCMS: (method E) 508.2 (M + +H), Rt. 3.42 min, 96.34% (max).

[0281] Intermediate 12 2-(((2-Amino-5-methoxyphenyl)thio)methyl)butanoic acid

[0282]

Chemical Structure

[0283] To a solution of 6-methoxybenzothiazol-2-amine (13 g, 0.072 mol) in water (260 mL) was added KOH (64.7 g, 1.15 mol), and the reaction mixture was heated at 120 °C for 48 h. The reaction mixture was then cooled to 10 °C. A solution of 2-(bromomethyl)butanoic acid (10.4 g, 0.06 mol) in THF (60 mL) was added dropwise, and the reaction mixture was heated at 70 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched and then acidified using concentrated HCl, and the aqueous layer was extracted with DCM (2 × 150 mL). The combined organic layers were washed with ice-cold water (150 mL) and brine (150 mL), and dried over anhydrous Na2SO4. The organic layer was filtered off, and the resulting solution was concentrated under vacuum to give the title compound. Yield: 14.5 g (crude, brown liquid). LCMS: (Method E) 256.2 (M + +H), Rt. 1.50 min, 50.35% (max).

[0284] Intermediate 13 3-Ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0285]

Chemical Structure

[0286] A solution of 2-(((2-amino-5-methoxyphenyl)thio)methyl)butanoic acid (Intermediate 12; 14.9 g, 0.06 mol) in EtOAc (150 mL) was added with triethylamine (16.3 mL, 0.12 mol), and the solution was cooled to 0 °C. Then, a solution of 1-propanephosphonic anhydride (50% in EtOAc; 26 g, 0.08 mol) was added dropwise, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with ice-cold water (50 mL) and diluted with EtOAc (100 mL). The organic portion was washed with ice-cold water (100 mL) and brine (100 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 18% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 44% (6.2 g, yellow solid). LCMS: (Method A) 238.1 (M + +H), Rt. 1.96 min, 87.42% (max).

[0287] Intermediate 14 7-Bromo-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0288] [Chemical formula]

[0289] A solution of 3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 13; 6.2 g, 0.03 mol) in DCM (30 mL) and ACN (30 mL) at 0 °C was added N-bromosuccinimide (5.58 g, 0.03 mol) portionwise, and the reaction mixture was stirred at room temperature for 4 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum and the resulting residue was dissolved in EtOAc (100 mL). The organic layer was washed with ice-cold water (70 mL) and brine (100 mL), and dried over anhydrous Na2SO4. The organic portion was filtered off and concentrated under vacuum. The obtained crude material was purified by Isolera column chromatography (eluent: 10% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 34% (2.8 g, light brown solid). 1 H NMR (400 MHz, DMSO-d6): δ 9.66 (s, 1H), 7.31 (s, 1H), 7.24 (s, 1H), 3.86 (s, 3H), 3.53 - 3.48 (m, 1H), 2.97 - 2.91 (m, 1H), 2.39 - 2.33 (m, 1H), 1.72 - 1.65 (m, 1H), 1.32 - 1.27 (m, 1H), 0.78 - 0.73 (m, 3H). LCMS: (Method E) 315.9 (M + ), Rt. 2.44 min, 50.63% (max).

[0290] Intermediate 15: 7-Bromo-3-ethyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3-ethyl-7-iodo-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0291]

Chemical Structure

[0292] To a degassed solution of 7-bromo-3-ethyl-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 14; 1.6 g, 5.05 mmol) in iodobenzene (16 mL) were added K2CO3 (1.4 g, 10.2 mmol), tris[2-(2-methoxyethoxy)ethyl]amine (0.32 g, 1.01 mmol) and CuI (0.1 g, 0.5 mmol), and the reaction mixture was heated at 135 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum and the resulting residue was diluted with EtOAc (50 mL). The organic layer was washed with water (50 mL) and brine (50 mL) and dried over anhydrous Na2SO4. The organic portion was filtered off and concentrated under vacuum, 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: 54% (1.9 g, brown gummy solid). LCMS: (Method A) For the 7-bromo substituted compound 394.0 (M + +2), Rt. 2.82 min, 34.92% (max) and for the 7-iodo substituted compound 439.9 (M + +H), Rt. 2.88 min, 31.38% (max).

[0293] Intermediate 16 7-Bromo-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3-Ethyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine

[0294]

Chemical Structure

[0295] A solution of a mixture of 7-bromo-3-ethyl-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one and 3-ethyl-7-iodo-8-methoxy-5-phenyl-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 15; 1.9 g, 4.84 mmol) in THF (20 mL) at 0 °C was added with borane dimethyl sulfide (2 M solution in THF, 7.2 mL, 14.5 mmol), and the reaction mixture was heated at 75 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0 °C. Then, methanol (10 mL) was added dropwise, and the mixture was heated at 60 °C for 2 h. Then, the reaction mixture was cooled to room temperature and concentrated under vacuum. The obtained residue was partitioned between water (30 mL) and EtOAc (30 mL), and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with water (50 mL) and brine (50 mL), and dried over anhydrous Na2SO4. The organic portion was filtered off and concentrated under vacuum to give the title compound. Yield: 2.5 g (crude, brown gum). LCMS: (Method A) 378.1 (M + ) for the 7-bromo-substituted compound, Rt. 3.50 min, 14.77% (max) and 426.1 (M + +H) for the 7-iodo-substituted compound, Rt. 3.62 min, 11.01% (max).

[0296] Intermediate 17 7-Bromo-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 3-ethyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0297]

Chemical formula

[0298] A solution of a mixture of 7-bromo-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine and 3-ethyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine (Intermediate 16; 2.5 g, 6.60 mmol) in a mixture of THF and water (8:2, 25 mL) was added with Oxone (20.3 g, 66.0 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 extracted with EtOAc (2×30 mL). The combined organic layers were washed with ice-cold water (50 mL) and brine (50 mL). The organic portion was dried over anhydrous Na2SO4 and concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 13% EtOAc / PE; silica gel: 230 - 400 mesh) to obtain the title compound. Yield: 55% (1.5 g, brown gum). LCMS: (Method A) For the 7-bromo-substituted compound, 410.0 (M + ), Rt. 2.76 min, 43.90% (max) and for the 7-iodo-substituted compound, 457.9 (M + +H), Rt. 2.81 min, 38.27% (max).

[0299] Intermediate 18 3-Ethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0300]

Chemical Structure

[0301] A solution of a mixture of 7-bromo-3-ethyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and 3-ethyl-7-iodo-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 17; 1.5 g, 3.6 mmol) in DMF (15 mL) was added with sodium thiomethoxide (1.3 g, 18.2 mmol), and the reaction mixture was stirred at 80 °C for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with ice-cold water (30 mL), and the aqueous layer was extracted with EtOAc (2 × 30 mL). The combined organic layers were washed with water (30 mL) and brine (30 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, 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: 68% (0.9 g, off-white solid). LCMS: (Method E) 364.0 (M + +H), Rt. 2.67 min, 78.14% (max).

[0302] Intermediate 19 Ethyl (Z)-3-((3-ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate

[0303]

Chem.

[0304] To a stirred solution of 3-ethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 18; 0.3 g, 0.82 mmol) in DMA (6 mL) at 0 °C was added 60% NaH (0.107 g, 2.68 mmol) portionwise and the reaction mixture was stirred at 0 °C for 30 minutes. Then a solution of ethyl 3-bromo-2,2-difluoropropanoate (0.26 mL, 2.06 mmol) in DMA (1 mL) was added and the reaction mixture was heated at 65 °C for 3 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0 °C, quenched with dilute HCl (1.5 N, pH ~4) and diluted with water (10 mL). The aqueous layer was extracted with EtOAc (2 × 15 mL). The combined organic layers were then washed with brine (10 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum and the resulting crude was purified by Isolera column chromatography (eluent: 16% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 33% (0.13 g, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.78-7.73 (m, 1H), 7.68 (s, 1H), 7.22-7.16 (m, 3H), 6.74 (t, J = 7.2 Hz, 1H), 6.65 (d, J = 8.0 Hz, 2H), 4.28 (q, J = 7.2 Hz, 2H), 3.53-3.49 (m, 2H), 3.30-3.24 (m, 2H), 2.43 (s, 3H), 2.25-2.23 (m, 1H), 1.49-1.39 (m, 2H), 1.37-1.26 (m, 3H), 1.01-0.97 (m, 3H). LCMS: (Method E) 480.0 (M + +H), Rt. 2.98 min, 94.13% (max).

[0305] Intermediate 20 tert-Butyl (E)-3-((3-ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

[0306]

Chem.

[0307] To a stirred solution of 3-ethyl-8-hydroxy-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 18, 300 mg, 0.82 mmol) in THF (5 mL) were added DABCO (0.01 g, 0.08 mmol) and tert-butyl propiolate (0.16 g, 1.23 mmol) at room temperature, and the reaction mixture was stirred at room temperature for 3 h. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water (20 mL), and the aqueous layer was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with water (2 × 10 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The obtained crude product was purified by Isolera column chromatography (eluent: 12% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 89% (0.36 g, white solid). LCMS: (Method E) 433.9 (M + - t Bu + H), Rt. 3.17 min, 89.46% (max).

[0308] Intermediate 21 2-((tert-Butoxycarbonyl)amino)hexanoic acid

[0309]

Chem.

[0310] A solution of 2-aminohexanoic acid (25 g, 0.19 mol) in water (250 mL) and THF (250 mL) was added with NaHCO3 (48 g, 0.57 mol) and Boc anhydride (52.2 mL, 0.23 mol), and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled, quenched, and acidified using 1.5 N HCl. The reaction mixture was extracted with EtOAc (2 × 150 mL). The combined organic layers were washed with ice-cold water (150 mL) and brine (150 mL), dried over anhydrous Na2SO4, and concentrated to give the title compound. Yield: 45.5 g (crude, white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 5.10-4.98 (m, 1H), 4.33-4.30 (m, 1H), 1.95-1.81 (m, 1H), 1.75-1.55 (m, 1H), 1.46 (s, 10H), 1.45-1.31 (m, 4H), 0.93 (t, J = 7.2 Hz, 3H). LCMS: (Method E) 132.2 (M + -Boc+H), Rt. 2.36 min, 99.98% (max).

[0311] Intermediate 22 tert-Butyl (1-oxo-1-(phenylamino)hexan-2-yl)carbamate

[0312]

Chemical Structure

[0313] To a stirred solution of 2-((tert-butoxycarbonyl)amino)hexanoic acid (Intermediate 21; 45.5 g, 0.196 mol) in DMF (150 mL) was added triethylamine (54.83 mL, 0.39 mol), and the reaction mixture was cooled to 0 °C. A solution of 1-propanephosphonic anhydride (50% in EtOAc; 75.1 g, 0.23 mol) and aniline (18 g, 0.196 mol) were added, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with ice-cold water (50 mL) and diluted with EtOAc (200 mL). The aqueous layer was washed with ice-cold water (200 mL) and brine (200 mL), then dried over anhydrous Na2SO4, filtered off. The organic portion was concentrated under vacuum to afford the title compound. Yield: 82% (49.7 g, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 9.93 (s, 1H), 7.60 (d, J = 7.6 Hz, 2H), 7.30 (t, J = 8.0 Hz, 2H), 7.06 - 6.99 (m, 2H), 4.07 - 4.02 (m, 1H), 1.63 - 1.56 (m, 2H), 1.46 - 1.28 (m, 13H), 0.86 (t, J = 6.8 Hz, 3H). LCMS: (Method E) 207.0 (M + -Boc+H), Rt. 2.69 min, 91.20% (max).

[0314] Intermediate 23 2-Amino-N-phenylhexanamide

[0315]

Chemical Structure

[0316] A solution of tert-butyl (1-oxo-1-(phenylamino)hexan-2-yl)carbamate (Intermediate 22; 49.7 g, 0.162 mol) in 1,4-dioxane (500 mL) at 0 °C was treated with a solution of HCl in 1,4-dioxane (4 M, 199 mL), and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum, and the resulting residue was quenched with saturated NaHCO3 solution. The aqueous layer was extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with brine (200 mL) and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated under vacuum to afford the title compound. Yield: 95% (32 g, colorless gum). 1 H NMR (400 MHz, DMSO-d6): δ 10.20-9.66 (m, 1H), 7.64 (dd, J = 4.8, 2.4 Hz, 2H), 7.32-7.27 (m, 2H), 7.06-7.02 (m, 1H), 3.29-3.26 (m, 1H), 1.67-1.61 (m, 1H), 1.46-1.27 (m, 5H), 0.89-0.85 (m, 3H). LCMS: (Method B) 207.2 (M + +H), Rt. 2.03 min, 84.36% (max).

[0317] Intermediate 24 N1-Phenylhexane-1,2-diamine

[0318]

Chemical formula

[0319] A solution of 2-amino-N-phenylhexanamide (Intermediate 23; 32 g, 0.15 mol) in THF (320 mL) at 0 °C was treated with borane dimethyl sulfide (2 M solution in THF, 117 mL, 0.23 mol), and the reaction mixture was heated at 75 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0 °C, quenched with methanol (150 mL), and then heated at 60 °C for 2 h. The reaction mixture was cooled to room temperature and concentrated under vacuum. The resulting residue was partitioned between water (50 mL) and EtOAc (50 mL). The aqueous layer was extracted with EtOAc (2 × 250 mL). The combined organic layers were washed with water (250 mL) and brine (250 mL). The organic portion was dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 10% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 93% (28 g, yellow oil). LCMS: (Method B) 193.3 (M + +H), Rt. 1.98 min, 76.9% (max).

[0320] Intermediate 25 2,4-Dibromo-5-methoxy-N-(1-(phenylamino)hexan-2-yl)benzenesulfonamide

[0321]

Chemical Structure

[0322] A solution of 2,4-dibromo-5-methoxybenzenesulfonyl chloride (3.2 g, 8.79 mmol) in THF (50 mL) at 0 °C was added to N1-phenylhexane-1,2-diamine (Intermediate 24; 1.3 g, 6.76 mmol) and triethylamine (2.8 mL, 20.3 mmol), and the reaction mixture was stirred at room temperature for 4 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum, and the resulting residue was dissolved in EtOAc (60 mL). The organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 18% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 97% (3.5 g, brown gum). LCMS: (Method E) 521.0 (M + +H), Rt. 3.14 min, 93.57% (max).

[0323] Intermediate 26 7-Bromo-3-butyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0324] [Chemical formula]

[0325] To a solution of 2,4-dibromo-5-methoxy-N-(1-(phenylamino)hexan-2-yl)benzenesulfonamide (Intermediate 25; 3.5 g, 6.72 mmol) in DMF (30 mL) were added K2CO3 (1.73 g, 12.5 mmol) and copper powder (0.42 g, 6.72 mmol). The reaction mixture was degassed under a N2 atmosphere for 5 minutes and then the reaction mixture was heated at 115 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with ice-cold water (25 mL) and the aqueous layer was extracted with a 1:1 mixture of EtOAc and PE (2 × 50 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. 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: 51% (1.5 g, brown gum). LCMS: (Method A) 439.0 (M + +H), Rt. 2.83 min, 82.06% (max).

[0326] Intermediate 27 7-Bromo-3-butyl-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0327] [Chemical formula]

[0328] To a solution of 7-bromo-3-butyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 26; 1.25 g, 2.84 mmol) in N-methyl-2-pyrrolidone (15 mL) was added Cs2CO3 (1.4 g, 4.27 mmol), and the reaction mixture was stirred at room temperature for 15 minutes. Then, p-methoxybenzyl bromide (0.5 mL, 3.41 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with ice-cold water (25 mL), and the precipitated solid was filtered off. The obtained crude material was purified by Isolera column chromatography (eluent: 30% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 90% (1.4 g, white solid). LCMS: (Method E) 558.9 (M + +H), Rt. 3.36 min, 90.75% (max).

[0329] Intermediate 28 3-Butyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0330]

Chemical Structure

[0331] To a stirred solution of 7-bromo-3-butyl-8-methoxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 27; 1.4 g, 2.57 mmol) in DMF (10 mL) was added sodium thiomethoxide (0.9 g, 12.8 mmol), and the reaction mixture was heated at 60 °C for 16 h. 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 × 30 mL) and brine (30 mL). The organic portion was dried over anhydrous Na2SO4 and concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 16% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 68% (0.9 g, off-white solid). LCMS: (Method E) 513.0 (M + +H), Rt. 3.12 min, 90.12% (max).

[0332] Intermediate 29 tert-Butyl (E)-3-((3-butyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

[0333] [Chemical Structure]

[0334] A stirred solution of 3-butyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 28; 0.2 g, 0.39 mmol) in THF (5 mL) at 0 °C was treated with DABCO (0.005 g, 0.04 mmol) and tert-butyl propiolate (0.08 g, 0.58 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 poured into ice-cold water (10 mL), and the aqueous layer was extracted with EtOAc (2 × 15 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 8 - 10% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 80% (0.2 g, white gum). LCMS: (Method E) 583.0 (M + - t Bu + H), Rt. 3.49 min, 86.23% (max)

[0335] Intermediate 30 Ethyl (Z)-3-((3-butyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0336]

Chem.

[0337] A stirred solution of 3-butyl-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 28; 0.5 g, 0.97 mmol) in DMA (5 mL) at 0 °C was treated portionwise with 60% NaH (0.127 g, 3.16 mmol), and the reaction mixture was stirred at 0 °C for 30 minutes. Then, a solution of ethyl 3-bromo-2,2-difluoropropanoate (0.47 g, 2.18 mmol) in DMA (2.5 mL) was added, and the reaction mixture was heated at 70 °C for 3 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0 °C, then quenched with dilute HCl (1.5 N, pH ~4) and diluted with water (10 mL). The aqueous layer was extracted with EtOAc (2 × 10 mL), the combined organic layers were washed with brine (10 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude product was purified by Isolera column chromatography (eluent: 15 - 20% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 50% (0.3 g, off-white solid). LCMS: (Method E) 629.2 (M + +H), Rt. 3.37 min, 90.09% (max).

[0338] Intermediate 31 (Z)-3-((3-Butyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0339]

Chemical Structure

[0340] A stirred solution of ethyl (Z)-3-((3-butyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 30; 0.3 g, 0.48 mmol) in a mixture of 1,4-dioxane and water (4:1, 5 mL) was treated with lithium hydroxide (0.06 g, 1.43 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH ca. 4) and diluted with ice-cold water (10 mL). The aqueous layer was extracted with EtOAc (2 × 15 mL), and the combined organic layers were washed with water (10 mL) and brine (10 mL). The organic portion was dried over anhydrous Na2SO4 and concentrated under vacuum. The crude material obtained was purified by Isolera column chromatography (eluent: 100% EtOAc; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 77% (0.22 g, brown solid). LCMS: (Method E) 600.8 (M + +H), Rt. 3.10 min, 41.73% (max).

[0341] Intermediate 32 7-Bromo-3-butyl-8-methoxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0342] [Chemical Structure]

[0343] To a stirred solution of 7-bromo-3-butyl-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 26; 4.5 g, 10.24 mmol) in N-methyl-2-pyrrolidone (20 mL) was added Cs2CO3 (6.7 g, 20.5 mmol), followed by iodomethane (3.2 mL, 51.2 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 quenched with water (25 mL), and the aqueous layer was extracted with a mixture of EtOAc and PE (30%, 2 × 75 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The crude obtained was carried on to the next step without further purification. Yield: 4.5 g (crude, light brown solid). LCMS: (Method E) 452.8 (M + +H), Rt. 3.19 min, 95.26% (max).

[0344] Intermediate 33 3-Butyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0345]

Chemical Structure

[0346] A solution of 7-bromo-3-butyl-8-methoxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 32; 4.5 g, 9.92 mmol) in DMF (50 mL) was added with sodium thiomethoxide (3.5 g, 49.6 mmol), and the reaction mixture was heated at 80 °C for 16 h. After completion of the reaction (monitored by UPLC), the reaction mixture was quenched with ice-cold water (25 mL), and the aqueous layer was extracted with EtOAc (4 × 50 mL). The combined organic layers were washed with water (2 × 100 mL) and brine (100 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 26% EtOAc in PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 77% (3.1 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.73 (s, 1H), 7.29 (s, 1H), 7.12 - 7.16 (m, 2H), 6.99 (s, 1H), 6.67 (t, J = 7.2 Hz, 1H), 6.54 (d, J = 8.0 Hz, 2H), 4.06 - 3.89 (m, 2H), 3.21 - 3.09 (m, 1H), 2.42 (s, 3H), 2.34 (s, 3H), 1.68 - 1.55 (m, 1H), 1.54 - 1.42 (m, 1H), 1.41 - 1.25 (m, 4H), 0.93 - 0.90 (m, 3H). LCMS: (Method A) 406.9 (M + +H), Rt. 2.65 min, 92.05% (max).

[0347] Intermediate 34 (S)-3-Butyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide and (R)-3-butyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0348] [Chemical]

[0349] The two enantiomers of racemic 3-butyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 33; 10.0 g, 24.59 mmol) were separated by an SFC instrument (Method E). The substance was concentrated at 40 °C under vacuum. The first elution fraction corresponded to Enantiomer 1 and the second elution fraction corresponded to Enantiomer 2. The absolute configurations of the two enantiomers are unknown. Enantiomer 1: Yield: 42% (4.3 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.69 (s, 1H), 7.28 (s, 1H), 7.13 (t, J = 10.8 Hz, 2H), 6.99 (s, 1H), 6.66 (t, J = 9.6 Hz, 1H), 6.53 (d, J = 10.8 Hz, 2H), 4.00 - 3.89 (m, 2H), 3.19 - 3.14 (m, 1H), 2.42 (s, 3H), 2.34 (s, 3H), 1.68 - 1.55 (m, 1H), 1.55 - 1.45 (m, 1H), 1.40 - 1.25 (m, 4H), 0.95 - 0.85 (m, 3H). LCMS: (Method A) 407.1 (M + +H), Rt. 2.56 min, 98.41% (max). HPLC: (Method E) Rt. 5.43 min, 98.58% (max). Chiral SFC: (Method D) Rt. 1.83 min, 100% (max). Enantiomer 2: Yield: 34% (4.2 g, off-white solid). 11H NMR (400 MHz, DMSO-d6): δ 10.58 (s, 1H), 7.27 (s, 1H), 7.13 (t, J = 10.0 Hz, 2H), 6.98 (s, 1H), 6.66 (t, J = 10.0 Hz, 1H), 6.53 (d, J = 10.8 Hz, 2H), 4.10 - 3.80 (m, 2H), 3.25 - 3.05 (m, 1H), 2.42 (s, 3H), 2.34 (s, 3H), 1.68 - 1.45 (m, 2H), 1.45 - 1.25 (m, 4H), 0.98 - 0.85 (m, 3H). LCMS: (Method A) 407.2 (M + +H), Rt. 2.56 min, 98.25% (max). HPLC: (Method E) Rt. 5.43 min, 97.49% (max). Chiral SFC: (Method D) Rt. 3.06 min, 99.76% (max).

[0350] Intermediate 35 Ethyl (Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0351] [Chemical Structure]

[0352] A stirred solution of 3-butyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 33; 0.5 g, 1.23 mmol) in DMA (7.5 mL) at 0 °C was treated portionwise with NaH (60%, 0.16 g, 3.99 mmol), and the reaction mixture was stirred at 0 °C for 30 min. Then a solution of ethyl 3-bromo-2,2-difluoropropanoate (0.6 g, 2.78 mmol) in DMA (2.5 mL) was added, and the reaction mixture was heated at 70 °C for 3 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to 0 °C, quenched with dilute HCl (1.5 N HCl, pH ca. 4), and diluted with water (10 mL). The aqueous layer was extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude was purified by Isolera column chromatography (eluent: 15 - 20% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 94% (0.6 g, off-white solid). 1 HNMR (400 MHz, DMSO-d6): δ 7.83 (m, 1H), 7.60 (s, 1H), 7.18 (t, J = 7.6 Hz, 3H), 6.74 (t, J = 10.0 Hz, 1H), 6.66 (d, J = 10.8 Hz, 2H), 4.28 (q, J = 9.2 Hz, 2H), 4.09 - 4.02 (m, 1H), 3.87 - 3.82 (m, 1H), 3.34 - 3.31 (m, 1H), 2.49 (s, 3H), 2.41 (m, 3H), 1.62 - 1.53 (m, 2H), 1.51 - 1.49 (m, 3H), 1.35 - 1.24 (m, 4H), 0.93 - 0.91 (m, 3H). LCMS: (Method E) 523.1 (M + +H), Rt. 3.18 min, 97.01% (max).

[0353] Intermediate 36 Ethyl (S)-(Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate and Ethyl (R)-(Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0354] [Chemical formula]

[0355] To a stirred solution of enantiomer 1 of 3-butyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (intermediate 34; 2.0 g, 4.91 mmol) in DMA (20 mL) at 0 °C was added NaH (60%; 640 mg, 15.98 mmol), and the reaction mixture was stirred at room temperature for 1 h. Then, ethyl 3-bromo-2,2-difluoropropanoate (2.66 g, 12.29 mmol) was added at 0 °C, and the resulting reaction mixture was stirred at 90 °C for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (1.5 N, 20 mL), diluted with water (50 mL), and the aqueous layer was extracted with EtOAc (2 × 100 mL). The combined organic layers were washed with water (50 mL) and brine (50 mL), and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 15 - 20% EtOAc / PE; silica gel: 230 - 400 mesh) to give enantiomer 1 of the title compound. Starting from enantiomer 2 of 2.0 g of intermediate 34, following the same procedure, enantiomer 2 of the title compound was obtained. The absolute configurations of the two enantiomers are unknown. Enantiomer 1: Yield: 76% (1.95 g, off-white solid). LCMS: (Method E) 523.0 (M+ +H), retention time 3.19 minutes, 97.85% (maximum). HPLC: (method B) retention time 6.25 minutes, 97.32% (maximum). Enantiomer 2: Yield: 59% (1.51 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.84 (d, J = 18.8 Hz, 1H), 7.60 (s, 1H), 7.20 - 7.16 (m, 3H), 6.77 - 6.65 (m, 3H), 4.31 - 4.25 (m, 2H), 4.08 (m, 1H), 3.95 - 3.70 (m, 1H), 3.30 - 3.15 (m, 1H), 2.51 (s, 3H), 2.42 (s, 3H), 1.70 - 1.50 (m, 2H), 1.50 - 1.32 (m, 7H), 0.95 - 0.85 (m, 3H). LCMS: (method E) 523.0 (M + +H), retention time 3.19 minutes, 98.36% (maximum). HPLC: (method B) retention time 6.26 minutes, 97.97% (maximum).

[0356] Intermediate 37 tert-Butyl (E)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

[0357]

Chemical Structure

[0358] A stirred solution of 3-butyl-8-hydroxy-2-methyl-7-(methylthio)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 33; 0.5 g, 1.23 mmol) in THF (7.5 mL) at 0 °C was treated with DABCO (0.02 g, 0.12 mmol) and tert-butyl propiolate (0.23 g, 1.84 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 poured into ice-cold water (15 mL), and the aqueous layer was extracted with EtOAc (2 × 15 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 8–10% EtOAc / PE; silica gel: 230–400 mesh) to afford the title compound. Yield: 92% (0.6 g, off-white gum).

[0359] Intermediate 38 3-Butyl-7-(ethylthio)-8-hydroxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0360]

Chem.

[0361] To a solution of 7-bromo-3-butyl-8-methoxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (0.3 g, 0.66 mmol) in N-methyl-2-pyrrolidone (5 mL) was added sodium ethanethiolate (0.27 g, 3.31 mmol), and the reaction mixture was stirred at 100 °C for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with ice-cold water (25 mL), and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with ice-cold water (50 mL) and brine (50 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 30% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 69% (0.2 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.63 (s, 1H), 7.42 (s, 1H), 7.23 - 7.21 (m, 2H), 6.83 (t, J = 7.6 Hz, 1H), 6.68 - 6.64 (m, 3H), 4.05 - 4.01 (m, 2H), 3.23 - 3.21 (m, 1H), 2.80 (q, J = 7.2 Hz, 2H), 2.60 (s, 3H), 1.57 - 1.31 (m, 6H), 1.27 (t, J = 7.20 Hz, 3H), 0.96 (t, J = 7.20 Hz, 3H). LCMS: (Method E) 421.0 (M + +H), Rt. 3.04 min, 95.23% (max).

[0362] Intermediate 39 tert-Butyl (E)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

[0363]

Chemical Structure

[0364] To a stirred solution of 3-butyl-7-(ethylthio)-8-hydroxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 38; 0.15 g, 0.36 mmol) in THF (5 mL) were added tert-butyl propiolate (67 mg, 0.53 mmol) and DABCO (20 mg, 0.18 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 concentrated under vacuum, and the resulting residue was partitioned between water (5 mL) and EtOAc (5 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), the combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The obtained crude material was purified by Isolera column chromatography (eluent: 20% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 70% (140 mg, white solid). LCMS: (Method E) 490.9 (M + - t Bu + H), Rt. 3.43 min, 98.33% (max).

[0365] Intermediate 40 tert-butyl (1-((4-fluorophenyl)amino)-1-oxohexan-2-yl)carbamate

[0366]

Chemical Structure

[0367] To a stirred solution of 2-((tert-butoxycarbonyl)amino)hexanoic acid (93.0 g, 402 mmol) in DMF (930 mL) at 0 °C was added dropwise triethylamine (112 mL, 804 mmol). Then, 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinan 2,4,6-trioxide (307 g, 483 mmol) was added and the reaction mixture was stirred at 0 °C for 10 minutes. Next, 4-fluoroaniline (44.7 g, 402 mmol) was added dropwise and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with ice-cold water (500 mL) to precipitate the solid product. The resulting solid was filtered, washed with water (2 × 100 mL) and dried under vacuum to give the title compound. The crude product obtained was sent directly to the next step without further purification. Yield: 69% (90 g, light yellow solid). LCMS: (Method K) 225.2 (M + +H-Boc), Rt. 2.66 min, 99.41% (max).

[0368] Intermediate 41 2-Amino-N-(4-fluorophenyl)hexanamide

[0369] [Chemical formula]

[0370] To a stirred solution of tert-butyl (1-((4-fluorophenyl)amino)-1-oxohexan-2-yl)carbamate (Intermediate 40; 90 g, 277 mmol) in 1,4-dioxane (900 mL) was added HCl in dioxane (4 M in 1,4-dioxane, 360 mL, 1280 mmol) at 0 °C, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum and the resulting residue was basified with 10% NaHCO3 solution (400 mL). The aqueous layer was extracted with EtOAc (2 × 500 mL), the combined organic layers were washed with water (2 × 100 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum and the resulting crude product was sent directly to the next step without further purification. Yield: 91% (60 g, light brown liquid). LCMS: (Method K) 225.1 (M + +H), Rt. 1.97 min, 94.23% (max).

[0371] Intermediate 42 N1-(4-Fluorophenyl)hexane-1,2-diamine

[0372]

Chemical Structure

[0373] To a stirred solution of 2-amino-N-(4-fluorophenyl)hexanamide (Intermediate 41; 60 g, 268 mmol) in THF (300 mL) was added borane dimethyl sulfide (1 M in THF, 401 mL, 401 mmol) at 0 °C, and the reaction mixture was heated at 75 °C for 24 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with methanol (200 mL) at 0 °C and the reaction mixture was heated at 75 °C for 2 h. The reaction mixture was concentrated under vacuum and the resulting residue was dissolved in DCM (500 mL). The organic layer was washed with water (2 × 200 mL), dried over anhydrous Na2SO4 and then concentrated under vacuum. The obtained crude product was purified by Isolera column chromatography (eluent: 5 - 20% MeOH in DCM; silica gel: 230 - 400 mesh) to give the title compound. Yield: 87% (53 g, brown liquid). LCMS: (Method K) 211.2 (M + +H), Rt. 1.97 min, 92.44% (max).

[0374] Intermediate 43 2,4-Dibromo-N-(1-((4-fluorophenyl)amino)hexan-2-yl)-5-methoxybenzenesulfonamide

[0375]

Chem.

[0376] To a stirred solution of N1-(4-fluorophenyl)hexane-1,2-diamine (Intermediate 42; 20 g, 95 mmol) in THF (200 mL) was added 2,4-dibromo-5-methoxybenzenesulfonyl chloride (41.6 g, 114 mmol) and triethylamine (39.8 mL, 285 mmol) at 0 °C, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (400 mL). The organic layer was washed with water (2 × 100 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude product was purified by Isolera column chromatography (eluent: 15% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 86% (45.5 g, brown solid). 1 1H NMR (400 MHz, DMSO-d6): δ 8.02 - 7.90 (m, 2H), 7.55 (s, 1H), 6.82 (m, 2H), 6.31 - 6.26 (m, 2H), 5.45 - 5.30 (m, 1H), 3.90 (s, 3H), 3.30 - 3.15 (m, 1H), 3.05 - 2.82 (m, 2H), 1.60 - 1.40 (m, 1H), 1.40 - 1.30 (m, 1H), 1.30 - 1.20 (m, 1H), 1.15 - 0.98 (m, 3H), 0.73 (t, J = 8.80 Hz, 3H). LCMS: (Method A) 538.9 (M + +H), Rt. 3.04 min, 85.57% (max).

[0377] Intermediate 44 7-Bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0378]

Chemical Structure

[0379] To a stirred solution of 2,4-dibromo-N-(1-((4-fluorophenyl)amino)hexan-2-yl)-5-methoxybenzenesulfonamide (Intermediate 43; 45 g, 84 mmol) in DMF (450 mL) were added potassium carbonate (23.11 g, 167 mmol) and copper(I) iodide (1.59 g, 8.36 mmol) at room temperature. The reaction mixture was degassed with nitrogen for 15 minutes. Then, tris[2-(2-methoxyethoxy)ethyl]amine (5.41 g, 16.72 mmol) was added at room temperature and the reaction mixture was heated at 130 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a pad of Celite and the Celite pad was washed with EtOAc (50 mL). The filtrate portion was concentrated under vacuum. The residue obtained was diluted with EtOAc (400 mL) and the organic layer was washed with water (2 × 100 mL). The organic portion was dried over anhydrous Na2SO4 and the crude obtained was purified by Isolera column chromatography (eluent: 12% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 55% (22.0 g, white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 7.57 (s, 1H), 7.50 - 7.42 (d, J = 6.6 Hz, 2H), 7.02 (t, J = 9.0 Hz, 2H), 6.70 - 6.50 (m, 2H), 4.38 - 4.22 (m, 1H), 3.95 (s, 3H), 3.33 (m, 1H), 2.95 - 2.80 (m, 1H), 1.65 - 1.15 (m, 6H), 0.89 (t, J = 9.2 Hz, 3H). LCMS: (Method A) 455.0 (M + -H), Rt. 2.71 min, 95.55% (max).

[0380] Intermediate 45 7-Bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-2-methyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0381]

Chem.

[0382] To a stirred solution of 7-bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 44; 5.8 g, 12.68 mmol) in N-methyl-2-pyrrolidone (24 mL) was added Cs2CO3 (8.26 g, 25.4 mmol) at 0 °C and the reaction mixture was stirred for 15 minutes. Then methyl iodide (1.58 mL, 25.4 mmol) was added dropwise at 0 °C and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with EtOAc (100 mL) and the organic layer was washed with water (50 mL). The organic portion was dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product obtained was purified by Isolera column chromatography (eluent: 15% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 75% (5.1 g, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.47 (s, 1H), 7.41 (s, 1H), 6.98 - 6.94 (m, 2H), 6.72 - 6.69 (m, 2H), 3.99 - 3.92 (m, 5H), 3.40 - 3.22 (m, 1H), 2.63 (s, 3H), 1.68 - 1.60 (m, 1H), 1.50 - 1.35 (m, 5H), 0.95 (t, J = 7.2 Hz, 3H). LCMS: (Method A) 471.0 (M + +H), Rt. 2.89 min, 88.26% (max).

[0383] Intermediate 46 3-Butyl-5-(4-fluorophenyl)-8-hydroxy-2-methyl-7-(methylthio)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0384]

Chemical Structure

[0385] To a stirred solution of 7-bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-2-methyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 45; 2.0 g, 4.24 mmol) in DMF (20 mL) was added sodium thiomethoxide (1.48 g, 21.21 mmol) at room temperature, and the reaction mixture was stirred at 100 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and quenched with water (20 mL). The aqueous layer was extracted with EtOAc (2 × 30 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 15 - 20% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 81% (1.6 g, pale yellow solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.72 (s, 1H), 7.27 (s, 1H), 6.97 (t, J = 11.2 Hz, 3H), 6.55 - 6.50 (m, 2H), 3.98 - 3.86 (m, 2H), 3.30 - 3.05 (m, 1H), 2.42 (s, 3H), 2.34 (s, 3H), 1.70 - 1.45 (m, 2H), 1.40 - 1.25 (m, 4H), 1.00 - 0.80 (m, 3H). LCMS: (Method A) 425.2 (M + +H), Rt. 2.56 min, 90.64% (max).

[0386] Intermediate 47 tert-Butyl (E)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

[0387]

Chemical Structure

[0388] To a stirred solution of 3-butyl-5-(4-fluorophenyl)-8-hydroxy-2-methyl-7-(methylthio)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 46; 50 mg, 0.118 mmol) in dry THF (2 mL) were added tert-butyl propiolate (22 mg, 0.18 mmol) and DABCO (6.61 mg, 0.06 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum and the resulting residue was partitioned between water (5 mL) and EtOAc (5 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), the combined organic layers were washed with brine (5 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum and the resulting crude material was purified by Isolera column chromatography (eluent: 9% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 80% (55 mg, colorless gum). 1 H NMR (400 MHz, CDCl3): δ 7.62 - 7.59 (m, 2H), 7.01 - 6.97 (m, 2H), 6.88 (s, 1H), 6.81 - 6.78 (m, 2H), 5.55 (d, J = 12.0 Hz, 1H), 3.97 - 3.93 (m, 2H), 3.55 - 3.30 (m, 1H), 2.67 (s, 3H), 2.31 (s, 3H), 1.70 - 1.60 (m, 2H), 1.50 (s, 9H), 1.45 - 1.35 (m, 4H), 1.00 - 0.90 (m, 3H). LCMS: (Method A) 495.0 (M + - t Bu + H), Rt. 3.18 min, 94.92% (max).

[0389] Intermediate 48 3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-8-hydroxy-2-methyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0390]

Chem.

[0391] To a stirred solution of 7-bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-2-methyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 45; 1.0 g, 2.12 mmol) in DMF (5 mL) was added sodium ethanethiolate (0.89 g, 10.61 mmol) at room temperature, and the reaction mixture was stirred at 100 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature and quenched with water (10 mL). The aqueous layer was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 15% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 74% (0.8 g, pale yellow solid). 1 H NMR (400 MHz, DMSO-d6): δ 10.66 (s, 1H), 7.28 (s, 1H), 7.10 - 6.90 (m, 3H), 6.62 - 6.40 (m, 2H), 4.02 - 3.80 (m, 2H), 3.00 - 3.10 (m, 1H), 3.00 - 2.78 (m, 2H), 2.43 (s, 3H), 1.70 - 1.45 (m, 2H), 1.45 - 1.28 (m, 4H), 1.20 (t, J = 9.60 Hz, 3H), 0.98 - 0.82 (m, 3H). LCMS: (Method K) 439.1 (M + +H), Rt. 3.07 min, 85.59% (max).

[0392] Intermediate 49 tert-Butyl (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

[0393] [Chemical formula]

[0394] To a stirred solution of 3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-8-hydroxy-2-methyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 48; 60 mg, 0.137 mmol) in dry THF (1 mL) were added tert-butyl propiolate (26 mg, 0.20 mmol) and DABCO (7.67 mg, 0.06 mmol), and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum, and the resulting residue was partitioned between water (5 mL) and EtOAc (5 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), and the combined organic layers were washed with brine (5 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, 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: 74% (60 mg, colorless gum). 11H NMR (300 MHz, DMSO-d6): δ 7.74 (d, J = 12.0 Hz, 1H), 7.49 (s, 1H), 7.17 (s, 1H), 7.04 (t, J = 8.4 Hz, 2H), 6.75 - 6.73 (m, 2H), 5.48 (d, J = 12.3 Hz, 1H), 4.06 - 4.01 (m, 1H), 3.79 (m, 1H), 2.94 - 2.92 (m, 3H), 2.58 (s, 3H), 1.60 - 1.50 (m, 2H), 1.50 - 1.40 (m, 10H), 1.40 - 1.30 (m, 3H), 1.20 - 1.15 (m, 3H), 0.95 - 0.85 (m, 3H). LCMS: (Method A) 509.1 (M + - t Bu + H), Rt. 3.17 min, 94.65% (max).

[0395] Intermediate 50 3-Butyl-7-chloro-5-(4-fluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one

[0396]

Chem.

[0397] A stirred solution of 3-butyl-7-chloro-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (6 g, 20.01 mmol) in 1-bromo-4-fluorobenzene (88 g, 500 mmol) was added with copper(I) iodide (0.38 g, 2.00 mmol) and K2CO3 (5.53 g, 40.0 mmol), and the solution was purged with nitrogen for 20 minutes for degassing. Then, tris[2-(2-methoxyethoxy)ethyl]amine (1.30 g, 4.00 mmol) was added under a nitrogen atmosphere, and the resulting reaction mixture was heated at 135 °C for 40 hours. After completion of the reaction (monitored by UPLC), the reaction mixture was filtered through Celite, and the Celite pad was washed with EtOAc (100 mL). The filtrate was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 15 - 20% EtOAc / PE; silica gel: 230 - 400 mesh) to obtain the title compound. Yield: 64% (5.5 g, off-white solid). LCMS: (Method E) 394.0 (M + +H), Rt. 3.19 min, 91.57%.

[0398] Intermediate 51 3-Butyl-7-chloro-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine

[0399]

Chemical formula

[0400] A stirred solution of 3-butyl-7-chloro-5-(4-fluorophenyl)-8-methoxy-2,3-dihydro-1,5-benzothiazepin-4(5H)-one (Intermediate 50; 5.5 g, 13.96 mmol) in THF (60 mL) at 0 °C was added dropwise with borane dimethyl sulfide (140 mL, 140 mmol), and the reaction mixture was refluxed at 75 °C for 40 h. After completion of the reaction (monitored by UPLC), the reaction mixture was cooled to 0 °C and quenched with methanol (100 mL). The resulting solution was heated at 65 °C for 2 h, then cooled to room temperature and concentrated under vacuum. The obtained crude material was purified by Isolera column chromatography (eluent: 15 - 20% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 77% (4.5 g, colorless liquid). LCMS: (Method A) 380.0 (M + ) Rt. 3.61 min, 90.76% (max).

[0401] Intermediate 52 3-Butyl-7-chloro-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0402]

Chem.

[0403] To a stirred solution of 3-butyl-7-chloro-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine (Intermediate 51; 4.5 g, 11.84 mmol) in THF (45 mL) and water (10 mL) was added Oxone (36.4 g, 59.2 mmol), and the reaction mixture was stirred at room temperature for 24 h. 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 × 200 mL). The combined organic layers were washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The crude material was purified by Isolera column chromatography (eluent: 10 - 13% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 84% (4.1 g, colorless solid). LCMS: (Method A) 412.1 (M + +H), Rt. 2.83 min, 96.78% (max).

[0404] Intermediate 53 3-Butyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide

[0405]

Chem.

[0406] To a stirred solution of 3-butyl-7-chloro-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 52; 1 g, 2.42 mmol) in DMF (12 mL) was added sodium thiomethoxide (0.85 g, 12.14 mol) at room temperature and the resulting mixture was stirred at 100 °C overnight. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with water (20 mL) and the aqueous layer was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The obtained crude material was purified by Isolera column chromatography (eluent: 30 - 35% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 30% (0.300 g, off-white solid). 1 1H NMR (400 MHz, CDCl3): δ 7.66 (s, 1H), 7.27 (s, 1H), 6.94 - 6.94 (m, 2H), 6.62 - 6.64 (m, 2H), 4.14 - 4.16 (m, 1H), 3.41 - 3.42 (m, 1H), 2.96 - 2.96 (m, 1H), 2.50 (s, 1H), 2.40 (s, 3H), 1.37 - 1.37 (m, 4H), 1.29 - 1.30 (m, 4H), 0.95 (t, J = 7.20 Hz, 3H). LCMS: (Method A) 408.2 (M +_ + H), Rt. 2.54 min, 98.39% (max).

[0407] Intermediate 54 tert-Butyl (E)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

[0408]

Chemical Structure

[0409] To a stirred solution of 3-butyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 53; 50 mg, 0.12 mmol) in dry THF (3 mL) were added tert-butyl propiolate (23 mg, 0.18 mmol) and DABCO (6.85 mg, 0.06 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 concentrated and the resulting residue was partitioned between water (5 mL) and EtOAc (5 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), the combined organic layers were washed with brine (5 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum and the resulting crude material was purified by Isolera column chromatography (eluent: 12% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 95% (65 mg, off-white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 7.72 (d, J = 12.0 Hz, 1H), 7.56 (s, 1H), 7.17 (s, 1H), 7.04 (t, J = 9.2 Hz, 2H), 6.71 - 6.67 (m, 2H), 5.46 (d, J = 12.0 Hz, 1H), 4.22 - 4.19 (m, 1H), 3.51 - 3.48 (m, 1H), 3.33 - 3.24 (m, 1H), 3.18 - 3.13 (m, 1H), 2.40 (s, 3H), 2.30 - 2.20 (m, 1H), 1.46 - 1.43 (m, 9H), 1.39 - 1.34 (m, 3H), 1.31 - 1.28 (m, 3H), 0.9 (t, J = 7.2 Hz, 3H). LCMS: (Method A) 480.1 (M + - t + Bu+H), Rt. 3.03 min, 95.09% (max).

[0410] Intermediate 55 7-Bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-2-(4-methoxybenzyl)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0411]

Chem.

[0412] To a stirred solution of 7-bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 44; 5 g, 10.93 mmol) in N-methyl-2-pyrrolidone (50 mL) was added Cs2CO3 (7.12 g, 21.86 mmol), and the reaction mixture was stirred at 0 °C for 30 minutes. Then 1-(bromomethyl)-4-methoxybenzene (3.30 g, 16.40 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a pad of Celite, and the Celite pad was washed with EtOAc (2 × 20 mL). The filtrate was concentrated under vacuum, and the crude product was purified by Isolera column chromatography (eluent: 15 - 20% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 73% (6 g, off-white solid). LCMS: (Method A) 578.1 (M + +H), Rt. 3.49 min, 75.44%.

[0413] Intermediate 56 3-Butyl-5-(4-fluorophenyl)-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide

[0414]

Chem.

[0415] To a stirred solution of 7-bromo-3-butyl-5-(4-fluorophenyl)-8-methoxy-2-(4-methoxybenzyl)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 55; 1 g, 1.23 mmol) in DMF (20 mL) was added sodium thiomethoxide (0.43 g, 6.14 mmol) at room temperature and the resulting mixture was stirred at 100 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with water (50 mL) and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4 and concentrated in vacuo. The crude obtained was purified by Isolera column chromatography (eluent: 10 - 15% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 66% (500 g, off-white solid). LCMS: (Method B) 529.1 (M +_ H), Rt. 3.20 min, 85.70% (max).

[0416] Intermediate 57 tert-Butyl (E)-3-((3-butyl-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

[0417]

Chemical Structure

[0418] To a stirred solution of 3-butyl-5-(4-fluorophenyl)-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 56; 60 mg, 0.113 mmol) in dry THF (2 mL) was added tert-butyl propiolate (21.40 mg, 0.17 mmol) and DABCO (6.34 mg, 0.06 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum and the resulting residue was partitioned between water (5 mL) and EtOAc (5 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), the combined organic layers were washed with brine (5 mL) and then dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum and the resulting crude material was purified by Isolera column chromatography (eluent: 20% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 81% (60 mg, colorless gum). LCMS: (Method A) 601.1 (M + - t Bu + H), Rt. 3.44 min, 94.98% (max).

[0419] Intermediate 58 3-Butyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiadiazepine 1,1-dioxide

[0420]

Chem.

[0421] To a solution of 3-butyl-7-chloro-8-methoxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (1 g, 2.54 mmol) in DMF (25 mL) was added sodium ethanethiolate (1.07 g, 12.69 mmol), and the reaction mixture was stirred at 120 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with ice-cold water (25 mL), and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with ice-cold water (50 mL) and brine (50 mL), and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 30% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 52% (0.56 g, off-white solid). LCMS: (Method A) 406.1 (M + +H), Rt. 3.11 min, 96.20% (max).

[0422] Intermediate 59 tert-Butyl (E)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate

[0423] [Chemical formula]

[0424] To a stirred solution of 3-butyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 58; 100 mg, 0.24 mmol) in dry THF (4 mL) were added tert-butyl propiolate (46.7 mg, 0.37 mmol) and DABCO (13.83 mg, 0.12 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum and the resulting residue was partitioned between water (5 mL) and EtOAc (5 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), the combined organic layers were washed with brine (5 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum and the resulting crude material was purified by Isolera column chromatography (eluent: 15 - 20% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 55% (72 mg, off-white solid). 1 H NMR (300 MHz, DMSO-d6): δ 7.72 (d, J = 12.30 Hz, 1H), 7.58 (s, 1H), 7.16 - 7.19 (m, 3H), 6.65 - 6.68 (m, 3H), 5.46 (d, J = 12.30 Hz, 1H), 4.25 (m, 1H), 3.47 (s, 1H), 3.15 (s, 1H), 2.96 (d, J = 7.50 Hz, 2H), 2.28 (m, 2H), 1.45 (s, 9H), 1.17 - 1.19 (m, 6H), 1.22 - 1.38 (m, 3H), 0.90 (t, J = 6.90 Hz, 3H). LCMS: (Method A) 476.1 (M + - t Bu), Rt. 3.18 min, 98.54% (max).

[0425] Intermediate 60 tert-butyl (E)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

[0426]

Chem.

[0427] To a stirred solution of 3-butyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (50 mg, 0.12 mmol) in THF (5 mL) was added tert-butyl propiolate (16 mg, 0.12 mmol) and DABCO (14 mg, 0.12 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 concentrated under vacuum and the resulting residue was partitioned between water (5 mL) and EtOAc (5 mL). The aqueous layer was extracted with EtOAc (2 × 5 mL), the combined organic layers were washed with brine (5 mL) and dried over anhydrous Na2SO4. The organic portion was concentrated under vacuum 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: 30% (20 mg, off-white solid). LCMS: (Method E) 477.1 (M + - t - Bu + H), Rt. 3.23 min, 99.30% (max).

[0428] Intermediate 61 tert-butyl (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiadiazepin-8-yl)oxy)acrylate

[0429]

Chem.

[0430] The title compound was prepared from 3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-8-hydroxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and tert-butyl propiolate according to the same procedure as described for Intermediate 60.

[0431] Intermediate 62 Ethyl (Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate

[0432]

Chem.

[0433] The title compound is prepared from 3-butyl-5-(4-fluorophenyl)-8-hydroxy-7-(methylthio)-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 53) and ethyl 3-bromo-2,2-difluoropropanoate according to the same procedure as described for Intermediate 10.

[0434] Intermediate 63 Ethyl (Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate

[0435]

Chem.

[0436] The title compound is prepared from 3-butyl-7-(ethylthio)-8-hydroxy-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide (Intermediate 58) and ethyl 3-bromo-2,2-difluoropropanoate according to the same procedure as described for Intermediate 10.

[0437] Intermediate 64 Ethyl (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate

[0438]

Chemical formula

[0439] The title compound is prepared from 3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-8-hydroxy-2,3,4,5-tetrahydro-1,5-benzothiazepine 1,1-dioxide and ethyl 3-bromo-2,2-difluoropropanoate by the same procedure as described for Intermediate 10.

[0440] Intermediate 65 Ethyl (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0441]

Chemical formula

[0442] The title compound is prepared from 3-butyl-5-(4-fluorophenyl)-8-hydroxy-2-methyl-7-(methylthio)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 46) and ethyl 3-bromo-2,2-difluoropropanoate by the same procedure as described for Intermediate 10.

[0443] Intermediate 66 Ethyl (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0444]

Chem.

[0445] The title compound is prepared from 3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-8-hydroxy-2-methyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 48) and ethyl 3-bromo-2,2-difluoropropanoate by the same procedure as described for Intermediate 10.

[0446] Intermediate 67 Ethyl (Z)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0447]

Chem.

[0448] The title compound is prepared from 3-butyl-7-(ethylthio)-8-hydroxy-2-methyl-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 38) and ethyl 3-bromo-2,2-difluoropropanoate by the same procedure as described for Intermediate 10.

[0449] Intermediate 68 Ethyl (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0450]

Chem.

[0451] The title compound is prepared from 3-butyl-5-(4-fluorophenyl)-8-hydroxy-2-(4-methoxybenzyl)-7-(methylthio)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide (Intermediate 56) and ethyl 3-bromo-2,2-difluoropropanoate by a procedure similar to that described for Intermediate 10.

[0452] Intermediate 69 (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0453]

Chem.

[0454] The title compound is prepared from ethyl (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 68) according to the procedure as described for Intermediate 31.

[0455] Intermediate 70 Ethyl (Z)-3-((3-butyl-7-(ethylthio)-2-(4-methoxybenzyl)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0456]

Chem.

[0457] The title compound is prepared from 3-butyl-7-(ethylthio)-8-hydroxy-2-(4-methoxybenzyl)-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide and ethyl 3-bromo-2,2-difluoropropanoate by the same procedure as described for Intermediate 10.

[0458] Intermediate 71 (Z)-3-((3-butyl-7-(ethylthio)-2-(4-methoxybenzyl)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0459]

Chem.

[0460] The title compound is prepared from ethyl (Z)-3-((3-butyl-7-(ethylthio)-2-(4-methoxybenzyl)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 70) according to the procedure as described for Intermediate 31.

[0461] Intermediate 72 Ethyl (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate

[0462] [Chemical formula]

[0463] The title compound is prepared from 3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-8-hydroxy-2-(4-methoxybenzyl)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide and ethyl 3-bromo-2,2-difluoropropanoate by the same procedure as described for Intermediate 10.

[0464] Intermediate 73 (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0465] [Chemical formula]

[0466] The title compound is prepared from ethyl (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 72) according to the procedure as described for Intermediate 31.

[0467] Intermediate 74 tert-Butyl (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate

[0468]

Chem.

[0469] The title compound is prepared from 3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-8-hydroxy-2-(4-methoxybenzyl)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine 1,1-dioxide and tert-butyl propiolate by the same procedure as described for Intermediate 57.

[0470] (Example 1) (E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiadiazepin-8-yl)oxy)acrylate

[0471]

Chem.

[0472] A stirred solution of ethyl (E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (Intermediate 9; 0.23 g, 0.45 mmol) in a mixture of 1,4-dioxane and water (4 mL; 3:1) was treated with lithium hydroxide (97 mg, 2.29 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 under vacuum and the residue obtained was acidified with dilute HCl (1.5 N, 2 mL). The aqueous portion was extracted with EtOAc (2 × 15 mL) and the combined organic layers were washed with water (15 mL) and brine (15 mL). The organic portion was dried over anhydrous Na2SO4 and concentrated under vacuum. The crude material obtained was purified by Isolera column chromatography (eluent: 2 - 3% MeOH / DCM; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 59% (125 mg, off-white solid). 1 1H-NMR (400 MHz, DMSO-d6): δ 12.29 (s, 1H), 7.76 (d, J = 12.0 Hz, 1H), 7.56 (s, 1H), 7.18 (s, 1H), 7.18 (t, J = 7.2 Hz, 2H), 6.73 (t, J = 7.2 Hz, 1H), 6.65 (d, J = 8.0 Hz, 2H), 5.52 (d, J = 12.4 Hz, 1H), 4.26-4.22 (m, 1H), 3.49-3.46 (m, 1H), 3.32-3.27 (m, 1H), 3.15-3.08 (m, 1H), 2.41 (s, 3H), 2.27-2.26 (m, 1H), 1.40-1.23 (m, 6H), 0.98-0.88 (m, 3H). LCMS: (Method A) 462.2 (M + +H), Rt. 2.56 min, 94.94% (max). HPLC: (Method B) Rt. 5.42 min, 95.08% (max).

[0473] (Examples 2 and 3) (R)-(E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid and (S)-(E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

[0474] [Chemical formula]

[0475] The two enantiomers of racemic (E)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid (Example 1; 0.12 g, 0.26 mmol) were separated by chiral preparative SFC (Method A); mobile phase: CO2: 0.5% isopropylamine in IPA (70:30); wavelength: 280 nm; cycle time: 5 minutes; back pressure: 100 bar. The substance was concentrated at 40 °C under vacuum. The first eluted fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configurations of the two enantiomers are unknown. Enantiomer 1: Yield: 10% (15 mg, off-white solid). 11H NMR (400 MHz, DMSO-d6): δ 12.36 (s, 1H), 7.74 (d, J = 12.0 Hz, 1H), 7.56 (s, 1H), 7.22 (s, 1H), 7.18 (t, J = 7.2 Hz, 2H), 6.73 (t, J = 7.2 Hz, 1H), 6.65 (d, J = 8.4 Hz, 2H), 5.53 (d, J = 12.4 Hz, 1H), 4.27-4.23 (m, 1H), 3.49-3.46 (m, 1H), 3.33-3.31 (m, 1H), 3.28-3.13 (m, 1H), 2.41 (s, 3H), 2.31-2.27 (m, 1H), 1.41-1.29 (m, 6H), 0.9 (t, J = 7.2 Hz, 3H). LCMS: (Method E) 462.1 (M + +H), Rt. 2.45 min, 97.2% (max). HPLC: (Method B) Rt. 5.42 min, 97.78% (max). SFC: (Method A) Rt. 2.79 min, 98.25%. Enantiomer 2: Yield: 12% (23 mg, off-white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 12.29 (s, 1H), 7.75 (d, J = 12.0 Hz, 1H), 7.57 (s, 1H), 7.22 (s, 1H), 7.18 (t, J = 7.2 Hz, 2H), 6.73 (t, J = 7.2 Hz, 1H), 6.65 (d, J = 8.4 Hz, 2H), 5.53 (d, J = 12.0 Hz, 1H), 4.27-4.21 (m, 1H), 3.49-3.33 (m, 1H), 3.33-3.31 (m, 1H), 3.28-3.13 (m, 1H), 2.41 (s, 3H), 2.33-2.28 (m, 1H), 1.42-1.31 (m, 6H), 0.97-0.95 (m, 3H). LCMS: (Method E) 462.1 (M ++H), Rt. 2.45 min, 96.57% (max). HPLC: (Method B) Rt. 5.42 min, 97.12% (max). SFC: (Method A) Rt. 4.51 min, 99.75 %.

[0476] (Example 4) (Z)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0477] [Chemical formula]

[0478] To a stirred solution of ethyl (Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 10; 170 mg, 0.33 mmol) in a mixture of 1,4-dioxane and water (5 mL, 4:1) was added lithium hydroxide (28 mg, 0.66 mmol), and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH about 4) and then diluted with water (5 mL). The aqueous layer was extracted with EtOAc (2 × 20 mL), and the combined organic layers were washed with water (10 mL) and brine (10 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The resulting crude material was purified by preparative HPLC (Method D) to give the title compound. Yield: 43% (70 mg, white solid). 11H NMR (400 MHz, DMSO-d6): δ 7.58 (s, 1H), 7.50 (d, J = 18.8 Hz, 1H), 7.19 - 7.16 (m, 3H), 6.72 (t, J = 7.2 Hz, 1H), 6.63 (d, J = 8.0 Hz, 2H), 4.24 (d, J = 14.8 Hz, 1H), 3.51 - 3.46 (m, 1H), 3.28 - 3.24 (m, 1H), 3.12 - 3.09 (m, 1H), 2.42 (s, 3H), 2.33 - 2.20 (m, 1H), 1.41 - 1.24 (m, 6H), 0.91 (t, J = 7.20 Hz, 3H). LCMS: (Method E) 480.1 (M + +H), Rt. 2.51 min, 96.23% (max). HPLC: (Method B) Rt. 5.54 min, 94.04% (max)

[0479] (Examples 5 and 6) (S)-(Z)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0480] [Chemical formula]

[0481] A stirred solution of enantiomer 1 of ethyl (Z)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 11; 650 mg, 1.28 mmol) in a mixture of 1,4-dioxane and water (10 mL, 4:1) was treated with lithium hydroxide (107 mg, 2.56 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH ca. 4) and then diluted with water (5 mL). The aqueous layer was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with water (10 mL) and brine (10 mL), dried over anhydrous Na2SO4, 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 afford the title compound. Starting from 600 mg of enantiomer 2 of intermediate 11 and following the same procedure, enantiomer 2 of the title compound was obtained. The absolute configuration of the two enantiomers is unknown. Enantiomer 1: Yield: 36% (224 mg, light brown solid). 1 H NMR (400 MHz, DMSO-d6): δ 13.60 (s, 1H), 7.69 (s, 1H), 7.64 (d, J = 3.6 Hz, 1H), 7.20-7.16 (m, 3H), 6.73 (t, J = 7.6 Hz, 1H), 6.64 (d, J = 8.0 Hz, 2H), 4.25 (d, J = 14.4 Hz, 1H), 3.51-3.47 (m, 1H), 3.30-3.26 (m, 1H), 3.13-3.07 (m, 1H), 2.43 (s, 3H), 2.34-2.21 (m, 1H), 1.44-1.28 (m, 6H), 0.91 (t, J = 7.20 Hz, 3H). LCMS: (method E) 480.0 (M ++(H), Retention time: 2.93 minutes, 99.97% (maximum). HPLC: (Method B) Retention time: 5.54 minutes, 97.21% (maximum). Chiral HPLC: (Method E) Retention time: 4.83 minutes, 100% (maximum) Enantiomer 2: Yield: 51% (290 mg, light brown solid). 1 1H NMR (400 MHz, DMSO-d6): δ 13.76 (s, 1H), 7.62 (t, J = 8.4 Hz, 2H), 7.20 - 7.16 (m, 3H), 6.73 (t, J = 7.2 Hz, 1H), 6.64 (d, J = 8.0 Hz, 2H), 4.25 (d, J = 15.2 Hz, 1H), 3.51 - 3.46 (m, 1H), 3.33 - 3.27 (m, 1H), 3.13 - 3.07 (m, 1H), 2.43 (s, 3H), 2.34 - 2.29 (m, 1H), 1.44 - 1.28 (m, 6H), 0.91 (t, J = 7.20 Hz, 3H). LCMS: (Method E) 480.0 (M + +(H), Retention time: 2.93 minutes, 96.76% (maximum). HPLC: (Method B) Retention time: 5.54 minutes, 98.69% (maximum). Chiral HPLC: (Method E) Retention time: 4.02 minutes, 100% (maximum)

[0482] (Example 7) (Z)-3-((3-Ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0483]

Chemical Structure

[0484] A stirred solution of ethyl-(Z)-3-((3-ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 19; 0.13 g, 0.27 mmol) in a mixture of 1,4-dioxane and water (4:1, 5 mL) was treated with lithium hydroxide (0.02 g, 0.54 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH ca. 4) and diluted with ice-cold water (25 mL). The aqueous layer was extracted with EtOAc (2 × 20 mL), and the combined organic layers were washed with water (15 mL) and brine (15 mL). The organic portion was dried over anhydrous Na2SO4 and concentrated in vacuo. The crude product thus obtained was purified by preparative HPLC (Method B) to give the title compound. Yield: 12% (15 mg, off-white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 7.46 (s, 1H), 7.19-7.15 (m, 3H), 7.10-7.06 (m, 1H), 6.71 (t, J = 7.2 Hz, 1H), 6.62 (d, J = 8.4 Hz, 2H), 4.26-4.23 (m, 1H), 3.51-3.41 (m, 2H), 3.11-3.09 (m, 1H), 2.41 (s, 3H), 2.33-2.23 (m, 1H), 1.48-1.36 (m, 2H), 1.01-0.99 (m, 3H). LCMS: (Method D) 452.0 (M + + H), Rt. 2.11 min, 99.01% (max). HPLC: (Method C) Rt. 4.50 min, 97.92% (max).

[0485] (Example 8) (E)-3-((3-Ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

[0486] [Chemical Structure]

[0487] To a stirred solution of tert-butyl (E)-3-((3-ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (Intermediate 20; 0.36 g, 0.73 mmol) in DCM (10 mL) at 0 °C was added TFA (2 mL), and then the reaction mixture was stirred at room temperature for 2 h. 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 × 20 mL). The combined organic layers were washed with water (10 mL) and brine (10 mL), and dried over anhydrous Na2SO4. The organic layer was concentrated under vacuum, and the resulting crude product was purified by preparative HPLC (Method B) to give the title compound. Yield: 23% (75 mg, white solid). 1 H NMR (400 MHz, DMSO-d6):): δ 12.30 (s, 1H), 7.77 (d, J = 12.4 Hz, 1H), 7.57 (s, 1H), 7.23-7.16 (m, 3H), 6.73 (t, J = 7.2 Hz, 1H), 6.66 (d, J = 8.0 Hz, 2H), 5.53 (d, J = 12.4 Hz, 1H), 4.28-4.24 (m, 1H), 3.51-3.48 (m, 1H), 3.39-3.33 (m, 1H), 3.15-3.09 (m, 1H), 2.42 (s, 3H), 2.28-2.22 (m, 1H), 1.48-1.37 (m, 2H), 1.01-0.97 (m, 3H). LCMS: (Method D) 434.1 (M + +H), Rt. 2.51 min, 99.14% (max). HPLC: (Method B) Rt. 4.47 min, 99.54 % (max).

[0488] (Example 9) (E)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0489]

Chem.

[0490] To a stirred solution of tert-butyl (E)-3-((3-butyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (Intermediate 29; 0.2 g, 0.31 mmol) in toluene (5 mL) at 0 °C were added triphenylamine (0.15 g, 0.62 mmol) and TFA (0.5 mL, 6.2 mmol). The reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated under vacuum and the resulting crude material was purified by Isolera column chromatography (eluent: 30 - 50% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 14% (20 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.26 (s, 1H), 7.74 (d, J = 12.40 Hz, 1H), 7.52 (s, 1H), 7.39 (d, J = 9.2 Hz, 1H), 7.20 - 7.16 (m, 3H), 6.75 (t, J = 7.20 Hz, 1H), 6.67 (d, J = 8.40 Hz, 2H), 5.46 (d, J = 12.40 Hz, 1H), 4.39 - 4.35 (m, 1H), 3.51 - 3.49 (m, 1H), 2.95 - 2.91 (m, 1H), 2.40 (s, 3H), 1.61 - 1.58 (m, 2H), 1.48 - 1.39 (m, 2H), 1.36 - 1.29 (m, 2H), 0.95 - 0.92 (m, 3H). LCMS: (Method E) 462.9 (M ++H), Rt. 2.76 min, 94.28% (max). HPLC: (Method B) Rt. 5.24 min, 94.22% (max)

[0491] (Example 10) (Z)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0492] [Chemical formula]

[0493] To a stirred solution of (Z)-3-((3-butyl-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Intermediate 31; 0.22 g, 0.37 mmol) in toluene (4 mL) were added triphenylamine (0.18 g, 0.73 mmol) and TFA (0.6 mL, 7.3 mmol) at 0 °C, and the resulting mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum, and the obtained crude material was purified by Isolera column chromatography (eluent: 30 - 50% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. The obtained compound was further purified by preparative HPLC (Method A) to give the title compound. Yield: 14% (25 mg, off-white solid). 11H NMR (400 MHz, DMSO-d6): δ 7.65 (s, 1H), 7.60 (d, J = 3.6 Hz, 1H), 7.39 (d, J = 9.2 Hz, 1H), 7.20 - 7.16 (m, 3H), 6.74 (t, J = 7.2 Hz, 1H), 6.66 (d, J = 8.0 Hz, 2H), 4.38 - 4.35 (m, 1H), 3.45 - 3.33 (m, 1H), 2.95 - 2.92 (m, 1H), 2.42 (s, 3H), 1.61 - 1.58 (m, 2H), 1.48 - 1.42 (m, 2H), 1.41 - 1.31 (m, 2H), 0.95 - 0.92 (m, 3H). LCMS: (Method E) 481.1 (M + +H), Rt. 2.81 min, 99.52% (max). HPLC: (Method B) Rt. 5.36 min, 98.40% (max)

[0494] (Example 11) (Z)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0495] [Chemical Structure]

[0496] A stirred solution of ethyl (Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 34; 0.6 g, 1.15 mmol) in 1,4-dioxane / water (4:1, 10 mL) was treated with lithium hydroxide (0.141 g, 3.44 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was acidified with dilute HCl (1.5 N, pH ~4) and diluted with ice-cold water (10 mL). The aqueous layer was extracted with EtOAc (2 × 15 mL), and then the combined organic layers were washed with water (10 mL) and brine solution (10 mL). The organic portion was dried over anhydrous Na2SO4 and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 1-5% MeOH / DCM; silica gel: 230-400 mesh). Yield: 27% (0.15 g, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 7.73 (d, J = 18.4 Hz, 1H), 7.56 (s, 1H), 7.18 (t, J = 8.4 Hz, 3H), 6.75 (t, J = 7.2 Hz, 1H), 6.67 (d, J = 8.4 Hz, 2H), 4.08-4.03 (m, 1H), 3.88-3.86 (m, 1H), 3.34-3.31 (m, 1H), 2.51 (s, 3H), 2.46 (m, 3H), 1.64-1.52 (m, 2H), 1.51-1.31 (m, 4H), 0.94-0.92 (m, 3H). LCMS: (Method E) 495.1 (M + +H), Rt. 2.88 min, 98.16% (max). HPLC: (Method B) Rt. 6.85 min, 96.53% (max).

[0497] (Examples 12 and 13) (S)-(Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0498]

Chem.

[0499] To a stirred solution of enantiomer 1 of ethyl (Z)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (intermediate 36; 1.95 g, 3.73 mmol) in a mixture of 1,4-dioxane and water (3:1, 20 mL) was added LiOH·H2O (313 mg, 7.47 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 1 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with dilute HCl (1.5 N, 20 mL), diluted with water (30 mL), and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with water (20 mL) and brine (20 mL), and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 30 - 50% EtOAc / PE; silica gel: 230 - 400 mesh) to afford enantiomer 1 of the title compound. Starting from 1.5 g of enantiomer 2 of intermediate 36 and following the same procedure, enantiomer 2 of the title compound was obtained. The crude material was purified by trituration with n-hexane (2 × 10 mL) to afford the title compound. The absolute configuration of the two enantiomers is unknown. Enantiomer 1: Yield: 93% (1.72 g, white solid). 11H NMR (400 MHz, DMSO-d6): δ 13.58 (broad singlet, 1H), 7.76 (doublet, J = 18.8 Hz, 1H), 7.56 (singlet, 1H), 7.20 - 7.16 (multiplet, 3H), 6.74 (triplet, J = 7.2 Hz, 1H), 6.65 (doublet, J = 8.4 Hz, 2H), 4.08 - 4.04 (multiplet, 1H), 3.87 - 3.86 (multiplet, 1H), 3.25 (broad singlet, 1H), 2.52 (singlet, 3H), 2.42 (singlet, 3H), 1.64 - 1.61 (multiplet, 1H), 1.53 - 1.50 (multiplet, 1H), 1.37 - 1.30 (multiplet, 4H), 0.92 (triplet, J = 6.80 Hz, 3H). LCMS: (Method E) 495.0 (M + +H), retention time 2.94 minutes, 99.38% (maximum). HPLC: (Method B) retention time 5.53 minutes, 97.59% (maximum). Chiral SFC: (Method H) retention time 1.83 minutes, 99.93% (maximum). Enantiomer 2: Yield: 60% (850 mg, white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 13.63 (broad singlet, 1H), 7.77 (doublet, J = 18.8 Hz, 1H), 7.57 (singlet, 1H), 7.20 - 7.16 (multiplet, 3H), 6.75 (triplet, J = 7.2 Hz, 1H), 6.66 (doublet, J = 8.0 Hz, 2H), 4.06 - 4.03 (multiplet, 1H), 3.88 - 3.86 (multiplet, 1H), 3.24 (broad singlet, 1H), 2.53 (singlet, 3H), 2.46 (singlet, 3H), 1.66 - 1.61 (multiplet, 1H), 1.53 - 1.51 (multiplet, 1H), 1.39 - 1.31 (multiplet, 4H), 0.92 (triplet, J = 6.80 Hz, 3H). LCMS: (Method E) 495.0 (M + +H), retention time 2.94 minutes, 99.54% (maximum). HPLC: (Method B) retention time 5.55 minutes, 99.08% (maximum). Chiral SFC: (Method H) retention time 2.76 minutes, 100% (maximum).

[0500] (Example 14) (E)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0501] [Chemical formula]

[0502] To a stirred solution of tert-butyl (E)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (Intermediate 37; 0.6 g, 1.13 mmol) in DCM (6 mL) at 0 °C was added TFA (1.0 mL), and the resulting mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum, and the resulting crude material was purified by Isolera column chromatography (eluent: 2 - 3% MeOH / DCM; silica gel: 230 - 400 mesh). The obtained residue was purified by preparative HPLC (Method A) to give the title compound. Yield: 28% (150 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.30 (s, 1H), 7.81 (d, J = 12.4 Hz, 1H), 7.50 (s, 1H), 7.20 - 7.17 (m, 3H), 6.75 (t, J = 7.6 Hz, 1H), 6.68 (d, J = 8.0 Hz, 2H), 5.54 (d, J = 12.0 Hz, 1H), 4.09 - 4.05 (m, 1H), 3.87 - 3.85 (m, 1H), 3.34 - 3.31 (m, 1H), 2.51 (s, 3H), 2.40 (s, 3H), 1.66 - 1.53 (m, 2H), 1.52 - 1.30 (m, 4H), 0.92 (t, J = 6.80 Hz, 3H). LCMS: (Method E) 476.9 (M ++H), Rt. 2.87 min, 98.40% (max). HPLC: (Method B) Rt. 5.41 min, 99.27% (max).

[0503] (Examples 15 and 16) (S)-(E)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid and (R)-(E)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0504] [Chemical formula]

[0505] The two enantiomers of racemic (E)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid (Example 14; 120 mg, 0.25 mmol) were separated by chiral preparative SFC (Method B). The substance was concentrated at 40 °C under vacuum. The first elution fraction corresponded to enantiomer 1 and the second elution fraction corresponded to enantiomer 2. The absolute configurations of the two enantiomers are unknown. Enantiomer 1: Yield: 14% (20 mg, off-white solid). 11H NMR (400 MHz, DMSO-d6): δ 7.76 (d, J = 12.0 Hz, 1H), 7.48 (s, 1H), 7.21 - 7.17 (m, 3H), 6.75 (t, J = 7.2 Hz, 1H), 6.68 (d, J = 8.4 Hz, 2H), 5.55 (d, J = 12.0 Hz, 1H), 4.08 - 4.04 (m, 1H), 3.86 - 3.85 (m, 1H), 3.34 - 3.31 (m, 1H), 2.51 (s, 3H), 2.40 (s, 3H), 1.64 - 1.53 (m, 2H), 1.52 - 1.30 (m, 4H), 0.95 - 0.92 (m, 3H). LCMS: (Method E) 477.1 (M + +H), Rt. 2.85 min, 99.84% (max). HPLC: (Method B) Rt. 5.48 min, 98.34% (max). SFC: (Method H) Rt. 1.71 min, 99.79%. Enantiomer 2: Yield: 14% (20 mg, off - white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 7.75 (d, J = 12.0 Hz, 1H), 7.48 (s, 1H), 7.21 - 7.17 (m, 3H), 6.75 (t, J = 7.2 Hz, 1H), 6.68 (d, J = 8.0 Hz, 2H), 5.55 (d, J = 12.0 Hz, 1H), 4.08 - 4.05 (m, 1H), 3.87 - 3.86 (m, 1H), 3.34 - 3.31 (m, 1H), 2.51 (s, 3H), 2.41 (s, 3H), 1.64 - 1.53 (m, 2H), 1.52 - 1.30 (m, 4H), 0.95 - 0.92 (m, 3H). LCMS: (Method E) 477.1 (M + +H), Rt. 2.85 min, 99.88% (max). HPLC: (Method B) Rt. 5.48 min, 99.0% (max). SFC: (Method H) Rt. 3.47 min, 99.41%.

[0506] (Example 17) (E)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0507]

Chem.

[0508] To a stirred solution of tert-butyl (E)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (Intermediate 39; 0.14 g, 0.25 mmol) in DCM (5 mL) at 0 °C was added TFA (0.39 mL, 5.12 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 quenched with cold water (5 mL), and the aqueous layer was extracted with DCM (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL), and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated under vacuum. The resulting crude material was purified by preparative HPLC (Method A). The obtained fractions were concentrated under vacuum, dissolved in ethyl acetate (5 mL), and the organic portion was washed with water (2 × 5 mL) and brine (5 mL). The organic portion was dried over anhydrous Na2SO4 and concentrated under vacuum to afford the title compound. Yield: 45% (60 mg, white solid). 11H NMR (400 MHz, DMSO-d6): δ 12.27 (s, 1H), 7.80 (d, J = 12.4 Hz, 1H), 7.50 (s, 1H), 7.23 - 7.18 (m, 3H), 6.77 (t, J = 6.8 Hz, 1H), 6.69 (d, J = 8.0 Hz, 2H), 5.53 (d, J = 12.0 Hz, 1H), 4.09 - 4.05 (m, 1H), 3.90 - 3.80 (m, 1H), 2.98 - 2.93 (m, 2H), 2.51 (s, 3H), 2.50 - 2.90 (m, 1H), 1.70 - 1.50 (m, 2H), 1.40 - 1.25 (m, 4H), 1.20 (t, J = 7.20 Hz, 3H), 0.95 - 0.85 (m, 3H). LCMS: (Method E) 490.9 (M + +H), Rt. 3.02 min, 97.10% (max). HPLC: (Method E) Rt. 5.66 min, 95.37% (max).

[0509] (Examples 18 and 19) (S)-(E)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid and (R)-(E)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0510] [Chemical Structure]

[0511] The two enantiomers of racemic (E)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid (Example 17; 50 mg, 0.102 mmol) were separated by chiral SFC (Method S). The substance was concentrated under vacuum at 40 °C. The first eluted fraction corresponded to enantiomer 1 and the second eluted fraction corresponded to enantiomer 2. The absolute configurations of the two enantiomers are unknown. Enantiomer 1: Yield: 40% (20 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.26 (s, 1H), 7.80 (d, J = 12.0 Hz, 1H), 7.50 (s, 1H), 7.23 - 7.17 (m, 3H), 6.76 (t, J = 7.2 Hz, 1H), 6.69 (d, J = 8.4 Hz, 2H), 5.52 (d, J = 12.4 Hz, 1H), 4.09 - 4.05 (m, 1H), 3.90 - 3.75 (m, 1H), 2.98 - 2.92 (m, 2H), 2.51 (s, 3H), 2.50 - 2.40 (m, 1H), 1.75 - 1.50 (m, 2H), 1.40 - 1.32 (m, 4H), 1.20 (t, J = 7.60 Hz, 3H), 0.9 (t, J = 6.8 Hz, 3H). LCMS: (Method A) 491.0 (M + +H), Rt. 2.71 min, 99.40% (max). HPLC: (Method E) Rt. 5.61 min, 99.72% (max). Chiral SFC: (Method S) Rt. 2.99 min, 98.37% (max). Enantiomer 2: Yield: 40% (30 mg, white solid). 11H NMR (400 MHz, DMSO-d6): δ 12.29 (s, 1H), 7.81 (d, J = 12.0 Hz, 1H), 7.50 (s, 1H), 7.24 - 7.17 (m, 3H), 6.76 (t, J = 7.2 Hz, 1H), 6.69 (d, J = 8.0 Hz, 2H), 5.53 (d, J = 12.0 Hz, 1H), 4.07 (m, 1H), 3.92 - 3.80 (m, 1H), 3.42 - 3.38 (m, 1H), 2.98 - 2.93 (m, 2H), 2.51 (s, 3H), 1.75 - 1.48 (m, 2H), 1.48 - 1.30 (m, 4H), 1.20 (t, J = 7.20 Hz, 3H), 0.9 (t, J = 6.8 Hz, 3H). LCMS: (Method A) 491.2 (M + +H), Rt. 2.71 min, 98.99% (max). HPLC: (Method E) Rt. 5.61 min, 99.18% (max). Chiral SFC: (Method S) Rt. 3.73 min, 98.28% (max).

[0512] (Example 20) (E)-3-((3-Butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0513] [Chemical Structure]

[0514] A stirred solution of tert-butyl (E)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (Intermediate 47; 40 mg, 0.07 mmol) in DCM (2 mL) at 0 °C was treated with TFA (0.16 mL, 2.18 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with cold water (5 mL), and the aqueous layer was extracted with DCM (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL), and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 35 - 40% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 91% (32 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.30 (s, 1H), 7.80 (d, J = 12.4 Hz, 1H), 7.48 (s, 1H), 7.15 (s, 1H), 7.03 (t, J = 8.8 Hz, 2H), 6.72 - 6.69 (m, 2H), 5.53 (d, J = 12.4 Hz, 1H), 4.05 - 4.01 (m, 1H), 3.83 - 3.82 (m, 1H), 3.30 - 3.25 (m, 1H), 2.51 (s, 3H), 2.40 (s, 3H), 1.70 - 1.45 (m, 2H), 1.45 - 1.20 (m, 4H), 0.91 (t, J = 6.80 Hz, 3H). LCMS: (Method A) 495.1 (M + +H), Rt. 2.50 min, 93.89% (max). HPLC: (Method E) Rt. 5.56 min, 94.80% (max).

[0515] (Examples 21 and 22) (S)-(E)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid and (R)-(E)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0516]

Chem.

[0517] The two enantiomers of racemic (E)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid (Example 20; 35 mg, 0.07 mmol) were separated by chiral SFC (Method S). The substance was concentrated at 40 °C under vacuum. The first elution fraction corresponded to enantiomer 1 and the second elution fraction corresponded to enantiomer 2. The absolute configurations of the two enantiomers are unknown. Enantiomer 1: Yield: 29% (10 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.27 (s, 1H), 7.80 (d, J = 12.0 Hz, 1H), 7.49 (s, 1H), 7.15 (s, 1H), 7.03 (t, J = 8.8 Hz, 2H), 6.73-6.70 (m, 2H), 5.53 (d, J = 12.0 Hz, 1H), 4.10-3.98 (m, 1H), 3.90-3.75 (m, 1H), 3.30-3.25 (m, 1H), 2.53 (s, 3H), 2.40 (s, 3H), 1.68-1.45 (m, 2H), 1.42-1.30 (m, 4H), 0.91 (t, J = 6.80 Hz, 3H). LCMS: (Method A) 495.2 (M ++H), retention time (Rt.) 2.46 minutes, 93.75% (maximum). HPLC: (Method E) Rt. 5.43 minutes, 94.52% (maximum). Chiral SFC: (Method S) Rt. 2.29 minutes, 98.89% (maximum). Enantiomer 2: Yield: 28% (10 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.27 (s, 1H), 7.80 (d, J = 12.4 Hz, 1H), 7.49 (s, 1H), 7.15 (s, 1H), 7.03 (t, J = 8.4 Hz, 2H), 6.80 - 6.50 (m, 2H), 5.53 (d, J = 12.0 Hz, 1H), 4.10 - 3.95 (m, 1H), 3.90 - 3.75 (m, 1H), 3.35 - 3.30 (m, 1H), 2.58 (s, 3H), 2.40 (s, 3H), 1.78 - 1.45 (m, 2H), 1.42 - 1.30 (m, 4H), 0.95 - 0.88 (m, 3H). LCMS: (Method A) 495.1 (M + +H), Rt. 2.472 minutes, 97.27% (maximum). HPLC: (Method E) Rt. 5.434 minutes, 97.10% (maximum). Chiral SFC: (Method S) Rt. 4.79 minutes, 97.64% (maximum).

[0518] (Example 23) (E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0519] [Chemical Structure]

[0520] A stirred solution of tert-butyl (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (Intermediate 49; 60 mg, 0.10 mmol) in DCM (1.5 mL) at 0 °C was treated with TFA (0.16 mL, 2.12 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with cold water (5 mL), and the aqueous layer was extracted with DCM (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL), and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 35 - 40% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 37% (20 mg, off-white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 12.27 (s, 1H), 7.79 (d, J = 12.4 Hz, 1H), 7.49 (s, 1H), 7.18 (s, 1H), 7.04 (t, J = 8.8 Hz, 2H), 6.75 - 6.71 (m, 2H), 5.51 (d, J = 12.0 Hz, 1H), 4.05 - 4.01 (m, 1H), 3.82 - 3.80 (m, 1H), 3.30 (m, 1H), 2.96 - 2.90 (m, 2H), 2.46 (s, 3H), 1.61 - 1.51 (m, 2H), 1.34 - 1.29 (m, 4H), 1.21 (m, 3H), 0.92 - 0.89 (m, 3H). LCMS: (Method A) 509.2 (M + +H), Rt. 2.62 min, 95.13% (max). HPLC: (Method E) Rt. 5.62 min, 96.69% (max).

[0521] (Examples 24 and 25) (S)-(E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid and (R)-(E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0522]

Chem.

[0523] The two enantiomers of racemic (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid (Example 23; 35 mg, 0.078 mmol) were separated by chiral SFC (Method S). The substance was concentrated at 40 °C under vacuum. The first elution fraction corresponded to enantiomer 1 and the second elution fraction corresponded to enantiomer 2. The absolute configuration of the two enantiomers is unknown. Enantiomer 1: Yield: 40% (14 mg, off-white solid). δ 12.28 (s, 1H), 7.79 (d, J = 12.0 Hz, 1H), 7.49 (s, 1H), 7.18 (s, 1H), 7.04 (t, J = 8.8 Hz, 2H), 6.73 - 6.70 (m, 2H), 5.52 (d, J = 12.0 Hz, 1H), 4.05 - 4.01 (m, 1H), 3.90 - 3.72 (m, 1H), 3.45 - 3.38 (m, 1H), 2.94 (q, J = 6.40 Hz, 2H), 2.55 (s, 3H), 1.68 - 1.45 (m, 2H), 1.42 - 1.25 (m, 4H), 1.19 (t, J = 7.60 Hz, 3H), 0.9 (t, J = 6.8 Hz, 3H). LCMS: (Method A) 509.1 (M ++H), Rt. 3.03 min, 95.96% (max). HPLC: (Method E) Rt. 5.75 min, 97.05% (max). Chiral SFC: (Method S) Rt. 2.75 min, 100% (max). Enantiomer 2: Yield: 37% (13 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.28 (s, 1H), 7.79 (d, J = 12.0 Hz, 1H), 7.49 (s, 1H), 7.18 (s, 1H), 7.04 (t, J = 8.8 Hz, 2H), 6.74 - 6.70 (m, 2H), 5.52 (d, J = 12.0 Hz, 1H), 4.1 - 4.0 (m, 1H), 3.88 - 3.72 (m, 1H), 3.45 - 3.35 (m, 1H), 2.94 (q, J = 6.8 Hz, 2H), 2.55 (s, 3H), 1.70 - 1.45 (m, 2H), 1.45 - 1.28 (m, 4H), 1.19 (t, J = 7.20 Hz, 3H), 0.91 (t, J = 6.40 Hz, 3H). LCMS: (Method B) 508.9 (M + +H), Rt. 2.09 min, 96.57% (max). HPLC: (Method E) Rt. 5.76 min, 94.92% (max). Chiral SFC: (Method S) Rt. 3.42 min, 97.83% (max).

[0524] (Example 26) (E)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

[0525] [Chemical formula]

[0526] A stirred solution of tert-butyl (E)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (Intermediate 54; 65 mg, 0.12 mmol) in DCM (3 mL) at 0 °C was treated with TFA (1 mL, 12.98 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with cold water (5 mL), and the aqueous layer was extracted with DCM (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The resulting crude material was purified by Isolera column chromatography (eluent: 35 - 40% EtOAc / PE; silica gel: 230 - 400 mesh) to afford the title compound. Yield: 69% (42 mg, white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 12.28 (s, 1H), 7.76 (d, J = 12.0 Hz, 1H), 7.56 (s, 1H), 7.17 (s, 1H), 7.04 (t, J = 8.8 Hz, 2H), 6.70 - 6.68 (m, 2H), 5.52 (d, J = 12.4 Hz, 1H), 4.22 - 4.19 (m, 1H), 3.50 - 4.47 (m, 1H), 3.21 - 3.10 (m, 2H), 2.41 (s, 3H), 2.30 - 2.25 (m, 1H), 1.40 - 1.28 (m, 6H), 0.90 (t, J = 7.20 Hz, 3H). LCMS-Q-TOF: (Method J) 480.1 (M + +H), Rt. 2.88 min, 97.00% (max). HPLC: (Method E) Rt. 5.35 min, 96.21% (max).

[0527] (Examples 27 and 28) (S)-(E)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid and (R)-(E)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

[0528]

Chem.

[0529] The two enantiomers of racemic (E)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid (Example 26; 38 mg, 0.079 mmol) were separated by chiral SFC (Method T). The substance was concentrated at 40 °C under vacuum. The first elution fraction corresponded to enantiomer 1 and the second elution fraction corresponded to enantiomer 2. The absolute configuration of the two enantiomers was unknown. Enantiomer 1: Yield: 34% (13 mg, white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.30 (s, 1H), 7.75 (d, J = 12.4 Hz, 1H), 7.56 (s, 1H), 7.17 (s, 1H), 7.03 (t, J = 9.2 Hz, 2H), 6.69 - 6.66 (m, 2H), 5.52 (d, J = 12.0 Hz, 1H), 4.21 (d, J = 15.2 Hz, 1H), 3.51 - 3.47 (m, 1H), 3.34 - 3.28 (m, 1H), 3.20 - 3.05 (m, 1H), 2.41 (s, 3H), 2.30 - 2.18 (m, 1H), 1.50 - 1.20 (m, 6H), 0.90 (t, J = 7.20 Hz, 3H). LCMS: (Method A) 480.1 (M ++H), Rt. 2.43 min, 96.73% (max). HPLC: (Method E) Rt. 5.34 min, 96.74% (max). Chiral SFC: (Method T) Rt. 3.53 min, 100% (max). Enantiomer 2: Yield: 32% (12 mg, off-white solid). 1 H NMR (400 MHz, DMSO-d6): δ 12.23 (s, 1H), 7.75 (d, J = 12.4 Hz, 1H), 7.56 (s, 1H), 7.17 (s, 1H), 7.06 - 7.01 (m, 2H), 6.71 - 6.68 (m, 2H), 5.52 (d, J = 12.0 Hz, 1H), 4.21 (d, J = 14.4 Hz, 1H), 3.51 - 3.46 (m, 1H), 3.40 - 3.30 (m, 1H), 3.22 - 3.12 (m, 1H), 2.41 (s, 3H), 2.30 - 2.20 (m, 1H), 1.45 - 1.35 (m, 4H), 1.32 - 1.28 (m, 2H), 0.9 (t, J = 8.4 Hz, 3H). LCMS: (Method A) 480.1 (M + +H), Rt. 2.43 min, 97.29% (max). HPLC: (Method E) Rt. 5.34 min, 97.07% (max). Chiral SFC: (Method T) Rt. 5.07 min, 99.37% (max).

[0530] (Example 29) (E)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0531] [Chemical formula]

[0532] A stirred solution of tert-butyl (E)-3-((3-butyl-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (Intermediate 57; 60 mg, 0.09 mmol) in toluene (3 mL) at 0 °C was treated with triphenylamine (67.2 mg, 0.27 mmol) and TFA (0.11 mL, 1.46 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 quenched with cold water (5 mL), and the aqueous layer was extracted with ethyl acetate (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL), and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (Method D) to afford the title compound. Yield: 50% (22 mg, off-white solid). 1 1H NMR (400 MHz, DMSO-d6) δ 12.3 (s, 1H), 7.73 (d, J = 12.0 Hz, 1H), 7.51 (s, 1H), 7.41 (d, J = 9.2 Hz, 1H), 7.15 (s, 1H), 7.03 (t, J = 8.8 Hz, 2H), 6.72 - 6.69 (m, 2H), 5.41 (d, J = 12.4 Hz, 1H), 4.31 (m, 1H), 3.27 (m, 1H), 2.50 (m, 1H), 2.33 (s, 3H), 1.57 - 1.56 (m, 1H), 1.43 - 1.32 (m, 3H), 1.29 - 1.24 (m, 2H), 0.91 - 0.88 (m, 3H). LCMS: (Method A) 479.0 (M + -H), Rt. 2.80 min, 95.41% (max). HPLC: (Method E) Rt. 5.30 min, 97.48% (max).

[0533] (Example 30) (E)-3-((3-Butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

[0534]

Chem.

[0535] To a stirred solution of tert-butyl (E)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (Intermediate 59; 72 mg, 0.135 mmol) in DCM (5 mL) at 0 °C was added TFA (1 mL, 12.98 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with cold water (5 mL), and the aqueous layer was extracted with DCM (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL), and dried over anhydrous Na2SO4. The organic portion was filtered and concentrated under vacuum. The resulting crude material was purified by Isolera column chromatography (eluent: 30 - 35% EtOAc / PE; silica gel: 230 - 400 mesh) to give the title compound. Yield: 46% (30 mg, off-white solid). 11H NMR (400 MHz, DMSO-d6): δ 12.26 (s, 1H), 7.75 (d, J = 12.40 Hz, 1H), 7.58 (s, 1H), 7.26 (s, 1H), 7.17 - 7.19 (m, 2H), 6.75 (t, J = 7.20 Hz, 1H), 6.67 (d, J = 8.00 Hz, 2H), 5.51 (d, J = 12.40 Hz, 1H), 4.25 (m, 1H), 3.49 (m, 1H), 3.41 (m, 1H), 3.15 (m, 1H), 2.94 - 2.96 (m, 2H), 2.34 - 2.34 (m, 1H), 1.36 - 1.38 (m, 5H), 1.30 - 1.32 (m, 1H), 1.18 - 1.20 (m, 3H), 0.05 - 0.89 (m, 3H). LCMS: (Method A) 476.1 (M + +H), Rt. 2.68 min, 98.13% (max). HPLC: (Method B) Rt. 5.65 min, 99.73% (max).

[0536] (Example 31) (E)-3-((3-Butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0537] [Chemical Structure]

[0538] A stirred solution of tert-butyl (E)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylate (Intermediate 60; 40 mg, 0.07 mmol) in DCM (5 mL) at 0 °C was treated with TFA (0.11 mL, 1.50 mmol), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with cold water (5 mL), and the aqueous layer was extracted with DCM (2 × 5 mL). The combined organic layers were washed with water (5 mL) and brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (Method A) to afford the title compound. Yield: 56% (10 mg, off-white solid). 1 1H NMR (400 MHz, DMSO-d6): δ 12.28 (s, 1H), 7.74 (d, J = 12.0 Hz, 1H), 7.53 (s, 1H), 7.41 (d, J = 9.2 Hz, 1H), 7.25 (s, 1H), 7.19 (t, J = 8.4 Hz, 2H), 6.75 (t, J = 7.2 Hz, 1H), 6.67 (d, J = 8.4 Hz, 2H), 5.44 (d, J = 12.4 Hz, 1H), 4.39 - 4.35 (m, 1H), 3.45 - 3.35 (m, 2H), 2.98 - 2.92 (m, 2H), 1.65 - 1.55 (m, 1H), 1.50 - 1.25 (m, 5H), 1.19 (t, J = 7.20 Hz, 3H), 0.9 (t, J = 7.6 Hz, 3H). LCMS: (Method B) 474.9 (M + -H), Rt. 2.03 min, 94.04% (max). HPLC: (Method E) Rt. 5.36 min, 95.06% (max).

[0539] (Example 32) (E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid

[0540]

Chem.

[0541] The title compound was prepared from tert-butyl (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylate (Intermediate 61) according to the same procedure as described in Example 29.

[0542] (Example 33) (Z)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0543]

Chem.

[0544] The title compound is prepared from ethyl (Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 62) according to the procedure as described in Example 4.

[0545] (Examples 34 and 35) (S)-(Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0546]

Chem.

[0547] The two enantiomers of racemic (Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 33) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to enantiomer 1 and the second elution fraction corresponds to enantiomer 2. The absolute configuration of the two enantiomers is unknown.

[0548] (Example 36) (Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0549]

Chem.

[0550] The title compound is prepared from ethyl (Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 63) according to the procedure as described in Example 4.

[0551] (Examples 37 and 38) (S)-(Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0552] [Chemical formula]

[0553] The two enantiomers of racemic (Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 36) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to enantiomer 1 and the second elution fraction corresponds to enantiomer 2. The absolute configurations of the two enantiomers are unknown.

[0554] (Example 39) (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0555] [Chemical formula]

[0556] The title compound is prepared from ethyl (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 64) according to the procedure as described in Example 4.

[0557] (Examples 40 and 41) (S)-(Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0558] [Chemical formula]

[0559] The two enantiomers of racemic (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 39) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to enantiomer 1, and the second elution fraction corresponds to enantiomer 2. The absolute configuration of the two enantiomers is unknown.

[0560] (Example 42) (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0561] [Chemical formula]

[0562] The title compound is prepared from ethyl (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 65) according to the procedure as described in Example 4.

[0563] (Examples 43 and 44) (S)-(Z)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0564] [Chemical formula]

[0565] The two enantiomers of racemic (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 42) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to Enantiomer 1 and the second elution fraction corresponds to Enantiomer 2. The absolute configuration of the two enantiomers is unknown.

[0566] (Example 45) (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0567]

Chem.

[0568] The title compound is prepared from ethyl (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 66) according to the procedure as described in Example 4.

[0569] (Examples 46 and 47) (S)-(Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0570]

Chem.

[0571] The two enantiomers of racemic (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 45) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to Enantiomer 1 and the second elution fraction corresponds to Enantiomer 2. The absolute configuration of the two enantiomers is unknown.

[0572] (Example 48) (Z)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0573]

Chem.

[0574] The title compound is prepared from ethyl (Z)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylate (Intermediate 67) according to the procedure as described in Example 4.

[0575] (Examples 49 and 50) (S)-(Z)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0576]

Chem.

[0577] The two enantiomers of racemic (Z)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 48) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first eluted fraction corresponds to enantiomer 1 and the second eluted fraction corresponds to enantiomer 2. The absolute configurations of the two enantiomers are unknown.

[0578] (Example 51) (Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0579] [Chemical formula]

[0580] The title compound is prepared from (Z)-3-((3-butyl-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Intermediate 69) according to the procedure as described in Example 10.

[0581] (Examples 52 and 53) (S)-(Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0582] [Chemical formula]

[0583] The two enantiomers of racemic (Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 51) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to enantiomer 1, and the second elution fraction corresponds to enantiomer 2. The absolute configuration of the two enantiomers is unknown.

[0584] (Example 54) (Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0585] [Chemical formula]

[0586] The title compound is prepared from (Z)-3-((3-butyl-7-(ethylthio)-2-(4-methoxybenzyl)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Intermediate 71) according to the procedure as described in Example 10.

[0587] (Examples 55 and 56) (S)-(Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0588] [Chemical formula]

[0589] The two enantiomers of racemic (Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 54) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to enantiomer 1 and the second elution fraction corresponds to enantiomer 2. The absolute configuration of the two enantiomers is unknown.

[0590] (Example 57) (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0591] [Chemical formula]

[0592] The title compound is prepared from (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Intermediate 73) according to the procedure as described in Example 10.

[0593] (Examples 58 and 59) (S)-(Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid and (R)-(Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid

[0594] [Chem.]

[0595] The two enantiomers of racemic (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid (Example 57) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to enantiomer 1 and the second elution fraction corresponds to enantiomer 2. The absolute configuration of the two enantiomers is unknown.

[0596] (Example 60) (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0597] [Chem.]

[0598] The title compound is prepared from tert-butyl (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-(4-methoxybenzyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid (Intermediate 74) according to the procedure as described in Example 29.

[0599] (Examples 61 and 62) (S)-(E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid and (R)-(E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid

[0600]

Chemical formula

[0601] The two enantiomers of racemic (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid (Example 60) are separated by chiral SFC. The substance is concentrated at 40 °C under vacuum. The first elution fraction corresponds to enantiomer 1, and the second elution fraction corresponds to enantiomer 2. The absolute configuration of the two enantiomers is unknown.

[0602] Biological assay IBAT (h / m) assay protocol 10,000 cells (human or mouse IBAT overexpressing cells) are seeded into 200 μL of MEM-alpha medium (Gibco 12571-063) supplemented with 10% FBS (Gibco 10438026) containing puromycin (Gibco A1113803) (10 μg / mL) in a 96-well plate (Corning CLS3809) and incubated at 37 °C in 5% CO2 for 48 hours. After incubation, the medium is decanted from the wells and the cells are washed twice with 300 μL of basal MEM-alpha medium (without FBS). After decanting the basal MEM-alpha medium each time, the plate is gently tapped towards a paper towel to ensure that the remaining medium is removed as much as possible. A dilution of the test inhibitor prepared with DMSO (Sigma D2650) (highest test concentration 10 μM, 3-fold serial dilution, 10 points) was added to an incubation mixture containing 0.25 μM of 3H-taurocholic acid (ARC ART-1368) and 5 μM of cold taurocholic acid (Sigma T4009) (maintaining a final DMSO concentration of 0.2%). Next, 50 μL of the incubation mixture containing the test inhibitor was added to the wells (in duplicate), and the plate was incubated at 37 °C in a CO2 incubator for 20 minutes. After incubation, the reaction was stopped by keeping the plate on an ice-water mixture for 2 - 3 minutes, and then the incubation mixture was completely aspirated from the wells. The wells were washed twice with 250 μL of cooled unlabeled 1 mM taurocholic acid dissolved in HEPES (Gibco 15630080) buffered (10 mM) HBSS (Gibco 14175079) (pH 7.4). After each wash, the plate was gently tapped against a paper towel to ensure complete removal of the blocking buffer. 100 μL of MicroScint-20 (PerkinElmer 6013621) was added to the wells, kept at room temperature overnight, and then the plate was read in a PerkinElmer TopCount NXT (trademark) Microplate Scintillation and Luminescence Counter under the 3H Test protocol (set to a read time of 120 seconds per well).

[0603] LBAT (h / m) assay protocol 20,000 cells (human or mouse LBAT overexpressing cells) were seeded in 100 μL of MEM-alpha medium (Gibco 12571-063) supplemented with 10% FBS (Gibco 10438026) containing Geneticin (Gibco 10131-027) (1 mg / mL) in a 96-well plate (Corning CLS3809) and incubated at 37 °C in 5% CO2 for 24 hours. After incubation, the medium was decanted from the wells and the cells were washed twice with 300 μL of basal MEM-alpha medium (without FBS). After decanting the basal MEM-alpha medium each time, the plate was gently tapped towards a paper towel so that the remaining medium was surely removed to the maximum extent. For human LBAT, the incubation mixture was prepared by adding a dilution of the test inhibitor (3-fold serial dilution in DMSO (Sigma D2650), 10 points) to MEM-alpha (without FBS) containing 0.3 μM of 3H-tauroursodeoxycholic acid (ARC ART-1368) and 7.5 μM of cold tauroursodeoxycholic acid (Sigma T4009) (maintaining a final DMSO concentration of 0.2%). For mouse LBAT, the incubation mixture was prepared by adding a dilution of the test inhibitor (3-fold serial dilution in DMSO, 10 points) to MEM-alpha (without FBS) containing 0.3 μM of 3H-tauroursodeoxycholic acid and 25 μM of cold tauroursodeoxycholic acid (maintaining a final DMSO concentration of 0.2%). Then, 50 μL of the incubation mixture containing the test inhibitor was added to the wells (in duplicate), and the plate was incubated at 37 °C in a CO2 incubator for 20 minutes. After incubation, the reaction was stopped by keeping the plate on an ice-water mixture for 2-3 minutes, and then the incubation mixture was completely aspirated from the wells. The wells were washed twice with 250 μL of cooled unlabeled 1 mM tauroursodeoxycholic acid dissolved in HEPES (Gibco 15630080) buffered (10 mM) HBSS (Gibco 14175079) (pH 7.4). After each wash, the plate was gently tapped towards a paper towel so that the blocking buffer was surely removed to the maximum extent. 100 μL of MicroScint-20 (PerkinElmer 6013621) was added to the wells and kept at room temperature overnight, and then the plate was read in a PerkinElmer TopCount NXT (trademark) Microplate Scintillation and Luminescence Counter under a 3H Test protocol (set to a reading time of 120 seconds per well in the normal plate orientation).

[0604] Bidirectional permeability assay (Caco-2 cells) Caco-2 cells (Evotec) were seeded at a density of 70,000 cells / well in a Millicell (registered trademark) 24-well cell culture insert plate and maintained in an incubator (37 °C, 5% CO2, 95% RH) for 21 days while changing the medium every other day. Stock solutions (10 mM) of the test compound, atenolol (low permeability marker), propranolol (high permeability marker), and digoxin (substrate for the P-gp transport pathway) were prepared in dimethyl sulfoxide (DMSO). Intermediate stock solutions (1 mM) were prepared by diluting 10 μL of the 10 mM master stock solution with 90 μL of neat DMSO. Working stock solutions (10 μM) were prepared by diluting 50 μL of the 1 mM with 4950 μL of FaSSIF buffer. After addition of the compound to FaSSIF, the samples were subjected to sonication for 2 hours and centrifuged at 4000 RPM for 30 minutes at 37 °C. 4 mL of the resulting supernatant was used directly in the assay. The final DMSO concentration in the transport experiment was 1%. On the day of the assay, the Caco-2 monolayer was washed twice with transport buffer (HBSS, pH 7.4) and pre-incubated in an incubator for 30 minutes (37 °C, 5% CO2, 95% RH). The electrical resistance of the monolayer was measured using a Millicell (registered trademark)-ERS system. 350 ohm·cm 2 Monolayers with trans-epithelial electrical resistance (TEER) values above this were selected for the assay. The assays were performed in the absorption direction (A2B) and the secretion direction (B2A). The transport experiment was initiated by adding a transport assay buffer (FaSSIF buffer prepared in HBSS) consisting of the compound into the donor compartment (apical chamber A - B; basolateral chamber B - A) in duplicate (n = 2) wells. A drug - free HBSS buffer (pH 7.4) containing 1% bovine serum albumin (BSA) was introduced into the receiver (A - B - basolateral; B - A - apical) compartment. The volumes of the apical and basolateral compartments were 0.4 and 0.8 mL, respectively. After adding the dosing solution, the plate was incubated at 37 °C for 120 minutes in an incubator. After 120 minutes, donor and receiver samples were collected and matrix - matched with the buffer on the opposite side (1:1, 30 μL of test sample+30 μL of blank buffer). The dosed samples were matrix - matched with the buffer on the opposite side (1:1, 30 μL of test sample+30 μL blank buffer). Samples were processed by adding acetonitrile containing an internal standard (60 μL of test sample+200 μL of acetonitrile containing the internal standard - tolbutamide, 500 ng / mL). The samples were vortexed and centrifuged at 4000 rpm for 10 minutes. The resulting supernatant (100 μL) was diluted with 100 μL of water and transferred to a new 96 - well plate. The concentration of the compound in the sample was analyzed by liquid chromatography tandem mass spectrometry (LC - MS / MS) method using a discovery grade biological assay method when applicable. The mean apparent permeability (P app , ×10 -6 cm / s) of the test compounds, atenolol, propranolol, and digoxin was calculated as follows:

[0605]

Equation

[0606] (where dq / dt = transport rate (transport rate of the compound in the receiver compartment), C0 = initial concentration in the donor compartment, A = surface area of the effective filtration membrane).

[0607] HepaRG-based assay protocol A cryopreserved vial of differentiated HepaRG cells (Biopredic International HPR116080) is thawed in HepaRG Thawing / Plating / General Purpose Medium (Biopredic International ADD670C) supplemented with 200 mM Glutamax (Gibco 35050061) according to the protocol provided by Biopredic International. 70,000 cells per well are seeded into 100 μL of HepaRG Thawing / Plating / General Purpose Medium supplemented with 200 mM Glutamax in a 96-well plate (Corning CLS3809) and incubated at 37 °C in 5% CO2 for 24 hours. After incubation, the seeding medium is replaced with HepaRG Maintenance / Metabolism Medium (Biopredic International ADD620C), and fresh HepaRG Maintenance / Metabolism Medium is replenished every 48 hours for 6 days of incubation. After 7 days of incubation post-seeding, the incubation medium is decanted from the wells and the cells are washed twice with 250 μL of William's E Basal Media (Gibco 12551032). After decanting William's E Basal Media each time, the plate is gently tapped towards a paper towel to ensure that the remaining medium is removed maximally. The incubation mixture is prepared by adding a dilution of the test inhibitor (serial 3-fold dilutions in DMSO (Sigma D2650)) to William's E medium (basal) containing 0.3 μM of 3H-tauroursodeoxycholic acid (ARC ART-1368) and 7.5 μM of cold tauroursodeoxycholic acid (Sigma T4009) (maintaining a final DMSO concentration of 0.2%). Next, 50 μL of the incubation mixture containing the test inhibitor is added to the wells (in duplicate), and the plate is incubated at 37 °C for 30 min in a 5% CO2 incubator. After incubation, the reaction is stopped by keeping the plate on an ice-water mixture for 2 - 3 min, and then the incubation mixture is completely aspirated from the wells. The wells are washed twice with 250 μL of cooled unlabeled 1 mM tauroursodeoxycholic acid dissolved in HEPES (Gibco 15630080) buffered (10 mM) HBSS (Gibco 14175079) (pH 7.4). After each wash, the plate is gently tapped against a paper towel to ensure maximum removal of the blocking buffer. 100 μL of MicroScint-20 (PerkinElmer 6013621) is added to the wells, kept at room temperature overnight, and then the plate is read in a PerkinElmer TopCount NXT (trademark) Microplate Scintillation and Luminescence Counter under the 3H Test protocol (set at a read time of 120 s per well in the normal plate orientation).

[0608] Preparation of Dilutions of Test Compounds All test compounds were prepared in powder form at room temperature. A 10 mM DMSO stock solution of the test compound was prepared, aliquoted, and stored at -20 °C. From the 10 mM DMSO stock solution of the compound, serial 3-fold dilutions in DMSO were prepared to obtain dilutions of a total of 10 test compounds. 0.5 μL of this dilution in DMSO was added to 250 μL of serum-free basal medium containing 3H-tauroursodeoxycholic acid and cold tauroursodeoxycholic acid to prepare the incubation mixture.

[0609] Bioavailability Test Male mice (C57BL / 6 or CD1) or Wistar rats at 8 - 9 weeks of age were used. For each test compound, two groups consisting of three animals each were used. One group was administered a single intravenous dose of 1 mg / kg (vehicle 100% DMSO) through the tail vein, and the other group was administered a single oral dose of 10 mg / kg through a gavage needle. The group administered the oral dose was 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 hours after oral administration. The blood samples were taken from the supine vein. 0.2% EDTA was used as an anticoagulant. The samples were analyzed using an LC - MS / MS system by a discovery grade biological assay developed to estimate the test compound in plasma.

[0610] Results The biological data for the compounds of the examples are shown in Table 8 below.

[0611]

Table 8A

[0612]

Table 8B

[0613] Effect of LBAT - selective compounds on serum bile acid levels in OATP KO mice Male OATP KO mice (Oatp1a / 1b cluster knockout mice, Taconic, USA) at 8 - 9 weeks of age were used in the study. After completion of the quarantine and acclimation period, the animals were randomly divided into experimental groups (n = 4 mice in each) based on body weight. On the day of the experiment, baseline blood samples were collected after oral administration of vehicle (0.5% CMC, 0.1% Tween 80) or the compounds of Examples 3, 6, 13, or 15 at the indicated doses. Subsequently, blood samples were collected at various time points after administration (shown in each graph). Blood was allowed to clot and serum was collected by centrifugation at 3000 g for 15 minutes at 4 °C and stored at -80 °C for further analysis. The total bile acid concentration in each serum sample was determined using the CELL BIOLABS Total Bile Acid Kit (STA631) according to the manufacturer's instructions. Mean ± SEM, n = 4.

[0614] The results are shown in Figures 1 - 4. Serum bile acid levels before dosing and during vehicle treatment were in the range of 1 - 5 μmol / L. Oral administration of the compound of Example 13 increased serum bile acids in a dose - dependent manner. Within 1 hour after dosing, serum bile acid levels increased to approximately 100 μmol / L in response to the 30 mg / kg dose of the compound of Example 13. The compounds of Examples 3, 6, and 15 also increased serum bile acid levels.

[0615] PD model: Evaluation of test compounds for total bile acid levels in male C57BL6 mice Using 8- to 9-week-old C57BL / 6N Tac mice, the effect of a bile acid modulator on bile acid levels is tested. After completion of the quarantine and acclimation period, the animals are randomly divided based on body weight into x experimental groups: (i) vehicle control, and (ii) test compound y mg / kg orally once daily. The animals are treated with the test compound for 7 days. On the 5th day of the test, the animals are individually housed in new cages. On the 7th day, feces are collected from each cage, and then blood is collected from each animal via the retro-orbital route. The animals are euthanized and the liver and terminal ileum are collected from each animal for further analysis. Body weight and food intake are measured twice a week. Serum lipid profile is analyzed with serum samples on the 7th day. Total bile acids in serum are measured with serum samples on the 7th day. Bile excretion in feces is measured with fecal samples on the 7th day. Expression of CYP7A1 and SHP in the liver is quantified with liver samples on the 7th day. Liver triglyceride and total cholesterol are analyzed with liver samples on the 7th day.

[0616] Urinary bile acid model: Evaluation of a test compound on urinary bile acid levels in male C57BL / 6N mice Using 8- to 9-week-old C57BL / 6N Tac mice, the effect of a bile acid modulator on bile acid levels is tested. After completion of the quarantine and acclimation period, the animals are randomly divided based on body weight into x experimental groups: (i) vehicle control, and (ii) test compound y mg / kg orally once daily. The animals are treated with the test compound for 7 days. On the 6th day of the test, the animals are transferred to metabolic cages. On the 7th day, feces and urine are collected from each metabolic cage, and then blood is collected from each animal via the retro-orbital route. The animals are euthanized and the kidneys are collected from each animal for further analysis. Body weight is measured twice a week. Total bile acids in serum are measured with serum samples on the 7th day. Bile acid excretion in feces is measured with fecal samples on the 7th day. Urinary excretion of bile acids is measured with samples on the 7th day. Expression of ASBT, OSTa, OSTAb, and MRP2 in the kidney is quantified with samples on the 7th day.

Claims

1. The compound of formula (I) 【Chemical 1】 (In the formula, M is selected from -CH 2 - and -NR 7 -; R 1 is C 1～4 alkyl; R 2 is independently 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, C 3～6 cycloalkyloxycarbonylamino, C 1～4 alkylsulfonamide, and C 3～6 cycloalkylsulfonamide; n is an integer of 1, 2, or 3; R 3 is 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; R 4 and R 5 one of which is carboxyl, R 4 and R 5 On the other hand, it is selected from the group consisting of hydrogen, fluoro, C 1～4 alkyl, and C 1～4 haloalkyl; R 6 is selected from the group consisting of hydrogen and C 1～4 alkyl; R 7 is selected from the group consisting of hydrogen and C 1～4 alkyl) or a pharmaceutically acceptable salt thereof.

2. R 1 is C 2～4 alkyl, the compound according to claim 1.

3. R 1 is n-butyl, the compound according to claim 1 or 2.

4. R 2 is independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, amino, methylamino, dimethylamino, isopropylcarbonylamino, tert-butylcarbonylamino, tert-butylaminocarbonyl, tert-butoxycarbonylamino, methylsulfonamide, and cyclopropylsulfonamide, the compound according to any one of claims 1 to 3.

5. R 3 is selected from the group consisting of hydrogen, fluoro, chloro, bromo, methyl, cyclopropyl, methoxy, ethoxy, methylthio, ethylthio, amino, methylamino, and dimethylamino, the compound according to any one of claims 1 to 4.

6. R 4 is hydrogen or fluoro, the compound according to any one of claims 1 to 5.

7. R 5 is carboxyl, the compound according to any one of claims 1 to 6.

8. R 6The compound according to any one of claims 1 to 7, wherein the compound is hydrogen. **Claim 9** (E)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (Z)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (S)-(Z)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (R)-(Z)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E)-3-((3-Ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (Z)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (S)-(Z)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (R)-(Z)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (E)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-Butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-Butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-Butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (S)-(E)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (R)-(E)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; (E)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)acrylic acid; (Z)-3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-2-methyl-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; (Z)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-fluoroacrylic acid; and (E)-3-((3-Butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)acrylic acid; The compound according to claim 1 or a pharmaceutically acceptable salt thereof, selected from the group consisting of.

10. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to any one of claims 1 to 9 and one or more pharmaceutically acceptable excipients.

11. A medicament comprising the compound according to any one of Claims 1 to 9.

12. A cardiovascular disease including hypercholesterolemia, a disorder of fatty acid metabolism or a disorder of glucose utilization; a disorder of fatty acid metabolism; type 1 and type 2 true diabetes; cataract, microvascular and macrovascular diseases, retinopathy, neuropathy, nephropathy, and delayed wound healing, tissue ischemia, diabetic foot lesions, arteriosclerosis, myocardial infarction, acute coronary syndrome, unstable angina, stable angina, stroke, peripheral arterial occlusive disease, cardiomyopathy, heart failure, cardiac arrhythmia, and vascular restenosis, which are complications of diabetes; insulin resistance (disorder of glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, obesity, dyslipidemia, hypertriglyceridemia, which is a hyperlipidemia, metabolic syndrome (syndrome X), atherosclerotic arteriosclerosis, and hypertension, which are diabetes-related diseases; and the medicament according to Claim 11 for use in the treatment or prevention of an increase in high density lipoprotein level.

13. A gastrointestinal disease or disorder including constipation (including chronic constipation, functional constipation, chronic idiopathic constipation (CIC), intermittent / sporadic constipation, constipation secondary to true diabetes, 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, constipation-predominant irritable bowel syndrome (IBS-C), mixed irritable bowel syndrome (IBS-M), pediatric functional constipation, and opioid-induced constipation); Crohn's disease; primary bile acid malabsorption; irritable bowel syndrome (IBS); inflammatory bowel disease (IBD); inflammation of the ileum; and the medicament according to Claim 11 for use in the treatment or prevention of reflux and its complications including Barrett's esophagus, bile reflux esophagitis, and bile reflux gastritis.

14. Liver diseases or disorders including hereditary metabolic disorders of the liver; congenital abnormalities of bile acid synthesis; congenital anomalies of bile duct course; biliary atresia; biliary atresia after Kasai operation; biliary atresia after liver transplantation; neonatal hepatitis; neonatal cholestasis; genetic forms of cholestasis; cerebrotendinous xanthomatosis; secondary defects in BA synthesis; Zellweger syndrome; liver diseases associated with cystic fibrosis; alpha1-antitrypsin deficiency; Alagille syndrome (ALGS); Byler syndrome; primary defects in bile acid (BA) synthesis; progressive familial intrahepatic cholestasis (PFIC) including PFIC-1, PFIC-2, PFIC-3, and unspecified PFIC, PFIC after bile diversion, and PFIC after liver transplantation; benign recurrent intrahepatic cholestasis (BRIC) including BRIC1, BRIC2, and unspecified 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; cholestasis associated with parenteral nutrition (PNAC); cholestasis associated with low phospholipids; lymphedema cholestasis syndrome 1 (LCS1); primary sclerosing cholangitis (PSC); cholangitis associated with immunoglobulin G4; primary biliary cholangitis; cholelithiasis (gallstones); biliary lithiasis; common bile duct stones; gallstone pancreatitis; Caroli disease; malignant tumors of the bile duct; malignant tumors causing obstruction of the biliary tree; bile duct stricture; AIDS cholangiopathy; ischemic cholangiopathy; pruritus due to cholestasis or jaundice; pancreatitis; chronic autoimmune liver diseases leading to progressive cholestasis; hepatic steatosis; alcoholic hepatitis; acute fatty liver; fatty liver of pregnancy; drug-induced hepatitis; iron overload; congenital bile acid metabolism disorder type 1 (BAS disorder type 1); drug-induced liver injury (DILI); liver fibrosis; congenital hepatic fibrosis; cirrhosis; Langerhans cell histiocytosis (LCH); neonatal ichthyosis sclerosing cholangitis (NISCH); erythropoietic protoporphyria (EPP); idiopathic adult ductopenia (IAD); idiopathic neonatal hepatitis (INH); non-symptomatic intrahepatic ductopenia (NS PILBD); autosomal recessive hereditary intrahepatic cholestasis (North American Indian childhood cirrhosis) (NAIC);Hepatic sarcoidosis; amyloidosis; necrotizing enteritis; cardiac arrhythmias (including atrial fibrillation) in the situation of abnormal serum bile acid profile, cardiomyopathy associated with cirrhosis ("cholecardia"), and toxicity caused by serum bile acids including skeletal muscle wasting associated with cholestatic liver disease; polycystic liver disease; viral hepatitis (including hepatitis A, hepatitis B, hepatitis C, hepatitis D, and hepatitis E); hepatocellular carcinoma (hepatocellular tumor); cholangiocarcinoma; gastrointestinal cancer related to bile acids; and for use in the treatment or prevention of cholestasis caused by tumors and neoplasms of the liver, biliary tract, and pancreas; or for use in enhancing corticosteroid therapy in liver diseases; the pharmaceutical according to claim 11. ;

15. Malabsorption syndromes (including abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicron retention disease (CRD), and sitosterolemia); vitamin excess and marble bone disease; hypertension; glomerular hyperfiltration; polycystic kidney disease (PKD) including autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD); and for use in the treatment or prevention of pruritus in renal insufficiency; or for use in protecting against kidney injury related to liver or metabolic diseases; the pharmaceutical according to claim 11.

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