Treatment of hepatitis

Abstract

A method for treating hepatitis B and / or D with an inhibitor of the sodium / taurocholic acid cotransporting polypeptide (NTCP), such as a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof, is provided herein. Such methods can include reducing the concentration of hepatitis B DNA, reducing the concentration of hepatitis D DNA, reducing hepatitis B surface antigen, and reducing hepatitis B core antigen (HBcAg).

Description

Technical Field

[0001] Cross - reference to related applications This application claims priority to U.S. Provisional Application No. 63 / 350,693, filed on June 9, 2022, which is hereby incorporated by reference in its entirety.

[0002] The present disclosure relates to methods for treating hepatitis B and / or D using one or more NTCP inhibitors, such as 1,5 - benzothiazepine and 1,2,5 - benzothiadiazepine derivatives, or pharmaceutically acceptable salts thereof. The present disclosure also relates to methods for using such inhibitors not only to reduce the entry of hepatitis B virus particles and / or hepatitis D virus particles into hepatocytes but also to reduce hepatitis B and / or D replication in hepatocytes.

Background Art

[0003] Hepatitis B virus (HBV) infection is a major global public health problem (see, e.g., World Health Organization Hepatitis B fact sheet, 2021, available at: who.int / news - room / fact - sheets / detail / hepatitis - b). Worldwide, HBV is estimated to infect 296 million people, and HDV is estimated to infect 48 - 60 million people. HBV causes nearly 1 million deaths annually. HBV and HDV infect hepatocytes, and chronic HBV and HDV infections can cause severe liver diseases. HDV replicates dependently on HBV and thus grows only when co - infected with HBV.

[0004] Certain 1,5-benzothiazepine and 1,2,5-benzothiadiazepine derivatives are potent inhibitors of the apical sodium-dependent bile acid transporter (ASBT) and / or the Na+ / taurocholate cotransporting polypeptide (NTCP; also known as the liver bile acid transporter (LBAT)). The Na+-taurocholate cotransporting polypeptide (NTCP) is a bile acid (BA) transporter in the hepatocyte sinusoidal membrane. NTCP mediates the uptake of bile acids into hepatocytes. NTCP also acts as a host receptor for hepatitis B and D viruses (HBV / HDV). Pharmacological inhibition of NTCP could be a means to prevent HBV and HDV infections.

Prior Art Documents

Patent Documents

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Non-licensed literature

[0006] [Non-licensed document 1] World Health Organization Hepatitis B fact sheet, 2021, available below: who.int / news-room / fact-sheets / detail / hepatitis-b [Non-licensed document 2] Yan Hら, eLife. 2012;1:e00049 [Non-licensed document 3] Lempp FA, Urban S. Viruses. 2017;9:172

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Summary of the Invention

[0007] A method for treating hepatitis B virus (HBV) in a subject in need thereof, comprising the step of orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, is provided herein.

[0008] A method for preventing or reducing the entry of hepatitis B virus particles into hepatocytes in a subject in need thereof, comprising the step of administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, is also provided herein.

[0009] A method for reducing hepatitis B virus replication in hepatocytes in a subject in need thereof, comprising the step of administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, is also provided herein.

[0010] In some embodiments, the subject has hepatitis D.

[0011] Also provided herein is a method for preventing hepatitis D infection in a subject having hepatitis B, the method comprising orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

[0012] Also provided herein is a method for treating hepatitis D (HDV) in a subject in need of treatment for hepatitis D (HDV), the method comprising orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

[0013] Also provided herein is a method for preventing or reducing the entry of hepatitis D virus particles into hepatocytes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

[0014] A method for reducing hepatitis D virus replication in hepatocytes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, is also provided herein.

[0015] In some embodiments, the method further comprises administering an additional antiviral agent.

[0016] A method for treating HBV in a subject in need thereof, the method comprising orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, and an additional antiviral agent, is also provided herein.

[0017] A method for treating HDV in a subject in need thereof, the method comprising orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, and an additional antiviral agent, is also provided herein.

[0018] In some embodiments, the additional antiviral agent is selected from the group consisting of entecavir, tenofovir, tenofovir disoproxil, tenofovir alafenamide, lamivudine, adefovir, adefovir dipivoxil, telbivudine, brequinar, interferon, and combinations thereof. In some embodiments, the interferon is pegylated interferon, interferon alpha, or a combination thereof. In some embodiments, the additional antiviral agent is tenofovir disoproxil.

[0019] In some embodiments, the subject has hepatitis B. In some embodiments, the subject has chronic hepatitis B. In some embodiments, the subject has chronic hepatitis D.

[0020] In some embodiments, the concentration of one or more biomarkers selected from HBV DNA, hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg), hepatitis B e antigen (HBeAg), HDV DNA, and hepatitis D antigen (HDAg) in the serum of the subject is decreased after administration of (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, or a pharmaceutically acceptable salt thereof.

[0021] In some embodiments, the concentration of HBV DNA in a subject's serum decreases after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBV DNA is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBV DNA in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, decreases as compared to a reference concentration of HBV DNA. In some embodiments, the reference concentration of HBV DNA is the level of HBV DNA in a serum sample obtained from the subject prior to administration of (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, or a pharmaceutically acceptable salt thereof.

[0022] In some embodiments, the concentration of HBV DNA in the serum of the subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBV DNA in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBV DNA in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

[0023] In some embodiments, the concentration of hepatitis B surface antigen (HBsAg) in a subject's serum decreases after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBsAg is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBsAg in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, decreases as compared to a reference concentration of HBsAg. In some embodiments, the reference concentration of HBsAg is the concentration of HBsAg in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

[0024] In some embodiments, the concentration of HBsAg in the serum of the subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBsAg in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBsAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

[0025] In some embodiments, the concentration of hepatitis B core antigen (HBcAg) in a subject's serum decreases after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBcAg is determined in a sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBcAg in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, decreases as compared to a reference concentration of HBcAg. In some embodiments, the reference concentration of HBcAg is the concentration of HBcAg in a serum sample obtained from the subject prior to administration of (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, or a pharmaceutically acceptable salt thereof.

[0026] In some embodiments, the concentration of HBcAg in the serum of the subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBcAg in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBcAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

[0027] In some embodiments, the concentration of hepatitis B e antigen (HBeAg) in a subject's serum decreases after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBeAg is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBeAg in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, decreases as compared to a reference concentration of HBeAg. In some embodiments, the reference concentration of HBeAg is the concentration of HBeAg in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

[0028] In some embodiments, the concentration of HBeAg in the serum of a subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBeAg in the serum of a subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HBeAg in the serum of a subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

[0029] In some embodiments, the concentration of HDV DNA in the serum of a subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HDV DNA is determined in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HDV DNA in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, decreases as compared to a reference concentration of HDV DNA. In some embodiments, the reference concentration of HDV DNA is the concentration of HDV DNA in a serum sample obtained from the subject prior to administration of (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, or a pharmaceutically acceptable salt thereof.

[0030] In some embodiments, the concentration of HDV DNA in the serum of a subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HDV DNA in the serum of a subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HDV DNA in the serum of a subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

[0031] In some embodiments, the concentration of hepatitis D antigen (HDAg) in a subject's serum decreases after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HDAg is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HDAg in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, decreases as compared to a reference concentration of HDAg. In some embodiments, the reference concentration of HDAg is the concentration of HDAg in a serum sample obtained from the subject prior to administration of (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, or a pharmaceutically acceptable salt thereof.

[0032] In some embodiments, the concentration of HDAg in the subject's serum is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HDAg in the subject's serum is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. In some embodiments, the concentration of HDAg in the subject's serum is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

[0033] In some embodiments, the subject is administered (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, or a pharmaceutically acceptable salt thereof, within 18 hours of exposure to hepatitis B. In some embodiments, the subject is administered (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, or a pharmaceutically acceptable salt thereof, within 18 hours of exposure to hepatitis D.

[0034] In some embodiments, a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, is administered to a subject.

[0035] In some embodiments, a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, is administered to a subject.

[0036] Details of one or more embodiments of the present disclosure are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will become apparent from the description and drawings, and from the claims. Unless otherwise defined, all scientific and technical terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents, applications, published applications, and other publications referred to throughout this specification and the accompanying appendices are hereby incorporated by reference in their entirety.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0038] HBV and HDV use sodium taurocholate cotransporting polypeptide (NTCP, also known as liver bile acid transporter (LBAT); gene symbol SLC10A1) to enter and infect human hepatocytes. See Yan H et al., eLife. 2012;1:e00049. NTCP and other members of the SLC10 family of solute transporter proteins, such as apical sodium-dependent bile acid transporter (ASBT, also known as ileal bile acid transporter (IBAT), ISBT, ABAT or NTCP2; gene symbol SLC10A2), control the transport of bile acids in the human body. In the liver, bile acids are efficiently extracted from portal blood by the liver bile acid transporter (LBAT) and are resecreted across the canalicular membrane by the bile salt export pump (BSEP; gene symbol ABCB11). The reabsorption of bile acids in the ileum is handled by the apical sodium-dependent bile acid transporter (ASBT), which is generally called the ileal bile acid transporter (IBAT) in the ileum. Both NTCP and ASBT function as electrogenic sodium-solute cotransporters that move more than two Na+ ions per molecule of solute.

[0039] LBAT also functions as a cellular receptor for viral entry of hepatitis B virus (HBV) and hepatitis D virus (HDV), which is, in turn, a major cause of liver disease and hepatocellular carcinoma. NTCP interacts with an important region within the pre-S1 domain of the HBV envelope L protein. Yan H et al., eLife. 2012;1:e00049. Residues 157-165 of NTCP are important for binding to the receptor-binding region of the preS1 domain of the L protein of HBV, and these residues have been suggested to contribute to HBV- and HDV-mediated infection by NTCP. Yan H et al., eLife. 2012;1:e00049.

[0040] A method of treating hepatitis B virus (HBV) and / or hepatitis D virus (HDV) in a subject in need of treatment for HBV and / or HDV, the method comprising administering to the subject one or more NTCP inhibitors (e.g., any of the NTCP inhibitors described herein). Also provided herein is a method for preventing or reducing the entry of hepatitis B virus particles and / or hepatitis D virus particles into hepatocytes in a subject in need of preventing or reducing such entry, the method comprising administering to the subject one or more NTCP inhibitors (e.g., any of the NTCP inhibitors described herein). Also provided herein is a method for reducing hepatitis B and / or hepatitis D virus replication in hepatocytes in a subject in need of reducing such replication in hepatocytes, the method comprising administering to the subject one or more NTCP inhibitors (e.g., any of the NTCP inhibitors described herein).

[0041] As used herein, "NTCP inhibitor" includes any compound that exhibits NTCP inactivating activity (e.g., inhibits or reduces). In some embodiments, the NTCP inhibitor reduces NTCP activity. In some embodiments, the NTCP inhibitor prevents or reduces the production and / or function of NTCP. In some embodiments, the NTCP inhibitor is a dual inhibitor, i.e., it inhibits NTCP and IBAT. In some embodiments, the NTCP inhibitor is more selective for NTCP than other members of the SLC10 family of solute transporter proteins, e.g., IBAT. In some embodiments, the NTCP inhibitor has a molecular weight of less than about 1,000 g / mol.

[0042] The ability of a compound to act as an inhibitor of NTCP can be demonstrated by assays known in the art. The activity of the compounds and compositions provided herein as NTCP inhibitors can be assayed in vitro, in vivo, or in cell lines. In vitro assays include assays that determine the inhibition of bile salt transport by NTCP. The assays can include, for example, those described in U.S. Patent No. 11,180,465.

[0043] NTCP inhibitors as described herein include compounds of formula (I):

[0044]

Chemical formula

[0045] (wherein, M is selected from -CH2- 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 selected from the group consisting of (alkyl)amino; R 4 and R 5 is carboxyl, and R 4 and R 5 The other is hydrogen, fluoro, C 1~4 Alkyl and C 1~4 haloalkyl; R 6 is hydrogen and C 1~4 selected from the group consisting of alkyl; R 7 is hydrogen and C 1~4 alkyl) or a pharmaceutically acceptable salt thereof.

[0046] In some embodiments, R 1 is C 2~4 In some embodiments, R 1 is n-propyl. In some embodiments, R 1 is n-butyl.

[0047] 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, methylsulfonamido, and cyclopropylsulfonamido. In some embodiments, n is 1, i.e., the phenyl ring contains only one substituent R2 is replaced. In some embodiments, R 2 is in the para position.

[0048] 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.

[0049] In some embodiments, R 4 is hydrogen or fluoro.

[0050] In some embodiments, R 5 is carboxyl.

[0051] In some embodiments, R 6 is hydrogen.

[0052] In some embodiments, R 7 is hydrogen or methyl.

[0053] In some embodiments, the compound of formula (I) is a compound of formula (I-a):

[0054]

Chemical formula

[0055] (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 1~6Alkylcarbonylamino, 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 Selected from the group consisting of cycloalkylsulfonamide; n is an integer of 1 or 2; R 3 Is hydrogen, halogen, C 1~4 Alkyl, C 3~6 Cycloalkyl, 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.

[0056] In some embodiments, the compound of formula (I) is a compound of formula (I-b):

[0057]

Chemical formula

[0058] (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 2is 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.

[0059] In some embodiments, the compound of formula (I) is a compound of formula (I-b) as defined above, wherein M and R 1 ~R 4 are as shown in Table 1 below, or a pharmaceutically acceptable salt thereof:

[0060]

Table 1A

[0061]

Table 1B

[0062]

Table 1C

[0063]

Table 1D

[0064]

Table 1E

[0065]

Table 1F

[0066]

Table 1G

[0067]

Table 1H

[0068] In some 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-benzothiadiazepin-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-benzothiadiazepin-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-benzothiadiazepin-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 is selected from the group consisting of a pharmaceutically acceptable salt thereof.

[0069] In some embodiments, the compound of formula (I) is (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:

[0070]

Chemical formula

[0071] or a pharmaceutically acceptable salt thereof. (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 is also referred to herein as "Compound 1".

[0072] In some embodiments, the compound of formula (I) is (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, or a pharmaceutically acceptable salt thereof, and is administered to a subject. In some embodiments, Compound 1 is (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.

[0073] In some embodiments, the compound of formula (I) is (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, or a pharmaceutically acceptable salt thereof, and is administered to a subject. In some embodiments, Compound 1 is (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.

[0074] The compound of formula (I) can be prepared as described in U.S. Patent No. 11,180,465.

[0075] NTCP inhibitors as described herein include compounds of formula (II)

[0076]

Chemical formula

[0077] (wherein, M is selected from -CH2- and -NR 5 -; 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, C 1~4 haloalkoxy, cyano, nitro, 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 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 4A and R 4B are each independently selected from the group consisting of hydrogen, halogen, hydroxy, C 1~4 alkyl and C 1~4 alkoxy; or R 4A and R 4B together with the carbon atom to which they are attached form a 3- to 5-membered saturated carbocyclic ring; R 4C and R 4D are each independently selected from the group consisting of hydrogen and C 1~4 alkyl; R5 is selected from the group consisting of hydrogen and C 1~4 alkyl) or a pharmaceutically acceptable salt thereof is also included.

[0078] In some embodiments, R 1 is C 2~4 alkyl. In some embodiments, R 1 is n-propyl. In some embodiments, R 1 is n-butyl.

[0079] In some embodiments, R 2 is selected from the group consisting of hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, amino, methylamino and dimethylamino. In some embodiments, n is 1, that is, the phenyl ring is substituted with only one substituent R 2 . In some embodiments, R 2 is in the para position.

[0080] In some embodiments, R 3 is selected from the group consisting of fluoro, chloro, bromo, methyl, cyclopropyl, methoxy, ethoxy, methylthio, ethylthio, amino, methylamino and dimethylamino.

[0081] In some embodiments, R 4A and R 4B are each independently selected from the group consisting of hydrogen, halogen, hydroxy, C 1~4 alkyl and C 1~4 alkoxy, or R 4A and R 4B together with the carbon atom to which they are attached form a cyclopropyl ring. In some embodiments, R 4A and R 4B are each independently fluoro, methyl or methoxy, or together with the carbon atom to which they are attached form a cyclopropyl ring.

[0082] In some embodiments, R 4Cand R 4D are each independently hydrogen or methyl. In some embodiments, R 4C and R 4D are each hydrogen.

[0083] In some embodiments, R 5 is hydrogen. In some embodiments, R 5 is methyl.

[0084] In some embodiments, the compound of formula (II) is a compound of formula (II-a):

[0085]

Chemical formula

[0086] (wherein, M is selected from the group consisting of -CH2-, -NH-, and -NCH3-; R 1 is C 2~4 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, C 1~4 haloalkoxy, amino, N-(C 1~4 alkyl)amino, N,N-di(C 1~4 alkyl)amino; n is an integer of 1 or 2; R 3 is selected from the group consisting of halogen, C 1~4 alkyl, C 3~6 cycloalkyl, C 1~4 alkoxy, C 1~4 alkylthio, amino, N-(C 1~4 alkyl)amino and N,N-di(C 1~4 alkyl)amino; R 4A and R 4B are each independently hydrogen, halogen, hydroxy, C 1~4 alkyl and C1~4 is selected from the group consisting of alkoxy, or R 4A and R 4B together with the carbon atom to which they are attached form a cyclopropyl ring) or a pharmaceutically acceptable salt thereof.

[0087] In some embodiments, the compound of formula (II) is a compound of formula (II-b):

[0088]

Chemical formula

[0089] (wherein, M is selected from the group consisting of -CH2-, -NH-, and -N(CH3)-; 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, and dimethylamino; R 3 is selected from the group consisting of fluoro, chloro, bromo, methyl, cyclopropyl, methoxy, ethoxy, methylthio, ethylthio, amino, methylamino, and dimethylamino; R 4A and R 4B are each independently hydrogen, fluoro, methyl, methoxy, or ethoxy, or together with the carbon atom to which they are attached form a cyclopropyl ring) or a pharmaceutically acceptable salt thereof.

[0090] Compounds of formula (II-b) as defined above, in which M, R 1 , R 2 , R 3 , R 4A and R 4B are as shown in Table 2 below, or a pharmaceutically acceptable salt thereof, are also included:

[0091]

Table 2A

[0092]

Table 2B

[0093]

Table 2C

[0094] In some embodiments, the compound of formula (II) is 3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; (S)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; (R)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; 1-(((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (S)-1-(((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (R)-1-(((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; (S)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; (R)-3-((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; 1-(((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (S)-1-(((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (R)-1-(((3-butyl-7-(ethylthio)-2-methyl-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 3-((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiadiazepin-8-yl)oxy)-2,2-dimethylpropanoic acid; 1-(((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 1-(((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (S)-1-(((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (R)-1-(((3-butyl-5-(4-fluorophenyl)-2-methyl-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 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)-2,2-dimethylpropanoic acid; (S)-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)-2,2-dimethylpropanoic acid; (R)-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)-2,2-dimethylpropanoic acid; 1-(((3-butyl-7-(ethylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 1-(((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (S)-1-(((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (R)-1-(((3-butyl-5-(4-fluorophenyl)-7-(methylthio)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 1-(((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (S)-1-(((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-1,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; (R)-1-(((3-butyl-7-(ethylthio)-5-(4-fluorophenyl)-l,1-dioxide-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)methyl)cyclopropane-1-carboxylic acid; 3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-difluoropropanoic acid; (S)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-difluoropropanoic acid; (R)-3-((3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2,2-difluoropropanoic acid; 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-methoxy-2-methylpropanoic acid; (S)-3-(((R)-3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxy-2-methylpropanoic acid; (S)-3-(((S)-3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxy-2-methylpropanoic acid; (R)-3-(((R)-3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxy-2-methylpropanoic acid; (R)-3-(((S)-3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxy-2-methylpropanoic acid; 3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiadiazepin-8-yl)oxy)propanoic acid; (S)-3-((3-butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiadiazepin-8-yl)oxy)propanoic acid; (R)-3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)propanoic acid; 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-ethoxypropanoic acid; 3-((3-Butyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)-2-hydroxypropanoic acid; 3-((3-Ethyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,5-benzothiazepin-8-yl)oxy)propanoic acid; 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-methoxypropanoic acid; 3-(((S)-3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxypropanoic acid; 3-(((R)-3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxypropanoic acid; (S)-3-(((R)-3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxypropanoic acid; (R)-3-(((R)-3-Butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxypropanoic acid; (S)-3-(((S)-3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxypropanoic acid; (R)-3-(((S)-3-butyl-2-methyl-7-(methylthio)-1,1-dioxide-5-phenyl-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepin-8-yl)oxy)-2-methoxypropanoic acid; and 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-hydroxypropanoic acid; or a salt thereof acceptable as a medicament, selected from the group consisting of.

[0095] In some embodiments, the compound of formula (II) is 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)-2,2-dimethylpropanoic acid:

[0096]

Chemical formula

[0097] or a salt thereof acceptable as a medicament. (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 is also referred to herein as "Compound 2".

[0098] In some embodiments, the compound of formula (II) is (S)-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)-2,2-dimethylpropanoic acid, or a pharmaceutically acceptable salt thereof, and is administered to a subject. In some embodiments, Compound 2 is ((S)-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)-2,2-dimethylpropanoic acid.

[0099] In some embodiments, the compound of formula (II) is (R)-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)-2,2-dimethylpropanoic acid, or a pharmaceutically acceptable salt thereof, and is administered to a subject. In some embodiments, Compound 2 is (R)-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)-2,2-dimethylpropanoic acid.

[0100] The compound of formula (II) can be prepared as described in U.S. Application No. 17 / 813,152.

[0101] The NTCP inhibitors used in this specification may also include the compounds disclosed in International Publication Nos. WO 93 / 16055, WO 94 / 18183, WO 94 / 18184, WO 96 / 05188, WO 96 / 08484, WO 96 / 16051, WO 97 / 33882, 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 2004 / 006899, WO 2004 / 076430, WO 2007 / 009655, WO 2007 / 009656, WO 2008 / 058628, WO 2008 / 058630, WO 2011 / 137135, WO 2019 / 234077, WO 2020 / 161216, WO 2020 / 161217, WO 2021 / 110883, WO 2021 / 110884, WO 2021 / 110885, WO 2021 / 110886, WO 2021 / 110887, WO 2022 / 029101, WO 2022 / 253997, and WO 2022 / 117778; DE 19825804; EP 864582, EP 489423, EP 549967, EP 573848, EP 624593, EP 624594, EP 624595, EP 624596, EP 0864582, EP 1173205, EP 1535913, EP 3210977.

[0102] In some embodiments, the subject has hepatitis B. In some embodiments, the subject has acute hepatitis B. For example, subjects having acute hepatitis B include subjects having hepatitis B for less than 6 months. In some embodiments, the subject has chronic hepatitis B. For example, subjects having chronic hepatitis B include subjects having hepatitis B for 6 months or more.

[0103] In some embodiments, the subject has hepatitis D. In some embodiments, the subject has acute hepatitis D. For example, subjects having acute hepatitis D include subjects having hepatitis D for less than 6 months. In some embodiments, the subject has chronic hepatitis D. For example, subjects having chronic hepatitis D include subjects having hepatitis D for 6 months or more.

[0104] In some embodiments, the subject has hepatitis B and hepatitis D. In some embodiments, the subject has chronic hepatitis B and hepatitis D. In some embodiments, the subject has chronic hepatitis B and acute hepatitis D. In some embodiments, the subject has chronic hepatitis B and chronic hepatitis D.

[0105] HDV replicates dependently on HBV and thus grows only when co-infected with HBV. Lempp FA, Urban S. Viruses. 2017;9:172. Co-infection with HDV and HBV is considered the most severe form of chronic viral hepatitis because it can lead to a more rapid progression to hepatocellular carcinoma and liver-related death. World Health Organization Hepatitis D fact sheet, 2021, available at: who.int / news-room / fact-sheets / detail / hepatitis-d. Accordingly, provided herein is also a method for preventing hepatitis D infection in a subject having hepatitis B, the method comprising administering to the subject one or more NTCP inhibitors (e.g., any of the NTCP inhibitors described herein).

[0106] In some of the embodiments described above, the subject is administered an NTCP inhibitor (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) after exposure to hepatitis B and / or hepatitis D. For example, the NTCP inhibitor (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) can be administered within about 1 to about 30 hours from exposure to hepatitis B and / or hepatitis D. In some embodiments, the NTCP inhibitor (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) can be administered within about 1 to about 2, about 1 to about 5, about 1 to about 10, about 1 to about 15, about 1 to about 20, about 1 to about 25, about 5 to about 10, about 5 to about 15, about 5 to about 20, about 5 to about 25, about 5 to about 30, about 10 to about 15, about 10 to about 20, about 10 to about 25, about 10 to about 30, about 15 to about 20, about 15 to about 25, about 15 to about 30, about 20 to about 25, about 20 to about 30, or about 25 to about 30 hours from exposure to hepatitis B and / or hepatitis D. In some embodiments, the subject is administered an NTCP inhibitor (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) within about 18 hours from exposure to hepatitis B and / or hepatitis D.

[0107] In some embodiments of any of the methods described herein, the presence of hepatitis B and / or D in a subject can be determined by one or more biomarkers. Non-limiting examples of such biomarkers for hepatitis B include the concentration of HBV DNA, the concentration of hepatitis B surface antigen (HBsAg), the concentration of hepatitis B core antigen (HBcAg), and the concentration of hepatitis B e antigen (HBeAg). Non-limiting examples of such biomarkers for hepatitis D include the concentration of HDV DNA and the concentration of hepatitis D antigen (HDAg). See, for example, Coffin et al., New and Old Biomarkers for Diagnosis and Management of Chronic Hepatitis B Virus Infection. Gastroenterology. January 2019;156(2):355-368.e3. Such biomarkers can be detected in a sample from the subject (e.g., serum or biopsy sample).

[0108] In some embodiments of any of the methods described herein, assays used to determine whether a subject has hepatitis B and / or hepatitis D using a sample from the subject include, but are not limited to, next-generation sequencing, immunohistochemistry, Southern blotting, Western blotting, Northern blotting, and PCR-based amplification (e.g., RT-PCR and quantitative real-time RT-PCR). As is well known in the art, hepatitis antigens such as HBsAg, HBcAg, HBeAg, and HDAg can be detected using immunoassays. Non-limiting examples of such assays include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), enzyme immunoassay (EIA), electrochemiluminescence immunoassay (ECLIA), microparticle enzyme immunoassay (MEIA), and chemiluminescent microparticle immunoassay (CMIA). See, for example, Tyas et al., Recent Advances of Hepatitis B Detection towards Paper-Based Analytical Devices. ScientificWorldJournal. February 26, 2021;2021:6643573. HBV DNA and / or HDV DNA can be detected, for example, using PCR-based amplification (e.g., RT-PCR and quantitative real-time RT-PCR). See, for example, Coffin et al., Gastroenterology. January 2019;156(2):355~368.e3.

[0109] In some embodiments, a biomarker as described herein (e.g., HBV DNA, HBsAg, HBcAg, HBeAg, HDV DNA, or HDAg) can be used to monitor a subject's responsiveness to a particular therapy (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or an NTCP inhibitor such as a compound of formula (II), or a pharmaceutically acceptable salt thereof). For example, a sample can be obtained from a subject prior to initiation of treatment with an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof), and the level of the biomarker can be determined in the sample. This sample can be considered a reference sample. Subsequently, one or more doses of an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) can be administered to the subject, and the level of the biomarker can be monitored after administration. For example, the level of the biomarker can be monitored after the first dose, the second dose, the third dose, etc. or after 1 week, 2 weeks, 3 weeks, 4 weeks, etc. If the concentration of the biomarker has decreased (i.e., been reduced) relative to the reference sample, this indicates responsiveness to the therapy.In some embodiments, the level of the biomarker is reduced by about 1% to about 99%, about 1% to about 95%, about 1% to about 90%, about 1% to about 85%, about 1% to about 80%, about 1% to about 75%, about 1% to about 70%, about 1% to about 65%, about 1% to about 60%, about 1% to about 55%, about 1% to about 50%, about 1% to about 45%, about 1% to about 40%, about 1% to about 35%, about 1% to about 30%, about 1% to about 25%, about 1% to about 20%, about 1% to about 15%, about 1% to about 10%, about 1% to about 5%, about 5% to about 99%, about 10% to about 99%, about 15% to about 99%, about 20% to about 99%, about 25% to about 99%, about 30% to about 99%, about 35% to about 99%, about 40% to about 99%, about 45% to about 99%, about 50% to about 99%, about 55% to about 99%, about 60% to about 99%, about 65% to about 99%, about 70% to about 99%, about 75% to about 95%, about 80% to about 99%, about 90% to about 99%, about 95% to about 99%, about 5% to about 10%, about 5% to about 25%, about 10% to about 30%, about 20% to about 40%, about 25% to about 50%, about 35% to about 55%, about 40% to about 60%, about 50% to about 75%, about 60% to about 80%, about 65% to about 85%, about 70% to about 90%, or about 75% to about 95% relative to the reference sample. In some embodiments, the concentration of the biomarker is reduced to below the detection limit of the device, i.e., the level of the biomarker is "undetectable".

[0110] In some embodiments, the severity of hepatitis B and / or D is determined by one or more biomarkers or their scoring systems that indicate one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis. The concentration of such biomarkers can be determined, for example, by measuring, quantifying, and monitoring the expression levels of the gene or mRNA encoding the biomarker and / or the peptide or protein of the biomarker.Non-limiting examples of biomarkers and / or their scoring systems that indicate one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis include aspartate aminotransferase (AST) to platelet ratio index (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, e.g., McPherson et al., Gut 2010, Vol. 59(9), pp. 1265-1269); hyaluronic acid; pro-inflammatory cytokines; a panel of biomarkers consisting of α2-macroglobulin, haptoglobin, apolipoprotein A1, bilirubin, gamma-glutamyl transpeptidase (GGT) combined 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 transpeptidase, hyaluronic acid, α2-macroglobulin combined 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®); a panel of biomarkers consisting of tissue inhibitor of metalloproteinase 1 (TIMP-1), amino-terminal propeptide of type III procollagen (PIIINP), and hyaluronic acid (HA) (e.g., enhanced liver fibrosis (ELF) score, see, e.g., Lichtinghagen R et al., J Hepatol. August 2013;59(2):236-242).In some embodiments, the presence of fibrosis in a subject having hepatitis B and / or hepatitis D 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).

[0111] In some embodiments, the concentration of aspartate aminotransferase (AST) in a sample derived from a subject decreases or does not increase after administration of an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof). In some embodiments, the concentration of alanine aminotransferase (ALT) in a sample derived from a subject decreases or does not increase after administration of an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof).

[0112] In some embodiments, a decrease in the concentration of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis during or after administration of an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) as compared to before administration of the NTCP inhibitor indicates treatment of hepatitis B and / or D. 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 concentration of one or more biomarkers indicative of one or more of liver injury, inflammation, liver fibrosis, and / or cirrhosis indicates treatment of hepatitis B and / or D. In some embodiments, the decrease in concentration at 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 NTCP inhibitor 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%.

[0113] In some embodiments, the “concentration” of an enzyme or antigen refers to, for example, the concentration of the enzyme in blood or serum. For example, the levels of AST or ALT can be expressed in units / L.

[0114] During the diagnostic process, the monitoring process, and / or the treatment process (e.g., with an NTCP inhibitor as described herein), samples can be obtained from the subject at multiple time points, and one or more clinically relevant parameters can be determined, including but not limited to the progression of hepatitis B and / or hepatitis D and the effectiveness of treatment. For example, during the diagnostic process, the monitoring process, and / or the treatment process, a first sample can be obtained at a first time point and a second sample can be obtained at a second time point. In some embodiments, the first time point can be a time point before diagnosing the subject with hepatitis B and / or hepatitis D (e.g., when the subject is healthy), and the second time point can be a time point after the subject has developed hepatitis B and / or hepatitis D (e.g., the second time point can be used to diagnose the subject with the disease). In some embodiments, the first time point can be a time point before diagnosing the subject with hepatitis B and / or hepatitis D (e.g., when the subject is healthy), and then the subject is monitored, and the second time point can be a time point after the subject has been monitored. In some embodiments, the first time point can be a time point after diagnosing the subject with hepatitis B and / or hepatitis D, and then an NTCP inhibitor (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) is administered to the subject, and the second time point can be a time point after the NTCP inhibitor (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) has been administered; in such a case, the second time point can be used to evaluate the effectiveness of the NTCP inhibitor (e.g., if the concentration of one or more biomarkers detected at the second time point is reduced or undetectable compared to the first time point).

[0115] In some embodiments, the time difference between the first time point and the second time point is from about 1 day to about 1 year, from about 1 day to about 11 months, from about 1 day to about 10 months, from about 1 day to about 9 months, from about 1 day to about 8 months, from about 1 day to about 7 months, from about 1 day to about 6 months, from about 1 day to about 5 months, from about 1 day to about 4 months, from about 1 day to about 3 months, from about 1 day to about 10 weeks, from about 1 day to about 2 months, from about 1 day to about 6 weeks, from about 1 day to about 1 month, from about 1 day to about 25 days, from about 1 day to about 20 days, from about 1 day to about 15 days, from about 1 day to about 10 days, from about 1 day to about 5 days, from about 2 days to about 1 year, from about 5 days to about 1 year, from about 10 days to about 1 year, from about 15 days to about 1 year, from about 20 days to about 1 year, from about 25 days to about 1 year, from about 1 month to about 1 year, from about 6 weeks to about 1 year, from about 2 months to about 1 year, from about 3 months to about 1 year, from about 4 months to about 1 year, from about 5 months to about 1 year, from about 6 months to about 1 year, from about 7 months to about 1 year, from about 8 months to about 1 year, from about 9 months to about 1 year, from about 10 months to about 1 year, from about 11 months to about 1 year, from about 1 day to about 7 days, from about 1 day to about 14 days, from about 5 days to about 10 days, from about 5 days to about 20 days, from about 10 days to about 20 days, from about 15 days to about 1 month, from about 15 days to about 2 months, from about 1 week to about 1 month, from about 2 weeks to about 1 month, from about 1 month to about 3 months, from about 3 months to about 6 months, from about 4 months to about 6 months, from about 5 months to about 8 months, or from about 7 months to about 9 months.

[0116] In some embodiments provided herein, biomarkers as described herein (e.g., HBV DNA, HBsAg, HBcAg, HBeAg, HDV DNA, or HDAg) are detected in a serum sample from a subject. In some embodiments, the level of the biomarker in the serum sample is compared to the level of the biomarker in a reference sample, e.g., a sample obtained from the subject prior to the initiation of treatment with an NTCP inhibitor as described herein. In some embodiments, the reference sample is a serum sample from the subject (i.e., a serum sample obtained from the subject prior to the initiation of treatment with an NTCP inhibitor).

[0117] Some embodiments of the methods described herein further comprise administering one or more additional antiviral agents. Accordingly, also provided herein is a method of treating hepatitis B (HBV) and / or hepatitis D (HDV) in a subject in need of treatment for hepatitis B (HBV) and / or hepatitis D (HDV), the method comprising administering to the subject one or more NTCP inhibitors (e.g., any of the NTCP inhibitors described herein) and one or more additional antiviral agents. The one or more NTCP inhibitors (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) and the one or more additional antiviral agents can be administered simultaneously, sequentially, or separately.

[0118] Non-limiting examples of additional antiviral agents include nucleoside reverse transcriptase inhibitors, brecanavir (also known as hepcludex or myrcludex-B), and interferons. Non-limiting examples of nucleoside reverse transcriptase inhibitors include tenofovir (e.g., VIREAD®), tenofovir alafenamide (e.g., VEMLIDY®), tenofovir disoproxil, lamivudine (e.g., EPIVIR®, ZEFFIX®, or HEPTODIN®), entecavir (e.g., BARACLUDE®), abacavir (e.g., ZIAGEN®), stavudine (e.g., ZERIT®), didanosine (e.g., VIDEX®), telbivudine (e.g., TYZEKA® or SEBIVO®), zidovudine (e.g., RETROVIR®), zalcitabine (e.g., HIVID®), adefovir (e.g., HESPERA®), adefovir dipivoxil, and emtricitabine (e.g., EMTRIVA®). Non-limiting examples of interferons include pegylated interferon (e.g., PEGASYS®) and interferon alpha (e.g., Intron A).

[0119] A method for treating hepatitis B and / or hepatitis D, comprising administering to a subject in need thereof a compound of formula I, or a pharmaceutically acceptable salt thereof, and an additional antiviral agent (e.g., any of the additional antiviral agents described herein), wherein the amounts of the compound of formula I, or a pharmaceutically acceptable salt thereof, and the additional antiviral agent are effective together to treat hepatitis. Also provided herein is a method.

[0120] A method for treating hepatitis B and / or hepatitis D, comprising administering to a subject in need thereof a compound of formula II, or a pharmaceutically acceptable salt thereof, and an additional antiviral agent (e.g., any of the additional antiviral agents described herein), wherein the amounts of the compound of formula II, or a pharmaceutically acceptable salt thereof, and the additional antiviral agent are effective together to treat hepatitis. Also provided herein is a method.

[0121] In some embodiments, the additional antiviral agent is tenofovir disoproxil.

[0122] A method for treating hepatitis B and / or hepatitis D, comprising administering to a subject in need thereof a compound of formula I, or a pharmaceutically acceptable salt thereof, and tenofovir disoproxil, wherein the amounts of the compound of formula I, or a pharmaceutically acceptable salt thereof, and tenofovir disoproxil are effective together to treat hepatitis. Also provided herein is a method.

[0123] A method for treating hepatitis B and / or hepatitis D, comprising administering to a subject in need thereof a compound of formula II, or a pharmaceutically acceptable salt thereof, and tenofovir disoproxil, wherein the amounts of the compound of formula II, or a pharmaceutically acceptable salt thereof, and tenofovir disoproxil are effective together to treat hepatitis. Also provided herein is a method.

[0124] Also provided herein are NTCP inhibitors (compounds of formula (I), or pharmaceutically acceptable salts thereof, or compounds of formula (II), or pharmaceutically acceptable salts thereof, etc.) for use in the treatment of hepatitis B (HBV) and / or hepatitis D (HDV).

[0125] Also provided herein are NTCP inhibitors (compounds of formula (I), or pharmaceutically acceptable salts thereof, or compounds of formula (II), or pharmaceutically acceptable salts thereof, etc.) for use in preventing or reducing the entry of hepatitis B virus particles and / or hepatitis D virus particles into hepatocytes.

[0126] Also provided herein are NTCP inhibitors (compounds of formula (I), or pharmaceutically acceptable salts thereof, or compounds of formula (II), or pharmaceutically acceptable salts thereof, etc.) for use in reducing hepatitis B and / or hepatitis D virus replication in hepatocytes.

[0127] Also provided herein is the use of NTCP inhibitors (compounds of formula (I), or pharmaceutically acceptable salts thereof, or compounds of formula (II), or pharmaceutically acceptable salts thereof, etc.) in the manufacture of a medicament for the treatment of hepatitis B (HBV) and / or hepatitis D (HDV).

[0128] Also provided herein is the use of NTCP inhibitors (compounds of formula (I), or pharmaceutically acceptable salts thereof, or compounds of formula (II), or pharmaceutically acceptable salts thereof, etc.) in the manufacture of a medicament for preventing or reducing the entry of hepatitis B virus particles and / or hepatitis D virus particles into hepatocytes.

[0129] Also provided herein is the use of NTCP inhibitors (compounds of formula (I), or pharmaceutically acceptable salts thereof, or compounds of formula (II), or pharmaceutically acceptable salts thereof, etc.) in the manufacture of a medicament for reducing hepatitis B and / or hepatitis D virus replication in hepatocytes.

[0130] In some embodiments, the methods described herein involve administering an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) as a pharmaceutical composition comprising the NTCP inhibitor, one or more pharmaceutically acceptable excipients, and optionally one or more additional therapeutic agents as described herein. In some embodiments, the method involves orally administering an NTCP inhibitor (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) as a pharmaceutical composition.

[0131] In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) and one or more pharmaceutically acceptable excipients. Exemplary excipients include, but are not limited to, fillers, binders, disintegrants, glidants, and lubricants. Generally, the pharmaceutical composition can be prepared in a conventional manner using conventional excipients.

[0132] 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.

[0133] Examples of suitable binders include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (such as sucrose, glucose, dextrose, lactose, and sorbitol), polyethylene glycol, waxes, natural and synthetic gums (such as acacia gum and tragacanth gum), sodium alginate, cellulose derivatives (such as hydroxypropyl methylcellulose (or hypromellose), hydroxypropyl cellulose, and ethyl cellulose), and synthetic polymers (such as acrylic acid and methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid / polymethacrylic acid copolymers, and polyvinyl pyrrolidone (povidone)). In certain embodiments, the binder is hydroxypropyl methylcellulose (hypromellose).

[0134] Examples of suitable disintegrants include, but are not limited to, dry starch, modified starches ((partially) pregelatinized starch, sodium starch glycolate, and sodium carboxymethyl starch), alginic acid, cellulose derivatives (such as sodium carboxymethyl cellulose, hydroxypropyl cellulose, and low-substituted hydroxypropyl cellulose (L-HPC)), and cross-linked polymers (such as carmellose, croscarmellose sodium, carmellose calcium, and cross-linked PVP (crospovidone)). In certain embodiments, the disintegrant is croscarmellose sodium.

[0135] Examples of suitable glidants and lubricants include, but are not limited to, talc, magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, colloidal silica, hydrated silicon dioxide, synthetic magnesium silicate, microcrystalline silicon dioxide, starch, sodium lauryl sulfate, boric acid, magnesium oxide, waxes (such as carnauba wax), hydrogenated oils, polyethylene glycol, sodium benzoate, polyethylene glycol, and mineral oil. In certain embodiments, the glidant or lubricant is magnesium stearate or colloidal silica.

[0136] Pharmaceutical compositions may conventionally be coated with one or more coating layers. Enteric coating layers or coating layers for delayed or targeted release of the compounds of formula (I) or formula (II), or pharmaceutically acceptable salts thereof, are also contemplated. The coating layer may comprise one or more coating agents and may optionally comprise a plasticizer and / or a pigment (or colorant).

[0137] Examples of suitable coating agents include cellulose-based polymers (such as ethyl cellulose, hydroxypropyl methylcellulose (or hypromellose), hydroxypropyl cellulose, cellulose acetate phthalate, cellulose acetate succinate, hydroxypropyl methylcellulose acetate succinate, and hydroxypropyl methylcellulose phthalate, etc.), vinyl-based polymers (such as polyvinyl alcohol, etc.), and polymers based on acrylic acid and its derivatives (such as copolymers of acrylic acid and methacrylic acid, copolymers of methacrylic acid, copolymers of methyl methacrylate, copolymers of aminoalkyl methacrylate, copolymers of polyacrylic acid / polymethacrylic acid, etc.), but are not limited thereto. In certain embodiments, the coating agent is hydroxypropyl methylcellulose. In other embodiments, the coating agent is polyvinyl alcohol.

[0138] Examples of suitable plasticizers include triethyl citrate, glyceryl triacetate, tributyl citrate, diethyl phthalate, acetyl tributyl citrate, dibutyl phthalate, dibutyl sebacate, and polyethylene glycol, but are not limited thereto. In certain embodiments, the plasticizer is polyethylene glycol.

[0139] Examples of suitable pigments include titanium dioxide, iron oxides (such as yellow, brown, red, or black iron oxides, etc.), and barium sulfate, but are not limited thereto.

[0140] The pharmaceutical composition may be in a form suitable for oral administration, parenteral injection (including intravenous, subcutaneous, intramuscular and intra-arterial injection), topical administration, or rectal administration. In some embodiments, the pharmaceutical composition is in a form suitable for oral administration. Examples of pharmaceutical compositions formulated for oral administration include, for example, tablets and capsules.

[0141] The dosage required for a therapeutic or prophylactic treatment will depend on the route of administration, the severity of the disease, the age and weight of the patient, and other factors normally considered by the attending physician when determining the appropriate regimen and dosage level for a particular patient.

[0142] The amount of the NTCP inhibitor administered (for example, a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) will vary depending on the subject being treated and can vary 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. 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 regulator 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.

[0143] Some compounds of formula (I) or formula (II), or a pharmaceutically acceptable salt 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%.

[0144] Some compounds of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, may be excreted in urine. In some embodiments, the proportion 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%.

[0145] After absorption from the intestine, some compounds of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, may be circulated via the enterohepatic circulation. In some embodiments, the proportion 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%.

[0146] Some compounds of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, may cause renal excretion of bile salts. In some embodiments, the proportion 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%.

[0147] Some compounds of formula (I) or formula (II), or pharmaceutically acceptable salts thereof, may exhibit improved or optimal permeability. Permeability may 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 about 0.1×10 -6 cm / s greater than, for example, about 0.2×10 -6 cm / s greater than, for example, about 0.4×10 -6 cm / s greater than, for example, about 0.7×10 -6 cm / s greater than, for example, about 1.0×10 -6 cm / s greater than, for example, about 2×10 -6 cm / s greater than, for example, about 3×10 -6 cm / s greater than, for example, about 5×10 -6 cm / s greater than, for example, about 7×10 -6 cm / s greater than, for example, about 10×10 -6 cm / s greater than, for example, about 15×10 -6 cm / s greater than.

[0148] Some compounds of formula (I) or formula (II), or pharmaceutically acceptable salts thereof, may exhibit improved or optimal bioavailability. In some embodiments, the oral bioavailability is greater than about 5%, for example, greater than about 7%, for example, greater than about 10%, for example, greater than about 15%, for example, greater than about 20%, for example, greater than about 30%, for example, greater than about 40%, for example, greater than about 50%, for example, greater than about 60%, for example, greater than about 70% or for example, greater than about 80%. In other embodiments, the oral bioavailability is between about 10% and about 90%, for example, between about 20% and about 80%, for example, between about 30% and about 70% or for example, between about 40% and about 60%.

[0149] Some compounds of formula (I) or formula (II), or pharmaceutically acceptable salts thereof, may serve as substrates for relevant transporters in the kidney.

[0150] A method for preventing or reducing the entry of hepatitis B virus particles and / or hepatitis D virus particles into cells, the method comprising contacting the cells with an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) is also provided herein. A method for reducing hepatitis B and / or hepatitis D virus replication in cells, the method comprising contacting the cells with an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) is also provided herein. A method for preventing the binding of the preS1 domain of the HBV L protein to NTCP in cells, the method comprising contacting the cells with an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof) is also provided herein. In some embodiments, the contacting step is in vitro. In some embodiments, the contacting step is in vivo. In some embodiments, the cells are hepatocytes. In some embodiments, the contacting step is in vivo and the method comprises administering to the hepatocytes of a subject an effective amount of an NTCP inhibitor as described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a compound of formula (II), or a pharmaceutically acceptable salt thereof).

[0151] Definitions As used herein, the term "halo" refers to fluoro, chloro, bromo and iodo.

[0152] As used herein, "C 1~6 alkyl" refers to a straight or branched alkyl group having 1 to 6 carbon atoms, and "C 1~4 alkyl" refers to a straight 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.

[0153] As used herein, "C" 1~4 The term "haloalkyl" refers to a straight-chain 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.

[0154] As used herein, "C" 1~4 "alkoxy" and "C" 1~4 "alkylthio" refer to a straight-chain or branched C 1~4 alkyl group bonded through an oxygen or sulfur atom, respectively, to the remainder of the molecule.

[0155] 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.

[0156] 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.

[0157] 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-chain 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.

[0158] 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. The term "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. The term "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. The terms "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 a C 3~6 cycloalkylsulfonyl group, respectively.

[0159] Some compounds of formula (I) or formula (II), or pharmaceutically acceptable salts thereof, may have chiral centers and / or geometric isomeric centers (E- and Z-isomers). It should be understood that the present invention encompasses all such optical isomers, diastereoisomers and geometric isomers possessing ASBT and / or LBAT inhibitory activity. The present invention also encompasses all tautomers of the compounds of formula (I) or formula (II), or pharmaceutically acceptable salts thereof, possessing ASBT and / or LBAT inhibitory activity. Certain compounds of formula (I) or formula (II), or pharmaceutically acceptable salts thereof, may exist not only in the unsolvated form but also in solvated forms such as, for example, the hydrated form. It should be understood that the present invention encompasses all such solvated forms possessing ASBT and / or LBAT inhibitory activity.

[0160] 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.

[0161] Suitable pharmaceutically acceptable salts of the compounds disclosed herein are, for example, base addition salts of compounds 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 afford physiologically acceptable cations, such as salts with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.

[0162] As used herein, the terms "treating," "treat," and "treatment" refer to the regression, alleviation, delay in the onset, or inhibition of progression of a disease or disorder, or one or more symptoms thereof, as described herein. In some embodiments, the NTCP inhibitor is administered after one or more symptoms have developed. In other embodiments, the NTCP inhibitor may be administered in the absence of symptoms. For example, the NTCP inhibitor 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. Administration of the NTCP inhibitor may also be continued after symptoms have resolved, for example, to prevent or delay recurrence thereof.

[0163] As used herein, the terms "subject," "individual," or "patient" are used interchangeably and refer to any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, primates, and humans. In some embodiments, the subject is a human. In some embodiments, the subject has and / or exhibits at least one symptom of hepatitis B and / or hepatitis D.

[0164] As used herein, the term "contacting" refers to bringing together the indicated moieties in an in vitro or in vivo system. For example, "contacting" an NTCP with a compound provided herein includes administering the compound provided herein to a subject having NTCP, such as a human, and, for example, introducing the compound provided herein into a sample containing cells or a purified preparation containing NTCP.

[0165] As used herein, the term "about" refers to a value or parameter in the present specification that includes (and describes) embodiments directed to the value or parameter itself. For example, a description referring to "about 20" includes a description of "20". Numerical ranges include the numbers defining the ranges. Generally speaking, the term "about" refers to all values of a variable that are either greater than or within the experimental error (e.g., within the 95% confidence interval of the mean) of the indicated value or within 10 percent of the indicated value.

Example

[0166] (Example 1) Inhibition of NTCP and IBAT in vitro. The effect of (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 (Compound 1) on bile acid transport mediated by NTCP and IBAT was evaluated using overexpressing cell line systems.

[0167] Method NTCP assay procedure. 20,000 cells (human and mouse NTCP overexpressing cells) were seeded in a 96-well plate in 100 μL of MEM-alpha medium supplemented with 10% FBS containing geneticin (1 mg / mL) 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 250 μL of basal MEM-alpha medium (without FBS). After each wash, the plate was tapped against a paper towel to ensure maximum removal of residual medium. For human NTCP (hNTCP), test inhibitor dilutions (3-fold serial dilutions in DMSO, 10 concentrations) were made at 0.3 μM 3An incubation mix was prepared by adding H - taurocholic acid and 7.5 μM cold taurocholic acid to MEM - alpha (without FBS) (maintaining a final DMSO concentration of 0.2%). For mouse NTCP, test inhibitor dilutions (3 - fold serial dilutions in DMSO, 10 concentrations) were made at 0.3 μM 3 An incubation mix was prepared by adding H - taurocholic acid and 25 μM cold taurocholic acid to MEM - alpha (without FBS) (maintaining a final DMSO concentration of 0.2%). Then, 50 μL of the incubation mix containing the test inhibitor was added to the wells (in duplicate), and the plate was incubated at 37 °C for 20 minutes in a CO2 incubator. The wells were washed twice with 250 μL of cold, non - labeled 1 mM taurocholic acid dissolved in HEPES - buffered (10 mM) HBSS (pH 7.4).

[0168] After each wash, the plate was blotted on a paper towel and tapped to ensure maximum removal of the blocking buffer. 100 μL of MicroScint - 2 was added to the wells and held overnight at room temperature. Then, the plate was read in a PerkinElmer MicroBeta2 liquid Scintillation Counter under the I / L - BAT 3 H Test protocol (set at a reading time of 120 seconds per well in the normal plate orientation).

[0169] Assay procedure for IBAT. 10,000 cells (human and mouse IBAT overexpressing cells) were seeded in 96-well plates in 100 μL of MEM-alpha medium supplemented with 10% FBS containing puromycin (10 μg / mL) and incubated at 37 °C in 5% CO2 for 48 hours. After incubation, the medium was decanted from the wells and the cells were washed twice with 250 μL of basal MEM-alpha medium (without FBS). After each wash, the plate was tapped against a paper towel to ensure maximum removal of the residual medium. Test inhibitor dilutions prepared in DMSO (highest test concentration is 10 μM, 3-fold serial dilution, 10 concentrations) were added to the incubation mix containing 0.25 μM 3 H-taurocholic acid and 5 μM of cold taurocholic acid (maintaining a final DMSO concentration of 0.2%). Then, 50 μL of the incubation mix 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.

[0170] After incubation, the reaction was stopped by holding the plate on an ice-water mix for 2 - 3 minutes and then the incubation mix was completely aspirated from the wells. The wells were washed twice with 250 μL of cold unlabeled 1 mM taurocholic acid dissolved in HEPES-buffered (10 mM) HBSS (pH 7.4). The plate was tapped against a paper towel after each wash to ensure maximum removal of the blocking buffer. 100 μL of MicroScint-20 was added to the wells and after holding overnight at room temperature, the plate was read in a PerkinElmer MicroBeta2 liquid Scintillation Counter under the 3 I / L-BAT H Test protocol (set at a reading time of 120 seconds per well in the normal plate orientation).

[0171] Statistical Analysis The percent inhibition was calculated relative to the assay control. Further data analysis was performed using validated statistical software (GraphPad Prism) to determine the IC 50The value was calculated.

[0172] Result The IC 50 value of compound 1 in human NTCP was 0.53 ± 0.081 nM. The positive control, Myrcludex B (MyrB), behaved as predicted.

[0173] The IC 50 value of compound 1 in mouse NTCP was 1.82 ± 0.25 nM. The positive control, MyrB, behaved as predicted.

[0174] The IC 50 value of compound 1 in human IBAT was 535 ± 83.3 nM. The positive control, AS0075, behaved as predicted.

[0175] The IC 50 value of compound 1 in mouse IBAT was 40.8 ± 4.6 nM. The positive control, AS0075, behaved as predicted.

[0176] (Example 2) Inhibition of NTCP and HBV in vitro. The in vitro potency of compound 1, which inhibits bile acid transport, was evaluated.

[0177] Method Assay procedure in CHO cells. Stably hNTCP-transfected CHO cells were seeded on 24-well plates. After incubation overnight, the CHO cells were treated with 10 mM sodium butyrate to induce NTCP transporter expression. After 24 hours, the medium was aspirated and the cells were washed with warm Hank's balanced salt solution (HBSS) containing sodium (+Na). The cells were incubated at 37 °C for < 10 minutes with tracer doses of 5 μM, 10 μM, or 25 μM 3 [H] taurocholic acid in the presence of increasing doses of compound 1 (e.g., 0, 10 nM, 100 nM, 1 μM, 10 μM). The cells were processed for radioactivity. The apparent K iAnd the kinetic type (competitive, non-competitive, uncompetitive, mixed) was determined by complementary Dixon / Cornish-Bowden plot analysis and Lineweaver-Burk plot analysis.

[0178] In vitro evaluation of the time course of dissociation. This study was 3 conducted using the 5 [[H]]taurocholic acid uptake assay protocol. Stable transfected CHO cells expressing human NTCP (hNTCP) were seeded on 24-well plates in CHO cell medium (DMEM / F12 medium supplemented with 10% FBS, penicillin / streptomycin) with Geneticin (G418) added to maintain selection during culture at a density of 1.5 - 2.0×10

[0179] 3 For the 3 [[H]]taurocholic acid uptake assay, at the end of each washout period, the medium was aspirated and the cells were washed three times with warm (37°C) Hank's balanced salt solution (HBSS) containing sodium (Na+). The cells were then incubated on a slide warmer at 37°C for 10 minutes in HBSS containing 5 μM 3 [[H]]taurocholic acid. For the 0-minute time point, 1 μM of the test inhibitor was present during the 10-minute​3 The taurocholic acid + inhibitor mix was removed by aspiration. Cells were washed with ice-cold HBSS + choline (instead of Na+), and lysed with 0.1 N NaOH. Duplicate 100 μL aliquots of the lysate were transferred to vials containing scintillation fluid, neutralized, quenched overnight, and then counted. Cell-associated proteins were measured in duplicate using the BCA protein assay, and cell-associated radioactivity was normalized to protein content.

[0180] Results Compound 1 acted as a competitive inhibitor of transporter activity, and the inhibition constant values (K i ) were 15 nmol / L and 2410 nmol / L for human NTCP (Figure 2A) and ASBT (Figure 2B), respectively. In vitro experiments demonstrated that Compound 1 exhibited a long-lasting inhibition of BA transport activity of human NTCP (>50% inhibition at 6 hours after washout) (Figure 3A (Mylotarg) and Figure 3B (Compound 1)).

[0181] (Example 3) Binding of HBV / HDV-derived preS1 peptide to NTCP. The myristoylated preS1 domain containing amino acids 2 - 48 of the large HBV envelope is essential for viral binding to NTCP and can be used as a surrogate parameter for HBV / HDV viral binding to NTCP. In this in vitro study, the taurocholic acid transport via human NTCP and cynomolgus monkey (Macaca fascicularis) Ntcp, the binding of HBV / HDV preS1 peptide to human NTCP, and the efficacy and selectivity of Compound 1 in inhibiting HDV and HBV infection in vitro in NTCP-HepG2 cells were investigated.

[0182] Methods NTCP-HEK293 and NTCP-HepG2 cell lines. Human embryonic kidney (HEK293) cells were stably transfected with human NTCP tagged with a FLAG epitope at the C-terminus (herein referred to as NTCP-HEK293 cells). The cells were maintained at 37 °C, 5% CO2 and 95% humidity in DMEM / F-12 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA), 4 mM L-glutamine, and penicillin / streptomycin. HepG2 cells stably transfected with NTCP-FLAG (herein referred to as NTCP-HepG2) were cultured under the same conditions in DMEM containing all the supplements listed above except L-glutamine. For induction of the transgene, the medium was supplemented with 1 μg / mL tetracycline for NTCP-HEK293 cells or 2 μg / mL doxycycline for NTCP-HepG2 cells.See also Konig A et al. (2014) Kinetics of the bile acid transporter and hepatitis B virus receptor Na+ / taurocholate cotransporting polypeptide (NTCP) in hepatocytes. Journal of Hepatology 61:867-875; Kirstgen M et al. (2021) Hepatitis D Virus Entry Inhibitors Based on Repurposing Intestinal Bile Acid Reabsorption Inhibitors. Viruses 13(4):666; Kirstgen M et al. (2021) Identification of novel HBV / HDV entry inhibitors by pharmacophore- and QSAR-guided virtual screening. Viruses 13:1489; and Grosser G et al. (2021) Substrate Specificities and Inhibition Pattern of the Solute Carrier Family 10 Members NTCP, ASBT and SOAT. Front Mol Biosci 8:689757.

[0183] HEK293 cells transfected with cynomolgus NTCP. GripTite 293 MSR cells (hereafter referred to as HEK293 cells, Invitrogen), a modified HEK293 cell line expressing the human macrophage scavenger receptor for stronger adhesion, were maintained in DMEM / F-12 medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (Sigma), 4 mM L-glutamine (PAA), and penicillin / streptomycin at 37 °C, 5% CO2, and 95% humidity. For transport and peptide binding assays, HEK293 cells were transiently transfected with cynomolgus macaque Ntcp cDNA constructs. Transfection was performed using Lipofectamine 2000 (Thermo Fisher Scientific). See also Muller SF et al. (2018) Characterisation of the hepatitis B virus cross-species transmission pattern via Na+ / taurocholate co-transporting polypeptides from 11 New World and Old World primate species. PLoS One 13(6):e0199200.

[0184] 3 H]TC transport and 3 H]preS1 binding inhibition concentration (IC 50 ). In NTCP-HEK293 cells, bile acid transport measurements were performed using tritiated taurocholic acid (hereafter referred to as 3 H]TC) (20 Ci / mmol, 0.09 mCi / mL, Perkin Elmer, Waltham, USA). In parallel, a tritiated myr-preS12-48 lipopeptide-HBV genotype D3- purchased from Pharmaron (hereafter referred to as 3 ​Using [³H]preS1 (120 Ci / mmol, 1 mCi / mL, Cardiff, UK), a peptide binding experiment was carried out. Briefly, cells were seeded on 96-well plates coated with polylysine, induced with 1 μg / mL of tetracycline, and grown to confluence at 37 °C for 72 h. The cells were then washed once with phosphate buffered saline (PBS, 137 mM NaCl, 2.7 mM KCl, 1.5 mM KH₂PO₄, 7.3 mM Na₂HPO₄, pH 7.4) adjusted to 37 °C and pre-incubated with 80 μL of DMEM at 37 °C for 5 min. The medium was replaced with 80 μL of DMEM containing only each inhibitor concentration or solvent (100% uptake / binding control), and the cells were incubated for an additional 5 min at 37 °C. After this pre-incubation, the bile acid transport experiment was initiated by adding 20 μL of DMEM containing 5 μM [³H]TC (final concentration: 1 μM). The binding of [³H]preS1 was initiated by adding 20 μL of DMEM containing 25 nM [³H]preS1 (final concentration: 5 nM). After 10 min, the experiment was stopped by washing twice with ice-cold PBS. For the 0% uptake / binding control, NTCP-HEK293 cells were not induced with tetracycline (-tet). 3 The cellular associated radioactivity of [³H]TC or 3 [³H]preS1 was quantified by liquid scintillation counting in a Packard Microplate Scintillation Counter TopCount NXT (Packard Instrument Company, Meriden, USA). The transport rate and 3 [³H]preS1 binding were determined in counts per minute (cpm). Subtracting the mean of the 0% control, the net 3 [³H]TC transport rate and the net 3 [³H]preS1 binding rate were expressed as % relative to the control, respectively. The IC from quadruplicate determinations was calculated by GraphPad Prism 6 (GraphPad, San Diego, CA, USA). 3 [³H]preS1 binding was determined in counts per minute (cpm). Subtracting the mean of the 0% control, the net 3 [³H]TC transport rate and the net 3 [³H]preS1 binding rate were expressed as % relative to the control, respectively. The IC from quadruplicate determinations was calculated by GraphPad Prism 6 (GraphPad, San Diego, CA, USA). 50Values were calculated. See also Kirstgen et al., 2020, 2021, Grosser et al., 2021.

[0185] MTT cell cytotoxicity assay. Using the In Vitro Toxicology Assay Kit (Sigma-Aldrich), a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay was performed to measure the cytotoxicity of the test article according to the manufacturer's protocol. Briefly, NTCP-HepG2 cells were incubated with 100 μL of the indicated concentration of the test article dissolved in hepatocyte growth medium (HGM) at 37 °C for 6 h. After 6 h, the medium was replaced with inhibitor-free HGM and the cells were cultured for an additional 24 h. The medium was then removed, 100 μL of DMEM containing 0.5 mg / mL MTT was added, and the cells were incubated at 37 °C for 1 h. Finally, the medium was replaced with 100 μL of isopropyl alcohol (Sigma-Aldrich) and the samples were measured by the Glo-Max-Multi Detection System (Promega, Madison, WI, USA). See also Lowjaga KAA et al. (2020). Long-term trans-inhibition of the hepatitis B and D virus receptor NTCP by taurolithocholic acid. Am J Physiol Gastrointest Liver Physiol 320(1):G66–G80; and Kirstgen et al., 2021.

[0186] HDV infection in vitro. HDV (HDV genotype 1, covered by the HBV surface protein of genotype D, see the following HBV) production was performed in vitro as described in the literature (Rasche A et al. (2019) Highly diversified shrew hepatitis B viruses corroborate ancient origins and divergent infection patterns of mammalian hepadnaviruses. Proc Natl Acad Sci U S A 116(34):17007~17012; De Carvalho Dominguez Souza BF et al. (2018) A novel hepatitis B virus species discovered in capuchin monkeys sheds new light on the evolution of primate hepadnaviruses. Journal of Hepatology 68:1114~1122). RT-qPCR was performed to determine the HDV RNA genome equivalents. NTCP-HepG2 cells were pre-incubated for 5 minutes with compound 1 at concentrations of 100 nM, 200 nM, and 400 nM in HGM. Infection experiments were performed in NTCP-HepG2 cells. The cells were cultured in HGM consisting of William's E medium (Thermo Fisher Scientific) containing 2% bovine serum albumin (BSA, Roth), 2 mM L-glutamine, 100 μg / mL gentamicin, 10 nM dexamethasone, 1 mM sodium pyruvate, 1× insulin-transferrin-selenium, 2% DMSO, 4% polyethylene glycol, and 2 μg / mL doxycycline in 48-well plates. The HDV stock solution was diluted with HGM (HDV: 2×10 9GE / Well), and added to the cells for 6 hours. Then, the cells were washed with DMEM and cultured in HGM supplemented with 2% DMSO, 2% BSA, and 2 μg / mL doxycycline. The medium was changed every 3 days until the cells were fixed with 3% paraformaldehyde in PBS at room temperature (RT) for 30 minutes on the 9th day after infection. The cells were permeabilized with 0.2% Triton X 100 in PBS at room temperature for 30 minutes and blocked by incubating with 5% bovine serum albumin in PBS at room temperature for 30 minutes. Next, the cells were immunostained with purified human anti-HDV positive serum at 37 °C for 1 hour (1:400 dilution). For the detection of delta hepatitis antigen (HDAg), a goat anti-human IgG secondary antibody conjugated to an Alexa Fluor fluorophore (1:400 dilution, Thermo Fisher Scientific) was added at 37 °C for 1 hour. The nuclei were stained with Hoechst 33342 (1 μg / mL, Thermo Fisher Scientific). The number of infected cells per well was determined by fluorescence microscopy.

[0187] HBV infection in vitro. In the first study, NTCP-HepG2 cells were inoculated with the same HBV genome equivalents / cell for 16 hours. HBV (genotype D) was generated in vitro as described in the literature (Konig et al., 2014). Infection was carried out in HGM supplemented with 2% DMSO and 4% polyethylene glycol (5×10 9GE / Well's HBV). Subsequently, the cells were washed twice with HGM and cultured in HGM supplemented with 2% DMSO and 2% FCS until day 10 post-infection. At 10 days post-infection, fixation was performed with 3.7% formaldehyde and 1% methanol at 4°C for 30 minutes. The cells were permeabilized with 0.2% Triton X100 in PBS at room temperature for 20 minutes. Nonspecific binding epitopes were blocked by incubating with 10% fetal bovine serum in PBS at 37°C for 45 minutes. For the detection of HBV core (HBc) protein expression, the cells were incubated with polyclonal rabbit anti-HBcAg antiserum (1:500 dilution, Dako, Hamburg, Germany) in PBS at 37°C for 2 hours, and then incubated with anti-rabbit IgG AlexaFluor594 (1:200 dilution in PBS, Immuno Jackson) at 37°C for 1 hour. The nuclei were stained with DAPI (10 μg / mL) in PBS. The number of infected cells per well was determined by fluorescence microscopy. In addition, HBeAg was quantified in the supernatant from day 7 to day 11 post-infection.

[0188] In the second study, recombinant HBV (genotype D) was generated in vitro as described in the literature (Konig et al., 2014). For the infection experiment, NTCP-HepG2 cells (2×10 5 cells) were pre-incubated with compound 1 or milk thistle extract B at concentrations of 1, 3, 10, 30, 100, 300, and 1000 nM in HGM for 5 minutes. See Figures 7D - 7F. The myr-preS12-48 peptide (500 nM, genotype D) was used as a control inhibitor. All infection experiments were performed in NTCP-HepG2 cells cultured in HGM in the presence of the corresponding compounds with different concentrations as the final concentration, 4% PEG-8000 (Sigma-Aldrich, Darmstadt, Germany), 2% DMSO (Carl Roth, Karlsruhe, Germany), and 100 ng / ml EGF (PeproTech, Hamburg, Germany). Recombinant HBV was added to HGM at 2.5×10 10 HBV genome copies (7×10 per well)8 It was diluted to the final concentration of IU) and added to the cells for 6 hours. Subsequently, the cells were washed with HGM and cultured in HGM supplemented with 2% DMSO. The medium was changed every 2 - 3 days until the cells were fixed with 3.8% paraformaldehyde (Sigma - Aldrich) in PBS at room temperature for 30 minutes on the 9th day after infection. The cells were immunostained at 37°C for 1 hour (dilution 1:400). For the detection of HBV core (HBc) protein expression, polyclonal anti - HBc antiserum derived from immunized guinea pigs (dilution 1:500, Eurogentec, Seraing, Belgium) was used as the primary antibody, and Alexa Fluor 488 - conjugated goat anti - guinea pig IgG antibody (dilution 1:400, Thermo Fisher Scientific) was used as the secondary antibody. The nuclei were stained with DAPI (0.5 μg / ml, Thermo Fisher Scientific). The analysis of immunofluorescence was performed using an ImageXpress Pico automated cell imaging system (Molecular Devices, San Jose, USA). HBV - infected cells secrete soluble HBeAg, which is a widely recognized quantitative marker for productive HBV infection in cell culture experiments. In this study, the Architect HBeAg assay (Abbott Laboratories, Wiesbaden, Germany), an automated in vitro diagnostic system, was used to determine the HBeAg secreted from infected cells into the cell culture supernatant for the supernatants collected from the 5th day to the 9th day after infection.

[0189] Statistics. IC 50 The determination of the IC value was performed by non - linear regression analysis using the equation log(inhibitor) vs. response setting of GraphPad Prism 6.0 software (GraphPad). 3 H]TC transport and 3Data on preS1 binding are presented as mean ± SD from quadruplicate determinations. Infection studies were performed in triplicate, and data are presented as mean ± SD. Statistical analysis of HBV / HDV infection experiments was performed by two-way ANOVA followed by Dunnett's multiple comparison test using GraphPad Prism 9.0, and p < 0.01 was considered statistically significant.

[0190] Results Inhibition of human NTCP by compound 1. Compound 1 3 H]taurocholic acid uptake (half-maximal inhibitory concentration [IC 50 , 186 nmol / L; Figure 4A) and 3 H]preS1 peptide binding (IC 50 , 149 nmol / L; Figure 4C) in a concentration-dependent manner. As a control inhibitor, 500 nM preS1 peptide was used in both assays, which significantly inhibited both assays as predicted and as reported in the literature (see König et al., 2014, Müller et al., 2018, Lowjaga et al., 2021; Figure 4B and Figure 4D).

[0191] Inhibition of cynomolgus Ntcp by compound 1. In addition, compound 1 was used as an inhibitor against cynomolgus NTCP. This transporter was transiently transfected into HEK293 cells, 3 and the effects on [H]TC transport inhibition were analyzed at increasing concentrations of 10, 25, 63, 158, 398, and 1000 nM. As shown in Figure 5A and Figure 5B, compound 1 potently inhibited bile acid transport via cynomolgus Ntcp in a concentration-dependent manner, with an IC 50 of 200 nM.

[0192] Cytotoxicity evaluation of compound 1 in NTCP-HepG2 cells. The cytotoxicity evaluation of compound 1 was performed in HepG2 cells in the concentration range of 1 nM to 100 μM. As shown in Figure 6, compound 1 did not show a cytotoxic effect in HepG2 cells.

[0193] Inhibition of HBV / HDV infection in vitro by Compound 1. The in vitro inhibition of HBV / HDV infection by Compound 1 was analyzed in human NTCP-HepG2 hepatoma cells, which represent a well-established cell culture model for in vitro HBV / HDV infection (see König et al., 2014).

[0194] In the first study, different concentrations (100 nM, 200 nM, and 400 nM) of Compound 1 were used before and during incubation of the viral inoculum with the cells. Compound 1 reduced the number of HBcAg-positive cells by 75% - 90% (Figure 7A), the number of HDAg-positive cells by 40% - 60% (Figure 7B), and the HBeAg levels in the supernatant by 83% - 87% (Figure 7C). A similar approach using the preS1 peptide (amino acids 2 - 48, genotype D, 500 nM) resulted in almost complete inhibition of HDV / HBV infection.

[0195] In the second study, NTCP-HepG2 cells were pre-incubated for 5 minutes with Compound 1 at concentrations of 1, 3, 10, 30, 100, 300, and 1000 nM. HBeAg secretion, even at the highest inhibitor concentration of 1000 nM for Compound A2342, exceeded the cut-off value (Figures 7D - 7G). Nevertheless, IC 50 values could be calculated. Compound 1 inhibited in vitro HBV infection with IC 50 values of 19.1 nM (HBeAg) (Figures 7D and 7E) and 16.1 nM (HBc) (Figures 7F and 7G).

[0196] Discussion The dual 3 H]TC transport and 3 H]preS1 peptide binding assays described in this example have been used to characterize inhibitors of human NTCP with respect to their inhibitory potency and selectivity against viral binding (see Kirstgen et al., 2020, 2021). Since the same assay with the same parameters was used in this example, the measured IC of Compound 150 The value can be directly compared with the published data. Compound 1 had the same potency as the viral preS1 peptide as an inhibitor and was much more potent than troglitazone, a well-established NTCP inhibitor.

[0197] At the selected concentrations, Compound 1 showed a strong inhibitory effect against HBV and HDV infections. Compound 1 potently blocked the entry of HBV and HDV viral particles in NTCP-HepG2 cells in the nanomolar range.

[0198] (Example 4) Effect of Compound 1 on HBV infection in human hepatocytes in vitro. The antiviral effect of Compound 1 was evaluated using HBV-infected human hepatocytes in vitro.

[0199] Method Incubation of hepatocytes. A single vial of primary human hepatocytes (PHH) was seeded onto 48-well collagen-coated plates according to the protocol provided by the hepatocyte provider (BioIVT). The cells were seeded at 0.14×10 6 cells per well in a total volume of 200 μL. The cells were incubated overnight at 37 °C in an environment of 5% CO2 using InVitroGro HI medium containing Torpedo antibiotic mix. Unless otherwise described, the diluted compound was added to the cells 18 hours before infection (standard conditions).

[0200] The multiplicity of infection (MOI) is defined as the number of HBV genome equivalents added per cell in each growth format as determined by qPCR quantification. The overnight medium was removed from the cells and replaced with 190 μL of fresh serum-free HI medium / 4% PEG8000 and 10 μL of diluted virus stock solution (HepG2 AD38 genotype D) such that the MOI was 500. For the parallel cytotoxicity assessment, medium only was added to the cells containing the compound. Eighteen hours after virus infection, the cells were gently washed 5 times with DPBS, and subsequently, 200 μL of HI medium containing the compound was freshly added on days 1, 4, and 7 post-infection. In one set of experiments (study number 01), the effect of giving the compound 18 hours before infection was studied as opposed to 18 hours after infection. The supernatant removed from the cytotoxicity plates at each medium change was stored at -20 °C for shipment to the customer. To quantitatively determine the total extracellular HBV DNA copy number by qPCR, the supernatant was collected at 10 days post-infection and stored at -20 °C. For the cytotoxicity plates, cell viability was measured with XTT tetrazolium dye on day 10.

[0201] Quantitative PCR detection of total HBV DNA in the supernatant. On the 10th day, 10 microliters (10 μL) of cell culture supernatant from the collected sample was diluted in 90 μL of buffer consisting of 10 mM Tris and 40 μg / mL sheared salmon sperm DNA and boiled for 15 minutes. In a 384-well plate, quantitative PCR (qPCR) was performed using the Bio-Rad CFX384 Touch Realtime-PCR Detection System and the supporting CFX Manager software. For each sample, 5 microliters (5 μL) of the diluted and boiled cell culture supernatant and 10-fold serial dilutions of the quantitative DNA standard were subjected to real-time qPCR using Platinum Quantitative PCR SuperMix-UDG (Invitrogen) and specific DNA oligonucleotide primers (IDT, Coralville, IA) HBV-AD38-qF1 (5'-CCG TCT GTG CCT TCT CAT CTG-3'), HBV-AD38-qR1 (5'-AGT CCA AGA GTY CTC TTA TRY AAG ACC TT-3'), and HBV-AD38-qP1 (5'-FAM-CCG TGT GCA / ZEN / CTT CGC TTC ACC TCT GC-3'BHQ1) at a final concentration of 0.2 μmol / L for each primer in a total reaction volume of 25 μL. The amount of HBV DNA in each sample was interpolated from the standard curve, and the data was imported into an Excel spreadsheet for analysis.

[0202] Cytotoxicity. After incubating for 10 days at 37 °C in a 5% CO2 incubator, the test plates were stained with the tetrazolium dye XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide). XTT-tetrazolium was metabolized to a soluble formazan product by mitochondrial enzymes of metabolically active cells. The XTT solution was prepared daily as a 1 mg / mL stock solution in RPMI1640. The phenazine methosulfate (PMS) solution was prepared at 0.15 mg / mL in PBS and stored at -20 °C in the dark.

[0203] Just before use, an XTT / PMS stock solution was prepared by adding 40 μL of PMS per 1 mL of XTT solution. 50 microliters of XTT / PMS was added to each well of the plate, and the plate was reincubated at 37 °C for 4 hours. The plate was sealed with an adhesive plate sealer and gently shaken or inverted several times to mix the soluble formazan product, and the plate was read spectrophotometrically at 450 / 650 nm using a Molecular Devices Vmax plate reader.

[0204] Experimental design. The experiment was designed as shown in Table 3.

[0205] [Table 3]

[0206] Data analysis. Each concentration of inhibitor and control was analyzed in triplicate for HBV DNA copy and cytotoxicity. From the control wells without inhibitor, the average of these three replicates was calculated, setting the HBV DNA average to 100% virus control and the XTT average to 100% cell viability. The effect of the compound was then expressed as the % of virus control and % of cell viability, respectively.

[0207] In two sets of experiments, the effect of combining compound 1 with tenofovir was studied. The effect of the drug combination was calculated based on the activities of the two compounds when tested alone. The predicted additive antiviral protection (based on the dose - response curves of the individual drugs used alone) was subtracted from the experimentally determined antiviral activity at each combination concentration, resulting in a positive value (synergy), a negative value (antagonism), or zero (additivity). Assuming that the compounds inhibited HBV replication by acting at the same site, the data were analyzed by the most rigorous statistical means. The results of the combination assay were expressed as the synergy volume μM 2 expressed as a percentage using the MacSynergy II template at the 95% confidence interval.

[0208] For these studies, synergy was defined as a drug combination that resulted in a synergy volume of more than 50 μM 2 %. Slightly synergistic activity and highly synergistic activity were defined as resulting in synergy volumes of 50 - 10 μM 2 % and > 100 μM 2 %, respectively.

[0209] Synergy volumes between - 50 and 50 μM 2 % were considered additive, and synergy volumes less than - 50 μM 2 % were considered antagonistic.

[0210] Results Study number 01. The EC 50 values of tenofovir and compound 1 when the compounds were given 18 hours before infection (standard conditions) are summarized in Table 4 and illustrated in Figure 8A. Tenofovir demonstrated a predicted potency (EC 50 : 43 nmol / L) consistent with past values. Milk thistle B (EC 50 : 0.4 nmol / L) and compound 1 (EC 50: 4 nmol / L) was more potent than tenofovir. None of the compounds induced cytotoxicity at all concentrations tested (highest concentration: 1 μmol / L). See Figure 8C.

[0211] The EC 50 values of tenofovir, compound 1, and milciclib when the compounds were given 18 hours after infection are summarized in Table 4 and illustrated in Figure 8B. The potency of tenofovir was maintained when the compound was given 18 hours after infection and was consistent with the inhibition of HBV replication. In contrast, when compound 1 and milciclib were given 18 hours after infection, antiviral activity was lost, which was consistent with the anti-entry mechanism.

[0212] Study number 02. The effect of combining compound 1 with tenofovir was also studied. The EC 50 values of tenofovir and compound 1 are summarized in Table 4. Tenofovir demonstrated a predicted potency (EC 50 : 64 nmol / L) consistent with past values. Compound 1 was more potent than tenofovir, and there was 58% inhibition at the lowest concentration tested (6.25 nmol / L), so the exact EC 50 could not be determined. None of the compounds induced cytotoxicity at all concentrations tested (highest concentration: 0.1 - 0.2 μmol / L). The combination of the two drugs resulted in a very potent antiviral effect. Since compound 1 was very effective by itself, it was a challenge to determine whether the combined effect was additive or synergistic, and further studies were conducted.

[0213] Study number 03. The effect of combining tenofovir with a lower concentration of compound 1 than in study number 02 was studied. The EC 50 values of tenofovir and compound 1 when given alone are summarized in Table 4 and illustrated in Figure 8D. Tenofovir demonstrated a predicted potency (EC 50: 59 nmol / L) was demonstrated. However, in contrast to previous experiments, the maximum inhibitory effect of tenofovir was approximately 60%. Compound 1 was more potent, and the EC 50 was 11 nmol / L. The maximum inhibitory effect of Compound 1 was also approximately 60%. When combined, the data suggested that Compound 1 and tenofovir exerted an additive effect.

[0214]

Table 4

[0215] Compound 1 consistently inhibited HBV infection in primary human hepatocytes in a concentration-dependent manner. Compound 1 did not affect cell viability at all concentrations tested (up to 100 μmol / L). When given together with tenofovir disoproxil fumarate, Compound 1 produced an additive antiviral effect.

[0216] The antiviral effects of Compound 1 from Study No. 01, Study No. 02, and Study No. 03 are summarized in Figure 9. The data fit well to a non-linear four-parameter regression curve (R 2 : 0.93). From the fitted curve, an EC 50 of 4.9 nmol / L was determined.

[0217] (Example 5) Effect of Compound 1 in HBV-infected PXB mice The plasma exposure after oral administration of Compound 1 to humanized non-infected PXB mice and the prophylactic anti-HBV effect of Compound 1 after oral administration to PXB mice before HBV infection were evaluated.

[0218] Method Pharmacokinetic phase study design. Using Compound 1, a study was conducted according to the experimental design as shown in Table 5.

[0219]

Table 5

[0220] Research design of the main study The study was conducted according to the experimental design shown in Table 6.

[0221]

Table 6

[0222] Blood sample collection in the pharmacokinetic phase From all animals, 30 microliters (30 μL) of blood was collected under isoflurane anesthesia via the retro-orbital plexus / venous sinus using a heparin sodium-coated graduated pipette according to the collection schedule shown in Table 7.

[0223]

Table 7

[0224] Individual animal blood samples were transferred to labeled microtubes and stored under refrigerated conditions until centrifugation at 3500 × g for 10 minutes at 5 °C to obtain plasma. Individual plasma samples were transferred to separately labeled microtubes and snap-frozen in liquid nitrogen.

[0225] Blood sample collection in the main study From all surviving animals, the target volume of blood was collected under isoflurane anesthesia via the retro-orbital plexus / venous sinus using Intramedic (trademark) Polyethylene Tubing at each time point. 2 microliters (2 μL) of the collected blood was used for human albumin measurement. The remaining blood was centrifuged to separate the serum used for the analysis of the indicated biomarkers. Blood samples were collected according to Table 8.

[0226]

Table 8

[0227] Individual blood samples from the animals were allowed to clot at room temperature for at least 5 minutes and then centrifuged at 13,200 × g at 4 °C for 3 minutes to obtain serum. The serum was aliquoted into separate labeled microtubes and stored at -80 °C until analysis.

[0228] Necropsy and tissue sampling. The entire liver was excised from all animals and weighed at necropsy. Liver samples from the left lateral lobe were immersed in RNAlater solution (registered trademark), incubated overnight, and stored at -80 °C until use.

[0229] Bioanalysis. The plasma exposure of compound 1 from the pharmacokinetic phase was measured by LC / MS-MS. The lower limit of quantification was 1 ng / mL.

[0230] Plasma human albumin concentration. The plasma human albumin concentration was measured using latex agglutination immunoturbidimetry with a clinical chemistry analyzer (BioMajesty (trademark) series JCA-BM6050, JEOL Ltd., Tokyo, Japan).

[0231] Serum HBV DNA. Using real-time PCR detection, the HBV DNA concentration in serum was measured using the KUBIX HBV qPCR Kit (KUBIX Inc.) and the CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad Laboratories, Inc., Hercules, CA, USA). 10 microliters (10 μL) of the HBV 2× PCR solution was added to 10 μL of the heated sample. The initial activation was performed at 95 °C for 2 minutes. Subsequent PCR amplification consisted of 45 cycles of denaturation at 95 °C for 5 seconds and annealing and extension at 54 °C for 30 seconds per cycle in the CFX96 Touch™ Real-Time PCR Detection System. The average serum HBV DNA level was calculated from the values of two separate wells. The primers and probes are shown in Table 9 (Table 9).

[0232]

Table 9

[0233] The lower limit of quantification of this assay was 4×10 4 copies / mL of serum.

[0234] HBsAg in serum. The HBsAg concentration in serum was determined by SRL, Inc. (Tokyo, Japan) based on the Chemiluminescent Enzyme Immuno Assay (CLEIA) (LUMIPULSE HBsAg-HQ, LUMIPULSE® PrestoII) developed by Fujirebio. The dilution factors were 30 and 10, and the measurement range of this assay was between 0.005 and 150 IU / mL. For the 30-fold diluted samples, the measurement range was adjusted to be between 0.15 and 4500 IU / mL. Also, for the 10-fold diluted samples, the measurement range was adjusted to be between 0.05 and 1500 IU / mL. The lower limit of quantification of this assay was 0.15 IU / mL of serum.

[0235] HBeAg in serum. The HBeAg concentration in serum was determined by SRL, Inc. based on the Chemiluminescent Enzyme Immuno Assay (CLEIA) (LUMIPULSE HBeAg, LUMIPULSE® PrestoII) developed by Fujirebio. The dilution factors were 30 and 10, and the measurement range of this assay was between 0.1 and 1590 C.O.I. For the 30-fold diluted samples, the measurement range was adjusted to be between 3 and 47700 C.O.I. For the 10-fold diluted samples, the measurement range was adjusted to be between 1 and 15900 C.O.I. The lower limit of quantification of this assay was 3.0 C.O.I.

[0236] HBV DNA in the liver. HBV DNA was extracted from frozen RNALATER®-preserved liver tissue using DNEASY® Blood & Tissue Kits (Qiagen K.K., Tokyo, Japan). The liver HBV DNA concentration (expressed as copies / 100 ng DNA) was determined using TaqMan Fast Advanced Master Mix and CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The DNA was dissolved in 200 μL of nuclease-free water, and then the concentration of the DNA solution was determined using a SPECTRAMAX® ABS Plus microplate reader. The concentration of the DNA solution was adjusted to 20 ng / μL using nuclease-free water.

[0237] The standard substance for liver HBV DNA quantification was extracted from 5 μL of serum using the SMITEST EX-R&D Nucleic Acid Extraction Kit (MEDICAL & BIOLOGICAL LABORATORIES CO., LTD., Nagoya, Japan). The DNA was dissolved in 20 μL of nuclease-free water.

[0238] The liver HBV DNA concentration (expressed as copies / 100 ng DNA) was determined using TaqMan Fast Advanced Master Mix and the CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The PCR reaction mixture was added to 5 μL of the extracted DNA. The PCR reaction mixture was added to 5 μL of the extracted DNA. Uracil-N-glycosylase was first activated at 50°C for 2 minutes, followed by activation of the polymerase at 95°C for 20 seconds. Subsequent PCR amplification consisted of 53 cycles of denaturation at 95°C for 3 seconds and annealing and extension at 60°C for 32 seconds per cycle in the CFX96 Touch™ Real-Time PCR Detection System. The mean hepatitis HBV DNA level was calculated from the values of two separate wells.

[0239] The lower limit of quantification of this assay was 50 copies / 100 ng DNA in the extracted DNA solution.

[0240] Intrahepatic cccDNA. Using real-time detection PCR, the intrahepatic HBV cccDNA concentration was measured using TaqMan Fast Advanced Master Mix and the CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The PCR reaction mixture was added to 5 μL of the extracted DNA. The PCR reaction was carried out based on the conditions of Takkenberg. Uracil-N-glycosylase was first activated at 50 °C for 2 minutes, followed by activation of the polymerase at 95 °C for 20 seconds. The subsequent 55 cycles of PCR amplification were carried out in the CFX96 Touch™ Real-Time PCR Detection System at 95 °C for 3 seconds per cycle and annealing and extension at 60 °C for 32 seconds. The average HBV cccDNA level was calculated from the values of two separate wells. The primers and probes are shown in Table 10 (Table 10).

[0241]

Table 10

[0242] The lower limit of quantification of this assay was 100 copies / 100 ng DNA in the extracted DNA solution.

[0243] Results Exposure of compound 1 after oral administration to non-infected PXB mice. Table 11 (Table 11) summarizes the data from mice exposed to 10 mg / kg p.o., while Table 12 (Table 12) summarizes the data from mice exposed to 30 mg / kg p.o. The plasma vs. time profile is also shown in Figure 10. In both groups, compound 1 was detected in plasma throughout the 24-hour period. The plasma exposure of compound 1 increased with increasing dose.

[0244]

Table 11

[0245]

Table 12

[0246] Effect of Compound 1 in HBV-infected PXB mice On day 16, it was found that animal number 102 had died, and on day 23, animal number 104 had died. Both were from the vehicle-treated group. The cause of death was considered to be intestinal hemorrhage. On day 33, it was found that animal 203 from the 10 mg / kg Compound 1 group had died. The cause of death was considered to be metastatic thymoma, which has been reported as a spontaneous change in SCID mice. No notable observations regarding Compound 1 were found.

[0247] Body weight Body weight remained constant throughout the study. No differences were found between the groups.

[0248] Human albumin blood concentration During the treatment period, the blood concentration of human albumin was stable in animals treated with Compound 1 (Compound 1 at 10 mg / kg, baseline: 10.8 ± 1.0 mg / mL, day 14: 10.8 ± 0.9 mg / mL; Compound 1 at 30 mg / kg, baseline: 10.6 ± 1.5 mg / mL, day 14: 11.3 ± 1.2 mg / mL), while it tended to decrease in the vehicle-treated group (baseline: 10.7 ± 0.9 mg / mL, day 14: 8.1 ± 2.6 mg / mL). The decrease in the vehicle group was mainly due to the discovery that two animals had died on days 16 and 23 thereafter.

[0249] HBV DNA in serum The overview of HBV DNA concentration is illustrated in FIGS. 11A and 11B. HBV DNA was detected on day 7 and was lower in mice treated with 30 mg / kg of Compound 1 compared to the control. The lower HBV DNA levels in response to Compound 1 were maintained throughout the study until the planned end on day 42, despite the treatment being stopped on day 14 as planned.

[0250] HBsAg in serum The overview of HBsAg concentration is illustrated in FIGS. 12A and 12B. HBsAg was detected on day 7 and was lower in mice treated with 10 mg / kg and 30 mg / kg of Compound 1 compared to the control. The lower HBsAg levels in response to both doses of Compound 1 were maintained throughout the study until the planned end on day 42, despite the treatment being stopped on day 14 as planned.

[0251] HBeAg in serum The overview of HBeAg concentration is illustrated in FIG. 12C. HBeAg was detected on day 14 (the first measurement time point) and was lower in mice treated with 10 mg / kg and 30 mg / kg of Compound 1 compared to the control. The lower HBeAg levels in response to both doses of Compound 1 were maintained throughout the study until the planned end on day 42, despite the treatment being stopped on day 14 as planned.

[0252] Liver HBV DNA and HBV cccDNA levels The overview of liver HBV DNA and HBV cccDNA levels at the planned end on day 42 can be seen in Table 13. There was no difference between the treatment groups.

[0253]

Table 13

[0254] Discussion In non-infected PXB mice, compound 1 was detected in plasma throughout a 24-hour period when administered at 10 and 30 mg / kg (p.o.). The plasma exposure of compound 1 increased with increasing dose. Compound 1 had a prophylactic effect against HBV genotype C infection in PXB mice. The serum levels of HBsAg (Figures 12A and 12B), HBeAg (Figure 12C), and HBV DNA (Figures 11A and 11B) were lower, particularly during the 14-day treatment period, in mice treated with 10 mg / kg of compound 1 (n = 4) and in mice treated with 30 mg / kg of compound 1 (n = 4) compared to vehicle-treated mice (n = 2). After treatment, the markers of HBV infection remained consistently lower in mice treated with 30 mg / kg of compound 1 compared to those treated with 10 mg / kg of compound 1 and the vehicle control group.

[0255] (Example 6) Characterization of Compound 2 Sodium taurocholate cotransporting polypeptide (NTCP) is a hepatocyte sinusoidal membrane bile acid (BA) transporter. In addition to its major role in hepatocyte clearance of BA, NTCP is used by hepatitis B and D viruses (HBV, HDV) to enter and infect human hepatocytes. The effect of compound 2 on the inhibition of BA uptake and HBV entry was evaluated.

[0256] Methods Human-derived NTCP and the apical sodium-dependent BA transporter (ASBT), a related BA transporter, were expressed in cell lines. 3 Using [H]taurocholic acid as a substrate, inhibition by compound 2 was studied. Cryopreserved human hepatocytes infected with a clinical isolate of HBV were used in the infection experiment. Two female non-human primates (NHPs) were force-fed compound 2 once daily for 5 days at dose levels of compound 2 in the range of 3 - 30 mg / kg. Total serum, urinary, and fecal BAs were analyzed enzymatically, and the plasma concentration of compound 2 was evaluated by liquid chromatography-tandem mass spectrometry. The oral pharmacokinetics of compound 2 were also studied in mice, rats, and dogs.

[0257] Results Compound 2 had maximum half-maximal inhibitory concentration (IC 50 ) values of 3.4 and 193 nmol / L against human NTCP and ASBT, respectively. Compound 2 prevented HBV infection of human hepatocytes at an IC 50 of 13.5 nmol / L without affecting cell viability. In NHPs, the plasma exposure of compound 2 increased with dose, and serum BA increased dose-dependently in response to oral administration of compound 2 on both day 1 and day 5. Doses of 10 and 30 mg / kg of compound 2 induced an increase in serum BA that persisted for at least 8 hours on both day 1 and day 5. Serum BA returned to baseline 24 hours after administration in response to 10 mg / kg of A7387, while it remained elevated in response to 30 mg / kg of compound 2. Urinary excretion of BA tended to increase in response to compound 2 compared to vehicle, while fecal BA levels did not change. Levels of 7-alpha-hydroxy-4-cholesten-3-one (C4), a BA synthesis biomarker, did not change in response to compound 2 after 5 days of repeated dosing. Good oral bioavailability was demonstrated in pharmacokinetic studies across all species. Compound 2 is a very potent, selective, and orally administrable NTCP inhibitor with potential in HBV / HDV infection and cholestatic diseases.

[0258] Other embodiments Although the present disclosure has been described in conjunction with its detailed description, it should be understood that the foregoing description is intended to illustrate and not limit the scope of the disclosure, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. The compound (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, or a pharmaceutically acceptable salt thereof, for use in the treatment of hepatitis B (HBV) in a subject.

2. The compound (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, or a pharmaceutically acceptable salt thereof, for use in the prevention or reduction of the entry of hepatitis B virus particles into hepatocytes in a subject.

3. The compound (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, or a pharmaceutically acceptable salt thereof, for use in the reduction of hepatitis B virus replication in hepatocytes in a subject.

4. The compound for use according to any one of claims 1 to 3, wherein the subject has hepatitis D.

5. The compound (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, or a pharmaceutically acceptable salt thereof, for use in the treatment of hepatitis D (HDV) in a subject.

6. The compound (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, or a pharmaceutically acceptable salt thereof, for use in the prevention or reduction of the entry of hepatitis D virus particles into hepatocytes in a subject.

7. For use in reducing hepatitis D virus replication in hepatocytes in a subject, the compound (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, or a pharmaceutically acceptable salt thereof.

8. A compound for use according to any one of claims 1 to 7, wherein the use further comprises administering a further antiviral agent.

9. For use in treating HBV in a subject, the compound (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, or a pharmaceutically acceptable salt thereof, and a further antiviral agent.

10. For use in treating HDV in a subject, the compound (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, or a pharmaceutically acceptable salt thereof, and a further antiviral agent.

11. A compound for use according to any one of claims 8 to 10, wherein the further antiviral agent is selected from the group consisting of entecavir, tenofovir, tenofovir disoproxil, tenofovir alafenamide, lamivudine, adefovir, adefovir dipivoxil, telbivudine, brequinar, interferon, and combinations thereof.

12. A compound for use according to claim 11, wherein the interferon is pegylated interferon, interferon alpha, or a combination thereof.

13. A compound for use according to any one of claims 8 to 11, wherein the further antiviral agent is tenofovir disoproxil.

14. A compound for use according to any one of claims 5 to 8 and 10 to 13, wherein the subject has hepatitis B.

15. A compound for use according to any one of claims 1 to 4 and 14, wherein the subject has chronic hepatitis B.

16. A compound for use according to any one of claims 5 to 8 and 10 to 15, wherein the subject has chronic hepatitis D.

17. A compound for use according to any one of claims 1 to 16, wherein the concentration of one or more biomarkers selected from HBV DNA, hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg), hepatitis B e antigen (HBeAg), HDV DNA, and hepatitis D antigen (HDAg) in the serum of a subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof.

18. A compound for use according to claim 17, wherein the concentration of HBV DNA in the serum of a subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof.

19. A compound for use according to claim 17, wherein the concentration of HBV DNA is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

20. A compound for use according to claim 18, wherein the concentration of HBV DNA in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, is decreased compared to a reference concentration of HBV DNA.

21. The reference concentration of HBV DNA is the level of HBV DNA in a serum sample obtained from a subject prior to administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to claim 19.

22. The concentration of HBV DNA in the serum of the subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to any one of claims 17 to 20.

23. The concentration of HBV DNA in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to any one of claims 17 to 21.

24. The concentration of HBV DNA in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to any one of claims 17 to 23.

25. The concentration of hepatitis B surface antigen (HBsAg) in the serum of the subject is reduced after administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to any one of claims 17 to 24.

26. The compound for use according to claim 25, wherein the concentration of HBsAg is determined in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

27. (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, or a pharmaceutically acceptable salt thereof, wherein the concentration of HBsAg in a serum sample derived from a subject obtained after administration thereof is decreased as compared with the reference concentration of HBsAg, the compound for use according to claim 26.

28. The reference concentration of HBsAg is the concentration of HBsAg in a serum sample obtained from a subject before administration of (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, or a pharmaceutically acceptable salt thereof, the compound for use according to claim 27.

29. The compound for use according to any one of claims 25 to 28, wherein the concentration of HBsAg in the serum of the subject is decreased by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

30. The compound for use according to any one of claims 25 to 29, wherein the concentration of HBsAg in the serum of the subject is decreased by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

31. A compound for use according to any one of claims 25 to 30, wherein the concentration of HBsAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

32. A compound for use according to any one of claims 17 to 31, wherein the concentration of hepatitis B core antigen (HBcAg) in the serum of the subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof.

33. A compound for use according to claim 32, wherein the concentration of HBcAg is determined in a sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

34. A compound for use according to claim 33, wherein the concentration of HBcAg in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, is decreased compared to the reference concentration of HBcAg.

35. A compound for use according to claim 34, wherein the reference concentration of HBcAg is the concentration of HBcAg in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

36. A compound for use according to any one of claims 32 to 35, wherein the concentration of HBcAg in the serum of the subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

37. A compound for use according to any one of claims 32 to 36, wherein the concentration of HBcAg in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

38. A compound for use according to any one of claims 35 to 37, wherein the concentration of HBcAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

39. A compound for use according to any one of claims 17 to 38, wherein the concentration of hepatitis B e antigen (HBeAg) in the serum of the subject is reduced after administration of (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, or a pharmaceutically acceptable salt thereof.

40. A compound for use according to claim 39, wherein the concentration of HBeAg is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

41. (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, or a compound for use according to claim 40, wherein the concentration of HBeAg in a serum sample derived from a subject obtained after administration of a pharmaceutically acceptable salt thereof is decreased as compared to the reference concentration of HBeAg.

42. The reference concentration of HBeAg is the concentration of HBeAg in a serum sample obtained from a subject before administration of (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, or a pharmaceutically acceptable salt thereof, a compound for use according to claim 41.

43. A compound for use according to any one of claims 39 to 42, wherein the concentration of HBeAg in the serum of the subject is decreased by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

44. A compound for use according to any one of claims 39 to 43, wherein the concentration of HBeAg in the serum of the subject is decreased by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

45. A compound for use according to any one of claims 39 to 44, wherein the concentration of HBeAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

46. A compound for use according to any one of claims 17 to 45, wherein the concentration of HDV DNA in the serum of the subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof.

47. A compound for use according to claim 46, wherein the concentration of HDV DNA is determined in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

48. (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, or a pharmaceutically acceptable salt thereof, a compound for use according to claim 47, wherein the concentration of HDV DNA in a serum sample derived from a subject obtained after administration is reduced as compared to the reference concentration of HDV DNA.

49. A compound for use according to claim 48, wherein the reference concentration of HDV DNA is the concentration of HDV DNA in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

50. A compound for use according to any one of claims 46 to 49, wherein the concentration of HDV DNA in the serum of the subject decreases by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

51. A compound for use according to any one of claims 46 to 50, wherein the concentration of HDV DNA in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

52. A compound for use according to any one of claims 46 to 51, wherein the concentration of HDV DNA in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

53. A compound for use according to any one of claims 17 to 52, wherein the concentration of hepatitis D antigen (HDAg) in the serum of the subject is reduced after administration of (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, or a pharmaceutically acceptable salt thereof.

54. A compound for use according to claim 53, wherein the concentration of HDAg is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

55. A compound for use according to claim 54, wherein the concentration of HDAg in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, is reduced as compared to the reference concentration of HDAg.

56. The reference concentration of HDAg is the concentration of HDAg in a serum sample obtained from a subject prior to administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to claim 55.

57. The concentration of HDAg in the serum of the subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to any one of claims 53 to 56.

58. The concentration of HDAg in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to any one of claims 53 to 57.

59. The concentration of HDAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof. A compound for use according to any one of claims 53 to 58.

60. The subject is administered (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, or a pharmaceutically acceptable salt thereof, within 18 hours of exposure to hepatitis B. A compound for use according to any one of claims 1 to 59.

61. A compound for use according to any one of claims 1 to 60, wherein the subject is administered (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, or a pharmaceutically acceptable salt thereof, within 18 hours of exposure to hepatitis D.

62. A compound for use according to any one of claims 1 to 61, wherein the compound is (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, or a pharmaceutically acceptable salt thereof.

63. A compound for use according to any one of claims 1 to 61, wherein the compound is (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, or a pharmaceutically acceptable salt thereof.

64. A method for treating hepatitis B (HBV) in a subject in need thereof, the method comprising orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

65. A method for preventing or reducing the entry of hepatitis B virus particles into hepatocytes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

66. A method for reducing hepatitis B virus replication in hepatocytes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

67. The method according to any one of claims 1 to 3, wherein the subject has hepatitis D.

68. A method for treating hepatitis D (HDV) in a subject in need thereof, the method comprising orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

69. A method for preventing or reducing the entry of hepatitis D virus particles into hepatocytes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

70. A method for reducing hepatitis D virus replication in hepatocytes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof.

71. The method according to any one of claims 1 to 7, further comprising the step of administering an additional antiviral agent.

72. A method for treating HBV in a subject in need of treatment for HBV, the method comprising the step of orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, and a further antiviral agent.

73. A method for treating HDV in a subject in need of treatment for HDV, the method comprising the step of orally administering to the subject a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, and a further antiviral agent.

74. The method according to any one of claims 8 to 10, wherein the further antiviral agent is selected from the group consisting of entecavir, tenofovir, tenofovir disoproxil, tenofovir alafenamide, lamivudine, adefovir, adefovir dipivoxil, telbivudine, brequinar, interferon, and combinations thereof.

75. The method according to claim 11, wherein the interferon is pegylated interferon, interferon alpha, or a combination thereof.

76. The method according to any one of claims 8 to 11, wherein the further antiviral agent is tenofovir disoproxil.

77. The method according to any one of claims 5 to 8 and 10 to 13, wherein the subject has hepatitis B.

78. The method according to any one of claims 1 to 4 and 14, wherein the subject has chronic hepatitis B.

79. The method according to any one of claims 5 to 8 and 10 to 15, wherein the subject has chronic hepatitis D.

80. The method according to any one of claims 1 to 16, wherein the concentration of one or more biomarkers selected from HBV DNA, hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg), hepatitis B e antigen (HBeAg), HDV DNA, and hepatitis D antigen (HDAg) in the serum of the subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof.

81. The method according to claim 17, wherein the concentration of HBV DNA in the serum of the subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof.

82. The method according to claim 17, wherein the concentration of HBV DNA is determined in a serum sample from the subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

83. The method according to claim 18, wherein the concentration of HBV DNA in a serum sample from the subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, decreases as compared to the reference concentration of HBV DNA.

84. The method according to claim 19, wherein the reference concentration of HBV DNA is the level of HBV DNA in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

85. The method according to any one of claims 17 to 20, wherein the concentration of HBV DNA in the serum of the subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

86. The method according to any one of claims 17 to 21, wherein the concentration of HBV DNA in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

87. The method according to any one of claims 17 to 23, wherein the concentration of HBV DNA in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

88. The method according to any one of claims 17 to 24, wherein the concentration of hepatitis B surface antigen (HBsAg) in the serum of the subject is reduced after administration of (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, or a pharmaceutically acceptable salt thereof.

89. The method according to claim 25, wherein the concentration of HBsAg is determined in a serum sample from the subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

90. The method according to claim 26, wherein the concentration of HBsAg in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, is decreased as compared to the reference concentration of HBsAg.

91. The method according to claim 27, wherein the reference concentration of HBsAg is the concentration of HBsAg in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

92. The method according to any one of claims 25 to 28, wherein the concentration of HBsAg in the serum of the subject is decreased by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

93. The method according to any one of claims 25 to 29, wherein the concentration of HBsAg in the serum of the subject is decreased by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

94. The method according to any one of claims 25 to 30, wherein the concentration of HBsAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

95. The method according to any one of claims 17 to 31, wherein the concentration of hepatitis B core antigen (HBcAg) in the serum of the subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof.

96. The method according to claim 32, wherein the concentration of HBcAg is determined in a sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

97. The method according to claim 33, wherein the concentration of HBcAg in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, is decreased as compared to the reference concentration of HBcAg.

98. The method according to claim 34, wherein the reference concentration of HBcAg is the concentration of HBcAg in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

99. The method according to any one of claims 32 to 35, wherein the concentration of HBcAg in the serum of the subject decreases by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

100. The method according to any one of claims 32 to 36, wherein the concentration of HBcAg in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

101. The method according to any one of claims 35 to 37, wherein the concentration of HBcAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

102. The method according to any one of claims 17 to 38, wherein the concentration of hepatitis B e antigen (HBeAg) in the serum of the subject is reduced after administration of (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, or a pharmaceutically acceptable salt thereof.

103. The method according to claim 39, wherein the concentration of HBeAg is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

104. The method according to claim 40, wherein the concentration of HBeAg in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof is reduced as compared to the reference concentration of HBeAg.

105. The method according to claim 41, wherein the reference concentration of HBeAg is the concentration of HBeAg in a serum sample obtained from a subject prior to administration of (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 or a pharmaceutically acceptable salt thereof.

106. The method according to any one of claims 39 to 42, wherein the concentration of HBeAg in the serum of the subject is reduced by about 10% to about 99% after administration of (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 or a pharmaceutically acceptable salt thereof.

107. The method according to any one of claims 39 to 43, wherein the concentration of HBeAg in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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 or a pharmaceutically acceptable salt thereof.

108. The method according to any one of claims 39 to 44, wherein the concentration of HBeAg in the serum of the subject is undetectable after administration of (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 or a pharmaceutically acceptable salt thereof.

109. The method according to any one of claims 17 to 45, wherein the concentration of HDV DNA in the serum of the subject is reduced after administration of (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 or a pharmaceutically acceptable salt thereof.

110. The method according to claim 46, wherein the concentration of HDV DNA is determined in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

111. The method according to claim 47, wherein the concentration of HDV DNA in a serum sample derived from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, is decreased as compared to the reference concentration of HDV DNA.

112. The method according to claim 48, wherein the reference concentration of HDV DNA is the concentration of HDV DNA in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

113. The method according to any one of claims 46 to 49, wherein the concentration of HDV DNA in the serum of the subject is decreased by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

114. The method according to any one of claims 46 to 50, wherein the concentration of HDV DNA in the serum of the subject is decreased by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

115. The method according to any one of claims 46 to 51, wherein the concentration of HDV DNA in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

116. The method according to any one of claims 17 to 52, wherein the concentration of hepatitis D antigen (HDAg) in the serum of the subject decreases after administration of (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, or a pharmaceutically acceptable salt thereof.

117. The method according to claim 53, wherein the concentration of HDAg is determined in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof.

118. The method according to claim 54, wherein the concentration of HDAg in a serum sample from a subject obtained after administration of (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, or a pharmaceutically acceptable salt thereof, is decreased as compared to the reference concentration of HDAg.

119. The method according to claim 55, wherein the reference concentration of HDAg is the concentration of HDAg in a serum sample obtained from the subject before administration of (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, or a pharmaceutically acceptable salt thereof.

120. The method according to any one of claims 53 to 56, wherein the concentration of HDAg in the serum of the subject is reduced by about 10% to about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

121. The method according to any one of claims 53 to 57, wherein the concentration of HDAg in the serum of the subject is reduced by about 5%, about 10%, about 25%, about 50%, about 75%, or about 99% after administration of (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, or a pharmaceutically acceptable salt thereof.

122. The method according to any one of claims 53 to 58, wherein the concentration of HDAg in the serum of the subject is undetectable after administration of (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, or a pharmaceutically acceptable salt thereof.

123. The method according to any one of claims 1 to 59, wherein the subject is administered (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, or a pharmaceutically acceptable salt thereof within 18 hours after exposure to hepatitis B.

124. The method according to any one of claims 1 to 60, wherein the subject is administered (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, or a pharmaceutically acceptable salt thereof within 18 hours after exposure to hepatitis D.

125. The method according to any one of claims 1 to 61, wherein a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, is administered to a subject.

126. The method according to any one of claims 1 to 61, wherein a therapeutically effective amount of (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, or a pharmaceutically acceptable salt thereof, is administered to a subject.

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