Treatment methods for chronic hepatitis B

Abstract

This disclosure relates to methods for treating a person having chronic hepatitis B. These methods comprise administering a therapeutically effective amount of antisense oligonucleotide (ASO) (e.g., bepirovirsen) and a therapeutically effective amount of interferon to a person in need, who has a baseline HBsAg level below the threshold level. This disclosure further relates to specific dosing regimens.

Description

Technical Field

[0001] The present disclosure relates to methods for treating humans having chronic hepatitis B. These methods comprise administering to a human in need thereof a therapeutically effective amount of an antisense oligonucleotide (ASO) (e.g., bepirovirsen) and a therapeutically effective amount of interferon, wherein the human has a baseline level of HBsAg below a threshold level. The present disclosure further relates to specific dosing regimens.

Background Art

[0002] Hepatitis B virus (HBV) is highly liver-directed and is a partially double-stranded DNA virus. Although DNA is the genetic material, the replication cycle includes a reverse transcription step in which the pregenomic RNA is copied into DNA. Initial infection with HBV causes acute hepatitis, accompanied by symptoms such as inflammation of the organs, fever, jaundice, and an increase in serum liver transaminases. Patients who cannot overcome this acute viral infection develop a chronic disease over several years, with a high risk of developing cirrhosis and liver cancer.

[0003] HBV infection results in the production of two different types of particles: 1) the HBV virus itself (or Dane particles), which are assembled from the HBV core antigen protein (HBcAg) and are covered with hepatitis B surface antigen (HBsAg) and have the ability to reinfect cells, and 2) non-infectious subviral particles (or SVP), which are high-density lipoprotein-like particles composed of lipids, cholesterol, cholesterol esters, and small and medium-sized hepatitis B surface antigen (HBsAg). For every Dane particle produced, 1,000 to 10,000 SVP are released into the blood. Thus, SVP (and the HBsAg proteins they carry) make up the overwhelming majority of viral proteins in the blood. HBV-infected cells also secrete a soluble proteolytic product of a precore protein called hepatitis B e antigen (HBeAg). The presence of HBeAg in a patient's serum can serve as a marker of active replication in chronic hepatitis.

[0004] Currently, the American Association for the Study of Liver Diseases (AASLD) and the European Association for the Study of the Liver (EASL) recommend treatments for chronic HBV infection, including interferon-alpha (IFN-alpha), pegylated interferon-alpha-2a, entecavir, and tenofovir. However, typical interferon therapy lasts 48 weeks and is associated with tolerability issues due to side effects, with HBeAg serum conversion rates ranging from 27% to 36% 24 weeks after treatment discontinuation. Serum conversion rates for HBsAg are even lower, starting at approximately 3% immediately after treatment discontinuation but rising to over 12% after 5 years.

[0005] While entecavir and tenofovir, as nucleoside and nucleotide therapies, have successfully reduced viral load (HBV DNA), HBeAg serum conversion rates and HBsAg clearance rates are even lower than those seen with IFNα therapy. Other similar therapies, including lamivudine (3TC), terbivudine (LdT), and adefovir, are also used, but the emergence of resistance limits the therapeutic effect of nucleoside / nucleotide therapies in general.

[0006] Antisense oligonucleotides (ASOs) include bepirovirsen, an experimental antisense oligonucleotide under development for the treatment of chronic hepatitis B (CHB) infection. Bepirovirsen directly targets all HBV messenger ribonucleic acid (mRNA) and reduces viral proteins, including HBsAg, through ribonuclease H (RNase H) degradation.

[0007] A phase 2b clinical trial (B-Clear) was conducted to evaluate the efficacy and safety of bepirovirsen treatment in patients with chronic hepatitis B. See Yuen et al. “Efficacy and safety of bepirovirsen in chronic hepatitis B infection,” N. Engl. J. Med. 2022; 387:1957-68. Administration of bepirovirsen at a dose of 300 mg per week for 24 weeks resulted in sustained cleavage of HBsAg and HBV DNA in 9–10% of participants with chronic HBV infection. [Overview of the project]

[0008] There is still a need to discover and develop new anti-HBV therapies, or more specifically, combination therapies, that can achieve higher functional cure rates in patients with chronic hepatitis B (CHB).

[0009] overview In a first aspect of the present invention, a method is provided for treating chronic hepatitis B (CHB) in a person in need, comprising administering to the person a therapeutically effective amount of antisense oligonucleotide (ASO) and a therapeutically effective amount of interferon, wherein the person has an HBsAg baseline level below a threshold level.

[0010] In a second aspect of the present invention, a method is provided for treating chronic hepatitis B (CHB) in a person in need, comprising administering to the person a therapeutically effective amount of bepirovirsen and a therapeutically effective amount of interferon, wherein the person has an HBsAg baseline level below the threshold level.

[0011] In a third aspect of the present invention, a method for treating CHB in a person in need, (a) Determining that the person has an HBsAg baseline level below the threshold level; and (b) Administering to the person a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon. A method is provided that includes the following:

[0012] A fourth aspect of the present invention, a method for treating CHB in a person in need, (a) Measuring the baseline level of HBsAg in the aforementioned human; (b) comparing the HBsAg baseline level with a threshold level; and (c) If the HBsAg baseline level is below the threshold level, administer to the person a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon. A method is provided that includes the following:

[0013] In a fifth aspect of the present invention, a combination of ASO (e.g., bepirovirsen) and PegIFNα-2a is provided for the treatment of CHB in humans. (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The ASO is administered to the person at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human subjects at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of ASO.

[0014] In a sixth aspect of the present invention, a combination of ASO (e.g., bepirovirsen) and PegIFNα-2a is provided for achieving functional HBV remission in a human with CHB. (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The ASO is administered to the person at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human subjects at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of ASO.

[0015] In a seventh aspect of the present invention, there is provided the use of an ASO (e.g., bepirovirsen) in the manufacture of a medicament for the treatment of CHB in humans, (a) the human to be treated has a baseline HBsAg of 3000 IU / mL or less; (b) the ASO is administered weekly to the human at a dose of about 300 mg for 12 or 24 weeks; (c) PegIFNα-2a is administered weekly to the human at a dose of about 180 μg for 12 to 24 weeks after discontinuation of the ASO.

[0016] In an eighth aspect of the present invention, there is provided the use of PegIFNα-2a in the manufacture of a medicament for the treatment of CHB in humans, (a) the human to be treated has a baseline HBsAg of 3000 IU / mL or less; (b) the ASO is administered weekly to the human at a dose of about 300 mg for 12 or 24 weeks; (c) PegIFNα-2a is administered weekly to the human at a dose of about 180 μg for 12 to 24 weeks after discontinuation of the ASO.

Brief Description of the Drawings

[0017] [Figure 1] Figure 1 shows the study design of a Phase 2b clinical trial (B-Together trial) to evaluate the efficacy and safety of treatment with bepirovirsen and PegIFN in CHB patients. [Figure 2] Figure 2 shows the proportion of participants who achieved HBsAg < 0.05 IU / mL and HBV DNA < LLOQ at the end of the trial without initiating a new antiviral therapy (rescue therapy), shown overall and at baseline HBsAg levels (B-Together trial). [Figure 3] Figure 3 shows the study design of a Phase 3b clinical trial (B-Sustained trial) to evaluate the efficacy and safety of sequential therapy with bepirovirsen and PegIFN in nucleoside / nucleotide analogue-treated CHB patients. Detailed Description of the Invention

[0018] Detailed explanation definition Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. For example, the specific terms used in this specification are defined as described in "A multilingual glossary of biotechnological terms: (IUPAC Recommendations)", Leuenberger, H.G.W, Nagel, B. and Klbl, H. eds. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland).

[0019] Throughout this specification and the following claims, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", are to be understood to mean including the stated element, integer, or step, or group of elements, integers, or steps, but not to exclude any other element, integer, or step, or group of elements, integers, or steps.

[0020] Unless otherwise specified, the following terms have the following meanings.

[0021] "About", as used in this specification, is intended to qualify the numerical value it modifies and indicates a value that varies within the range of error. When no specific error such as a standard deviation with respect to an average value given in a graph or table of data is mentioned, the term "about" should be understood to mean a range encompassing ±10% of the recited value, and that range is included.

[0022] An "antisense oligonucleotide (ASO)" refers to a single-stranded oligonucleotide having a nucleic acid base sequence that enables hybridization with a corresponding region or segment of a target nucleic acid.

[0023] Baseline levels of specific substances (e.g., HBsAg, HBeAg, HBsAb, HBeAb, HBV DNA, ALT) are measured in a blood sample taken from the patient before the first dose of treatment.

[0024] Bepirovirsen is an ASO currently in clinical evaluation for the treatment of chronic HBV infection. Bepirovirsen is compound ISIS No. 505358, disclosed in International Publication No. 2012 / 145697. Bepirovirsen has 20 linked nucleosides and has the nucleic acid sequence 5'-GCAGAGGTGAAGCGAAGTGC-3' (SEQ ID NO: 1). A gap segment consisting of 10 linked deoxynucleosides, A 5' wing segment consisting of five linked nucleosides, and A 3' wing segment consisting of five linked nucleosides. The compound contains a gap segment located between the 5' and 3' wing segments, each nucleoside in each wing segment contains a 2'-O-methoxyethyl sugar, each internucleoside bond is a phosphorothioate bond, and each cytosine is 5-methylcytosine. The CAS registry number for bepirobircene is 1403787-62-1.

[0025] AHB-137 is a non-conjugated ASO thought to have the same nucleic acid base sequence as bepilobircene. AHB-137 was developed by Ausper Biopharma Co., Ltd. and is currently being investigated in clinical studies for the treatment of HBV infection. See International Publication No. 2023 / 131098; Qiu, X. et al., “AHB-137, a novel and potent hepatitis B virus antisense oligonucleotide with a favorable preclinical pharmacokinetics and safety profile,” EASL 2023, Abstract No. SAT-178 (concludes that the novel HBV ASO AHB-137 showed a favorable preclinical pharmacokinetic and safety profile); Wu, X. et al., “AHB-137, a novel hepatitis B virus antisense oligonucleotide with substantially enhanced in vitro and in vivo antiviral activity,” EASL 2023, Abstract No. SAT-173 (concludes that AHB-137 has potent serum HBsAg-reducing activity compared to BPV [bepirovirsen] by reducing hepatic HBV RNA and HBsAg in multiple in vitro and in vivo HBV models).

[0026] A "bicyclic sugar" refers to a furanose ring modified by a bridge between two non-geminal carbon atoms. Bicyclic sugars are modified sugars.

[0027] Chronic hepatitis B (CHB) infection occurs when an individual initially develops an acute infection but subsequently becomes unable to fight off the infection. Approximately 90% of infants infected at birth progress to chronic disease. However, the risk of chronic infection decreases with age, with 20-50% of infected children and less than 10% of infected adults progressing from acute to chronic infection. The terms "chronic hepatitis B infection," "chronic hepatitis B virus infection," "chronic hepatitis B," "chronic HBV infection," "CHB infection," and "CHB" are used interchangeably in this specification.

[0028] "Dose" refers to a specific amount of an active pharmaceutical agent to be administered in a single dose or within a specific period of time. In certain embodiments, a dose may be administered in two or more boluses, tablets, or injections. For example, in certain embodiments, where subcutaneous administration is desired, the desired dose requires a volume that cannot be easily accommodated in a single injection. In such embodiments, two or more injections may be used to achieve the desired dose. In certain embodiments, a dose may be administered in two or more injections to minimize injection site reactions in an individual.

[0029] A "dosage plan" is a combination of doses designed to achieve one or more desired effects.

[0030] "Period" refers to the duration for which a certain activity or event persists. In certain embodiments, the treatment period is the period during which the medicinal agent is administered.

[0031] "Functional cure" refers to the sustained suppression of HBV DNA (for 24 weeks or more) (<lower limit of quantification or "LLOQ") after all HBV treatments, and the disappearance of HBsAg (<0.05 IU / mL) after a finite period of therapy, or HBsAg negativity regardless of the presence or absence of HBsAb.

[0032] "HBV" refers to mammalian hepatitis B viruses, including human hepatitis B virus. This term encompasses geographical genotypes of hepatitis B viruses, particularly human hepatitis B virus, as well as variants of geographical genotypes of hepatitis B virus. Human geographical genotypes of HBV include, but are not limited to, genotypes A (Northwestern Europe, North America, Central America); B (Indonesia, China, Vietnam); C (East Asia, South Korea, China, Japan, Polynesia, Vietnam); D (Mediterranean region, Middle East, India); E (Africa); F (Native Americans, Polynesia); G (United States, France); and H (Central America) genotypes.

[0033] "HBV antigen" refers to any hepatitis B virus antigen or protein, including core proteins such as "hepatitis B core antigen" or "HBcAg" and "hepatitis B E antigen" or "HBeAg," as well as envelope proteins such as "HBV surface antigen" or "HBsAg."

[0034] "Hepatitis B E antigen" or "HBeAg" is a secreted non-particulate HBV core protein. Because the HBV antigens HBeAg and HBcAg share a primary amino acid sequence, they exhibit cross-reactivity at the T-cell level. While HBeAg is not necessary for viral aggregation or replication, research suggests it may be required for the establishment of chronic infection.

[0035] "HBV surface antigen," "HBsAg," or "HBs" is the envelope protein of infectious HBV virus particles (Dane particles), but it is also secreted as non-infectious particles (subviral particles, SVPs) at serum levels 1000 times higher than those of HBV virus particles. Serum levels of HBsAg in infected individuals or animals can reach as high as 1000 μg / mL (Kann and Gehrlich (1998) Topley & Wilson's Microbiology and Microbial Infections, 9th edition, 745).

[0036] "HBV-related condition" means any disease, biological condition, medical condition, or event that is caused by, related to, associated with, or attributable to hepatitis B virus infection, exposure, or exacerbation of the disease. The term hepatitis B-related condition includes chronic HBV infection, inflammation, fibrosis, cirrhosis, liver cancer, serum hepatitis, jaundice, hepatitis, hepatic fibrosis, cirrhosis, hepatic failure, diffuse hepatocellular inflammatory disease, hemophagocytic syndrome, HBV viremia, transplant-related liver disease, and conditions having symptoms that may include any or all of the following: flu-like illness, weakness, pain, headache, fever, loss of appetite, diarrhea, nausea and vomiting, pain in the liver area, clay-colored or gray stools, generalized itching, or dark-colored urine, when these are associated with a positive test result for the presence of hepatitis B virus, hepatitis B virus antigen, or antibodies specific to hepatitis B virus antigen.

[0037] Terms such as "induce," "inhibit," "enhance," "increase," "boost," and "decrease" generally indicate a quantitative difference between two states. Such terms may refer to a statistically significant difference between two states. For example, "an amount effective in inhibiting HBV activity or expression" refers to the level of HBV activity or expression in treated cells, which may be quantitatively different and statistically significant from the level of HBV activity or expression in untreated cells. Such terms can be applied, for example, to expression levels and activity levels.

[0038] "Modified nucleoside bonds" refer to substitutions or alterations from naturally occurring nucleoside bonds (i.e., phosphodiester nucleoside bonds).

[0039] "Modified nucleic acid bases" refers to any nucleic acid base other than adenine, cytosine, guanine, thymidine, or uracil.

[0040] "Modified sugar" means substitution and / or any alteration from the natural sugar portion.

[0041] A "nucleic acid base" refers to a heterocyclic portion that can pair with a base of another nucleic acid.

[0042] "Nucleic acid base sequence" refers to the sequence of nucleic acid bases that are consecutive and independent of sugars, bonds, and / or modifications of the nucleic acid bases.

[0043] In this specification, "interferon" means interferon appropriate for the treatment of chronic hepatitis B in humans; this includes interferon-alpha, interferon-λ and their pegylated forms (see Ye et al., Front. Immunol. 2021, 12: 733364).

[0044] "PegIFN," "pegylated interferon," "pegylated interferon," and "pegylated interferon α" are used interchangeably herein unless otherwise specified and refer to the PEGylated form of human interferon α. ​​PegIFN includes α-2a and PegIFNα-2b. PEG-IFNα-2a is synthesized by covalent bonding of a branched bis-[monomethoxypolyethylene glycol (PEG)] polymer with a molecular weight of approximately 44,000 to interferon α-2a, which is produced using recombinant DNA technology. PegIFNα-2a is marketed under the trademark PEGASYS. PegIFNα-2b includes PEGINTRON and PEGBERON.

[0045] A "pharmaceutically acceptable salt" refers to a physiologically acceptable salt of a compound, that is, a salt that retains the desired biological activity of the parent active ingredient and does not impart undesirable toxicological effects.

[0046] A "phosphorothioate bond" refers to a bond between nucleotides in which a phosphodiester bond has been modified by replacing one of the non-bridged oxygen atoms with a sulfur atom. A phosphorothioate bond is a modified nucleoside bond.

[0047] "Region" is defined, for example, as a portion of a nucleic acid having at least one distinguishable structure, function, or characteristic.

[0048] "Stable nucleoside or nucleotide analog (NA) therapy" is defined as having no change in the nucleoside or nucleotide analog regimen for at least six months prior to the therapy disclosed in the present disclosure and no planned change in the regimen during the treatment period.

[0049] "Segment" can refer, for example, to a smaller region or sub-region within a nucleic acid.

[0050] "Serum clearance" refers to the clearance or removal of an antigen (such as HBsAg and / or HBV DNA levels) in the blood of a CHB patient after treatment (or a measurement below the lower limit of quantification by the laboratory (i.e., <LLOQ)). In the examination of human samples, when measuring the serum HBsAg level by a sandwich immunoassay using COBAS HBsAg quant II (Roche), the LLOQ is 0.05 IU / mL. In the examination of human samples, when measuring the serum HBV DNA level by COBAS Ampliprep / COBAS Taqman HBV test v.2.0 (Roche), the LLOQ is 20 IU / mL. When a qualitative assay is used to measure the serum HBsAg level, a binary term (positive / detected, negative / not detected) is used.

[0051] "Subject" refers to a human or non-human animal selected for treatment or therapy. In one embodiment, the subject is human.

[0052] "Therapeutically effective amount" refers to the administration of a pharmaceutical agent, alone or as part of a pharmaceutical composition, in a single dose or as part of a series of doses, to a subject in an amount that can have any detectable positive effect on any symptom, aspect, or characteristic of a disease or medical condition when administered to the subject.

[0053] "Treatment" means administering a medicinal agent to a subject to influence a change or improvement in a disease or condition. When used herein in connection with chronic hepatitis B infection, the term "treat" means administering a suitable composition intended to alleviate the symptoms of CHB, prevent the progression of CHB, or reduce the level of one or more detectable markers of CHB.

[0054] method This disclosure provides a method for treating chronic hepatitis B (CHB) in a human subject of need, comprising administering to the subject a therapeutically effective amount of an antisense oligonucleotide (ASO) (e.g., bepirovirsen) and a therapeutically effective amount of interferon, wherein the subject has a baseline HBsAg level below a threshold level.

[0055] In some embodiments, the ASO comprises 20 linked nucleosides and has the nucleic acid base sequence of Sequence ID No. 1. In some embodiments, the ASO comprises 20 linked nucleosides and has the nucleic acid base sequence of Sequence ID No. 1. In some embodiments, the ASO comprises at least one modified sugar. In some embodiments, the at least one modified sugar is a bicyclic sugar or comprises a 2'-O-methoxyethyl (2'-MOE) group.

[0056] In some embodiments, the ASO comprises at least one modified nucleoside bond. In some embodiments, the at least one modified nucleoside bond is a phosphorothioate bond. In some embodiments, each nucleoside bond of the ASO is a phosphorothioate bond.

[0057] In some embodiments, the ASO comprises at least one modified nucleic acid base. In some embodiments, the modified nucleic acid base is 5-methylcytosine.

[0058] In some embodiments, ASO is A gap segment consisting of linked deoxynucleosides, A 5' wing segment consisting of linked nucleosides, and 3' wing segment consisting of linked nucleosides The system comprises the gap segment, which is positioned between the 5' wing segment and the 3' wing segment, and each nucleoside in each wing segment contains a modified sugar.

[0059] In some embodiments, the ASO includes separator segments positioned between gap segments, as disclosed in International Publication No. 2023 / 131098, which is entirely part of this specification by reference. In one embodiment, the ASO is selected from compounds AUS1233 / AUS1138, AUS1444, AUS1458, AUS1459, AUS1460, AUS1427 / AUS1461, AUS1462, AUS1463, AUS1464, AUS1465, AUS1463, AUS1467, AUS1468, AUS1470, AUS1471, AUS1472, AUS1473, AUS1474, AUS1475, AUS1476 / AUS1493, AUS1478, AUS1479, AUS1489, AUS1490, AUS1443, and AUS1322, as disclosed in International Publication No. 2023 / 131098. In one embodiment, the ASO is selected from compounds AUS1233 / AUS1138, AUS1463, and AUS1476 / AUS1493, as disclosed in International Publication No. 2023 / 131098. In one embodiment, the ASO is compound AUS1493, as disclosed in International Publication No. 2023 / 131098.

[0060] In one embodiment, ASO is bepilobircene. In another embodiment, ASO is AHB-137.

[0061] In another respect, this disclosure relates to a method for treating chronic hepatitis B in a person in need, (a) Determining that the person has an HBsAg baseline level below the threshold level; and (b) Administering to the person a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon. We provide a method that includes the following:

[0062] The baseline level of HBsAg in the blood of CHB patients is measured by methods known in the art before the initiation of treatment. As used herein, the step of “determining” can be achieved by measuring the patient’s baseline HBsAg level and comparing that baseline level to a predetermined threshold level. The “determining” step can also be achieved by receiving the baseline HBsAg level from the patient or laboratory and comparing that baseline HBsAg level to a predetermined threshold level.

[0063] In another respect, this disclosure relates to a method for treating chronic hepatitis B in a person in need, (a) Measuring the baseline level of HBsAg in the aforementioned human; (b) comparing the HBsAg baseline level with a threshold level; and (c) If the HBsAg baseline level is below the threshold level, administer to the person a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon. We provide a method that includes the following:

[0064] Low baseline levels of HBsAg have been reported to be a predictor of sustained HBsAg loss in HBeAg-negative patients treated with pegylated interferon-α2a and adefovir. (See Takkenberg, et al., “Baseline hepatitis B surface antigen (HBsAg) as predictor of sustained HBsAg loss in chronic hepatitis B patients treated with pegylated interferon-α2a and adefovir,” Antivir. Ther. 2013;18(7):895-904). In particular, baseline HBsAg levels of less than 400 IU / mL were identified as a good predictor for treating HBeAg-negative patients with pegylated interferon-α2a and adefovir. However, low baseline HBsAg levels did not predict HBsAg loss in HBe antigen-positive patients in the same study.

[0065] In the methods disclosed herein, ASO (e.g., bepirovirsen) may be administered as a free acid, a pharmaceutically acceptable salt thereof (e.g., a sodium salt), or a combination thereof. In some embodiments, ASO (e.g., bepirovirsen) is administered as a free acid. In some embodiments, ASO (e.g., bepirovirsen) is administered as a pharmaceutically acceptable salt thereof (e.g., a sodium salt). In some embodiments, ASO (e.g., bepirovirsen) is administered as a combination of a free acid and a sodium salt. In some embodiments, ASO (e.g., bepirovirsen) is administered by subcutaneous injection.

[0066] As used herein, “a therapeutically effective amount of ASO (e.g., bepirovirsen)” refers to the amount of free ASO (e.g., bepirovirsen). In some embodiments, the therapeutically effective amount of ASO (e.g., bepirovirsen) is about 150 mg to 450 mg once per week. In some embodiments, the therapeutically effective amount of ASO (e.g., bepirovirsen) is about 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, or 450 mg once per week, or a range between any two of the above values. In some embodiments, the therapeutically effective amount of ASO (e.g., bepirovirsen) is about 150 mg once per week. In some embodiments, the therapeutically effective amount of ASO (e.g., bepirovirsen) is about 300 mg once per week. In some embodiments, the ASO (e.g., bepirovirsen) is administered weekly with additional loading doses on days 4 and 11 after the initial two-week dose (also known as "two loading doses"). In some embodiments, the ASO (e.g., bepirovirsen) is administered once a week at a dose of approximately 300 mg, with additional loading doses on days 4 and 11 after the initial two-week dose. In certain embodiments, the loading dose is 300 mg.

[0067] In some embodiments, ASO (e.g., bepirovirsen) is administered for approximately 12 to 48 weeks. In some embodiments, ASO (e.g., bepirovirsen) is administered for 12 weeks, 24 weeks, or 48 weeks, or within a range of any two of the above periods. In one embodiment, ASO (e.g., bepirovirsen) is administered for 12 weeks. In one embodiment, ASO (e.g., bepirovirsen) is administered for 24 weeks. In one embodiment, ASO (e.g., bepirovirsen) is administered for 48 weeks. In one embodiment, ASO (e.g., bepirovirsen) is administered for 12 weeks or 24 weeks, with additional loading doses on days 4 and 11 after the initial dose.

[0068] In some embodiments, the ASO (e.g., bepirovirsen) is administered at a dose of approximately 300 mg once a week for 24 weeks. In some embodiments, the ASO (e.g., bepirovirsen) is administered at a dose of approximately 300 mg once a week for 12 weeks. In some embodiments, the ASO (e.g., bepirovirsen) is administered at a dose of approximately 300 mg once a week for 12 weeks, followed by a dose of approximately 150 mg once a week for 12 weeks.

[0069] In some embodiments, the ASO (e.g., bepirovirsen) is administered at a dose of approximately 300 mg once a week for 24 weeks, with additional loading doses of 300 mg on days 4 and 11 after the initial dose. In some embodiments, the ASO (e.g., bepirovirsen) is administered at a dose of approximately 300 mg once a week for 12 weeks, with additional loading doses of 300 mg on days 4 and 11 after the initial dose.

[0070] In some embodiments, the Disclosure provides a method for treating CHB in a person in need, comprising administering to the person a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon, the subject having an HBsAg baseline level below a threshold level. In some embodiments, the HBsAg baseline threshold level is in the range of about 1,000 IU / mL to 3,000 IU / mL.

[0071] In some embodiments, the HBsAg baseline threshold level is approximately 1000, 1500, 2000, 2500, or 3000 IU / mL, or in the range of any two of the above numbers. In some embodiments, the HBsAg baseline threshold level is approximately 1000, 2000, or 3000 IU / mL. In some embodiments, the HBsAg baseline threshold level is approximately 1000 IU / mL. In some embodiments, the HBsAg baseline threshold level is approximately 3000 IU / mL.

[0072] In one embodiment, the disclosure provides a method for treating CHB in a person in need, comprising administering to the person a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon, wherein the person has an HBsAg baseline level of about 1000 IU / mL or less. In one embodiment, the disclosure provides a method for treating CHB in a person in need, comprising administering to the person a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon, wherein the person has an HBsAg baseline level of about 3000 IU / mL or less. In one embodiment, the interferon is interferon-α or interferon-λ. In another embodiment, the interferon is PegIFN.

[0073] In one embodiment, the present disclosure provides a method for treating CHB in a person in need, comprising administering an ASO (e.g., bepirovirsen) to the person at a dose of 300 mg once weekly for 24 weeks, wherein the person has an HBsAg baseline level not exceeding about 1000 IU / mL. In one embodiment, the present disclosure provides a method for treating CHB in a person in need, comprising administering an ASO (e.g., bepirovirsen) to the person at a dose of 300 mg once weekly for 24 weeks, wherein the person has an HBsAg baseline level of about 3000 IU / mL or less.

[0074] In one embodiment, the present disclosure provides a method for treating CHB in a person in need, comprising administering an ASO (e.g., bepirovirsen) to the person at a dose of 300 mg once weekly for 24 weeks, wherein the person has an HBsAg baseline level of about 1000 IU / mL or less and is HBeAg negative.

[0075] In one embodiment, the present disclosure provides a method for treating CHB in a person in need, comprising administering an ASO (e.g., bepirovirsen) to the person at a dose of 300 mg once weekly for 24 weeks, wherein the person has an HBsAg baseline level of about 1000 IU / mL or less, is HBeAg negative, and is under stable nucleoside or nucleotide analog (NA) therapy.

[0076] In one embodiment, ASO (e.g., bepirovirsen) and interferon are administered sequentially. In one embodiment, ASO (e.g., bepirovirsen) is administered first, followed by interferon. In one embodiment, the human is evaluated for interferon eligibility based on their level prior to interferon treatment. In one embodiment, interferon is administered after discontinuation of ASO (e.g., bepirovirsen). In one embodiment, the interferon is interferon α or interferon λ. In another embodiment, the interferon is PegIFN. In one embodiment, the interferon is PegIFNα-2a. In one embodiment, the interferon is PegIFNα-2b.

[0077] In one embodiment, HBsAg levels in a person with CHB infection are lower than baseline after ASO (e.g., bepirovirsen) treatment and before interferon, particularly PEGIFNα-2a treatment. In one embodiment, HBsAg levels in a person with CHB infection are below the lower limit of quantification (LLOQ) after ASO (e.g., bepirovirsen) treatment and before interferon, particularly PEGIFNα-2a treatment.

[0078] In another aspect of the present invention, a combination of ASO (e.g., bepirovirsen) and interferon, particularly PegIFNα-2a, is provided for the treatment of CHB in humans, wherein (a) the human being treated has a baseline HBsAg of ≤ 3000 IU / mL; (b) the ASO (e.g., bepirovirsen) is administered to the human at a dose of approximately 300 mg weekly for 12 or 24 weeks; and (c) the interferon is administered to the human at a dose of approximately 180 μg weekly for 12 to 48 weeks after discontinuation of the ASO (e.g., bepirovirsen). In one embodiment, the human being treated has a baseline HBsAg of ≤ 1000 IU / mL. In one embodiment, the HBsAg level of the human having CHB is below LLOQ after ASO (e.g., bepirovirsen) treatment and before interferon treatment. In one embodiment, ASO (e.g., bepirovirsen) is administered to the human at a dose of approximately 300 mg weekly for 12 weeks. In one embodiment, ASO (e.g., bepirovirsen) is administered to the human at a dose of approximately 300 mg weekly for 24 weeks. In one embodiment, PegIFNα-2aj is administered to the human at a dose of approximately 180 μg weekly for 12 weeks. In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 24 weeks. In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 48 weeks.

[0079] In one aspect of the present invention, a combination of ASO (e.g., bepirovirsen) and interferon, particularly PegIFNα-2a, is provided to achieve functional HBV cure in a person with CHB, wherein (a) the person to be treated has a baseline HBsAg of 3000 IU / mL or less; (b) the person is administered ASO (e.g., bepirovirsen) at a dose of about 300 mg for 12 or 24 weeks; and (c) the person is administered interferon weekly at a dose of about 180 μg for 12 to 48 weeks after discontinuation of ASO (e.g., bepirovirsen). In one embodiment, the person to be treated has a baseline HBsAg of 1000 IU / mL or less. In one embodiment, the HBsAg level of the person with CHB is below LLOQ after ASO (e.g., bepirovirsen) treatment and before interferon treatment. In one embodiment, ASO (e.g., bepirovirsen) is administered to the human at a dose of approximately 300 mg weekly for 12 weeks. In one embodiment, ASO (e.g., bepirovirsen) is administered to the human at a dose of approximately 300 mg weekly for 24 weeks. In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 12 weeks. In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 24 weeks. In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 48 weeks.

[0080] In some embodiments, the functional cure rate is greater than 15%, greater than 20%, greater than 25%, greater than 30%, greater than 35%, or greater than 40% in patients with a baseline HBsAg of 3000 IU / mL or less. In some embodiments, the functional cure rate is greater than 30% in patients with a baseline HBsAg of 3000 IU / mL or less. In some embodiments, the functional cure rate is greater than 35% in patients with a baseline HBsAg of 3000 IU / mL or less. In some embodiments, the functional cure rate is greater than 40% in patients with a baseline HBsAg of 3000 IU / mL or less. In some embodiments, the functional cure rate is greater than 35% in patients with a baseline HBsAg of 1000 IU / mL or less. In some embodiments, the functional cure rate is greater than 40% in patients with a baseline HBsAg of 1000 IU / mL or less. In some embodiments, the functional cure rate is greater than 45% in patients with a baseline HBsAg of 1000 IU / mL or less. In some embodiments, the functional cure rate exceeds 50% in patients with baseline HBsAg levels of 1000 IU / mL or less. In some embodiments, the functional cure rate exceeds 55% in patients with baseline HBsAg levels of 1000 IU / mL or less.

[0081] In one embodiment, a person receiving a combination of ASO (e.g., bepirovirsen) and interferon is under stable NA therapy. In one embodiment, the NA therapy is lamivudine, adefovir, adefovir dipivoxil, terbivudine, entecavir, tenofovir, tenofovir disoproxil fumarate, or tenofovir alafenamide, or any pharmaceutically acceptable salt thereof. In one embodiment, the person discontinues NA therapy after discontinuation of interferon. In one embodiment, the person discontinues NA therapy 12 weeks after discontinuation of interferon. In one embodiment, the person discontinues NA therapy 24 weeks after discontinuation of interferon.

[0082] In one embodiment, humans are administered ASO (e.g., bepirovirsen) from week 1 to week 12, then interferon from week 13 to week 36, and NA throughout week 1 to week 60.

[0083] In another embodiment, humans are administered ASO (e.g., bepirovirsen) from week 1 to week 24, then interferon from week 25 to week 48, and NA throughout week 1 to week 72.

[0084] In one aspect of the present invention, the use of an ASO (e.g., bepirovirsen) in the manufacture of a pharmacopoeia for the treatment of CHB in humans is provided, wherein (a) the human to be treated has a baseline HBsAg of 3000 IU / mL or less; (b) the ASO (e.g., bepirovirsen) is administered to the human at a dose of about 300 mg weekly for 12 or 24 weeks; and (c) interferon, in particular PegIFN, is administered to the human at a dose of about 180 μg weekly for 12 to 48 weeks after discontinuation of the ASO (e.g., bepirovirsen). In one embodiment, the human to be treated has a baseline HBsAg of 1000 IU / mL or less. In one embodiment, the HBsAg level of the human with CHB is below LLOQ after ASO (e.g., bepirovirsen) treatment and before interferon treatment. In one embodiment, ASO (e.g., bepirovirsen) is administered to the human at a dose of approximately 300 mg weekly for 12 weeks. In one embodiment, ASO (e.g., bepirovirsen) is administered to the human at a dose of approximately 300 mg weekly for 24 weeks. In one embodiment, interferon is administered to the human at a dose of approximately 180 μg weekly for 12 weeks. In one embodiment, interferon is administered to the human at a dose of approximately 180 μg weekly for 24 weeks. In one embodiment, interferon is administered to the human at a dose of approximately 180 μg weekly for 48 weeks.

[0085] In a further aspect of the present invention, the use of interferon, in particular PegIFN, in the manufacture of a pharmacopoeia for the treatment of CHB in humans is provided, wherein (a) the human to be treated has a baseline HBsAg of 3000 IU / mL or less; (b) an ASO (e.g., bepirovirsen) is administered to the human at a dose of about 300 mg weekly for 12 or 24 weeks; and (c) interferon is administered to the human at a dose of about 180 μg weekly for 12 to 48 weeks after discontinuation of the ASO (e.g., bepirovirsen). In one embodiment, the human to be treated has a baseline HBsAg of 1000 IU / mL or less. In one embodiment, the HBsAg level of the human having CHB is below LLOQ after ASO (e.g., bepirovirsen) treatment and before interferon treatment. In one embodiment, the ASO (e.g., bepirovirsen) is administered to the human at a dose of about 300 mg for 12 weeks. In one embodiment, ASO (e.g., bepirovirsen) is administered to the human at a dose of approximately 300 mg weekly for 24 weeks. In one embodiment, interferon is administered to the human at a dose of approximately 180 μg weekly for 12 weeks. In one embodiment, interferon is administered to the human at a dose of approximately 180 μg weekly for 24 weeks. In one embodiment, interferon is administered to the human at a dose of approximately 180 μg weekly for 48 weeks.

[0086] In some embodiments, the interferon is interferon-α or interferon-λ. In some embodiments, the interferon is PegIFN. In some embodiments, the PegIFN is PEGASYS. In some embodiments, the PegIFN is PEGINTRON or PEGBERON. In some embodiments, the human is not treated with another HBsAg-reducing agent or another immunomodulator. That is, ASO (e.g., bepirovirsen) and interferon are used as sequential ditherapy for treating CHB. The HBsAg-reducing agent may be a small molecule / short interfering RNA (siRNA), a second ASO, a nucleic acid polymer (NAP), an HBV RNA destabilizer, an HBV-specific neutralizing mAb, or a combination thereof. The immunomodulator may be a therapeutic vaccine, a PD-1 / PD-L1 inhibitor, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, or a combination thereof. In some embodiments, the human is treated with another HBsAg-reducing agent or another immunomodulator.

[0087] In one embodiment, the human is under the therapy of a stable nucleoside or nucleotide analog (NA) (e.g., tenofovir disoproxil, tenofovir alafenamide, or entecavir). In some embodiments, the NA therapy is lamivudine, adefovir, adefovir dipivoxil, terbivudine, entecavir, tenofovir, tenofovir disoproxil fumarate (TDF), or tenofovir alafenamide (TAF), or a pharmaceutically acceptable salt thereof. In some embodiments, the NA therapy is entecavir, tenofovir, tenofovir disoproxil fumarate, or tenofovir alafenamide. In some embodiments, the NA therapy is entecavir. In some embodiments, the NA therapy is tenofovir. In some embodiments, the NA therapy is tenofovir disoproxil fumarate. In some embodiments, the NA therapy is tenofovir alafenamide.

[0088] In another embodiment, the subjects are not under NA therapy. In some embodiments, the subjects are untreated.

[0089] In some embodiments, individuals with HBsAg baseline levels below the threshold level are more likely to have HBsAg below LLOQ at the end of ASO (e.g., bepilovirsen) and interferon treatment compared with individuals with HBsAg baseline levels above the threshold level. In some embodiments, such individuals are at least 5%, 10%, 15%, 20%, 25%, or 30% more likely to have HBsAg below LLOQ at the end of ASO (e.g., bepilovirsen) and interferon treatment.

[0090] In some embodiments, individuals with HBsAg baseline levels below the threshold level are more likely to have HBV DNA below LLOQ at the end of ASO (e.g., bepilovirsen) and interferon treatment compared with individuals with HBsAg baseline levels above the threshold level. In some embodiments, such individuals are at least 5%, 10%, 15%, 20%, 25%, or 30% more likely to have HBV DNA below LLOQ at the end of ASO (e.g., bepilovirsen) and interferon treatment.

[0091] In some embodiments, individuals with HBsAg baseline levels below the threshold level are more likely than individuals with HBsAg baseline levels above the threshold level to have HBsAg and HBV DNA below LLOQ at the end of ASO (e.g., bepirovirsen) and interferon treatment. In some embodiments, individuals with HBsAg baseline levels below the threshold level are at least 5%, 10%, 15%, 20%, 25%, or 30% more likely to have HBsAg and HBV DNA below LLOQ at the end of ASO (e.g., bepirovirsen) and interferon treatment.

[0092] In some embodiments, individuals with HBsAg baseline levels below the threshold level are at least 5%, 10%, 15%, 20%, 25%, or 30% more likely to achieve functional recovery than individuals with HBsAg baseline levels above the threshold level.

[0093] In some embodiments, hepatitis B virus infection is caused by any of the following human geographical genotypes, but are not limited to: A (Northwestern Europe, North America, Central America); B (Indonesia, China, Vietnam); C (East Asia, South Korea, China, Japan, Polynesia, Vietnam); D (Mediterranean region, Middle East, India); E (Africa); F (Native Americans, Polynesia); G (United States, France); or H (Central America).

[0094] In some embodiments, the human is HBeAg-negative or HBeAg-positive before treatment. In some embodiments, the human is HBeAg-negative before treatment. In some embodiments, the human is HBeAg-positive before treatment.

[0095] In some embodiments, human HBV DNA and HBsAg levels fall below the lower limit of quantification (LLOQ) 24 weeks after discontinuation of all HBV therapy. In some embodiments, the human achieves functional remission.

[0096] In another aspect, the disclosure provides an ASO (e.g., bepirovirsen) and an interferon for use in a method of treating chronic hepatitis B in a person having a baseline HBsAg level below a threshold level.

[0097] In another aspect, this disclosure provides ASOs (e.g., bepirovirsen) and interferons, particularly PegIFN, for use in methods of treating chronic hepatitis B in humans where such methods are needed. (a) Determining that the person has an HBsAg baseline level below the threshold level; and (b) Administering to the person a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon. It consists of including.

[0098] composition ASOs (e.g., bepirovirsen) and interferons can each be incorporated into a pharmaceutical composition for use in the treatment of CHB, as described herein. In one embodiment, the pharmaceutical composition comprises an ASO (e.g., bepirovirsen) in combination with one or more pharmaceutically acceptable carriers and / or excipients. In another embodiment, the pharmaceutical composition comprises an interferon, in particular PEGIFN, in combination with one or more pharmaceutically acceptable carriers and / or excipients.

[0099] Such compositions comprise pharmaceutically acceptable carriers and / or excipients, as are known and required by acceptable pharmaceutical practices.

[0100] The pharmaceutical composition may be administered by injection or serial infusion (examples, but not limited to, intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intraocular, and portal vein). In one embodiment, the composition is suitable for subcutaneous administration.

[0101] In one embodiment, the composition comprising ASO (e.g., bepirobircene) also comprises injectable water, which is adjusted to pH 8.0 with an acid or base.

[0102] In one embodiment, the composition comprising PegIFN also comprises sodium chloride, polysorbate 80, benzyl alcohol, sodium acetate, acetic acid, and water for injection. In one embodiment, PegIFN is PEGASYS. In one embodiment, PegIFN is PEGINTRON or PEGBERON.

[0103] This disclosure provides compositions comprising an ASO (e.g., bepirovirsen) and an interferon for treating chronic hepatitis B virus infection in subjects requiring treatment, the treatment method comprising administering to a subject a therapeutically effective amount of the ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon, the subject having an HBsAg baseline level below the threshold level.

[0104] The disclosure also provides an ASO (e.g., bepirovirsen) and interferon for treating chronic hepatitis B virus infection in a subject of interest, the method comprising administering to the subject a therapeutically effective amount of the ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon, the subject having a baseline HBsAg level below a threshold level.

[0105] The Disclosure also provides a composition comprising an ASO (e.g., bepirovirsen), or a composition comprising an ASO and an interferon, or an interferon, for use in a method of treating chronic hepatitis B in a person in need, the method comprising (a) determining that the person has an HBsAg baseline level below a threshold level; and (b) administering to the person a therapeutically effective amount of an ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon.

[0106] In other embodiments, the ASO (e.g., bepirobircene) composition or ASO, the interferon composition or interferon, the method of administering the ASO composition or ASO, and the interferon composition or interferon may be according to any of the methods described in detail above.

[0107] For example, in some embodiments, the HBsAg baseline threshold level is approximately 1000, 1500, 2000, 2500, or 3000 IU / mL, or in the range of any two of the above values. In some embodiments, the HBsAg baseline threshold level is approximately 1000, 2000, or 3000 IU / mL. In some embodiments, the HBsAg baseline threshold level is approximately 1000 IU / mL. In some embodiments, the HBsAg baseline threshold level is approximately 3000 IU / mL.

[0108] In one embodiment, the Disclosure provides an ASO (e.g., bepirovirsen) and interferon for use in a method of treating CHB in a person in need, the method comprising administering a therapeutically effective amount of ASO (e.g., bepirovirsen) to the person, the person having an HBsAg baseline level of about 1000 IU / mL or less. In one embodiment, the Disclosure provides an ASO (e.g., bepirovirsen) and interferon for use in a method of treating CHB in a person in need, the method comprising administering a therapeutically effective amount of ASO (e.g., bepirovirsen) and a therapeutically effective amount of interferon to the person, the person having an HBsAg baseline level of about 3000 IU / mL or less.

[0109] In another embodiment, the Disclosure provides an ASO (e.g., bepirovirsen) and interferon for use in a method of treating CHB in a person in need, the method comprising administering the ASO (e.g., bepirovirsen) to the person at a dose of 300 mg once weekly for 24 weeks, the person having an HBsAg baseline level of about 1000 IU / mL or less. In one embodiment, the Disclosure provides an ASO (e.g., bepirovirsen) and interferon for use in a method of treating CHB in a person in need, the method comprising administering the ASO (e.g., bepirovirsen) to the person at a dose of 300 mg once weekly for 24 weeks, the person having an HBsAg baseline level of about 3000 IU / mL or less. In one embodiment, PegIFNα-2a is administered to the person weekly at a dose of about 180 μg for 24 to 48 weeks after discontinuation of the ASO (e.g., bepirovirsen). In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 24 weeks after discontinuation of ASO (e.g., bepirovirsen). In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 36 weeks after discontinuation of ASO (e.g., bepirovirsen). In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 48 weeks after discontinuation of ASO (e.g., bepirovirsen).

[0110] In another embodiment, the Disclosure provides an ASO (e.g., bepirovirsen) and interferon for use in a method of treating CHB in a person in need, the method comprising administering the ASO (e.g., bepirovirsen) to the person at a dose of 300 mg once weekly for 12 weeks, the person having an HBsAg baseline level of about 1000 IU / mL or less. In one embodiment, the Disclosure provides an ASO (e.g., bepirovirsen) and interferon for use in a method of treating CHB in a person in need, the method comprising administering the ASO (e.g., bepirovirsen) to the person at a dose of 300 mg once weekly for 12 weeks, the person having an HBsAg baseline level of about 3000 IU / mL or less. In one embodiment, PegIFNα-2a is administered to the person weekly at a dose of about 180 μg for 24 to 48 weeks after discontinuation of the ASO (e.g., bepirovirsen). In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 24 weeks after discontinuation of ASO (e.g., bepirovirsen). In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 36 weeks after discontinuation of ASO (e.g., bepirovirsen). In one embodiment, PegIFNα-2a is administered to the human at a dose of approximately 180 μg weekly for 48 weeks after discontinuation of ASO (e.g., bepirovirsen).

[0111] kit In one aspect of the present invention, an ASO (e.g., bepirobircene) and an interferon, particularly PegIFN, are included in the kit along with instructions for use. In one embodiment, the kit comprises an ASO (e.g., bepirobircene) and an interferon in separate containers, as disclosed herein.

[0112] For convenience, the kit may contain predetermined amounts of ASO (e.g., bepirovirsen) and interferon, along with instructions for use. In one embodiment, the kit contains 300 mg of ASO (e.g., bepirovirsen); in one embodiment, the kit contains 180 μg of PegIFNα-2a; in one embodiment, the kit contains multiple unit doses of ASO (e.g., bepirovirsen) and interferon; in one embodiment, the interferon is interferon α or interferon λ; in another embodiment, the interferon is PegIFNα-2a; in one embodiment, the kit contains 1 mL of injectable PegIFNα-2a solution in a stoppered (rubber butyl) vial (Type I glass); in one embodiment, the kit contains 1 mL of injectable ASO (e.g., bepirovirsen) solution in a stoppered (rubber) and aluminum oversealed vial (Type I glass).

[0113] The kit may also include a device used for administering an ASO (e.g., bepirovirsen) and / or an interferon, in particular a device used for administering PegIFNα-2a. In one embodiment, each unit dose of ASO (e.g., bepirovirsen) is contained in a pre-filled syringe. In one embodiment, the pre-filled syringe is for subcutaneous injection. In one embodiment, each unit dose of interferon is contained in a pre-filled syringe. In one embodiment, the pre-filled syringe is for subcutaneous injection.

[0114] In further embodiments, the kit comprises one or more additional agents.

[0115] Embodiment 1. A method for treating chronic hepatitis B in a person in need, comprising administering to the person a therapeutically effective amount of bepirovirsen and a therapeutically effective amount of interferon, wherein the person has an HBsAg baseline level below the threshold level.

[0116] 2. A method for treating chronic hepatitis B in persons who require it, (a) Determining that the person has an HBsAg baseline level below the threshold level; and (b) Administering the person a therapeutically effective amount of bepirovirsen and a therapeutically effective amount of interferon. A method that includes the following:

[0117] 3. A method for treating chronic hepatitis B in people who need it, (a) Measuring the baseline level of HBsAg in the aforementioned human; (b) comparing the HBsAg baseline level with a threshold level; and (c) If the HBsAg baseline level is below the threshold level, administer a therapeutically effective amount of bepirovirsen and a therapeutically effective amount of interferon to the person. A method that includes the following:

[0118] 4. The method according to any one of paragraphs 1 to 3, wherein the threshold level is in the range of approximately 1000 IU / mL to 3000 IU / mL.

[0119] 5. The method according to any one of paragraphs 1 to 4, wherein the threshold level is approximately 1000 IU / mL.

[0120] 6. The method according to any one of paragraphs 1 to 5, wherein the threshold level is approximately 3000 IU / mL.

[0121] 7. The method according to any one of paragraphs 1 to 6, wherein the person is not treated with another HBsAg-lowering agent or another immunomodulator.

[0122] 8. The method according to any one of paragraphs 1 to 7, wherein bepilovirsen is administered by subcutaneous injection.

[0123] 9. The method according to any one of paragraphs 1 to 8, wherein bepirovirsen is administered once a week at a dose of approximately 150 mg or 300 mg.

[0124] 10. The method described in any one of paragraphs 1 to 9, wherein bepirovirsen is administered for 12 to 48 weeks.

[0125] 11. The method according to paragraph 9 or 10, wherein bepirovirsen is administered weekly, with additional loading doses on days 4 and 11 after the initial two-week dose.

[0126] 12. The method according to any one of paragraphs 1 to 11, wherein bepirovirsen is administered at a dose of 300 mg once a week for 12 weeks.

[0127] 13. The method according to any one of paragraphs 1 to 11, wherein bepirovirsen is administered at a dose of 300 mg once a week for 24 weeks.

[0128] 14. The method according to any one of paragraphs 1 to 13, wherein the interferon is interferon-alpha.

[0129] 15. The method according to any one of paragraphs 1 to 13, wherein the interferon is interferon λ.

[0130] 16. The method according to the 14th claim, wherein the interferon is peg-interferon α-2a.

[0131] 17. The method according to paragraph 16, wherein PegIFN is administered once a week at a dose of approximately 180 μg.

[0132] 18. The method according to any one of paragraphs 1 to 17, wherein interferon is administered by subcutaneous injection.

[0133] 19. The method according to any one of paragraphs 1 to 18, wherein interferon is administered after discontinuation of bepirovirsen.

[0134] 20. The method according to any one of paragraphs 1 to 19, wherein interferon is administered for 12 to 24 weeks after discontinuation of bepirovirsen.

[0135] 21. The method according to any one of paragraphs 1 to 20, wherein the human is under stable nucleoside or nucleotide analog (NA) therapy.

[0136] 22. The method according to claim 21, wherein the NA therapy is selected from the group consisting of lamivudine, adefovir, adefovir dipivoxil, terbivudine, entecavir, tenofovir, tenofovir disoproxil fumarate, tenofovir alafenamide, and any pharmaceutically acceptable salt thereof.

[0137] 23. The method according to paragraph 21 or 22, wherein the human discontinues NA therapy 24 weeks after discontinuing interferon.

[0138] 24. The method according to any one of paragraphs 2 to 20, wherein the human is not treated with either a nucleoside or a nucleotide analog (NA).

[0139] 25. The method according to any one of paragraphs 1 to 24, wherein the person does not have cirrhosis of the liver.

[0140] 26. The method according to any one of paragraphs 1 to 25, wherein the human is HBeAg negative prior to bepilovirsen treatment.

[0141] 27. The method according to any one of paragraphs 1 to 25, wherein the human is HBeAg positive prior to bepilovirsen treatment.

[0142] 28. The method according to any one of paragraphs 1 to 27, wherein the human HBV DNA and HBsAg levels are below the lower limit of quantification (LLOQ) 24 weeks after discontinuation of all HBV therapy.

[0143] 29. A combination of bepilovirsen and PegIFNα-2a for the treatment of chronic hepatitis B in humans, (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The person is given bepirovirsen at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human subjects at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of bepirovirsen. combination.

[0144] 30. A combination of bepirovirsen and PegIFNα-2a for achieving functional HBV cure in a person with chronic hepatitis B, (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The person is given bepirovirsen at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human subjects at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of bepirovirsen. combination.

[0145] 31. The use of bepirovirsen in the manufacture of a pharmaceutical product for the treatment of chronic hepatitis B in humans, (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The person is given bepirovirsen at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human subjects at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of bepirovirsen. use.

[0146] 32. The use of PegIFNα-2a in the manufacture of pharmaceuticals for the treatment of chronic hepatitis B in humans, (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The person is given bepirovirsen at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human subjects at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of bepirovirsen. use. [Examples]

[0147] While aspects of the disclosure presented herein have been described in more detail with reference to several embodiments, the following examples, which highlight specific features and characteristics of exemplary embodiments of the disclosure described herein, are for illustrative purposes only and are not intended to limit the disclosure described herein.

[0148] Example 1 Phase IIb trial of sequential treatment with bepilovirsen and pegylated interferon (PegIFN) in participants with chronic hepatitis B (B-Together trial) A. Experimental Design This study is a multicenter, randomized, open-label, phase IIb (B-Together) trial to evaluate the efficacy and safety of sequential treatment with bepilovirsen (300 mg / week, 12 or 24 weeks) followed by PEG-IFN (PEGASYS, 180 mcg / week, up to 24 weeks) in chronic hepatitis B patients receiving stable nasogabacterial therapy. During the bepilovirsen treatment period, a loading dose of 300 mg will be administered on days 4 and 11. Participants will continue nasogabacterial therapy throughout the study period.

[0149] Participants were randomly assigned to one of two parallel treatment groups (1:1) (Figure 1). 1,300 mg bepirovirsen, once a week for 24 weeks (with loading doses on days 4 and 11), followed by PEGIFN (180 mcg / week for up to 24 weeks). Participants will be followed up for another 24 weeks after the completion of PEGIFN treatment (non-treatment period). 2,300 mg bepirovirsen, once weekly for 12 weeks (with loading doses on days 4 and 11), followed by PEGIFN (180 mcg / week, up to 24 weeks). Participants will be followed up for an additional 36 weeks after the completion of PEGIFN treatment (non-treatment period).

[0150] The planned duration of bepirovirsen treatment is 24 weeks and 12 weeks for treatment groups 1 and 2, respectively. At the end of the planned bepirovirsen treatment period, participants' eligibility to initiate PegIFN treatment will be reassessed based on their labeling. 1. Participants who are immediately eligible for PegIFN treatment will begin treatment. 2. Participants who are deemed ineligible may have their eligibility re-evaluated, and the initiation of PegIFN treatment may be delayed by up to 12 weeks. 3. PegIFN eligibility assessment: Participants who fail to meet the eligibility criteria for PegIFN by week 13 (i.e., participants who are unable to receive at least 12 weeks of PegIFN treatment) will not receive PegIFN treatment. Note: Regardless of when treatment is initiated, all participants are expected to complete PegIFN treatment at the same time (Treatment Group 1: All participants are expected to complete treatment by week 48; Treatment Group 2: Treatment is expected to be completed by week 36). Therefore, the planned duration of PegIFN treatment may vary from a minimum of 12 weeks to a maximum of 24 weeks.

[0151] The population is stratified based on HBsAg levels (HBsAg ≤ 3 logIU / mL and > 3 logIU / mL) and whether participants are HBeAg positive or negative.

[0152] In the B-Clear trial to evaluate the efficacy of bepirovirsen monotherapy, the primary analysis was performed separately for two cohorts: participants stably maintaining nucleoside / nucleotide analog (NA) background therapy [On-NA], or participants not receiving NA therapy at the time [Not-on-NA]. See Yuen et al., N. Engl. J. Med. 2022; 387:1957-68. Participants were randomly assigned to receive weekly subcutaneous injections of bepirovirsen 300 mg for 24 weeks (Group 1), bepirovirsen 300 mg for 12 weeks followed by 150 mg for 12 weeks (Group 2), bepirovirsen 300 mg for 12 weeks followed by placebo for 12 weeks (Group 3), or placebo for 12 weeks followed by bepirovirsen 300 mg for 12 weeks (Group 4) (in a ratio of 3:3:3:1). The primary composite endpoint was the maintenance of HBsAg levels below the detection limit (0.05 IU / mL) and HBV DNA levels below the limit of quantification (LLOQ) (20 IU / mL) for 24 weeks from the scheduled end date of bepirovirsen treatment, without the use of drugs initiated to suppress HBV replication (Groups 1, 2, and 3).

[0153] In Group 1, the primary endpoint event occurred in 6 people (9%) who received NA therapy and in 7 people (10%) who did not. In Group 2, the primary endpoint event occurred in 6 people (9%) who received NA therapy and in 4 people (6%) who did not. In Group 1, the primary endpoint event occurred in a total of 16% of participants receiving NA therapy and 25% of participants not receiving NA therapy who had low baseline HBsAg levels (≤3 log10 IU per milliliter), compared to 6% and 7%, respectively, among participants with high baseline HBsAg levels (≥3 log10 IU per milliliter).

[0154] The B-Together trial was designed to evaluate whether sequential bitherapy with bepilovirsen and PEGIFN could improve upon the efficacy rates of bepilovirsen monotherapy observed in the B-Clear trial, and in particular whether up to 24 weeks of bepilovirsen treatment followed by up to 24 weeks of PEGIFN treatment could increase the hepatitis B virus surface antigen (HBsAg) clearance rate in participants receiving stable nucleoside / nucleotide analog (NA) therapy, and whether the virological response could persist after discontinuation of PEGIFN treatment.

[0155] This study was designed to answer several questions, including (1) when is the optimal time to add PegIFN, (2) can sequential addition of PegIFN convert partial HBsAg responders to bepirovirsen into complete responders, and (3) can the addition of PegIFN improve the duration of response after treatment completion. An innovative approach was used to address the designed challenges. The sequential design and eligibility period between bepirovirsen and PegIFN treatment were implemented to manage potential overlapping toxicity risks (such as thrombocytopenia). Frequent PegIFN eligibility assessments after the completion of bepirovirsen treatment ensured adequate safety monitoring and timely initiation of PegIFN treatment. At the time of the study design, safety data for bepirovirsen beyond 4 weeks were limited. To demonstrate therapeutic efficacy, the design relied heavily on indirect comparisons using historical PegIFN data and available bepirovirsen monotherapy data.

[0156] B. Objectives and Estimates The primary, secondary, and exploratory objectives, as well as the estimates and endpoints, are as follows:

[0157] [Table 1] JPEG2026519935000002.jpg214170JPEG2026519935000003.jpg44168JPEG2026519935000004.jpg191170JPEG20265199350 00005.jpg156170JPEG2026519935000006.jpg170170JPEG2026519935000007.jpg189170JPEG2026519935000008.jpg104170

[0158] C. Study population: Inclusion and exclusion criteria The inclusion criteria include the following: age 1. At least 18 to 75 years of age at the time of signing informed consent [If the age requirements for consent differ between countries / facilities, a stricter (e.g., higher age) restriction will be the requirement for that country / facility].

[0159] Participant types and disease characteristics 2. Participants eligible for PegIFN treatment 3. The patient was diagnosed with chronic HBV infection more than 6 months prior to screening and is currently receiving stable NA therapy excluding terbivudine (defined as having no changes to the NA regimen for at least 6 months prior to screening and no plans to change the stable regimen during the study period). 4. Plasma or serum HBsAg concentration > 100 IU / mL 5. Plasma or serum HBV DNA <90 IU / mL 6. Alanine transaminase (ALT) ≤ 2 × ULN

[0160] Exclusion criteria include the following: Medical condition 1. Clinically significant abnormalities in medical history other than chronic HBV infection (e.g., moderate to severe liver disease other than chronic HBV, acute coronary syndrome within 6 months prior to screening, major surgery within 3 months prior to screening, severe / unstable heart disease, poorly controlled diabetes, bleeding diathesis, autoimmune disease, or coagulation disorder) or physical examination 2. The following co-infections: Presence or history of hepatitis aC virus (HCV) b. Human immunodeficiency virus (HIV) Hepatitis cD virus (HDV) 3. A history of or suspected cirrhosis of the liver, and / or evidence of cirrhosis as determined by: a. Both aspartate aminotransferase (AST)-platelet index (APRI) > 2 and FibroSure / FibroTest result > 0.7 i. If either parameter (APRI or FibroSure / FibroTest) yields a positive result, consultation with a medical monitor is required before allowing enrollment in the study. b. Regardless of APRI or Fibrosure / FibroTest scores, participants will be excluded from the study if they meet any of the following medical history criteria. i. Liver biopsy (i.e., Metavir score F4) ii.Liver stiffness >12kPa 4. Diagnosis or suspicion of hepatocellular carcinoma based on the following evidence. a. Alpha-fetoprotein concentration ≥ 200 ng / mL b. If the alpha-fetoprotein concentration at screening is ≥50 ng / mL and <200 ng / mL, confirm the absence of liver masses by imaging within 6 months prior to randomization. 5. Participants with a history of malignant tumors within the past five years (excluding certain cancers cured by surgical resection, such as skin cancer), or those currently being evaluated for the possibility of malignant tumors, are excluded. 6. A history of vasculitis, or symptoms and signs suggestive of vasculitis [e.g., vasculitic rash, skin ulcers, recurrent hematuria of unknown cause], or a history / presence of other diseases that may be associated with vasculitic conditions [e.g., lupus erythematosus, rheumatoid arthritis, relapsing polychondritis, mononeuropathy]. 7. A history of extrahepatic disorders that may be related to HBV immunopathology (e.g., nephrotic syndrome, any type of glomerulonephritis, polyarteritis nodosa, cryoglobulinemia, uncontrolled hypertension) 8. Uncontrolled thyroid dysfunction or abnormal thyroid-stimulating hormone (TSH) levels 9. Positive (or borderline positive) for anti-neutrophil cytoplasmic antibody (ANCA) at screening: a. Participants who meet these criteria will be considered for inclusion in the study as follows: i. Analysis of MPO-ANCA [perinuclear anti-neutrophil cytoplasmic antibody (pANCA)] and PR3-ANCA [classical anti-neutrophil cytoplasmic antibody (cANCA)], and ii. In consultation with the medical monitor, re-examine the participant's complete medical history and confirm that there is no history of or current symptoms of vasculitis / inflammatory / autoimmune conditions. 10. Low C3 levels at screening, and evidence of a history or current symptoms of vasculitis / inflammatory / autoimmune conditions. a. All participants who show low C3 levels at screening will consult with a medical monitor about their medical history before enrollment. 11. History of alcohol or drug abuse / dependence a. Current alcohol use that the principal investigator determines may hinder participant compliance. b. A history or current status of substance abuse / dependence that the principal investigator determines may impede the participant's ability to comply with the trial. i. Refers to illegal drugs and substances with potential for abuse. Medications used by participants as directed, whether over-the-counter or prescription, are acceptable and do not fall under the exclusion criteria. 12. Pre-existing severe psychotic conditions or a history of severe mental disorders, including severe depression, suicidal ideation, and suicide attempts.

[0161] Past medical history / combination therapy 13. If you are currently taking an immunosuppressant (e.g., prednisone) or have taken one within the past three months prior to screening, excluding short-term therapy (≤2 weeks) or use of topical / inhaled corticosteroids. 14. Participants for whom immunosuppressive therapy, including therapeutic doses of steroids, is contraindicated. 15. Patients with a history of treatment with pegylated interferon or interferon will be excluded. 16. Participants requiring anticoagulation therapy (e.g., antiplatelet agents such as warfarin, factor Xa inhibitors, or clopidogrel). 17. Participants who are currently taking terbivudine or who have taken it within the past 6 months prior to screening.

[0162] Previous clinical trial experience with / in combination therapy 18. Participants must have participated in the clinical trial and received the investigational drug within the following periods prior to the first dose date of this study: five times the half-life (if known) or five times the duration (if known) of the biological effect of the investigational treatment (whichever is longer), or 90 days (if the half-life or duration is unknown). 19. If you have a history of treatment with any oligonucleotide or small interfering RNA (siRNA) within 12 months prior to the date of your first dose.

[0163] Diagnostic evaluation 20. Freedericia's QT correction formula (QTcF) ≥ 450 milliseconds (If QTcF ≥ 450 milliseconds is shown in a single ECG at the time of screening, use the mean of 3 replicates to confirm that the participant meets the exclusion criteria.) 21. The following test results: a. Serum albumin <3.5 g / dL b. Glomerular filtration rate (GFR) < 60 mL / min / 1.73 m² calculated using the Chronic Kidney Disease Epidemiology Collaborative Research Formula (in Japan, the Japanese Society of Nephrology-Chronic Kidney Disease Initiative Formula) 2 c. International Ratio of Standardization (INR) > 1.25 d. Platelet count <140×10 9 / L e. Absolute neutrophil count (ANC) < 1.5 × 10⁻⁶ 9 cells / L f. Baseline hemoglobin <10 g / dL g. Total bilirubin > 1.25 × ULN. Participants with benign unconjugated hyperbilirubinemia with total bilirubin > 1.25 × ULN should be consulted with the medical monitor regarding their inclusion in the study. h. Urinary albumin / creatinine ratio (ACR) ≥ 0.03 mg / mg (or ≥ 30 mg / g). In the case of ACR exceeding this threshold, eligibility can be confirmed by a second measurement. If the urinary albumin value and urinary creatinine value of the participant are low and the calculated urinary ACR is ≥ 0.03 mg / mg or more (or ≥ 30 mg / g), the principal investigator of the trial shall confirm that the participant has no history of diabetes, hypertension, or other risk factors that may affect renal function, and consult with the medical monitor or designee.

[0164] Other exclusion criteria 22. History of previous treatment with bepirovirsen or interferon-containing therapy, or allergy to its components, or drug allergy or other allergy history that is contraindicated for participation in the judgment of the principal investigator of the trial or the medical monitor

[0165] D. Evaluation and procedures of the trial The main objective measurement of efficacy in this trial is the evaluation of serum HBsAg and HBV DNA levels when PegIFN is administered after bepirovirsen treatment in CHB participants. The primary efficacy endpoint is to achieve serum HBsAg and HBV DNA levels < LLOQ.

[0166] If HBsAg exceeding LLOQ is detected after HBsAg seroclearance is achieved, confirmation by retesting is required within one week after receiving the test result. If the initial test is not confirmed, the retest result will be used.

[0167] For all safety evaluations, there are scheduled time points. To ensure appropriate safety monitoring, additional time points for safety tests (vital signs, physical examinations, and clinical safety tests, etc.) may be added during the trial period based on newly obtained data.

[0168] E. Intermediate results After all participants completed the treatment period of the trial, an interim analysis as specified in the protocol was conducted. See Table 1.

[0169]

Table 2

[0170] In this analysis, it was reported that 8 out of 53 participants (15%) in the group treated with bepirovirsen for 12 weeks + PegIFN for 24 weeks achieved the primary efficacy endpoints of HBsAg and HBV DNA <LLOQ that persisted for 24 weeks after completion of PegIFN treatment. The response rates in participants with baseline HBsAg values of ≤ 3000 IU / mL and ≤ 1000 IU / mL were 26% (8 / 31) and 41% (7 / 17), respectively. The point estimate of the primary efficacy endpoint rate observed in B-Together was numerically higher than the values observed in the 12-week bepirovirsen monotherapy in B-Clear (in participants under stable NA therapy, showing rates of 7% (3 / 41) and 11% (2 / 19) in the ≤ 3000 IU / mL population and the ≤ 1000 IU / mL population, respectively).

[0171] Regarding the interim analysis, the data of the second treatment group that received bepirovirsen for 24 weeks followed by PegIFN administration for another 24 weeks were incomplete, but in an additional 55 participants, a similar main evaluation rate at the end of the trial was predicted.

[0172] The relapse rates compared between the B-Together interim analysis and the B-Clear final data showed that the overall relapse rates in B-Clear groups 1 and 3 were 63% and 75%, respectively. In contrast, in B-Together, 4 participants (31%) who achieved a virological response at the end of the bepirovirsen treatment period relapsed by the end of the PegIFN period. Furthermore, at the interim analysis point, among the participants who showed a primary efficacy response at the end of PegIFN treatment, no relapses had been observed so far.

[0173] The additional effect of 24 weeks of PEGASYS in this regimen is thought to stem from a significant reduction in the relapse rate during the 6 months following the completion of bepilovirsen treatment, supporting the hypothesis that adding immunomodulatory therapy after suppression of viral antigens by bepilovirsen treatment increases the likelihood of a sustained response and enhances the chances of achieving functional remission after the completion of all treatment.

[0174] F. Results at the end of the exam Table 2 shows the virological response data analyzed at the end of the study. One HBeAg(+) participant in Group 1 achieved the primary endpoint (PE) (HBsAg ≤ 1000 IU / mL), while no HBeAg(+) participants in Group 2 achieved PE. Two participants in Group 1 maintained their response for most of the treatment interruption period (OT) from PEG-EOT, except for a mild HBsAg elevation at OT-W24. In both groups, participants with lower baseline HBsAg levels showed a higher response. See Figure 2. All responders had baseline HBsAg ≤ 3000 IU / mL. Only bepilovirsen responders benefited from PEG-IFN treatment by preventing relapse during treatment interruption (relapse rate: B-Together vs. B-Clear, Group 1: 58% vs. 63%, Group 2: 2.0% vs. 75%). Compared to HBeAg-positive participants (4 out of 28: 14.3%), a higher proportion (30 out of 80: 37.5%) of HBeAg-negative participants had low HBsAg levels (≤1000 IU / mL) at baseline.

[0175] [Table 3]

[0176] In contrast to the results of the B-Clear trial, the proportion of participants achieving the primary endpoint was higher in the 12-week treatment group (Group 2) than in the 24-week bepilovirsen treatment group (Group 1). However, the 95% confidence intervals (CIs) overlapped widely, indicating high levels of variability and uncertainty regarding the difference between the treatment groups. The difference in efficacy observed between the two groups is likely due to the higher relapse rate in Group 1 during the Peg-IFN treatment period and the follow-up period after treatment completion. Group 1 was disproportionately affected by COVID-19 lockdowns during the eligibility period and the treatment period for Peg-IFN therapy.

[0177] Logistic regression models were constructed, and variables associated with the primary efficacy response were determined using a forward-stepwise selection method, selecting from the following ordered variables based on AIC: age, race, sex, BMI, fibrosis score, duration of diagnosis, NA type used at baseline, ALT, urinary albumin, history of interferon use, and GFR. In the final subgroup logistic regression models, several variables were identified that were statistically significantly associated with the response, including low baseline HBsAg levels, high body mass index, and treatment with tenofovir alafenamide fumarate or tenofovir disoproxil fumarate (Table 3).

[0178] [Table 4]

[0179] The results of the B-Together trial demonstrate that sequential therapy with bepirovirsen followed by PEG-IFN is tolerable and effective in chronic HBV-infected participants with stable NA. Participants responded to bepirovirsen treatment similarly to that seen in the B-Clear trial, with increased response rates in participants with low baseline HBsAg. The addition of PEG-IFN treatment improved the maintenance of the bepirovirsen response after treatment completion by preventing relapse. These data support the use of PEG-IFN treatment to maintain HBsAg and HBV DNA clearance in participants who responded to bepirovirsen. These data also support the potential benefits of bepirovirsen in combination with other therapies. Overall, this trial highlights sequential therapy as an effective treatment strategy for finite treatment regimens for HBV infection.

[0180] Example 2 Meta-analysis based on models to predict the response rate (HBsAg clearance) of PegIFN-based regimens in a specific chronic hepatitis B patient population. To establish an absolute effect model for HBsAg clearing for PEGIFN-based regimens, a model-based meta-analysis (MBMA) was developed, enabling simulations to predict response rates (HBsAg clearing) for a given population. Using baseline HBsAg level data published between 2000 and 2022, a systematic re-examination of clinical trials and prospective and retrospective cohort studies examining HBsAg clearing with PEGIFN treatment was conducted. The MBMA used aggregate summary-level data from a total of 83 trial strata (63 trial strata from randomized controlled trials) to generate HBsAg clearing outcomes from 13,235 participants at treatment completion. Covariates predicting HBsAg clearing in PEGIFN-treated patients were treatment duration, baseline HBsAg level, and baseline HBeAg positivity. Additional covariates were evaluated, but were either not significant or lacked sufficient data.

[0181] Table 4 shows data from 100,000 simulations based on different patient characteristics regarding baseline HBsAg and HBeAg. HBeAg status (positivity rate) and baseline HBsAg levels (3.09 and 2.73log10IU / mL) are shown to allow comparison with B-Together results, as they represent the proportion of HBeAg-positive patients and the median baseline HBsAg level, respectively, within the patient population with baseline HBsAg ≤3000IU / mL and ≤1000IU / mL.

[0182] [Table 5]

[0183] Example 3 A Phase 3 trial evaluating the efficacy and safety of sequential therapy with bepilovirsen and PEGIFN in participants with chronic hepatitis B virus (persistent-onset B) who have a history of nucleoside / nucleotide analog therapy. A. Experimental Design This was a phase 3b, multicenter, randomized, double-blind trial to evaluate the efficacy, safety, PK profile, and duration of HBsAg suppression observed in chronic HBV-infected participants with a background of NA treatment (baseline HBsAg levels ≤ 3000 IU / mL) (participants stratified by HBsAg levels) who received 300 mg of bepirovirsen weekly for 24 weeks (two loading doses), followed by 180 μg of PEGIFN weekly for 24 weeks, compared to a placebo-controlled 48-week treatment (see Figure 3).

[0184] Exam details: • The trial period can be up to 117 weeks (including 45-60 days of screening). The test consists of five levels: o Investigational drug administration phase (divided into two phases): • Phase 1 = Investigational drug administration phase 1 (TS1) with bepirovirsen (or placebo) under background NA treatment. • Phase 2 = Investigational drug administration phase 2 (TS2) with PegIFN (or placebo) under background NA treatment. o Stage 3 = NA alone Stage 4 = NA discontinuation or NA continuation Stage 5 = Extension of observation period and persistence of response The total duration of the investigational drug administration phase will be a maximum of 48 weeks, with TS1 and TS2 each lasting 24 weeks. This study has two groups, and participants will be randomly assigned to either Group 1 or Group 2 of the study. Group 1: oTS1 is given bepirovirsen 300 mg weekly for 24 weeks (2 loading doses). oTS2 involves taking PEGIFN 180μg weekly for 24 weeks. Group 2: oTS1 involves receiving a placebo weekly for 24 weeks (2 loadings). oTS2 involves taking a placebo weekly for 24 weeks. • In the third stage (NA only), background NA is continued for 12 weeks. • Evaluate eligibility to discontinue background NA at week 60. Participants who meet the eligibility criteria for oNA cancellation can cancel their NA. These participants will be followed up for a further 48 weeks, including the following: • 24-week suspension of NA (Phase 4) • 24-week follow-up and persistence of response (Stage 5) For participants who qualify for NA discontinuation, the trial period will be up to approximately 117 weeks. Participants who do not meet the criteria for discontinuation of oNA will continue NA therapy for 24 weeks, after which the trial will end (the trial period is up to approximately 93 weeks). As the trial progresses, the frequency of hospital visits decreases. Hospital visits for administration are approximately weekly (until week 24), then approximately every other week to every other month in treatment phase 2 (weeks 25-48), then approximately weekly to every other week (weeks 48-60), and then gradually decrease to approximately monthly during the final trial period. In some countries / centers, participants have the option of using centrally managed home care services for most hospital visits.

[0185] B. Objectives and Estimates The objectives, endpoints, and expectations are as follows:

[0186] [Table 6]

[0187] Unless otherwise specified, all primary, secondary, and exploratory trial objectives are evaluated using estimates defined by the following common elements with respect to the population, treatment comparison, and complication event management approach. Eligibility assessment for NA discontinuation and responses after discontinuation of all HBV (or trial) treatment are based on the analysis period.

[0188] The primary clinical interest is to evaluate whether sequential bepirovirsen plus pegIFN therapy increases the FC response rate compared to placebo, without rescue medication, regardless of discontinuation, interruption, or non-compliance with bepirovirsen, pegIFN, or NA, and regardless of non-rescue medications, including oral medications with proven anti-HBV activity, that are not intended to manage virological breakthroughs, in chronic HBV-infected participants with baseline HBsAg ≤3000 IU / mL and receiving NA therapy.

[0189] Main Estimates

[0190] [Table 7]

[0191] Major secondary estimates Treatment and comorbidities are defined in the same way as the primary estimate for secondary estimates. For the three secondary objectives, the population and population summary are defined as follows:

[0192] Primary secondary efficacy objectives: To evaluate the therapeutic effect of sequential bepilovirsen + pegIFN therapy in chronic HBV virus infection participants with baseline HBsAg levels of ≤1000 IU / mL and receiving nasogabacterial therapy.

[0193] The clinical interest is to evaluate whether sequential bepilovirsen + pegIFN therapy increases FC compared to placebo, without rescue medication, regardless of discontinuation, interruption, or non-compliance with bepilovirsen or NA, and without the use of non-rescue medications, including oral medications with proven anti-HBV activity that are not intended to manage virological breakthroughs, in chronic HBV-infected participants with baseline HBsAg ≤ 1000 IU / mL who are receiving NA therapy.

[0194] [Table 8]

[0195] Primary secondary efficacy objectives: To evaluate the therapeutic effect of bepilovirsen + pegIFN sequential therapy on HBV DNA suppression at the time of treatment discontinuation after a finite treatment period in chronic HBV-infected participants with baseline HBsAg ≤ 3000 IU / mL who are receiving NA therapy.

[0196] The clinical interest is to evaluate whether sequential bepirovirsen plus pegIFN therapy improves HBV DNA suppression at treatment discontinuation after a finite treatment period compared to placebo, without the use of rescue medications, regardless of discontinuation, interruption, or non-compliance with bepirovirsen, pegIFN, or NA, and regardless of non-rescue medications, including oral medications with proven anti-HBV activity that are not intended to manage virological breakthroughs, in chronic HBV-infected participants with HBsAg ≤3000 IU / mL at baseline and receiving NA therapy.

[0197] [Table 9]

[0198] Primary secondary efficacy objectives: To evaluate the therapeutic effect of bepilovirsen + pegIFN sequential therapy on HBV DNA suppression at treatment discontinuation after a finite treatment period in chronic HBV-infected participants with baseline HBsAg ≤ 1000 IU / mL who are receiving NA therapy.

[0199] The clinical interest is to evaluate whether sequential bepirovirsen plus pegIFN therapy improves HBV DNA suppression at treatment discontinuation after a finite treatment period compared to placebo, without the use of rescue medication, regardless of discontinuation, interruption, or non-compliance with bepirovirsen, pegIFN, or NA, and regardless of non-rescue medications, including oral medications with proven anti-HBV activity that are not intended to manage virological breakthroughs, in chronic HBV-infected participants with HBsAg ≤1000 IU / mL at baseline and receiving NA therapy.

[0200] [Table 10]

[0201] Additional Stimand Additional estimates are defined to complement the primary estimate and primary secondary estimates with alternative strategies for comorbid events.

[0202] The initial additional estimates for the primary and primary secondary objectives address the clinical question of whether bepilovirsen + pegIFN sequential therapy, compared to placebo, increases FC and improves HBV DNA suppression without rescue medication, provided that no investigational discontinuation, interruption, or non-compliance caused by widespread disruptive events resulting in intervals of 21 days or more between doses occurs. The attributes of this additional estimate are the same as the primary estimate, except that a hypothetical strategy will be used for the concomitant event of investigational discontinuation, interruption, or non-compliance caused by widespread disruptive events resulting in intervals of 21 days or more between doses.

[0203] A second additional stimand for the primary and primary secondary objectives addresses the clinical question of whether bepirovirsen + pegIFN sequential therapy increases FC and improves HBV DNA suppression compared to placebo without rescue medication, in the event of discontinuation, interruption, and non-compliance with the clinical trial, and in the absence of use of a proven anti-HBV drug not intended for HBV DNA reversal management. The attributes of this additional stimand are the same as the primary stimand, except that a hypothetical strategy is used for the concomitant event of use of a proven anti-HBV drug not intended for HBV DNA reversal management.

[0204] Exploratory estimator: Eligibility for NA discontinuation and responses after discontinuation of all HBV (or experimental) treatments are based on the prescribed analysis period.

[0205] [Table 11] JPEG2026519935000019.jpg139170

[0206] C. Study population: Inclusion and exclusion criteria The inclusion criteria include the following: age 1. At least 18 years of age at the time of signing informed consent (if the age requirements for consent differ in a country / facility, a stricter [e.g., higher age] restriction may apply in that country / facility).

[0207] Participant types and disease characteristics 2. Participants who are HBeAg-negative or HBeAg-positive at the time of screening. 3. Chronic HBV infection was confirmed more than 6 months prior to screening, and • Currently receiving stable NA therapy (excluding terbivudine), defined as having no changes to the NA regimen for at least 6 months prior to screening and no plans to change that stable regimen during the study period. 4. Plasma or serum HBsAg concentration > 100 IU / mL and HBsAg concentration ≤ 3000 IU / mL 5. Plasma or serum HBV DNA concentration is sufficiently suppressed (defined as plasma or serum HBV DNA < 90 IU / mL) 6. Alanine transaminase (ALT) ≤ 2 × ULN 7. Willing to receive treatment with PegIFN and eligible participants 8. Willing to discontinue NA treatment according to the protocol and eligible participants who can discontinue

[0208] Informed Consent 9. Have the ability to sign the informed consent, which includes compliance with the ICF and the requirements and restrictions described in this protocol

[0209] Exclusion criteria are as follows: Medical condition 1. Clinically significant abnormalities other than chronic HBV infection in the past history (e.g., moderate to severe liver diseases other than chronic HBV, acute coronary syndrome within 6 months before screening, major surgery within 3 months before screening, severe / unstable heart disease, poorly controlled diabetes, bleeding tendency, or coagulation disorder) or physical examination 2. The following coinfections: a. Hepatitis C virus including past infected individuals with less than 12 months after cure at the time of screening (defined as having HCV RNA negative for 12 months) b. Hepatitis D virus defined as HDV antibody positive c. Human immunodeficiency virus (HIV) 3. Past history or suspicion of cirrhosis and / or evidence of cirrhosis determined by the following: a. Both APRI > 2 and FibroSure / FibroTest result > 0.7 · If only one parameter (APRI or FibroSure / FibroTest) result is positive, consultation with the medical monitor is required before allowing inclusion in the study b. Regardless of the APRI or Fibrosure / FibroTest score, if the participant's medical record has a history of either / both liver biopsy or liver stiffness measurement and meets one of the following criteria, they will be excluded from the trial · Liver biopsy (i.e., Metavir score F4) · Liver stiffness > 12 kPa 4. Diagnosis or suspicion of hepatocellular carcinoma evidenced by the following a. Alpha-fetoprotein concentration ≥ 200 ng / mL b. If the alpha-fetoprotein concentration at screening is ≥ 50 ng / mL and < 200 ng / mL, confirm the absence of liver tumors by imaging examination within 6 months before randomization 5. History of malignant tumors within the past 5 years (except for certain cancers cured by surgical resection, e.g., skin cancer). Participants being evaluated for the possibility of malignant tumors are excluded 6. History of vasculitis, or symptoms and signs indicating the possibility of vasculitis (e.g., vasculitic rash, skin ulcers, recurrent hematuria of unknown origin). Presence or history of autoimmune disorders, or history / presence of other diseases that may be associated with vasculitic conditions (e.g., systemic lupus erythematosus, rheumatoid arthritis, relapsing polychondritis, polyneuropathy) 7. History of extrahepatic disorders associated with HBV immune conditions (e.g., nephrotic syndrome, any type of glomerulonephritis, polyarteritis nodosa, cryoglobulinemia, poorly controlled hypertension) 8. Poorly controlled hypothyroidism or abnormal thyroid-stimulating hormone (TSH) levels 9. Existing severe psychopathy, or history of severe mental disorders including severe depression, suicidal thoughts, and suicide attempts 10. History of alcohol or drug abuse / dependence a. Current alcohol use that the principal investigator of the clinical trial determines may interfere with the participant's compliance b. History or current situation of drug abuse / dependence that the principal investigator of the clinical trial determines may interfere with the participant's compliance

[0210] Past medical history / combination therapy 11. If you are currently taking immunosuppressants (e.g., prednisone) or have taken them within the past three months prior to screening, excluding short-term therapy (≤2 weeks) or use of topical / inhaled steroids. 12. Participants for whom immunosuppressive therapy, including therapeutic doses of steroids, is contraindicated should not be considered for inclusion in the study. 13. You are currently taking or have taken interferon-containing therapy within the past three years of screening. 14. Participants requiring anticoagulant therapy (e.g., warfarin, factor Xa inhibitor) or antiplatelet agents (e.g., clopidogrel, aspirin), unless the treatment can be safely discontinued at the discretion of the principal investigator during the study period. Temporary use is permitted. 15. Participants must have participated in the clinical trial and received the investigational drug within the following periods prior to the first dose date of this study: five times the half-life (if known) or five times the duration (if known) of the biological effect of the investigational treatment (whichever is longer), or 90 days (if the half-life or duration is unknown). 16. If you have a history of treatment with any oligonucleotide or siRNA within 12 months prior to the date of your first dose. 17. History of treatment with bepilovirsen 18. Participants who are currently taking terbivudine or who have taken it within the past 6 months of screening.

[0211] Diagnostic evaluation 19. Freedericia's QT correction formula (QTcF) ≥ 450 milliseconds (If QTcF ≥ 450 milliseconds is shown in a single ECG at the time of screening, use the mean of 3 replicates to confirm that the participant meets the exclusion criteria.) 20. The following test results: a. Platelet count < 140 × 10 9 / L b. Total bilirubin > 1.25 × ULN • Participants with benign unconjugated hyperbilirubinemia (total bilirubin > 1.25 × ULN) should consult with the medical monitor regarding their inclusion in the study. Serum albumin <3.5 g / dL c. INR > 1.25 d. Serum albumin ≤3.5 g / dL e. Glomerular filtration rate (GFR) <60 mL / min / 1.73 m² calculated using the CKD EPI formula (JSN CKDI formula in Japan) 2 f. Urinary albumin / creatinine ratio (uACR) ≥ 0.3 mg / mg (or ≥ 300 mg / g). If the ACR exceeds this threshold, eligibility can be confirmed by a second measurement.

[0212] Other exclusion criteria 21. History of / hypersensitivity to bepirobircene or peg-IFN-containing therapy, or any of its components, or a history of drug allergies or other allergies that the principal investigator or medical monitor deems contraindicated for participation. 22. Participants who do not wish to discontinue NA therapy for chronic HBV infection.

[0213] Those skilled in the art will readily understand that this disclosure is well suited to achieving its purpose and obtaining the purposes and benefits described, as well as those specific thereto. The particular embodiments described herein are intended to be representative and illustrative and not to limit the scope of the invention. Modifications and other uses therein will be apparent to those skilled in the art in the scope of the invention as defined by the claims.

[0214] All patent applications, patents, and printed publications referenced herein are incorporated herein by reference in their entirety, except for definitions, waivers of subject matter, or denials, and to the extent that incorporated material conflicts with the express disclosure herein (in which case the language of this disclosure shall prevail).

Claims

1. A method for treating chronic hepatitis B in a person in need, comprising administering to the person a therapeutically effective amount of antisense oligonucleotide (ASO) and a therapeutically effective amount of interferon, wherein the person has an HBsAg baseline level below a threshold level.

2. A method for treating chronic hepatitis B in people who need it, (a) Determining that the person has an HBsAg baseline level below the threshold level; and (b) Administering the person a therapeutically effective amount of ASO and a therapeutically effective amount of interferon. A method that includes the following:

3. A method for treating chronic hepatitis B in people who need it, (a) Measuring the baseline level of the human HBsAg; (b) comparing the HBsAg baseline level with a threshold level; and (c) If the HBsAg baseline level is below the threshold level, administer a therapeutically effective amount of ASO and a therapeutically effective amount of interferon to the person. A method that includes the following:

4. The method according to any one of claims 1 to 3, wherein the threshold level is in the range of about 1000 IU / mL to 3000 IU / mL.

5. The method according to any one of claims 1 to 4, wherein the threshold level is approximately 1000 IU / mL.

6. The method according to any one of claims 1 to 4, wherein the threshold level is approximately 3000 IU / mL.

7. The method according to any one of claims 1 to 6, wherein the person is not treated with another HBsAg lowering agent or another immunomodulator.

8. The method according to any one of claims 1 to 6, wherein the human is treated with another HBsAg lowering agent or another immunomodulator.

9. The method according to any one of claims 1 to 8, wherein the ASO is administered by subcutaneous injection.

10. The method according to any one of claims 1 to 9, wherein the ASO is administered once a week in a dose of approximately 150 mg or 300 mg.

11. The method according to any one of claims 1 to 10, wherein the ASO is administered for 12 to 48 weeks.

12. The method according to claim 10 or 11, wherein the ASO is administered weekly with additional loading doses on the 4th and 11th days after the initial dose during the first two weeks.

13. The method according to any one of claims 1 to 12, wherein the ASO is administered at a dose of 300 mg once a week for 12 weeks.

14. The method according to any one of claims 1 to 12, wherein the ASO is administered at a dose of 300 mg once a week for 24 weeks.

15. The method according to any one of claims 1 to 14, wherein the interferon is interferon-alpha.

16. The method according to any one of claims 1 to 14, wherein the interferon is interferon λ.

17. The method according to claim 15, wherein the interferon is peg-interferon α-2a.

18. The method according to claim 17, wherein PEGIFN is administered once a week at a dose of approximately 180 μg.

19. The method according to any one of claims 1 to 18, wherein interferon is administered by subcutaneous injection.

20. The method according to any one of claims 1 to 19, wherein interferon is administered after discontinuation of ASO.

21. The method according to any one of claims 1 to 20, wherein interferon is administered for 12 to 24 weeks after discontinuation of ASO.

22. The method according to any one of claims 1 to 21, wherein the human is under stable nucleoside or nucleotide analog (NA) therapy.

23. The method according to claim 21, wherein the NA therapy is selected from the group consisting of lamivudine, adefovir, adefovir dipivoxil, terbivudine, entecavir, tenofovir, tenofovir disoproxil fumarate, tenofovir alafenamide, and any pharmaceutically acceptable salt thereof.

24. The method according to claim 22 or claim 23, wherein the human discontinues NA therapy 24 weeks after discontinuing interferon.

25. The method according to any one of claims 1 to 21, wherein the human is not treated with either a nucleoside or a nucleotide analog (NA).

26. The method according to any one of claims 1 to 25, wherein the human is HBeAg negative prior to ASO treatment.

27. The method according to any one of claims 1 to 25, wherein the human is HBeAg positive before ASO treatment.

28. The method according to any one of claims 1 to 27, wherein the human HBV DNA and HBsAg levels are below the lower limit of quantification (LLOQ) 24 weeks after discontinuation of all HBV therapy.

29. A combination of ASO and PEGIFNα-2a for the treatment of chronic hepatitis B in humans, (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The ASO is administered to the person at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of ASO. combination.

30. A combination of ASO and PEGIFNα-2a for achieving functional HBV cure in a person with chronic hepatitis B, (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The ASO is administered to the human being at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of ASO. combination.

31. The use of ASO in the manufacture of pharmaceuticals for the treatment of chronic hepatitis B in humans, (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The ASO is administered to the human being at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of ASO. use.

32. The use of PEGIFNα-2a in the manufacture of pharmaceuticals for the treatment of chronic hepatitis B in humans, (a) The person being treated has a baseline HBsAg of 3000 IU / mL or less; (b) The ASO is administered to the human being at a dose of approximately 300 mg weekly for 12 or 24 weeks; (c) PegIFNα-2a is administered to the aforementioned human at a dose of approximately 180 μg weekly for 12 to 24 weeks after discontinuation of ASO. use.

33. The method, combination, or use according to any one of claims 1 to 32, wherein the ASO comprises 20 linked nucleosides and has the nucleic acid base sequence of Sequence ID No.

1.

34. The method, combination, or use according to any one of claims 1 to 33, wherein the ASO consists of 20 linked nucleosides and has the nucleic acid base sequence of Sequence ID No.

1.

35. The aforementioned ASO A gap segment consisting of linked deoxynucleosides, A 5' wing segment consisting of linked nucleosides, and 3' wing segment consisting of linked nucleosides The method, combination, or use according to any one of claims 1 to 34, comprising, wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, and each nucleoside in each wing segment comprises a modified sugar.

36. The method, combination, or use according to claim 35, wherein the modified sugar comprises a 2'-O-methoxyethyl (2'-MOE) group or is a bicyclic sugar.

37. The method, combination, or use according to claim 35 or claim 36, wherein the internucleoside bonds of the ASO are phosphorothioate bonds.

38. The ASO is AUS1233 / AUS1138, AUS1444, AUS1458, AUS1459, AUS1460, AUS1427 / AUS1461, A US1462, AUS1463, AUS1464, AUS1465, AUS1463, AUS1467, AUS1468, AUS1470, AUS1471, A method, combination, or use according to any one of claims 1 to 32, selected from the group consisting of AUS1472, AUS1473, AUS1474, AUS1475, AUS1476 / AUS1493, AUS1478, AUS1479, AUS1489, AUS1490, AUS1443, and AUS1322.

39. The method, combination, or use according to any one of claims 1 to 32, wherein the ASO is bepilobirsene.

40. The method, combination, or use according to any one of claims 1 to 32, wherein the ASO is AUS1493.

41. The method, combination, or use according to any one of claims 1 to 32, wherein the ASO is AHB-137.

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