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Multivalent hepatitis B virus antigen-binding molecule and application thereof

A technology that combines molecules and antigens, applied in antiviral agents, antibody medical ingredients, medical preparations containing active ingredients, etc., can solve problems such as side effects

Active Publication Date: 2022-06-07
IGM BIOSCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Various monoclonal antibodies and combination therapies have been investigated for the treatment and / or cure of HBV infection, including chronic HBV infection, but none have been commercialized
[0019] Treatments for chronic infections, such as interferon and lamivudine, have limited effectiveness and are known to cause serious side effects

Method used

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  • Multivalent hepatitis B virus antigen-binding molecule and application thereof
  • Multivalent hepatitis B virus antigen-binding molecule and application thereof
  • Multivalent hepatitis B virus antigen-binding molecule and application thereof

Examples

Experimental program
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Embodiment approach

[0055] Furthermore, "and / or", as used herein, should be deemed to specifically disclose each of the two features or components, with or without the other. Thus, the term "and / or" as used herein in the phrase "A and / or B" is intended to include "A and B," "A or B," "A" (alone), and "B (alone)." Likewise, the term "and / or" as used in phrases such as "A, B, and / or C" is intended to include various embodiments of: A, B, and C; A, B, or C; A or C; A B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0056] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd Edition, 2002, CRC Press; The Dictionary of Cell and Molecular Biology (The Dictionary of Cell and Molecular Biology), 3rd ed., 1999, Academic Press; and Oxford Dictionary of Biochemistry...

Embodiment 1

[0275] Example 1: Surface antigen preparation

[0276] Surface antigens can be prepared as previously reported. (See, eg, Short et al., J. Mol. Biol. (2009) 390, 135-141). In one method, HBsAg is isolated from the blood of HBV carriers. Frozen serum containing HBV can be obtained, for example, from Diagnostics, Development and Research Division, National Blood Service, Colindale Centre, London, UK. Each package was then thawed and centrifuged (3700 x g, 20 min), the supernatant was layered in 0.5 ml of 20% sucrose TBS (in a 5.1 ml tube), and centrifuged (266,000 x g, 30 min). Particles were resuspended in 20 nM Tris chloride (pH 7.4) and 140 mM NaCl (Tris buffered saline) (wet at 4°C overnight to soften), pooled and equilibrated with CsCl (0.22 g / ml initial concentration) by centrifugation (266,000 x g, 72 hours). Fractions (250 [mu]l) were removed from the gradient and monitored by electron microscopy and DNA extraction, analyzing hepatitis B DNA. Fractions containing th...

Embodiment 2

[0277] Example 2: HBV capsid and virion isolation

[0278] Methods of isolation of capsids and virions are well known in the art and can be carried out by various known methods. (See, eg, Dryden, Molecular Cell, 22:843-850, Jun 23, 2006, Supplement). In one method of capsid isolation, freshly dissected whole livers are perfused with saline solution. The liver is then homogenized in lysis buffer (eg, 0.25M sucrose, 1 mM MgCl 2 , 5 mM Tris (pH 7.4)) and supplemented with complete protease inhibitor (Roche Applied Science, Indianapolis, IN). , the supernatant of each homogenate was obtained by centrifugation in a SW40 rotor (SW40rotor) at 4°C, 11,000 rpm for 15 minutes. Aliquots of the supernatant were taken for HBV-specific Southern blot analysis. The residue was then layered on a 30% sucrose pad (0.73M sucrose in phosphate buffered saline (PBS)) and the viral particles were pelleted by centrifugation in a SW40 rotor at 40,000 rpm for 5 hours at 4°C. Thereafter, the particl...

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Abstract

Disclosed herein is a multimerized hepatitis B virus (HBV) protein binding molecule, such as a bivalent IgA or pentameric or hexameric IgM binding molecule, comprising at least two bivalent binding units, or variants or Fragments, each comprising at least two antibody heavy chain constant regions or fragments thereof, wherein each heavy chain constant region or fragment thereof is associated with an HBV antigen binding domain. Also disclosed herein are compositions comprising multimerized binding molecules, polynucleotides encoding multimerized binding molecules, and methods of making and using multimerized binding molecules.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS [0002] This application claims priority to US Provisional Patent Application Serial No. 62 / 137,881, filed March 25, 2015, the disclosure of which is incorporated herein by reference in its entirety. Background technique [0003] Hepatitis B virus (HBV) belongs to the family of double-stranded DNA viruses and is currently known to include four major serotypes (adr, adw, ayr, and ayw) and eight major genotypes (A-H). Serotypes are based on changes in envelope protein sequences, while the eight genotypes are distinctly distributed in specific geographic regions around the world. Genotypic differences have been shown to correlate with disease severity and response to treatment (see Kramvis et al, Vaccine, 23(19):2409-23, 2005, and Magnius et al, Intervirology, 38(1-2):24- 34, 1995). [0004] The HBV genome consists of partially double-stranded circular DNA. That is, part of the genome is still single-stranded. The longer part of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/29A61P31/20A61P1/16G01N33/53
CPCA61K39/395A61K39/39541A61K39/40C07K16/2809C07K2317/24C07K2317/31C07K2317/56C07K2317/60C07K2317/734G01N2500/04G01N2500/10C07K16/082A61P1/16A61P31/20G01N2333/02G01N33/5761A61K2039/505C07K2317/35C07K2317/622C07K2317/76G01N33/502G01N33/56983
Inventor S·F·卡罗尔R·巴利加D·吴B·A·基特
Owner IGM BIOSCI INC
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