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Multi-valent hepatitis b virus antigen binding molecules and uses thereof

A hepatitis B virus, binding molecule technology, applied in the direction of antiviral agents, antibody medical ingredients, medical preparations containing active ingredients, etc., can solve problems such as side effects, and achieve the effect of strengthening virus clearance

Active Publication Date: 2018-04-17
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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  • Multi-valent hepatitis b virus antigen binding molecules and uses thereof
  • Multi-valent hepatitis b virus antigen binding molecules and uses thereof
  • Multi-valent hepatitis b virus antigen binding molecules and uses thereof

Examples

Experimental program
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Embodiment approach

[0055] Furthermore, use of "and / or" herein should be construed as specifically disclosing 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" used in phrases such as "A, B, and / or C" is intended to include the various embodiments of: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0056] Unless defined otherwise, 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" (Concise Dictionary of Biomedicine and Molecular Biology), Juo, Pei-Show, 2nd edition, 2002, CRC Press (CRCPress); "Dictionary of Cell and Molecular Biology" (The Dictionary of Cell and Molecular ...

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 minutes) and the supernatant was layered on 0.5 ml of 20% sucrose TBS (in 5.1 ml tubes) and centrifuged (266,000 x g, 30 minutes). The pellets were resuspended in 20nM Tris chloride (pH 7.4) and 140mM NaCl (Tris buffered saline) (wetted at 4°C overnight to soften), pooled and equilibrated with CsCl (0.22g / ml initial concentration) centrifuged (266,000 x g, 72 hours). Fractions (250 [mu]l) were removed from the gradient and monitored by electron microscopy and DNA extraction for analysis of Hepatitis B DNA. Fractions con...

Embodiment 2

[0277] Example 2: HBV capsid and virion isolation

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

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Abstract

This disclosure provides a multimeric hepatitis B virus (HBV) protein binding molecule, e.g., a dimeric IgA or a pentameric or hexameric IgM binding molecule, comprising at least two bivalent bindingunits, or variants or fragments thereof, 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. The disclosure also provides compositions comprising the multimeric binding molecules, polynucleotides encoding the multimeric binding molecules, and methods to make and use the multimeric binding molecules.

Description

[0001] Cross-references 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 class 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). The serotypes were based on variations in the envelope protein sequence, while the 8 genotypes were distinctly distributed in specific geographic regions of the world. Genotypic differences have been shown to be associated 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 leng...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P31/20A61K39/29G01N33/53
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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