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Antibodies for treatment of hepatitis B infection and related diseases

An antibody and antigen technology, applied in the direction of antibodies, introduction of foreign genetic material using vectors, antiviral agents, etc., can solve the problems of unstable nature, high price, and few sources of high-titer plasma

Active Publication Date: 2018-10-23
XIAMEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, direct use of HBIG in the treatment of HBV-infected patients (such as CHB patients) has no obvious curative effect, and it has many limitations such as fewer sources of high-titer plasma, high price, unstable properties, and potential safety issues.

Method used

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  • Antibodies for treatment of hepatitis B infection and related diseases
  • Antibodies for treatment of hepatitis B infection and related diseases
  • Antibodies for treatment of hepatitis B infection and related diseases

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0468] The invention will now be described with reference to the following examples, which are intended to illustrate the invention, but not to limit it.

[0469] Unless otherwise specified, the molecular biology experiment methods and immunoassay methods used in the present invention are basically with reference to J.Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, and F.M.Ausubel et al., Molecular Biology Experimental Guide, 3rd Edition, John Wiley & Sons, Inc., 1995. The method described in; restriction endonucleases were used in accordance with the conditions recommended by the product manufacturer. Those skilled in the art understand that the examples describe the present invention by way of example and are not intended to limit the scope of the claimed invention.

Embodiment 1

[0470] Example 1: Preparation of human-cynomolgus monkey chimeric monoclonal antibody specifically binding to HBsAg

[0471] 1.1 Immunization of cynomolgus monkeys

[0472] Using the recombinant HBV vaccine previously developed by the laboratory (the recombinant vaccine is described in detail in Chinese patent application 201710085194.3) to immunize cynomolgus monkeys older than 6 months and weighing 4±1kg intramuscularly, the injection dose is 20μg / monkey / time. The immunization time points were at 0, 2, 6, 10, 14, 18, 22, and 26 weeks respectively. After the serum titer of cynomolgus monkey reached the plateau at the 4th dose of immunization (10 weeks), we produced a large number of memory B cells at 13, 17, 25, and 29 weeks (that is, the third week after each immunization), The peripheral blood of cynomolgus monkeys was collected 10ml / monkey / time.

[0473] 1.2 Isolation of peripheral blood mononuclear cells from cynomolgus monkeys

[0474] The isolation of peripheral ...

Embodiment 2

[0534] Example 2: Analysis of properties of cynomolgus monkey-human chimeric monoclonal antibody specifically binding to HBsAg

[0535] According to the method of Example 1, three strains of cynomolgus monkey-human chimeric monoclonal antibodies specifically binding to HBsAg were prepared and named M1-23, M3-23, and M3-13 respectively. The VH and VL amino acid sequences of the three antibodies are shown in the table below (SEQ ID NO: 1-5). In addition, the CDR sequences of the three antibodies were determined by using IMGT, and the amino acid sequences of the CDRs of the heavy chain variable region and the light chain variable region are shown in Table 5 (SEQ ID NO: 6-20).

[0536] Table 4: M1-23 / M3-23 / M3-13 light and heavy chain variable region amino acid sequences

[0537]

[0538]

[0539] Table 5: Sequences of M1-23 / M3-23 / M3-13 light and heavy chain CDRs

[0540]

[0541] The inventors conducted a series of property analyzes on the three purified monoclonal antibo...

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Abstract

The present invention relates to antibodies (particularly humanized antibodies) against hepatitis B surface antigen (HBsAg), nucleic acid molecules encoding the antibodies, methods for preparing the antibodies, and pharmaceutical compositions containing the antibodies. Furthermore, the invention relates to use of the antibodies and the pharmaceutical compositions. The antibodies and pharmaceuticalcompositions are useful for preventing and / or treating HBV infection or hepatitis B infection and related diseases (such as hepatitis B), for neutralizing the virulence of HBV in a subject (such as ahuman), or for reducing serum levels of HBV DNA and / or HBsAg in the subject.

Description

technical field [0001] The invention relates to the field of molecular virology and immunology, especially the field of treating hepatitis B virus (Hepatitis Bvirus, HBV) infection. Specifically, the present invention relates to antibodies against hepatitis B surface antigen (HBsAg), nucleic acid molecules encoding them, methods for preparing them, and pharmaceutical compositions containing them. The pharmaceutical composition can be used to prevent and / or treat HBV infection or diseases related to HBV infection (such as hepatitis B), to neutralize the virulence of HBV in a subject (such as a human), or to treat Reduce the serum levels of HBV DNA and / or HBsAg in patients. Therefore, the present invention further relates to the use of said antibodies (especially humanized antibodies) and variants thereof in the preparation of pharmaceutical compositions for preventing and / or treating HBV infection or related to HBV infection diseases (such as hepatitis B), for neutralizing th...

Claims

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Application Information

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IPC IPC(8): C07K16/08A61K39/42A61P31/20A61P1/16
CPCA61P1/16A61P31/20C07K16/082A61K2039/505C07K2317/565C07K2317/56C07K2317/567C07K2317/51C07K2317/515A61K39/395A61K39/42C07K16/08C12N15/63C07K2317/24C07K2317/76
Inventor 罗文新王一文康赐明袁权张天英夏宁邵
Owner XIAMEN UNIV
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