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Therapeutic agent for anemia including renal anemia and cancer-induced anemia which contains anti-bmp9 antibody as active ingredient

一种抗体、贫血的技术，应用在含有效成分的医用配制品、抗体医疗成分、抗体等方向，能够解决难预料到抗BMP9抗体红细胞造血作用、尚未获知其他抗体等问题，达到肾性贫血改善的效果

Active Publication Date: 2015-03-11
KYOWA HAKKO KIRIN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, based on these reports, it is extremely difficult to expect that anti-BMP9 antibodies have erythrocyte hematopoiesis in vivo
[0006] In addition, as an anti-BMP9 monoclonal antibody, a monoclonal antibody (clone number 360107) with neutralizing activity against BMP9 has been commercially available from Andy Biotechnology Company, but no other antibodies have been known

Method used

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  • Therapeutic agent for anemia including renal anemia and cancer-induced anemia which contains anti-bmp9 antibody as active ingredient
  • Therapeutic agent for anemia including renal anemia and cancer-induced anemia which contains anti-bmp9 antibody as active ingredient
  • Therapeutic agent for anemia including renal anemia and cancer-induced anemia which contains anti-bmp9 antibody as active ingredient

Examples

Experimental program
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preparation example Construction

[0325] (1) Antigen preparation

[0326] BMP9 as an antigen or a tissue expressing BMP9 can be obtained by introducing an expression vector containing cDNA encoding full-length or partial-length BMP9 into Escherichia coli, yeast, insect cells, animal cells, or the like. In addition, BMP9 can be purified from human tissues expressing BMP9 in large quantities. Alternatively, the tissue or the like can be used as an antigen as it is. In addition, a synthetic peptide having a partial sequence of BMP9 can also be prepared by a chemical synthesis method such as the Fmoc method or the tBoc method and used as an antigen.

[0327] BMP9 used in the present invention can be obtained by using Molecular Cloning, A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press (1989) or Current Protocols Inmolecular Biology, John Wiley & Sons Publishing Company (1987-1997) and the like. Methods etc. The DNA encoding the BMP9 is expressed in a host cell and produced by, for example,...

Embodiment 1

[0456] Construction of Targeting Vector for Mouse BMP9 Gene Knockout

[0457] 1-1) Construction of expression cassette vector pBlueLAB-LoxP-Neo-DT-A

[0458] The expression cassette vector pBlueLAB-LoxP-Neo-DT-A, which is the basic vector for making a targeting vector for knockout (KO), is a neomycin resistance marker inserted after adding a restriction enzyme site to pBluescript The vector obtained by LoxP-Neo having the LoxP sequence at both ends of the gene expression unit and the diphtheria toxin A chain gene (DT-A) is the same as the vector described in Example 7 of International Publication No. 2006 / 078072. The outline of the carrier preparation is described below.

[0459] In order to add a new restriction site to the vector pBluescript II SK(-) (manufactured by Toyobo Co., Ltd.), the following oligo DNA was synthesized (LinkA1: SEQ ID NO: 1, LinkA2: SEQ ID NO: 2, LinkB1: SEQ ID NO: 3 and LinkB2: serial number 4).

[0460] Treat pBluescript II SK(-) with res...

Embodiment 2

[0484] Preparation of mouse BMP9 gene targeting vector

[0485] 60 μg of pBluemBmp9-Lox-Neo-DT-A-3'KO-5'KO obtained in Example 1-5 was added with spermidine (manufactured by Sigma) so that the final concentration was 1 mmol / L and Restriction enzymes adjusted to pH 7.0 were digested with NotI in H buffer (manufactured by Roche Diagnostics).

[0486] The carrier fragment was extracted from the reaction solution with phenol / chloroform and precipitated with ethanol. Add HBS solution with pH 7.05 [each 1 liter contains HEPES 5g, NaCl 8g, KCl 0.37g, Na 2 HPO 4 2H 2 (0.125 g, a solution of 1 g of dextrose (D-glucose)] so that the DNA concentration was 0.5 μg / μL, and stored at room temperature for 1 hour, thereby preparing the mouse BMP9 gene targeting vector pBluemBmp9 for electroporation -Lox-Neo-DT-A-3'KO-5'KO-NotI.

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Abstract

The present invention provides an anti-BMP9 (Bone morphogenetic protein-9) monoclonal antibody or an antibody fragment thereof binding to human BMP9, a hybridoma producing the antibody or the antibody fragment, a DNA encoding the antibody or the antibody fragment, a vector comprising the DNA, a transformant obtained by introduction of the vector, a method for preparing the antibody or the antibody fragment using the hybridoma or the transformant, and a therapeutic agent comprising the antibody or the antibody fragment as an active ingredient. Further, the present invention provides a pharmaceutical composition comprising the antibody or the antibody fragment as an active ingredient for the treatment of anemia such as renal anemia, cancer anemia or the like, and a method for treating anemia such as renal anemia, cancer anemia or the like using the same.

Description

technical field [0001] The present invention relates to an anti-BMP9 monoclonal antibody or an antibody fragment thereof binding to human BMP9 (Bone morphogenetic protein-9, bone morphogenetic protein-9), a hybridoma producing the antibody or the antibody fragment, encoding the antibody or the antibody fragment DNA, a vector containing the DNA, a transformant obtained by introducing the vector, a method for producing the antibody or the antibody fragment using the hybridoma or the transformant, a therapeutic agent containing the antibody or the antibody fragment as an active ingredient . [0002] In addition, the present invention relates to a pharmaceutical composition for treating anemia such as renal anemia and cancerous anemia containing the antibody or the antibody fragment as an active ingredient, and a method for treating anemia such as renal anemia and cancerous anemia using the pharmaceutical composition. treatment method. Background technique [0003] BMP9 is the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12P21/08
CPCC07K16/22C12N15/09C07K2317/34C07K2317/92A61K2039/505A61K39/3955C07K16/18C07K2317/24A61P7/06G01N33/53C07K2317/565C07K2317/50
Inventor 清水清山崎雄司久保田丽夫木村要
Owner KYOWA HAKKO KIRIN CO LTD
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