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Novel anti-human IL-23 receptor antibody

A technology of IL-23R and antibody, applied in the direction of anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-animal/human immunoglobulin, cells modified by introducing foreign genetic material, etc., can solve the problem Monoclonal antibodies are highly selective and not easy to achieve the effects of improving patient compliance, ensuring safety, and powerful immune cell suppression

Active Publication Date: 2015-05-13
ASTELLAS PHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in general, monoclonal antibodies are highly selective, and it is not easy to obtain antibodies that retain high activity and show species cross-reactivity

Method used

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  • Novel anti-human IL-23 receptor antibody
  • Novel anti-human IL-23 receptor antibody
  • Novel anti-human IL-23 receptor antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] (Example 1: Acquisition of human and monkey IL-23R-mouse Fc fusion protein)

[0124] In order to be used as an antigen and screening system material for the production of anti-IL-23R antibodies, the present inventors produced an extracellular partial sequence of the human IL-23R sequence (Non-Patent Document 1, Sequence No. 1 from 24th to 353th Amino acid) and the fusion protein of mouse immunoglobulin Fc. Specifically, the extracellular partial sequence of human IL-23R was amplified by PCR using primers ED14-1 (SEQ ID NO: 2) and ED14-2 (SEQ ID NO: 3), and inserted into a mouse Fc fusion protein expression vector The EcoRI and BglII sites of pFUSE-mIgG2A-Fc2 (InvivoGen) were made into a mouse Fc fusion IL-23R expression vector. Here, there are two EcoRI sites in the primer ED14-1 sequence and the human IL-23R gene sequence in the gene amplified by the PCR method, and the human IL-23R is cut out by partial restriction digestion and agarose gel. The gene fragment whose Eco...

Embodiment 2

[0125] (Example 2: Acquisition of 293 cells expressing human IL-23R)

[0126] In order to be used as a cell antigen for anti-IL-23R antibody binding test and antibody acquisition, the present inventors obtained full-length human IL-23R expressing cells. The full length of the human IL-23R gene (the full length of Non-Patent Document 1, SEQ ID NO: 1) was amplified by PCR using primer AA26-Fw (SEQ ID NO: 60) and primer AA10-4 (SEQ ID NO: 61), and this gene fragment Insert the cloning vector pCR2.1-TOPO vector (Invitrogen). After confirming the sequence, the gene fragment was recombined into the mammalian cell expression vector pcDNA3.1 vector (Invitrogen). Lipofectamine 2000 (Invitrogen) was used to introduce the vector gene into human established cultured 293 cells. After the cells were selectively cultured in RPMI1640 medium containing G418, they were cloned by the limiting dilution method. Then, human IL-23R-expressing cells were obtained by flow cytometry using a fluorescent...

Embodiment 3

[0127] (Example 3: Preparation of anti-IL-23R antibody-producing hybridoma)

[0128] In order to obtain anti-human IL-23R antibodies, the present inventors used the human IL-23R-Fc fusion protein or human IL-23R expressing cells obtained in Examples 1 and 2 together with an adjuvant that provokes an immune response to VelocImmune mice. Get immunized. The mice were immunized several times to confirm the increase in the antibody titer in the blood, and the final immunization was performed. The spleen, lymph nodes, etc. of the immunized mice were removed in accordance with a conventional method to collect lymphocytes, and cell-fused with mouse myeloma cells SP2 / 0 to produce hybridomas. A limited dilution sample of hybridomas was prepared and cloned. After expanding the culture of each clone, the medium was changed to a serum-free medium for CD hybridoma medium (Invitrogen) and cultured for about 5 days. The antibody was purified from the obtained culture supernatant using a prote...

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Abstract

The present invention provides anti-human IL-23R antibodies with superior activity and / or species cross-reactivity compared with existing anti-human IL-23R antibodies, and various diseases that use the antibodies to prevent or treat human IL-23R involved in pathogenesis. way of disease. An anti-human IL-23R antibody, which respectively comprises the heavy chain variable region composed of the amino acid sequence shown in SEQ ID NO: 5, 9, 13, 17, 21, 25 or 29 and the amino acid sequence represented by SEQ ID NO: 7, 11, 15, 19, 23, 27 or 31 of the amino acid sequence shown in the light chain variable region.

Description

Technical field [0001] The present invention relates to a novel anti-human IL-23 receptor antibody. In detail, the novel anti-human IL-23 receptor antibody of the present invention is an anti-human IL-23 receptor with better activity and / or species cross-reactivity than existing anti-human IL-23 receptor antibodies antibody. Background technique [0002] Interleukin-23 (also known as IL-23) is a cytokine produced by dendritic cells, etc. It is composed of 2 subunits (ie, p19 subunit unique to IL-23 and IL-12 also becomes its component The p40 subunit) is a heterodimeric cytokine (Non-Patent Document 1). IL-23 binds to IL-23 receptor (also referred to as IL-23R) to introduce signals into cells (Non-Patent Document 2). IL-23R is a heterodimeric receptor composed of the IL-23R subunit and the IL-12Rβ1 subunit which becomes a part of the IL-12 receptor. In addition, it is known that IL-12 receptor is a complex of IL-12Rβ1 subunit and IL-12Rβ2 subunit, and IL-23 does not bind to I...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C12N1/15C12N1/19C12N1/21C12N5/10C12N15/02C12N15/09C12P21/08
CPCC07K16/2866C07K2317/21C07K2317/33C07K2317/73C07K2317/76C07K2317/92C07K16/28C12N15/11
Inventor 大堀诚佐佐木孝则佐藤照田口克成
Owner ASTELLAS PHARMA INC
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