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Anti-il4 receptor antibodies for veterinary use

An antibody and chimeric antibody technology, applied in the direction of antibodies, antibody medical components, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve problems such as reducing signal transduction and reducing binding

Pending Publication Date: 2021-07-23
KINDRED BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, blocking the binding site on IL4R could potentially reduce the binding of IL4 and / or IL13 and reduce the signaling effects of these two cytokines

Method used

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  • Anti-il4 receptor antibodies for veterinary use
  • Anti-il4 receptor antibodies for veterinary use
  • Anti-il4 receptor antibodies for veterinary use

Examples

Experimental program
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preparation example Construction

[0511] The pharmaceutical composition can be stored in lyophilized form. Thus, in some embodiments, the method of preparation includes a lyophilization step. The lyophilized composition can then be reconstituted, typically as an aqueous composition suitable for parenteral administration, prior to administration to dogs, cats or horses. In other embodiments, particularly when the antibody is highly stable to thermal and oxidative denaturation, the pharmaceutical composition may be stored as a liquid, i.e., as an aqueous composition, which may be administered to the dog directly or at appropriate dilution. , cat or horse. Lyophilized compositions can be reconstituted with sterile water for injection (WFI). Bacteriostatic agents such as benzyl alcohol may be included. Accordingly, the present invention provides pharmaceutical compositions in solid or liquid form.

[0512] The pH of the pharmaceutical composition may range from about pH 5 to about pH 8 when administered. If t...

Embodiment 1

[0534] Preparation of IL4 and IL4R ECD reagents

[0535] Nucleotide sequences encoding fusion proteins comprising: (1) full-length canine IL4R (SEQ ID NO:94), canine, feline, equine, Mouse or human IL4R ECD (SEQ ID NO:99, 100, 101, 102 or 103), or canine, cat or horse IL4 (SEQ ID NO: 121, 122 or 123), (2) one or more His6, human Fc and / or a FLAG tag, (3) one or more linker sequences, and (4) a leader sequence. Plasmids were individually transfected into 293 cells, cultured, and supernatants containing secreted IL4R ECD or IL4 fusion polypeptides were collected and filtered individually. Use Ni-NTA column (GE Healthcare Life Sciences) to affinity purify poly-His fusion protein, and use Human Fc fusion proteins were affinity purified on protein A affinity resin (Repligen). Purified fusion proteins were confirmed by SDS-PAGE analysis (data not shown). The fusion proteins (before and after processing) are summarized in Table 3 below.

[0536] table 3..

[0537]

[0538] ...

Embodiment 2

[0540] Identification of mouse monoclonal antibodies that bind canine IL4R

[0541] Mouse monoclonal antibodies were identified following standard immunization with purified canine IL4R-ECD_C-His6 (SEQ ID NO: 107) as immunogen. Different adjuvants (Akesobio, Inc, China) were used during immunization, and monoclonal antibodies were obtained by standard hybridoma technology.

[0542] An enzyme-linked immunosorbent assay (ELISA) was developed to screen for clones producing IL4R-binding antibodies. First, biotinylated IL4R-ECD_C-His6 (SEQ ID NO: 107) was introduced into streptavidin-coated wells. Immune sera were then added to the wells, followed by washing and detection with HRP-conjugated anti-mouse antibodies. The presence of canine IL4R binding antibody revealed a positive signal. More than 121 ELISA positive top clones were identified.

[0543] The 121 antibody clones were screened by ELISA for their ability to block the interaction between canine IL4 and canine IL4R ECD....

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PUM

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Abstract

Provided are various embodiments relating to anti-IL4R antibodies that bind to canine IL4R. In various embodiments, such anti-IL4R antibodies can be used in methods to treat IL4 / IL13-induced conditions, such as atopic dermatitis, allergic dermatitis, pruritus, asthma, psoriasis, scleroderma and eczema, in companion animals, such as canines and felines. Also provided are various embodiments relating to variant IgG Fc polypeptides and variant light chain constant regions of companion animal species for the preparation of antibodies or bispecific antibodies.

Description

technical field [0001] The present invention relates to isolated anti-IL4 receptor (IL4R) antibodies that, for example, bind to canine or feline IL4R and reduce binding to IL4 or IL13; and the use of such antibodies, for example, to treat IL4 induction and and / or an IL13-induced condition or a method of reducing the IL4 or IL13 signaling function of a cell. Background technique [0002] Interleukin 4 (IL4) is a cytokine that induces differentiation of naive T helper cells into Th2 cells. IL4 can also stimulate the proliferation of activated B cells and T cells and induce B cell class switching to IgE. IL13 has a similar effect on immune cells as IL4. Both of these cytokines are associated with allergies. [0003] The IL4 receptor is also known as IL4Rα or IL4R. IL4R can pair with common gamma chain receptors and specifically bind IL4. IL4R can also pair with IL13Ra1, and together they can bind IL4 or IL13. Thus, blocking the binding site on IL4R could potentially reduc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61K39/00A61P37/00C07K14/715C07K16/28C07K16/22
CPCA61P37/00C07K16/2866C07K2317/24C07K2317/33C07K2317/34C07K2317/76C07K2317/565C07K2317/92A61K2039/55C07K14/7155C07K2317/90C07K2317/52A61P37/08A61P17/04A61K2039/505A61K39/3955A61K45/06C07K2317/31G01N33/6869G01N2333/7155
Inventor S·J·李L·阮R·陈H·詹
Owner KINDRED BIOSCI
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