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Anti-IL-13 antibodies and uses thereof

An antibody and antigen technology, applied in the direction of antibodies, anti-animal/human immunoglobulin, anti-cytokine/lymphokine/interferon immunoglobulin, etc.

Pending Publication Date: 2022-06-10
SHENZHEN FORWARD PHARMA LTD CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dupilumab monoclonal antibody targets IL-4Rα and can inhibit IL-4 and IL-13 signaling pathways at the same time; however, existing clinical trial data show that Tralokinumab and Lebrikizumab that only target IL-13 signaling pathways can obtain drugs similar to Dupilumab. effective, but the incidence of side effect conjunctivitis was significantly reduced

Method used

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  • Anti-IL-13 antibodies and uses thereof
  • Anti-IL-13 antibodies and uses thereof
  • Anti-IL-13 antibodies and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Preparation of Antibodies Using Genetic Engineering

[0052] Amino acid mutation modification of the CDR regions of the heavy chain and light chain, the amino acid sequences of the framework region and variable region of the heavy chain variable region and the framework region and variable region of the light chain variable region of the transformed 20 candidate antibodies are as follows As shown, all antibodies are human IgG4 S228P antibodies, that is, the 228th amino acid is proline.

[0053] Table 5. Sequences of the heavy chain CDRs and FRs of the antibodies of Example 1

[0054]

[0055]

[0056]

[0057] The heavy chain constant region CH of all antibodies is:

[0058] ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS...

Embodiment 2

[0065] Example 2 Screening Candidate Antibodies by In Vitro Experiments

[0066] 2.1 Determination of the affinity of anti-IL-13 antibodies to human, monkey and mouse IL-13 by ELISA method

[0067] Firstly, the 96-well ELISA plate was incubated with 50 μl of 1 μg / ml streptavidin overnight at 4° C., and incubated with 250 μl of blocking solution (3% BSA in TBST) at 37° C. for 2 hours, and then the plate was washed 3 times with PBST. 50 μl of biotin-labeled human IL-13 (obtained from Shanghai Ruizhi Chemical Research Co., Ltd., whose sequence is SEQ ID NO: 181: GPVPPS TALRELIEEL VNITQNQKAP LCNGSMVWSI NLTAGMYCAA LESLINVSGC SAIEKTQRMLSGFCPHKVSA GQFSSLHVRD TKIEVAQFVK DLLLHLKKLF REGRFN) was added to each ELISA experiment well ( 1 μg / ml), and incubated for 1 hour, then washed the plate 3 times with PBST. The 20 antibodies to be tested were serially diluted 5 times from 2 μg / ml, with a total of 8 concentration points (including the blank control), and the serially diluted antibodies ...

Embodiment 3

[0085] Example 3 Drug efficacy of anti-IL-13 antibody in mouse asthma model

[0086] This example evaluates the in vivo pharmacodynamic study of FWB13 on the Balb / c mouse respiratory response model induced by tracheal injection of human recombinant IL-13.

[0087] 29 mice were divided into 5 groups, respectively

[0088] 1. G1) Naive, n=5;

[0089] 2.G2) IL-13+human IgG4 group (antibodies that cannot specifically recognize and bind IL-13), 3mpk, n=8;

[0090] 3.G3) IL-13+tralokinumab group (Tralokinumab), 3mpk, n=8;

[0091] 4.G4) IL-13+FWB13 group, 3mpk, n=8;

[0092] Mice in groups G2, G3, and G4 were injected intraperitoneally with 0.2 mL of the antibody to be tested on the first and third days, at a concentration of 3 mg / kg. Human recombinant IL-13 was intubated on the second and fourth days, 25 μg per mouse, dissolved in PBS in a volume of 50 μL. The mice in the G1 Naive control group were administered with PBS by tracheal spray, without IL-13 and the antibody to be ...

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Abstract

The present application provides anti-IL-13 antibodies and their use in the diagnosis and treatment of IL-13 related disorders.

Description

technical field [0001] The present invention relates to the field of antibodies, in particular to human anti-IL-13 antibodies, especially those human antibodies that can neutralize the activity of IL-13. The present invention further relates to the use of the anti-IL-13 antibody in the preparation of a medicament for diagnosing or treating IL-13-related diseases or disorders, especially asthma and atopic dermatitis. Background technique [0002] Interleukin (IL)-13 is a cytokine that is mainly secreted by Th2 CD4+ lymphocytes, but can also be secreted by Th1 CD4+ T cells, CD8+ T lymphocytes, NK cells, and such as mast cells, basophils, Generation of non-T-cell populations of eosinophils, macrophages, monocytes, and airway smooth muscle cells. IL-13 consists of 114 amino acids and has a molecular weight of about 12 kDa. IL-13 and IL-4 belong to Th2 cytokines, have 30% sequence similarity at the amino acid level, and are closely related in function. The human IL-13 gene is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/24G01N33/68A61K39/395A61P11/06A61P17/00
CPCC07K16/244G01N33/6869A61P11/06A61P17/00C07K2317/565C07K2317/56G01N2800/122G01N2800/202G01N2800/24A61K2039/505
Inventor 朱程刚郭玉函徐良亮董洁娴李烨青彭方理
Owner SHENZHEN FORWARD PHARMA LTD CO
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