PD-l1 IgV affinity peptide S10 with antitumor activity

An anti-tumor activity and affinity technology, used in anti-tumor drugs, peptides, organic chemistry, etc.

Active Publication Date: 2016-06-08
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the use of PD-L1 blockers as tumor immunotherapy drugs or adjuvants has good application prospects and safety, but in the prior art, there is still a lack of better PD-L1 blocker products

Method used

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  • PD-l1 IgV affinity peptide S10 with antitumor activity
  • PD-l1 IgV affinity peptide S10 with antitumor activity
  • PD-l1 IgV affinity peptide S10 with antitumor activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The anti-tumor activity targeting PD-L1IgV affinity peptide S10 provided by the present invention specifically binds to the PD-L1IgV region, and is screened by phage display peptide library technology, and its amino acid sequence is:

[0021] Trp-Ser-His-Gly-Gly-His-Gln-His-Phe-Ile-Arg-Phe, namely W-S-H-G-G-H-Q-H-FI-R-F, has a molecular weight of 1507.7.

[0022] Due to the high cost of monoclonal antibodies and the inability of large-scale production, we selected the PD-L1IgV region protein purified by prokaryotic expression as the target, and screened the peptides that can specifically bind to PD-L1IgV by phage display technology. To block PD-1 / PD-L1 signaling pathway. For the convenience of those skilled in the art to implement the present invention, a brief description of its screening process is as follows:

[0023] The preparation instructions for the media and main solutions used in the screening process are as follows:

[0024] LB liquid medium: Weigh 10 g of ...

Embodiment 2

[0076] The affinity peptide S10 of PD-L1IgV with anti-tumor activity is synthesized by Fomc solid-phase peptide synthesis method, and the synthesis steps are briefly described as follows:

[0077] The main reagents used in the synthesis process are:

[0078] Heading liquid: acetic anhydride / pyridine solution (1:1, v / v);

[0079] Indene detection reagent: A. Ninhydrin / ethanol solution (5%, w / v)

[0080] B. Phenol / Ethanol (4:1, w / v)

[0081] C. Potassium cyanide / pyridine (2%, v / v)

[0082] Deprotection solution: piperidine / DMF solution (20%, v / v);

[0083] Cleavage reagent: by volume, TFA (82.5%), H 2 O (5%), phenol (5%), thioanisole (5%), ethanedithiol (2.5%).

[0084] The synthetic steps are briefly described as follows:

[0085] (1) Swell the resin, add the first amino acid

[0086] A. Swelling resin: Take 0.3~0.5g of Rink resin (the C-terminal amino acid of the peptide connected to the resin is an amide) and place it in a cleaned and dried peptide synthesizer, add an...

Embodiment 3

[0128] Taking the PD-L1IgV affinity peptide S10 with anti-tumor activity prepared in Example 2 as an example, the inventors conducted further in vivo experiments on tumor-bearing mice, and the specific experimental procedures were as follows:

[0129] (1) Affinity peptide S10 inhibits the growth of transplanted tumors in mice bearing CT26 colon cancer

[0130] Select 20 experimental Balb / c mice, adjust the cell concentration to 5×10 mouse colon cancer (CT26) cells with normal saline (NS) 6 cells / mL, 0.1mL cell suspension (containing 5×10 5 cells) were inoculated subcutaneously in the armpit of the right forelimb of each Balb / c mouse, and the growth of the subcutaneous tumor was continuously observed.

[0131] The affinity peptide S10 prepared in Example 2 was dissolved in physiological saline to prepare a polypeptide drug, which was subpackaged and stored at -20°C for future use.

[0132] Nine days after being inoculated with mouse-derived colon cancer (CT26) cells, the mice...

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Abstract

The invention belongs to the technical field of biopharmaceuticals, and in particular relates to an affinity peptide S10 product targeting PD-L1 IgV with anti-tumor activity and its preparation and application. The affinity peptide S10 specifically binds to the PD-L1 IgV region, its amino acid sequence is WSHGGHQHFIRF, and its molecular weight is 1507.7. The affinity peptide S10 is prepared by the Fomc solid-phase peptide synthesis method, and plays a role as the main active ingredient in the preparation of anti-colon cancer drugs. The affinity peptide S10 provided by the present invention is obtained by high-throughput screening with PD-L1 IgV as the target by using phage display peptide library screening technology. Through tumor-bearing experiments in mice, the inventors proved that the affinity peptide S10 has good anti-tumor activity and can significantly inhibit the growth of tumors in mice, thereby providing new ideas and ideas for the research and development of drugs based on PD-L1. theoretical basis.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, in particular to the screening, preparation and application of a polypeptide product with anti-tumor activity, and more particularly, the invention relates to an affinity peptide S10 product targeting PD-L1IgV with anti-tumor activity and its preparation and application. Background technique [0002] In recent years, the situation of tumor prevention and control is very serious. With the improvement of clinical diagnosis, surgical treatment, chemotherapy and radiotherapy, some patients can get early detection, early treatment and better prognosis. However, the search for new treatment methods and drugs has always been a global issue. Research hotspots. Compared with traditional treatment methods, tumor immunotherapy can activate or induce tumor patients to establish a specific immune response to tumor antigens, remove primary tumor cells, and establish immune memory to prevent tumor ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08A61K38/10A61P35/00C07K1/06C07K1/04
Inventor 高艳锋刘蓓媛祁元明李国栋周秀曼李雯雯周杨
Owner ZHENGZHOU UNIV
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