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Active polypeptide for targeting immune checkpoint LAG-3 and application of active polypeptide

A LAG-3, targeting technology, applied in the direction of medical preparations containing active ingredients, peptides, peptide/protein components, etc., can solve problems such as difficulties in tumor immunotherapy, achieve the elimination of immunosuppressive effects, simple polypeptide structure, and improve active effect

Inactive Publication Date: 2020-08-14
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although tumor immunotherapy is a current research hotspot, due to the diversity and complexity of tumor types, tumor immunotherapy has also become a research difficulty

Method used

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  • Active polypeptide for targeting immune checkpoint LAG-3 and application of active polypeptide
  • Active polypeptide for targeting immune checkpoint LAG-3 and application of active polypeptide
  • Active polypeptide for targeting immune checkpoint LAG-3 and application of active polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The polypeptides involved in this example were all obtained by solid phase synthesis, and then the purity was tested by high performance liquid phase HPLC.

[0034] Each polypeptide sequence is as follows: Val-Glu-Thr-Thr-Phe-Arg-Ser (VS-7-1), Val-Glu-Ala-Thr-Phe-Arg-Ser (VS-7-2), Val-Glu -Thr-Thr-Ala-Arg-Ser (VS-7-3), Val-Glu-Thr-Phe-Arg-Ser (VS-7-4), Val-Glu-Thr-Thr-Arg-Ser (VS -7-5) have a purity of 95% or more. See the test results figure 1 . Figure A, the purity of VS-7-1 is 98.98%. Figure B, the purity of VS-7-2 is 98.76%. Figure C, the purity of VS-7-3 is 99.76%. Figure D, the purity of VS-7-4 is 99.08 %, Figure E, the purity of VS-7-5 was 97.17%.

Embodiment 2

[0035] Example 2 The detection experiment of the specific binding between the target LAG-3 polypeptide and the target at the cellular level.

[0036] In this example, the VS-7-1 polypeptide is mainly used as the object, and its anti-tumor activity and / or anti-tumor activity are studied.

[0037]Flow cytometry is a kind of method that excites a single particle flowing in one direction by a laser beam, and detects the scattered light of the particle and the fluorescent markers carried by it, so as to complete the rapid detection and analysis of multiple physical characteristics of a single particle and cell sorting. technology. Widely used in scientific research and clinical medical testing, it is the most advanced cell quantitative analysis technology in the contemporary era. The main target of the active polypeptide for the prevention and / or treatment of tumors involved in the present invention is LAG-3. The present invention will design the VS-7-1 polypeptide to be connected...

Embodiment 3

[0047] The regulation of this series of targeted LAG-3 polypeptides on cytokines in the co-incubation system of tumor cells and T cells was detected. The ELISA kit was used to detect the secretion of IL-2 and IFN-γ in the co-incubation system. The related experiments are introduced as follows.

[0048] 2.1 Experimental materials

[0049] CD3 + T cells, regulating esophageal cancer cell KYSE-30, breast cancer cell MDA-MB-231, ovarian cancer cell A2780, colon cancer cell HCT-116, leukemia cell Jurkat, gastric cancer cell MGC-803, lung cancer cell A549, melanoma cell ME , kidney cancer cell A-498, classical Hodgkin's lymphoma cell L428, the above tumor cells were all derived from ATCC, Ficoll kit, CD3 sorting magnetic beads, CD3 / CD28 activation magnetic beads, IL-2 cytokines, IL -2 ELISA detection kit and IFN-γ detection kit, U-shaped 96-well plate, RPMI 1640 medium.

[0050] 2.2 Experimental Instruments

[0051] Inverted microscope, liquid nitrogen tank, horizontal centrifug...

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Abstract

The invention discloses LAG-3 targeting active polypeptide with antitumor activity and an application of the active polypeptide to tumors, belongs to the fields of drugs for treating the tumors, and particularly relates to the LAG-3 targeting active polypeptide and the application of the active polypeptide in the anti-tumor aspect. The polypeptide has the amino acid sequence represented as Val-Glu-Xa-Thr-Xb-Arg-Ser, or pharmaceutically acceptable salt of the polypeptide, and Xa and Xb are any amino acid substitution or amino acid deletion. A large number of experiments show that the target spot of the polypeptide is clear, experiments in vivo and vitro indicate that the polypeptide with the sequence can be specifically bonded with LAG-3 positive cells, a further in-vitro co-incubation testand a mouse tumor-bearing test indicate that the polypeptide has better application prospects in the aspects of tumor treatment and tumor-bearing mouse survival improvement, and new probability is provided for immunological therapy of biological tumors. The designed LAG-3 targeting polypeptide is scientific, reasonable, feasible and effective and can be used for preparing a treatment drug for human tumors.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and more specifically relates to an active polypeptide targeting LAG-3 and its application in antitumor. Background technique [0002] For tumor diseases, the previous treatment methods mainly include surgery, chemotherapy and radiotherapy. Although these methods have certain effects, they all have certain defects and deficiencies. With the deepening of scientific research, we have now entered a new era of immunotherapy. Immunotherapy is achieved by regulating the immune system, especially the function of T cells, which can activate or induce tumor patients to establish a specific immune response to tumor antigens, eliminate primary tumor cells, and establish immune memory to prevent tumor In addition, this therapy is not only effective, but also avoids damage to the body. In general, tumor immunotherapy is significantly different from previous tumor-targeted surgery, radiotherapy, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K38/08A61P35/00
CPCC07K7/06A61P35/00A61K38/00
Inventor 徐寒梅夏春磊刘晨
Owner CHINA PHARM UNIV
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