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Monoclonal antibody for treating triple-negative breast cancer

A triple-negative breast cancer, monoclonal antibody technology, applied in the direction of antibodies, anti-tumor drugs, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve the problem of inability to develop drugs and lack of targeted therapy methods And other issues

Inactive Publication Date: 2017-01-04
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For triple-negative breast cancer, due to the lack of specific cell surface molecular markers, targeted drug development cannot be carried out
At present, the first-line treatment drugs for triple-negative breast cancer are broad-based cytotoxic chemotherapy drugs, and there is still a lack of effective targeted therapies

Method used

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  • Monoclonal antibody for treating triple-negative breast cancer
  • Monoclonal antibody for treating triple-negative breast cancer
  • Monoclonal antibody for treating triple-negative breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] Example 1 Neutralizing antibodies can change the cell morphology of human triple-negative breast cancer cells

[0117] The human triple-negative breast cancer cell MDA-MB-231 (obtained from ATCC with high expression of PROCR. Product No. HTB-26 TM . ) were grown on coverslips at a relatively sparse density, and at the same time, 25ug / ml of neutralizing antibodies against PROCR (monoclonal antibody RCR-252, purchased from Abcam, catalog number ab81712) and anti-PROCR were added to the cell culture medium respectively. The control antibody of PROCR (purchased from Abcam, product number is ab56689), and Vim antibody (purchased from Cell Signaling Technology, product number is 5741P) was used for immunofluorescent staining after 12 hours.

[0118] The result is as figure 1 shown. The cell morphology of the MDA-MB-231 cells added with the control antibody did not change significantly, and was still long spindle-shaped, while the cell morphology of the cells added with t...

Embodiment 2

[0120] Example 2 Neutralizing antibodies inhibit the tumorigenicity of patient-derived triple-negative breast cancer (PDX) cells

[0121] Take female nude mice about 5 weeks old, and transfer patient-derived triple-negative breast cancer (PDX) cells to the same amount of cells (10 6 / only) were injected into the fourth pair of mammary glands on the right side of the mice. Nine days later, tumors began to form, and the mice were injected with control antibody and neutralizing antibody (monoclonal antibody RCR-252, purchased from Abcam, Cat. No. ab81712). The specific injection method is: intraperitoneal injection, the antibody dosage is 100ug / time. Inject every 2 days for a total of 3 injections per mouse. During this period, the size of the formed tumor was counted every 1 to 2 days.

[0122] The result is as figure 2 shown. The results showed that intraperitoneal injection of neutralizing antibodies was able to significantly inhibit the growth of patient-derived triple-...

Embodiment 3

[0123] Example 3 Comparison of the ability of the control antibody and the neutralizing antibody (RCR-252) to bind to the PROCR protein

[0124] 3.1 Western blot

[0125] experimental method:

[0126] Human 293T cells were transfected with a human PROCR overexpression plasmid (the overexpression fragment is the full-length PROCR protein, and the overexpression plasmid was purchased from addgene, plasma No. 18121) by transient transfection of PEI. After 48 hours, the untransfected 293T cells of the control group and the 293T cells overexpressing PROCR were harvested. After lysing with protein lysate, centrifuge at 12000rpm / min for 10 minutes to remove cell debris, add 0.1M DTT and protein loading buffer, and boil at 100°C for 10 minutes. Store protein frozen at -80°C for later use.

[0127] The protein samples obtained from the 293T cells of the control group and the 293T cells overexpressing the PROCR protein in the experimental group were subjected to SDS-PAGE gel electrop...

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Abstract

The invention discloses a monoclonal antibody for treating triple-negative breast cancer and application thereof for breast cancer treatment. Particularly, the monoclonal antibody provided by the invention has one or a plurality of characteristics as follows: (a) the monoclonal antibody specially inhibits the combination of protein C and protein C receptors (PROCR); (b) the monoclonal antibody is combined with the protein C receptors, and in addition, a combination region is a region generating combination with the protein C on the protein C receptors (PROCR); (c) the monoclonal antibody is (i) a monoclonal antibody RCR-252 or (ii) a competitive antibody of the RCR-252. The monoclonal antibody disclosed by the invention has the advantages that the growth of the breast cancer cells (particularly the triple-negative breast cancer cells) can be obviously inhibited, so that the monoclonal antibody can be used as a potential treatment agent for the breast cancer (particularly the triple-negative breast cancer).

Description

technical field [0001] The present invention relates to the field of antibody application, in particular to a monoclonal antibody for treating triple-negative breast cancer and its application in the treatment of breast cancer. Background technique [0002] Human breast cancer is roughly divided into four subtypes clinically. Three subtypes of breast cancer (Luminal A, Luminal B, Her2-positive) are known to be formed by mammary luminal cell lesions, and their specific cell surface molecular markers are estrogen receptor (Estrogen Receptor), pregnancy Ketone receptor (Progesterone Receptor) and EGF receptor (EGF Receptor, Her2). For these three types of breast cancer, there are now some targeted drugs for the treatment of breast cancer. [0003] Breast cancers that do not express any of the above three receptors are clinically diagnosed as triple-negative breast cancers. For triple-negative breast cancer, due to the lack of specific cell surface molecular markers, targeted...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395C07K16/30A61P35/00
Inventor 曾艺王代松
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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