Anti-tumor polypeptide and preparation method and application of anti-tumor polypeptide

An anti-tumor and anti-tumor drug technology, applied in the fields of peptide preparation, anti-tumor drugs, chemical instruments and methods, etc., can solve the problem of few peptide reports, and achieve stable process, high yield, low toxicity and anti-tumor activity. Effect

Inactive Publication Date: 2016-11-16
DALIAN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] A variety of peptides with anti-tumor effects have been discovered, and some have ente

Method used

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  • Anti-tumor polypeptide and preparation method and application of anti-tumor polypeptide
  • Anti-tumor polypeptide and preparation method and application of anti-tumor polypeptide
  • Anti-tumor polypeptide and preparation method and application of anti-tumor polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1 prepares polypeptide of the present invention

[0044] The Fmoc-Rink Linker-resin for preparing polypeptides, Fmoc protected amino acids, condensation reagents, cleavage reagents, ninhydrin detection reagents, anhydrous ether and acetonitrile were all purchased from domestic biochemical reagent companies.

[0045] 1.1 The synthesis of polypeptide resin of the present invention

[0046] Polypeptide resin of the present invention is:

[0047] Asp(otbu)-Lys(Boc)-Ser(tbu)-Ser(tbu)-Phe-Ile-Ser(tbu)-Val-Leu-Gln(trt)-Thr(tbu)-Ser(tbu)-Ser( tbu)-Ser(tbu)-Ser(tbu)-Leu-Arg(Pbf)-Met-Gly-Ala-Tyr(tbu)-NH-Rink Linker-resin.

[0048] Using the Fmoc-Rink Linker-resin as the starting carrier, the polypeptide resin of the present invention was prepared by de-protecting Fmoc and coupling reactions, followed by coupling with the protected amino acids shown in Table 1. The Fmoc protected amino acids corresponding to the 1st to 21st amino acids from the resin to the protected a...

Embodiment 2

[0065] Example 2 Study on the In vitro Migration of Tumor Cells Inhibited by the Polypeptide of the Present Invention

[0066] The polypeptide of the present invention is prepared by the method of Example 1. Dilute to 1mg / mL with 10mmol / L, pH7.4 PBS buffer and store at 0-4°C. Human colon cancer sw620 cell line, sw480 cell line and breast cancer MDA-MB-231 cell line were purchased from Shanghai Cell Bank of Chinese Academy of Sciences. Leibovitz's L-15 cell culture medium was purchased from Beijing Zhongke Maichen Technology Co., Ltd. DMEM cell culture medium was purchased from Gibco, USA.

[0067] 2.1 Using the scratch method to study the effect of the polypeptide of the present invention on the migration of different tumor cells in vitro

[0068] The human colon cancer sw620 and sw480 cell lines in the logarithmic growth phase were seeded in 12-well plates and cultured in Leibovitz's L-15 medium; the human breast cancer MDA-MB-231 in the logarithmic growth phase was placed...

Embodiment 3

[0071] Embodiment 3 The effect of the polypeptide of the present invention on the growth morphology of tumor cells

[0072] The human colon cancer sw620 and sw480 cell lines in the logarithmic growth phase were seeded in 12-well plates and cultured in Leibovitz's L-15 medium; the human breast cancer MDA-MB-231 in the logarithmic growth phase was placed in DMEM cultured in culture medium. The polypeptide of the present invention at a concentration of 60 μg / mL was added to each treatment group, and the blank control was not added (control). At 37°C, 5% CO 2 After culturing in the incubator for 48 hours, the effect of the polypeptide of the present invention on the growth morphology of different tumor cells was observed under an inverted microscope. The result is as Figure 4 ,in Figure 4 A, 4B and 4C respectively respectively 60 μ g / mL polypeptide of the present invention (on SW620, SW480 cell and MDA-MB-231 influence, as can be seen from the figure, after the polypeptide o...

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Abstract

The invention relates to polypeptide with an anti-tumor effect and a preparation method and application of the polypeptide. The anti-tumor polypeptide is composed of 21 amino acids, and the amino acid sequence is shown in SEQ ID No.1. The polypeptide is synthesized through a solid phase chemical synthesis method, the yield is high, the process is stable, and the molecular weight is 2265.15Da. The polypeptide has activity for restraining tumor cell proliferation, invasion and metastasis, is safe and effective for treating tumors and has important application value on clinical anti-tumor therapies.

Description

technical field [0001] The invention relates to a polypeptide and its preparation method and application, in particular to a polypeptide with anti-tumor effect and its preparation method and application, belonging to the field of biopharmaceuticals. Background technique [0002] Malignant tumors are a class of diseases that seriously threaten human health and life. Infiltration and metastasis are one of the main biological characteristics of malignant tumors, and are also the main reason why malignant tumors are difficult to cure and eventually lead to the death of patients. Tumor metastasis in vivo is a series of continuous cell migration processes, including the detachment of tumor cells from the primary tumor, the infiltration of tumor cells into surrounding tissues and migration in tissues, the penetration of tumor cells into the blood vessel wall, and the passage of tumor cells into the blood circulation. Blood circulation reaches the target organ of metastasis and the ...

Claims

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Application Information

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IPC IPC(8): C07K14/00C07K1/06C07K1/04A61K38/16A61P35/00
CPCA61K38/00C07K14/00Y02P20/55
Inventor 马克里黄晓华李颖栾明春王聪聪何欣马晓光张巧姝
Owner DALIAN MEDICAL UNIVERSITY
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