Polypeptide for inhibiting tumor cell diffusion transfer and preparation method and application thereof

A tumor cell and polypeptide chain technology, applied in the field of biopharmaceuticals, can solve the problem of few polypeptide reports, and achieve the effects of no toxic side effects, small molecular weight, and stable structure

Inactive Publication Date: 2017-09-22
DALIAN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] A variety of peptides with anti-tumor effects have been discovered, and some have entered the preclinical stage, but there are few reports on peptides that inhibit tumor metastasis

Method used

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  • Polypeptide for inhibiting tumor cell diffusion transfer and preparation method and application thereof
  • Polypeptide for inhibiting tumor cell diffusion transfer and preparation method and application thereof
  • Polypeptide for inhibiting tumor cell diffusion transfer and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 prepares polypeptide of the present invention

[0030] The following 2-CL resins for preparing polypeptides, Fmoc protected amino acids, condensation reagents, and cleavage reagents were all purchased from domestic biochemical reagent companies.

[0031] 1.1 The synthesis of polypeptide resin of the present invention

[0032] Polypeptide resin of the present invention is:

[0033] Lys(Boc)-Lys(Boc)-Lys(Boc)-Lys(Boc)-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) - 2-CL resin.

[0034] Using 2-CL resin as the initial carrier, the polypeptide resin of the present invention was prepared by de-Fmoc protection and coupling reaction, followed by coupling with the protected amino acids shown in Table 1. The protected amino acids used in this embodiment correspond to the 1st to 25th amino acids from the resin as shown in the following table:

[0035] Table 1...

Embodiment 2

[0054] Example 2 Detection of Polypeptide of the Present Invention Inhibiting Tumor Cell Migration in Vitro

[0055] The polypeptide of the present invention is prepared by the method of Example 1, and the serum-free medium is used to prepare 1 mg / mL, stored in a refrigerator at 4 degrees, and stored in a refrigerator at -20 degrees Celsius for a long time. Human colon cancer sw620 cell line, LLC tumor cells and breast cancer MDA-MB-231 cell line were purchased from Shanghai Cell Bank, Chinese Academy of Sciences. The medium used in sequence is 1640RPMI, DMEM.

[0056] 2.1 Use the scratch method to detect the effect of the polypeptide of the present invention on the migration of different tumor cells in vitro

[0057] Human breast cancer MDA-MB-231 cells in logarithmic growth phase and mouse lung cancer LLC tumor cells were inoculated in 12-well plates, and cultured in their respective culture media. Scratching was performed when the cells were 90% confluent. After scratchi...

Embodiment 3

[0060] Example 3 Effect of the polypeptide of the present invention on lung metastasis of mouse lung cancer LLC tumor cells in vivo

[0061] Congeneric Kunming mice of 25-35 g were divided into control group 1, control group 2 and treatment group, with 9 mice in each group. Control group 1 was injected with 0.1 mL of mouse lung cancer LLC tumor cells through the tail vein, about 1.5×10 6 cells; control group 2 was injected with 0.1 mL of mouse lung cancer LLC tumor cells through the tail vein, about 1.5×10 6 cells (containing low water-soluble natural polypeptide 50 μg / mL, amino acid sequence is: DKSSFISVLQTSSSSLRMGAY, has applied for a patent.); the treatment group was injected with 0.1 mL of mouse lung cancer LLC tumor cells through the tail vein, about 1.5 × 10 6 cells (containing 50 μg / mL of the polypeptide of the present invention). A mouse model of lung metastasis of lung cancer LLC tumor cells was established. The mice were killed four weeks after the injection, and ...

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Abstract

The invention discloses a polypeptide for inhibiting the in-vivo diffusion transfer effect of tumor cells and a preparation method and application thereof. The polypeptide consists of 25 amino acids, and its amino acid sequence is shown as SEQ ID No.1. The polypeptide is synthesized by utilizing a solid phase chemical synthesis method, the yield is high, the process is stable, and the molecular weight is 2,777.26 Da. The polypeptide has the activity of inhibiting the proliferation and metastasis of tumor cells and is safe and effective in tumor treatment. Therefore, the polypeptide has the important application value in clinical antitumor therapy.

Description

technical field [0001] The invention relates to a polypeptide and its preparation method and application, in particular to a polypeptide capable of inhibiting tumor cell diffusion and metastasis, its preparation method and application, and belongs to the field of biopharmaceuticals. Background technique [0002] Infiltration and metastasis is one of the main pathological features of malignant tumors, and it is also the main reason why malignant tumors are difficult to cure and eventually lead to the death of patients. According to clinical data and epidemiological investigation and analysis, when malignant tumors are clinically diagnosed, tumor cells in 40% of patients have spread throughout the body. While treating the primary tumor with surgery and radiotherapy, it will also lead to the extensive spread and metastasis of tumor cells in the body. Therefore, while treating the primary tumor, chemotherapy is often supplemented to kill these spreading tumor cells. However, c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00C07K1/36C07K1/30C07K1/20C07K1/06C07K1/04A61K38/16A61P35/04
CPCY02P20/55C07K14/00A61K38/00
Inventor 马克里何欣黄晓华马晓光
Owner DALIAN MEDICAL UNIVERSITY
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