Hydrolyzed profibrinolysin polypeptide and application thereof

A technology of hydrolyzing fiber and lysozyme, which is applied in the field of polypeptide for the treatment of malignant tumors, to achieve the effect of improving survival rate and inhibiting proliferation activity

Inactive Publication Date: 2015-06-10
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no well-developed hydrolyzed plas...

Method used

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  • Hydrolyzed profibrinolysin polypeptide and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] IC50 of hydrolyzed plasminogen polypeptide on the growth and survival of human vascular endothelial cells cultured in vitro.

[0016] The MTT colorimetric method was used. The logarithmic growth of human vascular endothelial cells HUVEC, 1.0 × 10 5 Add it to a 96-well culture plate and culture for 24 hours. Add different concentrations of the experimental drug hydrolyzed plasminogen polypeptide (Shanghai Sangong Synthetic) and the positive control drug paclitaxel to the experimental wells and positive drug control wells; add the same volume of solvent to the blank group. . Set up five duplicate wells per well, culture for 48h, add MTT to each well at 0h, 2h, 8h, 14h, 20h, 24h, 36h, and 48h respectively, add DMSO after 4h of action, incubate for 30min, and incubate at 620nm in a microplate reader Measure the absorbance A value, according to the formula HUVEC growth inhibition rate=(1-absorbance value of the experimental group / absorbance value of the control group)×100%...

Embodiment 2

[0018] In vivo activity of hydrolyzed plasminogen polypeptide was tested in a tumor model.

[0019] Uterine cancer tumor model was established, paclitaxel was used as a positive control drug; the same volume of solvent was added to the blank group, and three doses of polypeptide (Shanghai Sangong Synthetic) were set in the experimental group: 0.75, 1.5, 3 μM / Kg. After 21 days, the number of surviving mice was observed, and the survival rate was calculated. The results showed that the hydrolyzed plasminogen polypeptide can effectively protect mice and improve the survival rate of tumor-bearing mice, the survival rate reached 78.5%, paclitaxel 31.2%.

Embodiment 3

[0021] Inhibitory test of hydrolyzed plasminogen polypeptide on human cervical cancer cell HeLa xenograft tumor growth in nude mice

[0022] Take the human cervical cancer HeLa cell line in the logarithmic growth phase, and prepare 5 × 10 cells under sterile conditions. 6 / ml cell suspension, inoculated subcutaneously in the right armpit of nude mice with 0.1ml. Use a vernier caliper to measure the diameter of the transplanted tumor in nude mice, and wait until the tumor grows to 100-200mm 3 Animals were then randomly grouped. Using the method of measuring tumor diameter, dynamically observe the anti-tumor effect of the tested polypeptide. Tumor diameter was measured once every 2 days, and the weight of the mouse was also weighed at the same time as each measurement. The method of administration was by tail vein injection. The negative control group was injected with the same amount of normal saline once a day; the cisplatin group was given 10 mg / kg once a week and once a ...

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Abstract

The invention relates to the medicine field, in particular to polypeptide capable of promoting profibrinolysin to be hydrolyzed into angiostain and treating a malignant tumor. The sequence of the polypeptide is THELGHLSQKLYGKRSN and is a brand new sequence, the polypeptide can promote the hydrolyzation of the profibrinolysin in vitro and inhibit the proliferation of vascular endothelial cells, in a vivo test, the survival rate of tumor-bearing mice is improved, and the polypeptide has a potential new drug development value.

Description

technical field [0001] The invention relates to hydrolyzed plasminogen polypeptide and its application, in particular to the polypeptide capable of promoting the hydrolysis of plasminogen into angiostatin and treating malignant tumors. Background technique [0002] In 1971, Folkman proposed that tumor growth and metastasis depend on angiogenesis. It is also believed that after the formation of a solid tumor, its development can be divided into two stages: the avascular phase and the vascular phase. In the avascular stage, the existence of tiny tumors depends on the diffuse blood supply of the surrounding interstitium, the growth rate is linear, the tumor is limited within 1mm to 2mm, and the number of tumor cells is less than 10 5 ~10 6 One, forcing the tumor to be in a "dormant state"; after entering the vascular phase, the tumor tissue is perfused with blood supply, and the tumor grows exponentially, and can increase by 16,000 times after 2 weeks. New blood vessels not ...

Claims

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Application Information

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IPC IPC(8): C12N9/68A61K38/48A61P35/00
CPCA61K38/00C12N9/6435C12Y304/21007
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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