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Hydrolyzed plasminogen polypeptides

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: 2015-11-11
顾彩霞
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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 plasminogen polypeptide available for the treatment of malignant tumors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Activity of Hydrolyzed Plasminogen Polypeptides on Hydrolyzed Plasminogen in Vitro.

[0016] ELISA method was used to detect plasminogen hydrolyzate—angiostatin. With 1ml of 1mg / ml plasmin as substrate, 0.75, 1.5, 3.0 μg of hydrolyzed plasminogen polypeptide (Shanghai Sangong Synthetic) and 6 μg of positive control metalloproteinase were added to the substrate solution respectively. After reacting for 5 minutes at 37°C, centrifuge, take the supernatant, and use the ELISA method to determine the content of angiostatin in the solution. Statins were 14.76, 43.98, 63.98nM respectively.

Embodiment 2

[0018] IC50 of hydrolyzed plasminogen polypeptide on the growth and survival of human vascular endothelial cells cultured in vitro.

[0019] Using MTT colorimetric method. 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%. The ...

Embodiment 3

[0021] In vivo activity of hydrolyzed plasminogen polypeptide was tested in a tumor model.

[0022] 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, with the survival rate reaching 87.5%.

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Abstract

The invention provides hydrolyzed plasminogen polypeptides, relating to the field of drugs, particularly polypeptides capable of promoting hydrolysis of plasminogen into angiostatin and treating malignant tumors. The sequence EMQKTVDRLKALNHFDENMW is a brand-new sequence. The polypeptides are capable of promoting hydrolysis of plasminogen in vitro, inhibiting vessel endotheliocytes from proliferation and enhancing the survival rate of tumor bearing mice in an in-vivo test, and have potential new drug development value.

Description

technical field [0001] The invention relates to hydrolysis of plasminogen polypeptide, in particular to the polypeptide which can promote the hydrolysis of plasminogen into angiostatin and treat 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 only provide the nutrie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/68A61P35/00
CPCC12N9/50C12Y304/21007
Inventor 顾彩霞
Owner 顾彩霞
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