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Hydrolysis plasminogen polypeptide and application thereof

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

Inactive Publication Date: 2015-06-24
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 plasminogen polypeptide available for the treatment of malignant tumors

Method used

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  • Hydrolysis plasminogen polypeptide and application thereof
  • Hydrolysis plasminogen polypeptide and application thereof

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, add 2, 4, 8 μg of hydrolyzed plasminogen polypeptide (Shanghai Sangong Synthetic) and 6 μg of positive control drug metalloprotease in the substrate solution respectively. After reacting for 5 minutes at 37°C, centrifuge, take the supernatant, and use the ELISA method to react the content of angiostatin in the solution. The results show that 2, 4, 8 μg of hydrolyzed plasminogen polypeptide and angiostatin 9.76, 23.98, 41.98nM, respectively.

Embodiment 2

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

[0019] 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 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 56.4%.

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Abstract

The invention relates to the field of medicines, in particular to a polypeptide with the functions of promoting plasminogen to be hydrolyzed into angiostatin and treating malignant tumors. A sequence of the plasminogen polypeptide is THELGHLSQKLYGKRSN and is a bran-new sequence. The plasminogen polypeptide is capable of promoting the hydrolysis of the plasminogen in vitro, inhibiting proliferation of vascular endothelial cells, and improving the survival rate of tumor-bearing mice in an in vivo test, and has potential new medicine 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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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/68A61K38/48A61P35/00
CPCA61K38/484C12N9/6435C12Y304/21007
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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