Angiostatin activating agent polypeptide and application thereof

An angiostatin and activator technology, applied in the field of angiostatin activator polypeptides, 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 angiostatin activ

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Activity of angiostatin activator polypeptides on the hydrolysis of plasminogen in vitro.

[0016] Angiostatin, the hydrolyzate of plasminogen, was detected by ELISA. Using 1ml of 1mg / ml plasmin as substrate, add 2, 4, 8ng of angiostatin activator polypeptide (Shanghai Sangong Synthetic) and 10ng of positive control drug metalloprotease to the substrate solution respectively, at 37 ℃, after reacting for 5 minutes, centrifuge, take the supernatant, and use the ELISA method to react the content of angiostatin in the solution. The results show that the angiostatin produced by 2, 4, and 8 ng of angiostatin activator polypeptide and metalloprotease are respectively 24.87, 31.98, 53.98 μM.

Embodiment 2

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

[0019] Using the MTT colorimetric method. The logarithmic growth of human vascular endothelial cells HUVEC, with 1.0×10 5 Put it into a 96-well culture plate and incubate for 24 hours. Add different concentrations of the experimental drug angiostatin activator polypeptide (Shanghai Sangon 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, 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 calcu...

Embodiment 3

[0021] The in vivo activity of angiostatin activator polypeptide was detected by tumor model.

[0022] The prostate cancer tumor model was established, and the positive control drug paclitaxel was added; 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: 2, 4, and 8 mg / Kg. After 21 days, the number of surviving mice was observed, and the survival rate was calculated. The results showed that the angiostatin activator polypeptide can effectively protect mice and improve the survival rate of tumor-bearing mice, with the survival rate reaching 81.54%.

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PUM

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Abstract

The invention relates to the field of drugs, in particular to polypeptide for activating angiostatin and treating malignant tumors. The sequence of the polypeptide is EALALMYYHSQSLRKTYP and is a brand new sequence. By means of the polypeptide, the angiostatin can be activated out of a body, proliferation of vascular endothelial cells can be suppressed, the tumor-bearing mice survival rate can be increased in an in-vivo test, and the potential new drug development value is achieved.

Description

technical field [0001] The invention relates to angiostatin activator polypeptide and application thereof, in particular to a polypeptide capable of activating 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 only provide the nutrients and...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K38/10A61P35/00
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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