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Application of beta 3-endoxine protein factor as target molecule in regulation of neovascularization

A technology of protein factors and new blood vessels, applied in the field of biomedicine, can solve problems such as validity disputes and failures

Inactive Publication Date: 2018-11-16
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effectiveness of this mechanism has always been controversial. For example, the results of a clinical study released by Merck in 2013 showed that the previously highly anticipated integrin αvβ3 antagonist clengitides was effective in the treatment of glioblastoma Phase III. failed in clinical trials

Method used

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  • Application of beta 3-endoxine protein factor as target molecule in regulation of neovascularization
  • Application of beta 3-endoxine protein factor as target molecule in regulation of neovascularization
  • Application of beta 3-endoxine protein factor as target molecule in regulation of neovascularization

Examples

Experimental program
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Effect test

Embodiment 1

[0016]Example 1: Design and synthesis of control siRNA (siControl for short) and siRNA targeting β3-endonexin (siβ3-endo for short). These siRNAs were synthesized in Shanghai Gemma Company, and the sequence information of siControl and siβ3-endo are respectively

[0017] siControl:5'UUCUCCGAACGUGUCACGUTT3',

[0018] siRNA-β3-endo: 5'GAACUUGUCAAAUGAGUCU dTdT 3'

[0019] Design and synthesize mβ1CTP and mβ3CTP small molecule polypeptide compounds, and their respective amino acid sequences are:

[0020] mβ1CTP: YGRKKRRQRRRAVTTVVNPKYEG

[0021] mβ3CTP: YGRKKRRQRRRATSTFTNITYRG

Embodiment 2

[0022] Example 2: Human umbilical vein endothelial cells (HUVEC) blood vessel-like structure formation experiment

[0023] Human umbilical vein endothelial cells (HUVEC) cultured in vitro can self-assemble into a vessel-like pipeline structure on the basement membrane matrigel containing growth factors. siRNA (siβ3-endo) targeting β3-endonexin (siβ3-endo) and control siRNA (siControl) without targeting sequence were transfected into HUVEC respectively, and 48 hours later, HUVEC transfected with siβ3-endo and treated with siControl were collected and plated on pre-coated Matrigel cell culture on a 48-well plate, observe the formation of pipe-like structures in HUVEC under an inverted microscope, and count the number of formed pipe-like structures, see figure 1 , the results showed that compared with the control siControl treatment group, transfection of siβ3-endo group could effectively inhibit the formation of blood vessel-like structures in vascular endothelial cells, indicat...

Embodiment 3

[0024] Example 3: Adhesion experiment of human umbilical vein endothelial cells (HUVEC).

[0025] After the siβ3-endo targeting β3-endonexin and the control siControl without targeting sequence were transfected into HUVECs, the cells were incubated on a plate pre-coated with the β3-type integrin ligand fibrinogen (20 μg / ml) at 37°C for 30 minutes. Minutes, then fixed with 70% methanol, stained with 1% toluidine blue, and counted the number of adherent cells under a microscope. In addition, HUVECs were stimulated with vascular endothelial growth factor (VEGF) during the process of cell adhesion, and experiments and statistics were carried out in the aforementioned manner. see figure 2 a, The results showed that, compared with the control group, down-regulating the expression of endogenous β3-endonexin in HUVECs had no obvious effect on the adhesion ability of HUVECs.

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Abstract

The invention relates to an application of a beta 3-endoxine protein factor as a target molecule in regulation of neovascularization. When expression of beta 3-endoxine in vascular endothelial cells is down-regulated or small-molecule polypeptide compounds capable of being bound with beta 3-endoxine are used for acting on the vascular endothelial cells, expression of a VEGF (vascular endothelial growth factor) can be effectively inhibited, and proliferation of the vascular endothelial cells and the capacity of forming vascular-like structures by the vascular endothelial cells can also be reduced. Therefore, the beta 3-endoxine protein factor expressed by the vascular endothelial cells is an effective target for developing anti-angiogenic drugs, and two effective anti-angiogenic methods areprovided on the basis of the factor.

Description

technical field [0001] The invention belongs to the field of biomedicine, and particularly relates to the application of a β3-endonexin protein factor in vascular endothelial cells as a target molecule for regulating the formation of new blood vessels. Background technique [0002] β3-endonexin is a polypeptide protein factor that can specifically bind to the intracellular sequence of integrin β3 subunit, and is expressed in vascular endothelial cells, platelets and other cells. There are currently four splice forms of β3-endonexin: β3-endonexin-S (111 amino terminals, 12.6kDa), β3-endonexin-L (170 amino acids, 19.2kDa), NRIF3 (nuclear receptor coactivator), and a spliceosome theta-associated protein (TAP) 20 encoding a 62 amino acid sequence. β3-endonexin-S is mainly expressed in vascular endothelial cells. β3-endonexin herein refers to β3-endonexin-S. Previous studies have confirmed that β3-endonexin can interact with the intracellular sequence of integrin β3, but not w...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/7088A61K38/16A61P35/00C12Q1/686
CPCA61K31/7088A61K38/16A61K45/00A61P35/00C12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114C12Q2521/107
Inventor 马衍青锁昕凤褚羽丹徐舟鲍云曹钟元许祯高娟
Owner SHANGHAI UNIV
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