Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Polypeptide for promoting angiogenesis and pharmaceutical use of polypeptide

An angiogenesis and drug technology, applied in the field of biomedicine, can solve problems such as ischemic diseases that have not been reported yet

Pending Publication Date: 2021-09-10
SHANGHAI TENTH PEOPLES HOSPITAL
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The molecular weight of NLRC5 protein is 204kDa. Previous studies have shown that this gene mainly regulates immunity and inflammation, but there is no report about its role in promoting angiogenesis and its application in the treatment of ischemic diseases.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Polypeptide for promoting angiogenesis and pharmaceutical use of polypeptide
  • Polypeptide for promoting angiogenesis and pharmaceutical use of polypeptide
  • Polypeptide for promoting angiogenesis and pharmaceutical use of polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040]Example 1: NLRC5 Knockdown and Cell Function Detection in HUVEC

[0041] 1. NLRC5 knockdown in HUVEC: first put HUVEC into a 6-well plate, and when the confluence of the cells reaches 70%, transfect with siRNA (small interfering RNA, small interfering RNA). Use Lipofectamine 2000 (Cat#11668027, Invitrogen) liposomes for siRNA transfection, according to the product operation manual, transfect HUVEC with negative control siRNA and NLRC5 siRNA (SASI_Hs02_00359503, Sigma) at a concentration of 50 nM, and perform subsequent experiments after 48 hours of transfection .

[0042] 2. HUVEC function testing includes: tube formation, migration, proliferation ability testing:

[0043] (1) Tube formation: Add 50 μl of melted Matrigel matrigel to the pre-cooled 96-well cell culture plate, put it in the incubator for 30 minutes and solidify, spread the treated HUVEC into the 96-well plate at a density of 20,000 / well , and VEGFA-165 (50ng / ml) was added, placed in the incubator for 6-8...

Embodiment 2

[0047] Example 2: Establishment of mouse lower limb ischemia model, blood perfusion detection and lower limb ischemia score

[0048] 1. Establishment of lower limb ischemia model: 5 C57BL / 6J WT mice aged 8-10 weeks and 5 endothelial cell NLRC5-specific knockout mice were anesthetized with 2% isoflurane, and the skin was cut open. The femoral artery was separated and ligated to avoid damage to the nerve and vein, and the skin was sutured and sterilized.

[0049] 2. Lower limb blood perfusion detection: mice were anesthetized with 2% isoflurane, and lower limb blood perfusion was detected using a Doppler blood flow detector (moorLDILaser Doppler Perfusion Imager, England), and blood perfusion rate = ligated side blood flow Perfusion volume / contralateral sham operation blood perfusion volume. Mouse lower limb ischemia score: 28 days after ligation, the mouse lower limb ischemia score was scored, and the scoring rules were as follows: 0 points = no necrosis, 1 point = 1-3 toe tip...

Embodiment 3

[0051] Example 3: RNA sequencing and GSEA analysis after HUVEC knockdown of NLRC5

[0052] RNA-seq: After knocking down NLRC5 in HUVEC according to the steps in Example 1, stimulate VEGF (50ng / ml) for 12h, collect cellular RNA with Trizol, and use the Illumina TruSeq RNA Sample Prep Kit kit (Cat#FC-122- 1001) to build a library, then use the novaseq6000 platform for RNA sequencing, and finally use DESeq to screen differently expressed genes. GSEA analysis: according to the log of two groups of genes 2 Fold Change constructs RNK files, uses GSEA 4.1.0 version for analysis, and further screens according to p<0.05 and q<0.25.

[0053] The experimental flow and result figure of above-mentioned embodiment are as follows image 3 As shown, it was demonstrated that NLRC5 binds to STAT3 as a scaffolding protein. Among them, A is the experimental flow chart, B is the RNA sequencing map, and C is the GSEA analysis map. RNA sequencing was performed after knockdown of NLRC5 in HUVEC. ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention discloses a polypeptide for promoting angiogenesis and pharmaceutical use of the polypeptide. The polypeptide comprises an amino acid sequence as shown in SEQ ID No. 1. The polypeptide of the invention is a short fragment peptide chain of NLRC5 protein, and can promote tissue repair and prevent and treat ischemic diseases through angiogenesis; the polypeptide can play roles in combining with STAT3 and regulating and controlling, and remarkably inhibits an IL-6-STAT3 signal channel, so that angiogenesis is promoted; and the molecular weight of the peptide chain of the polypeptide is 60kDa and is 1 / 4 of the molecular weight of the NLRC5 protein, the protection effect of the NLRC5 protein in vivo is facilitated by adopting a short peptide form, tissue permeation can be better exerted without influencing the main activity, and the problems of weak absorption of in-vivo injection administration, easy generation of immune reaction and the like which are caused by large molecular weight of the protein are avoided.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an angiogenesis-promoting polypeptide and its pharmaceutical use. Background technique [0002] Angiogenesis mostly occurs in ischemic diseases, wound healing and tumor diseases, which can cause severe tissue necrosis, dysfunction, and even death, and the pathological process is often accompanied by tissue damage and repair. Angiogenesis refers to the process of forming a new vascular network by stimulating endothelial cells to sprout and grow new branches on the basis of the original vascular structure. It is a unique proliferation of mature and fully differentiated endothelial cells that already exist in blood vessels. , migration and remodeling results. [0003] Ischemic disease is a general term for a large group of common clinical diseases, including ischemic stroke, coronary heart disease, peripheral arteriosclerosis obliterans, and liver ischemia, which ma...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/705C12N15/12A61K38/17A61P17/02A61P9/10A61P9/00A61P1/16
CPCC07K14/705A61P17/02A61P9/10A61P9/00A61P1/16A61K38/00
Inventor 彭文辉石晔飞庄剑辉俞晴栾培培徐徐
Owner SHANGHAI TENTH PEOPLES HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com