Screening and application of targeted biological peptide for inhibiting autoimmune inflammatory response of eyes

An autoimmune and inflammatory technology, applied in the field of biomedicine, can solve problems such as endotoxin residues, aggravated immune disorders, and loss of biological activity

Pending Publication Date: 2020-09-01
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, most of these emerging targeted therapy drugs are biomacromolecules, which have the following disadvantages in specific applications: i) large molecular weight, complicated in vitro synthesis methods, cumbersome recombinant expression and purification processes in the preparation process, and residual endotoxins. Higher production cost; ii) Strong antigenicity, easy to induce host immune response and aggravate immune disorders; iii) Easy to lose biological activity due to changes in protein conformation and modification groups; iv) Due to the

Method used

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  • Screening and application of targeted biological peptide for inhibiting autoimmune inflammatory response of eyes
  • Screening and application of targeted biological peptide for inhibiting autoimmune inflammatory response of eyes
  • Screening and application of targeted biological peptide for inhibiting autoimmune inflammatory response of eyes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0164] Example 1: Screening of targeted biological peptides

[0165] 1.1 Peptide library construction:

[0166] 1.1.1 Amino acid sequence and secondary structure analysis

[0167] The HGF amino acid sequence was analyzed using the basic local alignment search tool (BLAST). Log on to the National Center for Biotechnology Information (NCBI) website (http: / / www.ncbi.nlm.nih.gov / ), and search for "Hepatocyte growth factor" in the NRDB protein sequence complex database. Select the BLAST search program to enter the Standard Protein-Protein BLAST (BLASTp) subroutine, and submit the searched HGF amino acid sequence and the secondary structure analysis of its Kringle 1 domain. Several candidate polypeptide sequences were screened out from Kringle 1, and their basic properties and biological characteristics were analyzed.

[0168] 1.1.2 Analysis of the basic properties of amino acid sequences

[0169] Through ExPASy (http: / / www.expasy.ch / tools / protparam.html), we have analyzed the b...

Embodiment 2

[0185] Example 2: Detection of effectiveness of targeted biological peptides in inhibiting ocular autoimmune inflammatory response:

[0186] Experimental autoimmune encephalomyelitis (EAE) mouse model is used to simulate MS and other human autoimmune nervous system inflammatory diseases, including related studies of autoimmune optic neuritis. As an animal model of autoimmune uveitis, the experimental autoimmune uveitis (EAU) mouse model is widely used in the study of the immunopathological mechanism of autoimmune uveitis and the development of preventive or therapeutic drugs in the process of.

[0187] In this example, EAE and EAU animal models were constructed, and the immunomodulatory and anti-inflammatory effects of biological peptide H-CN in these two models were studied

[0188] 2.1 Study on the effectiveness of H-CN in the treatment of EAE

[0189] 2.1.1 Establishment, treatment and observation of EAE mouse model

[0190] 1) Induction of EAE model ① Antigen emulsifica...

Embodiment 3

[0258] Example 3: Safety detection of targeted biological peptides:

[0259] 3.1 In vitro experiments

[0260] MTS Cytotoxicity Assay CellTiter 96AQueous One Solution Cell Proliferation Assay is a colorimetric assay used to detect the number of living cells in cell proliferation and cytotoxicity assays. Its main component contains a new type of tetrazole compound [3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS].

[0261] Its main working principle is that MTS can be bioreduced by cells into a colored formazan product, which is dissolved in the culture medium, and the formazan product detected at 490nm by spectrophotometry is proportional to the number of living cells. Take RAW264.7 cells (mouse macrophage cell line) and HUVEC cells (human umbilical vein endothelial cell line) in the logarithmic growth phase, and inoculate them on a 96-well plate at a concentration of 5×10^4 / ml. When the cells grow well to 80 When ab...

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Abstract

The invention provides a targeted biological peptide for inhibiting autoimmune inflammatory response of eyes and application of the targeted biological peptide. The invention also relates to a preparation method and application of the polypeptide and a pharmaceutical composition containing the polypeptide. The polypeptide disclosed by the invention has various advantages, such as small molecular weight, low production cost, good water solubility, small immunogenicity, low toxic and side effects, strong tissue penetrability and the like; moreover, researches on two models, namely EAE and EAU, show that the biological peptide has remarkable effects of immunoregulation, inflammation resistance and visual function improvement.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a targeted biopeptide for inhibiting ocular autoimmune inflammatory response and application thereof. Background technique [0002] With the improvement of the sanitation environment, the proportion of infectious diseases in ocular inflammatory diseases has gradually decreased, while the incidence of autoimmune eye diseases has increased year by year. [0003] Autoimmune eye diseases can be primary diseases of the eye, such as Koyanagi-Harada disease, white spot syndrome, etc., but are more often seen as ocular manifestations of systemic autoimmune diseases, such as neuromyelitis optica (NMO) and Multiple sclerosis (multiple sclerosis, MS). Ocular autoimmune diseases can affect multiple parts including the ocular surface, inner eye, and ocular appendages, among which the uvea and optic nerve are most likely to be involved. [0004] Due to their recurrent characteristics, ...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K14/00A61K38/10A61K38/16A61P37/02A61P29/00
CPCC07K7/08C07K14/001A61P37/02A61P29/00A61K38/00
Inventor 成璐许迅李京敬牛田王莉莉
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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