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

Anti-inflammatory peptide with membrane penetration effect

A short peptide, membrane-penetrating technology, applied in the field of medicine, can solve problems such as physiological dysfunction, and achieve the effect of short peptide sequence and easy synthesis

Inactive Publication Date: 2010-06-16
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as we all know, the application of NF-κB antagonists must face the following unavoidable problems: ①Safety: Since NF-κB is distributed in all cells in the body, indiscriminate inhibition of its activity will lead to extensive physiological dysfunction

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
  • Anti-inflammatory peptide with membrane penetration effect
  • Anti-inflammatory peptide with membrane penetration effect
  • Anti-inflammatory peptide with membrane penetration effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The operating steps of embodiment 1, Lys (D-Pro) ValCit preparation method

[0024]The main operation steps of solid-phase peptide synthesis are as follows: 1. Resin swelling: Put Wang-Resin resin into a reaction tube, add DMF (15ml / g) for 30min. 2. Deprotection: remove DMF, add 20% piperidine DMF solution (15ml / g) for 5min, remove and add 20% piperidine DMF solution (15ml / g) for 15min. 3. Detection: Take out the piperidine solution, take more than a dozen resins, wash them three times with ethanol, add ninhydrin, KCN, and phenol solution one drop each, heat at 105°C-110°C for 5 minutes, and turn dark blue to indicate a positive reaction. 4. Washing: DMF (10ml / g) twice, methanol (10ml / g) twice, DMF (10ml / g) twice. 5. Condensation: Add a three-fold excess of protected amino acids and a three-fold excess of activator HBTU, dissolve them with as little DMF as possible, add to a reaction tube, immediately add a ten-fold excess of NMM, and react for 30 minutes. 6. Washin...

Embodiment 2

[0025] Example 2, ELISA method to detect polypeptide binding to nuclear factor-κB p65 subunit and nuclear factor-κB cis-acting element Competitive Inhibition Experiment

[0026] The synthesis of the peptide sequence was synthesized by Shanghai Qiangyao Biotechnology Co., Ltd., with a purity of >95%. On a 96-well enzyme-linked plate coated with nuclear factor-κB cis-acting elements (purchased from Mercury TM Add 150 μl / well of transcription factor blocking solution to Transfactor p65kits), incubate at room temperature for 15 minutes; discard the transcription factor blocking solution, add 50 μl / well of detection samples diluted with transcription factor blocking solution, and the samples contain 10 ng / μl of p65 standard protein as positive Control, p65 standard protein containing 10ng / μl was added with serial concentrations of peptides at the same time, p65 standard protein containing 10ng / μl was added with the same series of negative peptides at the same time, incubated a...

Embodiment 3

[0028] Example 3. Inhibition experiment of polypeptide on expression of nuclear factor-κB responsive reporter gene.

[0029] RAW 264.7 cells were cultured in 1640 medium, and the cell density was adjusted to 0.5-2.5×10 the day before transfection 5 Cells / mL, overnight culture; take 1mL of overnight culture cells or 0.5×10 5 The amount of cells per well was added to a 24-well culture plate; 3 μL GeneJuice transfection reagent was added to 100 μL serum-free 1640 medium, vortexed to mix, and left at room temperature for 5 min; 1 μg p4kB-Luc plasmid was added to the GeneJuice / 1640 medium mixture, gently Mix by blowing and blowing, and place at room temperature for 5-15 minutes; add the above mixture dropwise to the complete medium, shake gently to mix the mixture and cells thoroughly; culture in a 5% CO2 incubator at 37°C; after 5 hours of transfection, use complete The culture medium was exchanged for 1 h, and then the cells were washed 3 times with PBS, and finally the cells ...

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

No PUM Login to View More

Abstract

The invention relates to an anti-inflammatory peptide with membrane penetration effect, in particular to a mutant peptide which is used for treatment and takes a nuclear factor-kappa B (NF-kappa B) rho65 subunit antagonistic peptide as a matrix. The mutant peptide has membrane penetration effect. More particularly, the invention relates to a small molecular peptide Lys(D-Pro)ValCit which not only can antagonize the activity of a nuclear factor-kappa B rho65 subunit, but also can act on other unknown targets. The invention also relates to the preparation and application of the anti-inflammatory peptide.

Description

technical field [0001] The invention relates to the field of medicine, in particular to a remodeled peptide for treatment, which uses nuclear factor-κB (NF-κB) p65 subunit antagonist peptide as a parent, and the restructured peptide has membrane-penetrating effect. More specifically, it relates to a small molecular peptide Lys(D-Pro)ValCit that can not only antagonize the activity of nuclear factor-κB p65 subunit, but also act on other unknown targets. The present invention also relates to this anti-inflammatory short peptide preparation and application. Background technique [0002] Since nuclear factor-κB (nuclear factor-κB, NF-κB) was accidentally discovered in 1986, its importance has been increasingly concerned and has become one of the most active research fields. Typical NF-κB is a heterodimer formed by two subunits of polypeptide chain p50 and p65. When cells are in a resting state, NF-κB mainly combines with κB inhibitory proteins (IκBs, of which IκBα and IκBβ pla...

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): C07K5/11A61K38/07A61P29/00A61P11/06A61P13/12A61P19/02A61P19/04A61P31/00A61P37/02
Inventor 范霞梁华平魏强王希杨雪
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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