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

Construction of a long-acting polypeptide and its application in anti-acute kidney injury and diabetic nephropathy

A technology for acute kidney injury and diabetes, which is applied in the field of biotechnology drugs, can solve the problems of weak binding ability between peptides and receptors, short half-life, lack of effective treatment methods for diabetic nephropathy, etc., and achieve the effect of prolonging plasma half-life and enhancing binding ability

Active Publication Date: 2021-06-11
HUAZHONG UNIV OF SCI & TECH
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention solves the technical problems of weak polypeptide-receptor binding ability and short half-life in the prior art
This solves the technical problem that the prior art lacks effective treatment means for acute kidney injury and diabetic nephropathy

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
  • Construction of a long-acting polypeptide and its application in anti-acute kidney injury and diabetic nephropathy
  • Construction of a long-acting polypeptide and its application in anti-acute kidney injury and diabetic nephropathy
  • Construction of a long-acting polypeptide and its application in anti-acute kidney injury and diabetic nephropathy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Solid-phase synthesis and purification methods for polypeptides used in the present invention.

[0054] Swell 1 g of Fmoc-rink AM resin (sample loading: 0.3 mmol / g) in 10 mL of DMF for 4 hours, remove the DMF, add DMF containing 20% ​​piperidine, react with shaking at room temperature for 45 min, and remove the Fmoc protecting group. The deprotected resin was washed with DMF to remove residual piperidine. Then add 15 mL of DMF solution containing Fmoc amino acid (3.0 equivalents), HBTU (2.9 equivalents), HOBt (3.0 equivalents), DIPEA (6.0 equivalents), shake at room temperature for 2 hours, remove the reaction solution, clean the resin with DMF, and perform Kaiser test Whether the reaction is complete (if the reaction is not complete, repeat the coupling once). According to the amino acid sequence of the polypeptide, the coupling of amino acids is carried out from the C-terminal to the N-terminal. After the polypeptide sequence is completed, use DMF, CH 2 Cl 2 , MeOH...

Embodiment 2

[0058] The in vitro plasma half-life of the K(pal)-E11 peptide, PEG-E11 peptide, Pal-E11 peptide and Dimer-E11 peptide of the present invention is longer than that of the E11 peptide.

[0059] Specific operation: 600 μL of fresh plasma, 60 μg of polypeptide, add 3000 μL of Tris-HCl buffer, incubate at 37 degrees Celsius, take 600 μ of reaction solution each time at the six time points of 0, 2, 4, 8, 12 and 24 hours, and transfer to the taken out Add 30 μL TFA to the reaction solution sample to terminate the reaction, then add 3000 to 4000 μL methanol and mix thoroughly - let it stand for 20 minutes, centrifuge at 10000 rpm for 5 minutes, take the supernatant, process it in a freeze concentrator, evaporate the liquid, and add methanol / water containing 0.1% TFA (1:1) 20 microliters, fully dissolved and centrifuged at 10,000 rpm for 5 minutes, the supernatant was taken for HPLC analysis, and the peak area was integrated to determine the half-life. The experimental results are sho...

Embodiment 3

[0063] The PEG-E11 peptide and Pal-E11 peptide of the present invention can inhibit renal dysfunction caused by renal I / R injury.

[0064] Specific operation: Before the formal experiment, we conducted a series of polypeptide injection dose pre-experiments of the present invention. After the injection of the series of polypeptides, the physiological indicators of the mice were normal in all aspects, and the series of polypeptides were considered to be safe. Mice weighing about 25g were divided into: control mouse group (CT group), renal ischemia-reperfusion mouse group (I / R group), and renal ischemia-reperfusion mouse group given E11 peptide (E11 group) , renal ischemia-reperfusion mouse group was given K(pal)-E11 peptide group (K(pal)-E11 group) peptide, renal ischemia-reperfusion mouse group was given Pal-E11 peptide group (Pal-E11 group), The renal ischemia-reperfusion mouse group was given PEG-E11 peptide group (PEG-E11 group) and the renal ischemia-reperfusion mouse group...

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 a variety of chemically modified long-acting polypeptides and the application of preparing drugs for acute kidney injury or diabetes complicated with nephropathy, belonging to the field of biopharmaceuticals. The first sequence: glutamic acid is modified on any amino acid of the amino acid sequence shown in SEQ ID NO:1, and the glutamic acid is connected to the fatty acid; the second sequence: any amino acid of the amino acid sequence shown in SEQ ID NO:1 Modified glutamic acid, glutamic acid is linked with water-soluble polymer; third sequence: fatty acid is modified on any amino acid of the amino acid sequence shown in SEQ ID NO:1, fatty acid is linked with glutamic acid, glutamic acid is linked with lysine Acid linkage; fourth sequence: the third sequence is linked to the N-terminal of the amino acid sequence shown in SEQ ID NO:1 through succinyl. The chemically modified long-acting polypeptide of the present invention can significantly enhance the binding ability with the receptor APJ, significantly relieve renal dysfunction caused by ischemia-reperfusion injury, relieve diabetes-induced nephropathy, and significantly relieve renal tubular necrosis, inflammation and fibrosis.

Description

technical field [0001] The invention belongs to the field of biotechnology drugs, and specifically relates to the construction of multiple long-acting polypeptides and their application in resisting acute kidney injury and diabetes complicated with nephropathy. Background technique [0002] Acute kidney injury is a disease with a high mortality rate worldwide, in which renal tubular cells are ischemic or toxically injured resulting in a sharp decline in renal function. Acute kidney injury is one of the important causes of end-stage renal disease. Acute kidney injury occurs to 133 million people worldwide every year and causes approximately 2 million deaths each year, and the number of deaths is increasing year by year. [0003] Renal ischemia-reperfusion injury is the leading cause of acute kidney injury and causes a high mortality rate worldwide. The pathological process of renal ischemia-reperfusion includes the occurrence of inflammation and apoptosis. Effective inhibi...

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): C07K7/06A61K38/08A61K47/54A61K47/60A61P13/12A61P3/10
CPCA61K38/00A61K47/542A61K47/60A61P3/10A61P13/12C07K7/06
Inventor 黄昆郑凌王超陈红熊明睿
Owner HUAZHONG UNIV OF SCI & TECH
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