Polypeptide capable of inhibiting beta-secretase digestion effect and application thereof

A technology for preparations and active ingredients, which is applied to polypeptides inhibiting β-secretase enzymatic cleavage and their application fields, and can solve problems such as allergy, large antibody molecules, and difficulty.

Active Publication Date: 2013-11-06
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the antibody molecule is relatively large, it is difficult to pass through the blood-brain barrier, and the antibody itself has good antigenicity, which is easy to cause side effects such as allergic reactions

Method used

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  • Polypeptide capable of inhibiting beta-secretase digestion effect and application thereof
  • Polypeptide capable of inhibiting beta-secretase digestion effect and application thereof
  • Polypeptide capable of inhibiting beta-secretase digestion effect and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1, the acquisition of polypeptide YS1 and its derivatives

[0044] 1. Obtaining the polypeptide YS1

[0045]According to the pathogenesis of Alzheimer's disease AD, the fundamental way to treat AD should be to reduce the production of β-amyloid (Aβ), inhibit the aggregation of Aβ or accelerate the clearance of Aβ. Using phage display technology, using the β-secretase cleavage site on APP and the partial amino-terminal sequence of Aβ as the screening substrate, 1×10 8 The heptapeptides were panned for four rounds, and a number of peptides that could bind to the β-secretase cleavage site and Aβ42 on APP were screened out by phage ELISA. The cleavage site of β-secretase and the binding characteristics of Aβ42 and the influence on Aβ aggregation and cytotoxicity finally select the polypeptide YS1 through the combination of peptide chains.

[0046] The amino acid sequence of YS1 is: Pro-Gln-Val-Gly-His-Leu (Sequence 1, PQVGHL).

[0047] Peptide YS1 is a hexapeptid...

Embodiment 2

[0055] Embodiment 2, the application of polypeptide YS1

[0056] 1. The specific binding of polypeptide YS1 and its derivatives to the β-secretase cleavage site on APP and Aβ42

[0057] 1. The specific binding of polypeptide YS1 to the β-secretase cleavage site on APP and Aβ42

[0058] The β-secretase cleavage site polypeptide (Glu-Val-Lys-Met-Asp-Glu-Phe) (synthesized by Shanghai Gil Polypeptide Co.), Aβ42 (American Peptide Co., catalog No. 62-0-80B) and negative control polypeptide (Lys-Thr-Asp-Met-Lys-His-Met-Ala-Gly-Ala-Ala-Ala-Ala-Gly-Ala-Val-Val-Gly-Gly -Leu-Gly) was coated with 1 μg / well on a 96-well aminolation microtiter plate, overnight at 4°C. Block the blank site on the microtiter plate that is not bound to the polypeptide with BSA, then add YS1 (1 μg / well) obtained in Example 1 with 6 histidines attached as tags, and act for 2 hours at room temperature (25°C). After washing, an HRP-coupled antibody capable of binding to the histidine tag (Beijing Boaosen Biotec...

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Abstract

The invention discloses a polypeptide capable of inhibiting a beta-secretase digestion effect, and application thereof. The polypeptide provided by the invention is characterized in that 1, the polypeptide has an amino acid sequence shown in the sequence 1 of the sequence table; or 2, the polypeptide has an amino acid sequence which is derived from the amino acid sequence shown in the sequence 1 through replacement and / or deletion and / or addition of one or more amino acid residues, and has functions the same as functions of the polypeptide having the amino acid sequence shown in the sequence 1. A result of an experiment shows that the polypeptide has a capability of simultaneous combination with a beta-secretase digestion site and A beta42 of an amyloid precursor protein (APP), has effects of inhibiting a beta-secretase digestion effect and A beta42 production, can inhibit A beta42 cytotoxicity, and can protect SH-SY5Y neurobiastoma cells from A beta42 cytotoxicity influences.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a polypeptide for inhibiting enzyme-cutting action of β-secretase and its application. Background technique [0002] Alzheimer's Disease Alzheimer's Disease, hereinafter referred to as AD, commonly known as senile dementia, is formed by the aggregation of β-amyloid monomer molecules (β-Amyloid 40 / 42 (Aβ40 / 42, hereinafter referred to as Aβ)) with toxic effects Aggregate-induced neurodegenerative diseases in the elderly are mainly characterized by memory loss and the formation of senile plaques in the brain. Medical statistics show that 5-6% of the elderly over the age of 60 in my country and European and American countries suffer from Alzheimer's disease. The disease has been ranked as the fourth leading cause of death after heart disease, cancer and stroke. my country's existing population over 60 years old is about 100 million. According to the estimation of 5% prevalence rate of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06A61K38/08A61P25/28
Inventor 刘瑞田杨世高王少伟
Owner TSINGHUA UNIV
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