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CN-121991161-A - Pilose antler polypeptide, virtual enzymolysis preparation method thereof and application thereof in anti-aging aspect

CN121991161ACN 121991161 ACN121991161 ACN 121991161ACN-121991161-A

Abstract

The invention belongs to the technical field of polypeptide preparation and biological medicine, and in particular relates to a pilose antler polypeptide, a virtual enzymolysis preparation method thereof and application thereof in anti-aging. The 8 pilose antler polypeptides provided by the invention are polypeptides with DRP1 inhibition activity obtained by screening pilose antler proteins, have higher anti-aging activity and definite action mechanism, are found to be nontoxic and non-sensitized, have strong binding capacity with DRP1 targets, have good drug forming property and application safety, can be used for developing anti-aging products, and provide a new lead compound for developing new drugs, health care products or cosmetics. Furthermore, the pilose antler polypeptide is obtained based on a virtual enzymolysis preparation method, a complete and efficient pilose antler anti-aging peptide computer screening process is established, virtual enzymolysis, activity prediction, safety evaluation and targeting molecule docking are organically combined, blindness of the traditional separation technology is overcome, and screening efficiency and success rate are remarkably improved.

Inventors

  • HE ZHONGMEI
  • LIU JINZE
  • ZHOU JIA
  • GENG JIANAN
  • CHEN WEIJIA
  • DU RUI

Assignees

  • 吉林农业大学

Dates

Publication Date
20260508
Application Date
20260126

Claims (10)

  1. 1. A pilose antler polypeptide, characterized by one or more of amino acid sequence PNG, ACAL, YR, AYG, GH, NGGF, YG and GR.
  2. 2. A method for preparing the pilose antler polypeptide of claim 1 based on virtual enzymolysis, comprising the steps of: Performing virtual enzymolysis on the deer antler-derived protein by using protease to obtain an enzymolysis peptide fragment library, wherein the protease comprises one or more of papain, bromelain, chymotrypsin A, trypsin, pepsin and proteinase K; Scoring the enzymolysis peptide fragment library by using a biological activity prediction tool, and screening peptide fragments with scores higher than a set threshold value to obtain a primary screening peptide fragment library; carrying out safety evaluation on the primary screening peptide fragment library through a toxicity prediction tool and a sensitization prediction tool, removing toxic and sensitization peptide fragments, and obtaining a safe peptide fragment library; And taking the power-related protein 1 as a molecular docking receptor, performing molecular docking virtual screening on the safe peptide fragment library, and screening out peptide fragments well combined with the power-related protein 1 according to a molecular docking score to obtain the pilose antler polypeptide.
  3. 3. The method of claim 2, wherein the papain, bromelain, chymotrypsin a, trypsin, pepsin and proteinase K have EC numbers EC 3.4.22.2, EC 3.4.22.32, EC 3.4.21.1, EC 3.4.21.4 and EC 3.4.23.1, respectively.
  4. 4. The method of claim 2, wherein the bioactivity prediction tool comprises PEPTIDERANKER, the set threshold being 0.5; the toxicity prediction means comprises ToxinPred and the sensitization prediction means comprises AlerTOP.
  5. 5. The method of claim 2, wherein the molecular docking acceptor is a protein crystal structure with a protein structure database accession number of 3W6P, and wherein the molecular docking score of the peptide fragment well-bound to the power-related protein 1 is less than-6 kcal/mol.
  6. 6. The use of a deer antler polypeptide of claim 1 or prepared by the method of any one of claims 2-5 in the preparation of anti-aging products and/or in food.
  7. 7. The use of claim 6, wherein the anti-aging product comprises a DRP1 inhibitor.
  8. 8. The use of claim 7, wherein the anti-aging comprises anti-brain aging.
  9. 9. The use according to any one of claims 6 to 8, wherein the anti-ageing product comprises a pharmaceutical and/or cosmetic product.
  10. 10. An anti-aging product, characterized in that the active ingredient comprises the pilose antler polypeptide of claim 1 or the pilose antler polypeptide prepared by the method of any one of claims 2-6.

Description

Pilose antler polypeptide, virtual enzymolysis preparation method thereof and application thereof in anti-aging aspect Technical Field The invention belongs to the technical field of polypeptide preparation and biological medicine, and in particular relates to a pilose antler polypeptide, a virtual enzymolysis preparation method thereof and application thereof in anti-aging. Background With the aging of the global population, the development of safe and effective anti-aging intervention products is slightly a research hotspot. Senescence is a complex biological process, one of which is mitochondrial dysfunction. Dynamic associated protein 1 (DRP 1) is a key protein regulating mitochondrial division, and excessive activation thereof leads to mitochondrial fragmentation, accelerating cell senescence. Therefore, DRP1 has become an anti-aging drug target with great potential. Natural active peptides are of great interest because of their high activity and low toxicity. The pilose antler is taken as a traditional rare traditional Chinese medicine material, is rich in protein and polypeptide, and is considered to have the effects of nourishing and strengthening as well as deferring senility. However, the traditional method for extracting the active peptide from the hairy antler is mostly dependent on physical and chemical separation and animal experiment verification, has the problems of complicated process, long period, high cost, certain blindness and the like, and is difficult to efficiently and accurately discover the active peptide with a specific targeting function. In recent years, with the development of bioinformatics, virtual screening technology provides a new solution for efficient discovery of active peptides. Through means of computer simulated enzymolysis, activity prediction, toxicity evaluation, molecular docking and the like, candidate peptides with specific targeting activity can be rapidly screened out from mass protein sequences, so that the research and development efficiency is greatly improved, and the research and development cost is reduced. At present, no related report of exploring peptides with anti-aging activity from pilose antler proteins through a systematic virtual screening method is seen. Disclosure of Invention The invention aims to provide a pilose antler polypeptide, a virtual enzymolysis preparation method thereof and application thereof in anti-aging, wherein the pilose antler polypeptide has better anti-aging activity, is nontoxic and non-sensitized, and has good drug forming property and application safety. The invention provides a pilose antler polypeptide, the amino acid sequence of which is one or more of PNG, ACAL, YR, AYG, GH, NGGF, YG and GR. The invention also provides a method for preparing the pilose antler polypeptide based on the technical scheme, which comprises the following steps: Performing virtual enzymolysis on the deer antler-derived protein by using protease to obtain an enzymolysis peptide fragment library, wherein the protease comprises one or more of papain, bromelain, chymotrypsin A, trypsin, pepsin and proteinase K; Scoring the enzymolysis peptide fragment library by using a biological activity prediction tool, and screening peptide fragments with scores higher than a set threshold value to obtain a primary screening peptide fragment library; carrying out safety evaluation on the primary screening peptide fragment library through a toxicity prediction tool and a sensitization prediction tool, removing toxic and sensitization peptide fragments, and obtaining a safe peptide fragment library; And taking the power-related protein 1 as a molecular docking receptor, performing molecular docking virtual screening on the safe peptide fragment library, and screening out peptide fragments well combined with the power-related protein 1 according to a molecular docking score to obtain the pilose antler polypeptide. Preferably, the EC numbers of papain, bromelain, chymotrypsin a, trypsin, pepsin and proteinase K are EC 3.4.22.2, EC 3.4.22.32, EC 3.4.21.1, EC 3.4.21.4 and EC 3.4.23.1, respectively. Preferably, the biological activity prediction tool comprises PEPTIDERANKER, the set threshold value is 0.5; the toxicity prediction means comprises ToxinPred and the sensitization prediction means comprises AlerTOP. Preferably, the molecular docking receptor is a protein crystal structure with a protein structure database accession number of 3W6P, and the molecular docking score of the peptide segment well combined with the power-related protein 1 is lower than-6 kcal/mol. The invention also provides application of the pilose antler polypeptide in the technical scheme or the pilose antler polypeptide prepared by the method in the technical scheme in preparation of anti-aging products and/or foods. Preferably, the anti-aging product comprises a DRP1 inhibitor. Preferably, the anti-aging includes anti-brain aging. Preferably, the anti-aging product comprises a pharmac