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CN-121974982-A - Polypeptide for inhibiting porcine epidemic diarrhea virus and application thereof

CN121974982ACN 121974982 ACN121974982 ACN 121974982ACN-121974982-A

Abstract

The invention belongs to the technical field of prevention and treatment of porcine epidemic diarrhea virus, and particularly relates to polypeptide for inhibiting porcine epidemic diarrhea virus and application thereof. The invention provides a polypeptide RTN4-P specifically combined with a C288 site of an RTN4 protein, wherein the combination energy is-7.354 kcal/mol, the number of amino acid residues is 19, the molecular weight is about 2.41kDa, the isoelectric point is about 11.0-11.5, and the polypeptide is mainly used for combining with 288 th cysteine residues of the RTN4 protein and upstream and downstream sequences thereof, blocking the infection process of PEDV mediated by RTN4 and achieving the effect of inhibiting replication of Porcine Epidemic Diarrhea Virus (PEDV). The results of the examples show that the polypeptide has strong specificity, good biocompatibility and low toxicity, and has wide application prospect.

Inventors

  • WU HAO
  • CHEN YINGXIAN
  • GUO YUBO
  • HE CHUAN

Assignees

  • 华中农业大学

Dates

Publication Date
20260505
Application Date
20260210

Claims (10)

  1. 1. A polypeptide for inhibiting porcine epidemic diarrhea virus is characterized in that the polypeptide is RTN4-P or RTN4-P with more than 75% of identity and specifically binds to a RTN4 protein C288 locus and upstream and downstream sequences thereof, and the sequence of the RTN4-P is shown as SEQ ID NO. 1.
  2. 2. The polypeptide of claim 1, wherein the sequence of the RTN4 protein is shown in SEQ ID NO. 2.
  3. 3. A nucleic acid molecule encoding the polypeptide of claim 1.
  4. 4. An agent that inhibits porcine epidemic diarrhea virus, comprising the polypeptide of claim 1.
  5. 5. The agent of claim 4, wherein the polypeptide is RTN4-P at a concentration of 3 μm.
  6. 6. The pharmaceutical agent of claim 5, wherein the pharmaceutical agent has a purity of RTN4-P of greater than or equal to 95%.
  7. 7. The medicament of claim 5, wherein the medicament is an injectable medicament.
  8. 8. Use of the polypeptide of claim 1 or the agent of any one of claims 4-7 for inhibiting replication of porcine epidemic diarrhea virus.
  9. 9. Use of the polypeptide of claim 1 or the agent of any one of claims 4-7 for inhibiting dimerization of RTN4 proteins caused by an epidemic diarrhea virus infection.
  10. 10. Use of the polypeptide of claim 1 for the preparation of a medicament for the prevention and treatment of porcine epidemic diarrhea.

Description

Polypeptide for inhibiting porcine epidemic diarrhea virus and application thereof Technical Field The invention belongs to the technical field of prevention and treatment of porcine epidemic diarrhea virus, and particularly relates to polypeptide for inhibiting porcine epidemic diarrhea virus and application thereof. Background Porcine Epidemic Diarrhea (PED) is a common disease in large-scale pig farms, and Porcine Epidemic Diarrhea Virus (PEDV) infection is a major cause of Porcine epidemic diarrhea. The virus first appeared in the uk and belgium in the late 1970 s and was first reported in the united states in 2013. Since 2010, PED has repeatedly exploded in china and the united states, current vaccines and antiviral drugs are not effective in preventing infection with newly evolved highly pathogenic PEDV variants. PEDV can infect all stages of pigs, with the greatest effect on suckling piglets, causing severe enteritis, watery diarrhea, vomiting, dehydration, and mortality rate of up to 100% for piglets within 3 days of age. The PEDV has high transmission speed and high infection rate, and causes great economic loss to the pig raising industry in China. Currently, the means of treatment and control for PEDV are relatively limited. Traditional prevention and control methods rely mainly on biosafety measures, vaccines and antiviral drugs. Due to the high variability and resistance of PEDV, traditional vaccines and drugs have limited efficacy. Therefore, there is an urgent need to find new, highly effective antiviral drugs to effectively control the spread of PEDV and epidemic spread. Disclosure of Invention The invention aims to provide a polypeptide for inhibiting porcine epidemic diarrhea virus and application thereof, and the polypeptide RTN4-P provided by the invention is a polypeptide inhibitor, has strong specificity, good biocompatibility and low toxicity, and has wide application prospect in the field of targeted treatment. In order to achieve the above object, the present invention provides the following technical solutions: The invention provides a polypeptide for inhibiting porcine epidemic diarrhea virus, which is RTN4-P or has more than 75% of identity with RTN4-P and specifically binds with a C288 locus of RTN4 protein and upstream and downstream sequences thereof, wherein the sequence of the RTN4-P is shown as SEQ ID NO. 1. Preferably, the sequence of the RTN4 protein is shown as SEQ ID NO. 2. The invention also provides a nucleic acid molecule which encodes the polypeptide. The invention also provides an agent for inhibiting porcine epidemic diarrhea virus, which comprises the polypeptide, preferably RTN4-P. Preferably, the concentration of RTN4-P is 3. Mu.M. Preferably, the purity of RTN4-P in the medicament is more than or equal to 95%. Preferably, the medicament is an injectable medicament. Preferably, the medicament further comprises a pharmaceutically acceptable carrier or adjuvant. The invention also provides the following applications: Use of the polypeptide or agent described above for inhibiting replication of Porcine Epidemic Diarrhea Virus (PEDV). Use of the polypeptide or agent described above for inhibiting dimerization of RTN4 proteins caused by PEDV infection. The application of the polypeptide in preparing a medicament for preventing and treating porcine epidemic diarrhea. The beneficial effects are that: the polypeptide has antiviral effect on PEDV, and RTN4-P has better antiviral effect at the concentration of 3 mu M. The polypeptide RTN4-P has the advantages of strong specificity, good biocompatibility, low toxicity and the like, and has wide application prospect in the field of targeted therapy. Drawings In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings that are needed in the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art. FIG. 1 shows the results of expression of PEDV-N protein in RTN4 knockout cells FIG. 2 is a three-dimensional structural analysis diagram of the interaction of RTN4-P with the sequence upstream and downstream of the C288 site of RTN4 protein; FIG. 3 RTN4-P inhibits PEDV infection in a time gradient; FIG. 4 fluorescence quantitative PCR assay for the inhibition of PEDV by RTN 4-P; FIG. 5 Western blot shows the effect of RTN4-P on inhibition of PEDV; FIG. 6 immunofluorescence assay of the inhibitory effect of RTN4-P on PEDV; FIG. 7 effect of RTN4-P on PEDV titer. Detailed Description The invention provides a polypeptide for inhibiting porcine epidemic diarrhea virus, which is RTN4-P and has the following sequence: YGRKKRRQRRRGHVNCTIK(SEQ ID NO:1) the binding energy is-7.354 kcal/mol, the number of amino acid residues is 19, the molecu