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CN-121991167-A - Antibacterial peptide derived from marine microorganisms, screening method and application thereof

CN121991167ACN 121991167 ACN121991167 ACN 121991167ACN-121991167-A

Abstract

The invention relates to an antibacterial peptide derived from marine microorganisms, a screening method and application thereof, wherein the antibacterial peptide is selected from a compound shown in a formula (I), or a stereoisomer, a prodrug, a crystal form, a pharmaceutically acceptable salt, a pharmaceutically acceptable ester or a pharmaceutically acceptable solvate of the compound; the screening method comprises the following steps of firstly obtaining a short open reading frame sequence based on marine source bacillus genome resources from marine environment sources, ensuring that a target sequence accords with the core structural characteristics and the length range of antimicrobial peptide precursor molecules, obtaining a non-redundant antimicrobial peptide sequence through sequence redundancy elimination optimization, multi-model collaborative verification and database construction, synthesizing polypeptide by adopting a solid phase synthesis method, reacting with acetic anhydride and pyridine, carrying out N-terminal acetyl end capping and C-terminal amino end capping, and obtaining the antimicrobial peptide, and the application of the antimicrobial peptide in preparation of antimicrobial drugs. The invention solves the problems of high cost, low antibacterial efficiency and high hemolysis of the existing antibacterial peptide manual screening, can efficiently inhibit various gram positive and negative pathogenic bacteria, and has broad-spectrum antibacterial activity and use safety.

Inventors

  • WEI BIN
  • LI MENGXIN
  • WANG HONG
  • Hu Gangao
  • YANG CAILING

Assignees

  • 浙江工业大学

Dates

Publication Date
20260508
Application Date
20260115

Claims (10)

  1. 1. An antibacterial peptide derived from marine microorganisms is characterized by being selected from a compound shown in the following formula (I), or a stereoisomer, a prodrug, a crystal form, a pharmaceutically acceptable salt, a pharmaceutically acceptable ester or a pharmaceutically acceptable solvate of the compound; 。
  2. 2. The method for screening antimicrobial peptides derived from marine microorganisms according to claim 1, wherein the method comprises the steps of obtaining a short open reading frame sequence based on marine environmental source bacillus genome resources, ensuring that a target sequence accords with the core structural characteristics and length range of antimicrobial peptide precursor molecules, obtaining a non-redundant antimicrobial peptide sequence through sequence redundancy elimination optimization, multi-model collaborative verification and database construction, synthesizing polypeptide RWWCRWR by adopting a solid-phase synthesis method, reacting with acetic anhydride and pyridine, and carrying out N-terminal acetyl end capping and C-terminal amino end capping to obtain antimicrobial peptide Ac-RWWCRWR-NH 2 shown in the formula (I).
  3. 3. The method for screening marine microorganism-derived antibacterial peptide according to claim 2, wherein in the process of obtaining the short open reading frame sequence, a bioinformatics tool EMBOSS is adopted to carry out genome-wide scanning on the marine bacillus genome resource, the short open reading frame sequence is specifically identified and predicted, the screening standard is set to be 18-150 base pairs in sequence length, and the target sequence is ensured to conform to the core structural characteristics and the length range of the antibacterial peptide precursor molecule.
  4. 4. The method for screening the antimicrobial peptide derived from the marine microorganism according to claim 2, wherein in the process of sequence redundancy elimination optimization, short open reading frame sequences obtained through prediction are combined in batches, efficient sequence duplication elimination treatment is carried out by adopting Seqkit tools, 100% similar redundant sequences are eliminated through accurate comparison screening, resource consumption and result deviation caused by repeated analysis are effectively avoided, and the efficiency and accuracy of subsequent prediction analysis are ensured.
  5. 5. The method for screening antimicrobial peptides derived from marine microorganisms according to claim 2, wherein in the process of multi-model collaborative verification, the smORF sequences after redundancy removal are input into 8 deep learning prediction models in three open source projects of c_amps-prediction, SMEP and AMP-potency-prediction-EvoGradient, collaborative prediction and cross verification of antimicrobial activity are performed, strict judgment standards are set, and only when all 8 deep learning prediction models are consistent in judging that the sequences have antimicrobial activity, the sequences can be included in candidate antimicrobial peptide categories, and reliability of prediction results are remarkably improved through multi-model cross verification.
  6. 6. The method for screening marine microorganism-derived antibacterial peptide according to claim 2, wherein in the process of constructing the database, non-redundant candidate antibacterial peptides are finally obtained through screening, a marine bacillus-derived antibacterial peptide comprehensive database is constructed, comprehensive evaluation is performed based on core indexes such as multi-model prediction confidence, sequence physicochemical properties and the like, and finally the antibacterial peptide Ac-RWWCRWR-NH 2 is selected.
  7. 7. The use of an antimicrobial peptide derived from a marine microorganism according to claim 1 for the preparation of an antimicrobial drug.
  8. 8. The method of claim 7, wherein the bacteria of the antimicrobial agent are E.coli, vibrio parahaemolyticus, acinetobacter baumannii, pseudomonas aeruginosa, bacillus subtilis, staphylococcus aureus and Bacillus anthracis.
  9. 9. The method of claim 8, wherein the antimicrobial agent has a minimum inhibitory concentration of 8. Mu.M, 16. Mu.M, 32. Mu.M, 4. Mu.M, 16. Mu.M, and 32. Mu.M for Escherichia coli, vibrio parahaemolyticus, acinetobacter baumannii, pseudomonas aeruginosa, bacillus subtilis, staphylococcus aureus, and Bacillus anthracis.
  10. 10. The marine microorganism-derived application of claim 7, wherein the active ingredient of the antibacterial agent is a compound represented by the above formula (I), or is selected from stereoisomers, prodrugs, crystal forms, pharmaceutically acceptable salts, pharmaceutically acceptable esters or pharmaceutically acceptable solvates of the compound.

Description

Antibacterial peptide derived from marine microorganisms, screening method and application thereof Technical Field The invention relates to the technical field of polypeptides, in particular to an antibacterial peptide derived from marine microorganisms, and a screening method and application thereof. Background The spread of drug-resistant pathogens worldwide has attracted considerable attention from the medical and scientific community. The widespread use of multiple antibiotics has led to the progressive development of bacteria from resistance to a single antibiotic to multiple resistance, even pan-resistance. The antibacterial peptide mainly plays an antibacterial role by destroying bacterial cell membranes, has broad-spectrum antibacterial activity and low drug resistance, and is expected to become a substitute drug of traditional antibiotics. The emergence and spread of antimicrobial drug resistance (AMR) is a global public health crisis, placing a heavy burden on healthcare systems, leading to high mortality and high economic costs. The traditional antibiotics have long development period and quick drug resistance generation, so that a novel antibacterial strategy is needed. Polypeptide antibacterial drugs, particularly antibacterial peptides (AMPs), are a hotspot in current research due to their high antibacterial activity, low drug resistance and broad biological activity. At present, the antibacterial activity of natural antibacterial peptides is relatively limited, and in actual drug development, antibacterial peptides with higher activity and stronger pertinence are required to exert antibacterial effects. Ocean is used as the most abundant ecological system of biodiversity on the earth, and a large amount of bioactive substances with unique structures and various functions are inoculated. Among them, marine-derived antibacterial peptides have been the focus of research in recent years in the biomedical field by virtue of their excellent antibacterial properties and unique action mechanisms. Compared with the traditional antibiotics, the marine antibacterial peptide has broad-spectrum antibacterial activity, can effectively inhibit bacteria such as gram-positive bacteria, gram-negative bacteria and the like, and also has obvious inhibition effect on fungi, viruses and certain tumor cells. In addition, the marine antibacterial peptide has the characteristics of small molecular weight, good thermal stability, high water solubility, excellent biocompatibility and the like, and has great application potential in the fields of clinical infection treatment, food preservation, aquaculture disease control and the like. Meanwhile, with the rise of artificial intelligence, the deep learning model can quickly learn a large number of samples of known medicines, the generated model after training can be directly sampled to produce totally new medicine molecules, and the discrimination model can screen and identify new potential patent medicine objects by predicting the attributes of the molecules, so that the medicine design process is effectively accelerated. AMPs databases and predictive models began to emerge, with the use of bioinformatics resources and computational tools to aid peptide design, predict peptide activity to screen for optimal candidate peptides. However, the wide range of antimicrobial peptides still face many challenges, mainly due to their low bacteriostatic activity and high hemolysis, which limit their wide range of applications in practical treatments. Disclosure of Invention Aiming at the defects of the prior art, the first object of the invention is to provide an antibacterial peptide from marine microorganism, which can inhibit various gram positive and negative pathogenic bacteria with high efficiency, broad-spectrum antibacterial activity and use safety. The second aim of the invention is to provide a screening method of antibacterial peptide from marine microorganism, which solves the problems of high cost, low antibacterial efficiency and high hemolysis of the existing antibacterial peptide manual screening. The third object of the present invention is to provide an application of antimicrobial peptides derived from marine microorganisms, which can efficiently inhibit various gram-positive and negative pathogenic bacteria, and has broad-spectrum antibacterial activity and use safety. In order to achieve the first object, the present invention provides the following technical solutions: An antibacterial peptide derived from a marine microorganism, selected from a compound represented by the following formula (I), or selected from stereoisomers, prodrugs, crystalline forms, pharmaceutically acceptable salts, pharmaceutically acceptable esters or pharmaceutically acceptable solvates of the compound; 。 Specifically, in the antibacterial peptide, the sequence of the antibacterial peptide is Ac-RWWCRWR-NH 2, the antibacterial peptide is synthesized by adopting a solid-phase synthesi