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Polypeptide suppressant for suppressing virus

A technology for viruses and uses, applied in the direction of antiviral agents, medical preparations containing active ingredients, peptides, etc., can solve the problem that mosquito-borne disease vectors cannot be effectively controlled.

Active Publication Date: 2013-10-30
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, ineffective control of mosquito vectors and increased long-distance migration have contributed to the increase and spread of dengue disease

Method used

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  • Polypeptide suppressant for suppressing virus
  • Polypeptide suppressant for suppressing virus
  • Polypeptide suppressant for suppressing virus

Examples

Experimental program
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preparation example Construction

[0084] According to the structures of the conotoxin MrIA and its variants provided by the present invention, various methods can be used to prepare them. Said methods include, but are not limited to: chemical synthesis and recombinant methods. Considering the short length of the polypeptide, chemical synthesis is preferred. A preparation method, for example, comprises the steps of:

[0085] (1) Synthesize crude peptides by chemical synthesis according to the provided amino acid sequence; and

[0086] (2) Purifying the crude peptide obtained in step (1), so as to obtain the conotoxin MrIA or its variants.

[0087] More preferably, when preparing the conotoxin MrIA or its variants, a protecting group is also set on the amino acid side chain of the polypeptide, which is a technique known in the art; making the polypeptide form a cyclic peptide is also known in the art Technology.

[0088] The present invention also includes nucleic acids encoding said conotoxin MrIA and varia...

Embodiment 1

[0101] Example 1. Screening of protease NS2B-NS3 inhibitors and identification of active components from conotoxins

[0102] 1. Preliminary screening of anti-protease NS2B-NS3 active peptides from conotoxins

[0103] Dengue protease NS2B-NS3 fusion protein was expressed (see NS2B and NS3 sequences identified in GenBank Accession No. NP056776). The constructed NS3-NS2B fusion protein expression plasmid PET15b-CF40-linker-NS3pro185 (TSV01) was obtained from the Novartis Institute for Tropical Diseases Pte Ltd, Singapore (see Li J., Lim S.P., et al., Functional Profiling of Recombinant NS3Proteases from All Four Serotypes of Dengue Virus Using Tetrapeptide and Octapeptide Substrate Libraries. J. Biol. Chem. 2005, 280(31): 28766-28774). The recombinant expression plasmid PET15b-CF40-linker-NS3pro185 was transformed into Escherichia coli BL21(DE3), induced by 0.5M IPTG, and cultured on a constant temperature shaker at 37°C at a speed of 200 rpm for 3 hours; then centrifuged to tak...

Embodiment 2

[0110] Example 2, artificial synthesis of antiviral active peptide MrIA and related mutants

[0111] The synthesis of the antiviral active peptide MrIA and related mutants (listed in Table 1) was completed on a 433A peptide synthesizer (ABI Applied Systems Biological Instruments, Inc.), using solid-phase synthesis technology. The resin used in this synthesis is RinkWang resin, and the amino group is Fmoc protected amino acid, both of which are produced by Advanced Chemtech Company of the United States. The Fmoc amino acid side chain protecting groups used are respectively: the side chain protecting groups of serine, tyrosine and threonine are tert-butyl (-tBu); histidine, cysteine, glutamine and asparagine The side chain protecting group of lysine and tryptophan is trityl (-Trt); the side chain protecting group of aspartic acid and glutamic acid is tert-butoxy (-OtBu); the side chain protecting group of lysine and tryptophan It is tert-butoxycarbonyl (-Boc); the side chain pr...

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Abstract

The invention relates to a polypeptide suppressant for suppressing virus, and discloses a small peptide which can efficiently suppress activity of virus protease NS2B-NS3 for the first time, and is derived from conotoxin MrIA of conus marmoreus. The inventor also analyzes and optimizes the amino acid sequence of the conotoxin MrIA so as to obtain a series of optimized small peptides. The optimized small peptides can suppress the duplication, processing and maturation of virus by suppressing the activity of NS2B-NS3, thus effectively resisting disease spreading and having a good medical prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology; more specifically, the invention relates to a polypeptide inhibitor for inhibiting viruses, and the inhibitor is purified from the crude venom of marbled cone snails and has the activity of inhibiting virus NS2B-NS3 protease Polypeptides and their variants. Background technique [0002] Dengue fever is an acute infectious disease caused by dengue virus and transmitted by Aedes mosquitoes. The clinical features are sudden onset, high fever, muscle, bone marrow and arthralgia all over the body, extreme fatigue, and some patients have rash, bleeding tendency and enlarged lymph nodes. Dengue is classified according to its clinical features: classic dengue, or the more severe form of dengue hemorrhagic fever syndrome (DHF), and dengue shock syndrome (DSS). Dengue virus is a member of the Flaviviridae family and includes four closely related virus serotypes: DEN-1, DEN-2, DEN-3 and DEN-4, recovery from i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/08A61K38/12C07K7/06A61P31/14
CPCY02A50/30
Inventor 王春光徐少琼邵晓霞
Owner TONGJI UNIV
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