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Novel coronavirus polypeptide vaccine coupled with TLR7 agonist and application of novel coronavirus polypeptide vaccine

A coronavirus and vaccine technology, applied in viral peptides, positive-sense single-stranded RNA viruses, viruses, etc., can solve the problems of virus immune escape, limited immune protection, and low immunogenicity

Active Publication Date: 2020-11-06
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the immune protection produced by the currently developed vaccines is limited, such as low immunogenicity, virus immune escape and other problems

Method used

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  • Novel coronavirus polypeptide vaccine coupled with TLR7 agonist and application of novel coronavirus polypeptide vaccine
  • Novel coronavirus polypeptide vaccine coupled with TLR7 agonist and application of novel coronavirus polypeptide vaccine
  • Novel coronavirus polypeptide vaccine coupled with TLR7 agonist and application of novel coronavirus polypeptide vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0190] Example 1 Antigen polypeptide screening based on the sequence and structure analysis of S protein RBD

[0191] In this example, the inventors took the RBD region interacting with human ACE2 in the S protein of SARS-CoV-2 as the analysis object, and determined the key interaction sites in the RBD, such as figure 1 and figure 2 shown.

[0192] The inventor used software to predict and analyze the CD8+ T cell epitope in the RBD sequence; used the AlleleFrequency Net Database database statistical analysis to obtain the main HLA class II molecular allelic types of the world population; further analyzed and predicted the HLA class II molecule in the RBD sequence The binding peptide was used as the predicted CD4+ T cell epitope; the linear and conformational B cell epitopes in the RBD sequence were predicted by using BepiPred and Discotope software; the glycosylation modification site in the RBD region was obtained by using the Uniprot database; ProtParam software predicts ...

Embodiment 2

[0206] The preparation of embodiment 2 polypeptide

[0207] In this example, polypeptides P-37, P-67, P-71 and LY54 were prepared using a polypeptide synthesizer.

[0208]

Embodiment 3

[0209] The preparation of embodiment 3 coupled peptides

[0210] The artificially synthesized polypeptides P-37, P-67 (including P-67-F1 and P-67-F2), P-71 and LY54 were coupled with TLR7 small molecule agonists to form corresponding coupling peptide.

[0211] The molecular structure of the TLR7 small molecule agonist used to couple the polypeptide is as follows:

[0212]

[0213]

[0214] 3.1 Preparation of coupled peptide P-37-S2

[0215] Preparation of conjugates of P-37 peptide (CYRLFRKSNLKPFERDISTEIYQAGS) and TLR7 small molecule agonist. Weigh 18 mg of P-37 peptide and dissolve in 270 μL H 2 In the mixed solution of O and 180 μL DMF, add 4.88 mg of SZU-101-Mal, and at the same time add NaHCO with a final concentration of 30 mM 3 , After reacting at room temperature for 1 hour, the product was separated by a C18 preparative column to obtain a total of 15.5 mg of SZU-101 and P-37 peptide coupling product P-37-S2, HRMS [M+5H] 5 +740.9976.

[0216] 3.2 Preparation...

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PUM

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Abstract

The invention discloses a novel coronavirus polypeptide vaccine coupled with a TLR7 agonist and application of the novel coronavirus polypeptide vaccine. Specifically, the invention provides a novel vaccine polypeptide for coronavirus pneumonia based on analysis and research of an RBD sequence and structural information of an S protein of SARS-CoV-2, the vaccine polypeptide has the following formula structure: Z-(J-U) n, and in the formula, Z, J, U, n and the like are as defined in the specification. The invention further provides a vaccine composition containing the vaccine polypeptide and application of the composition. Experiments show that the vaccine polypeptide can enable mice and cynomolgus monkeys to start strong cell and humoral immune effects, generates a neutralizing antibody for blocking combination of RBD and ACE2, and can be used for preventing and treating novel coronavirus pneumonia.

Description

technical field [0001] The present invention relates to the fields of polypeptide drugs and polypeptide vaccines, in particular to novel coronavirus polypeptide vaccines coupled with TLR7 small molecule agonists and applications thereof. Background technique [0002] The novel coronavirus pneumonia (Corona virus disease 2019, COVID-19) caused by the coronavirus SARS-CoV-2 is highly contagious and has caused severe outbreaks worldwide, endangering the lives of millions of people. However, there is still a lack of clear and effective preventive and therapeutic drugs and measures for COVID-19. Clinically, supportive and symptomatic treatments are mainly used. [0003] Establishing herd immunity against SARS-CoV-2 through vaccines is the ultimate way to control and block the COVID-19 epidemic. Various types of COVID-19 vaccines are currently in preclinical and clinical trials, including live attenuated vaccines, inactivated virus vaccines, recombinant virus vector vaccines, rec...

Claims

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Application Information

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IPC IPC(8): C07K14/165A61K39/215A61P31/14A61P37/04A61K31/522A61K39/39A61K47/55A61K47/65
CPCC07K14/005A61K39/12A61P31/14A61P37/04A61K31/522A61K39/39A61K47/55A61K47/65C12N2770/20022C12N2770/20034C12N2770/20071A61K2039/6031A61K47/646A61K39/215A61K2039/60A61K2039/627A61K2039/70
Inventor 宫丽崑任进靳广毅黄蔚秦秋平龙益如孙建华刘婷婷唐峰朱鹏
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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