Preparation method of viral vaccine, and pharmaceutical composition

A virus vaccine and virus technology, applied in the field of virus vaccine preparation method and pharmaceutical composition, can solve safety problems and other problems, achieve the effect of maintaining immunogenicity, avoiding serious side effects, and antagonizing virus infection

Active Publication Date: 2021-08-17
SHANGHAI JENOMED BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these experimental vaccines can induce protective responses in animals, the development of vaccines for SARS-CoV and MERS-CoV, which are similar to 2019-nCoV, suggest that there may be serious risks in using the full-length Spike protein and other viral proteins as vaccine candidates. Safety concerns, including immunopathology and contributing to disease progression, possibly due to induction of Th2-skewed immune responses and / or anti-spike protein non-neutralizing antibodies

Method used

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  • Preparation method of viral vaccine, and pharmaceutical composition
  • Preparation method of viral vaccine, and pharmaceutical composition
  • Preparation method of viral vaccine, and pharmaceutical composition

Examples

Experimental program
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Effect test

preparation example Construction

[0040] The preparation method of the virus vaccine comprises: providing at least one full-length or partial antigenic peptide or protein coding nucleic acid sequence of the virus; dividing the full-length or partial antigenic peptide or protein coding region into one or more 60 - Partially overlapping nucleic acid sequences of 150 nucleotides to encode 20-50 amino acid residues, each of said partially overlapping nucleic acid sequences encoding sequences with furin cleavage sites or non-immunogenic glycine or serine linkers Separation to make 5-15 amino acid residues overlap; and then connect multiple partially overlapping nucleic acids in series to form nucleic acid vaccine fragments.

[0041] In some embodiments of the present invention, the coding region of the full-length or partial antigenic peptide or protein is divided into one or more partially overlapping nucleic acid sequences consisting of 90-120 nucleotides to encode 30-40 amino acid residues base; each of the part...

Embodiment 1

[0085] Below with embodiment 1 DNA vaccine plasmid in vitro transfection 293T cell experiment, embodiment 2 mouse vaccine injection immunization experiment and embodiment 3 rhesus monkey vaccine injection experiment illustrate the beneficial effect of the present invention.

[0086] figure 1 It is a schematic diagram of the partially overlapping nucleic acid sequence structure encoding the novel coronavirus spike protein S1 subunit fragment including the receptor binding domain in the experimental process of some embodiments. Although the S1 subunit has good immunogenicity, it contains more than 650 amino acid residues and may cause serious side effects after entering the human body. refer to figure 1 , pJM908 circular DNA plasmid has S1 subunit NTD encoding nucleic acid fragment, containing 11 partially overlapping nucleic acid sequences, each encoding 36 amino acids, forming a partially overlapping 36 amino acid residue encoding nucleic acid sequence 13, its downstream and ...

Embodiment 3

[0100] Embodiment 3 of the present invention has carried out intramuscular injection to 3-6 years old, the male rhesus macaque of body weight 3.5-8 kilogram with reference to aforementioned mouse vaccine injection immunization experiment, has formed following experimental group:

[0101] Groups 6 and 7: 2 rhesus monkeys were injected with an empty pVax1 plasmid, and the injection volume was 2 mg / mouse;

[0102] Groups 8 and 9: 2 rhesus monkeys were injected with pJM909 vaccine separately, and the injection dose was 2 mg / monkey;

[0103] Group 10 and Group 11: 2 rhesus macaques were mixed with pJM908 vaccine and pJM909 vaccine with the same content, and the injection volume was 4 mg / monkey.

[0104] Specifically, immediately after the injection, the muscle cells were electroporated with a Tarisa electroporation device (from Shanghai Tarisa Health Technology Co., Ltd.).

[0105] Specifically, after repeated intramuscular injections of pJM909 and mixed plasmids for 8 weeks in rh...

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Abstract

The invention provides a preparation method of a viral vaccine. The preparation method comprises the following steps: providing a coding sequence of a full-length or partial antigen peptide or protein of at least one virus, dividing the coding region of the full-length or partial antigen peptide or protein into one or more partially overlapped nucleic acid sequences, and connecting the one or more partially overlapped nucleic acid sequences in series to form a nucleic acid vaccine fragment. After the nucleic acid vaccine prepared by using the preparation method of the viral vaccine is injected into animals and human bodies, small-fragment antigen peptide vaccines are formed through in-vivo cell treatment, so serious side effects possibly caused by full-length protein vaccines can be effectively avoided, but the immunogenicity of antigen peptides or proteins can be maintained, and therefore, virus infection can be effectively antagonized. Meanwhile, a small-fragment antigen peptide component can be rapidly and correspondingly adjusted according to the mutation of the S protein of the new coronavirus, so the nucleic acid vaccine can also be effective to new coronavirus mutant strains. The invention also provides a pharmaceutical composition obtained through the preparation method of the viral vaccine.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method and a pharmaceutical composition of a virus vaccine. Background technique [0002] On January 10, 2020, the first genome sequence data of the novel coronavirus was released. The researchers compared the new coronavirus with two coronaviruses known to clearly infect humans, namely the severe acute respiratory syndrome (Severe Acute Respiratory Syndrome, SARS) coronavirus and the Middle East Respiratory Syndrome (Middle East Respiratory Syndrome, MERS) coronavirus. Compared with the genome, it was found that the new coronavirus has an average sequence similarity of 70% and 40% compared with the SARS coronavirus and the MERS coronavirus, and the key gene for the interaction between different coronaviruses and host cells, namely the spike protein (spike protein) coding genes have greater differences. [0003] Currently there is no vaccine or specific treatment for ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165C12N15/50A61K39/215A61P31/14
CPCC07K14/005A61K39/12A61P31/14C12N2770/20022C12N2770/20034A61K2039/53Y02A50/30
Inventor 黄跃进
Owner SHANGHAI JENOMED BIOTECH CO LTD
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