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A recombinant novel coronavirus vaccine based on human replication-defective adenovirus

A coronavirus, recombinant adenovirus technology, applied in the direction of viruses/phages, viruses, viral peptides, etc., to achieve the effects of good immunogenicity, fast and simple preparation method, and good immune protection effect

Active Publication Date: 2020-09-11
ACADEMY OF MILITARY MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The types of new coronavirus vaccines under research include adenovirus vector vaccines, mRNA vaccines, DNA vaccines, recombinant protein vaccines and inactivated vaccines, and the results of the research have not yet been reported

Method used

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  • A recombinant novel coronavirus vaccine based on human replication-defective adenovirus
  • A recombinant novel coronavirus vaccine based on human replication-defective adenovirus
  • A recombinant novel coronavirus vaccine based on human replication-defective adenovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1. Preparation of recombinant novel coronavirus vaccine with human replication defective adenovirus as carrier

[0044] 1. S protein gene optimization and synthesis

[0045] The target antigen of the recombinant novel coronavirus vaccine is the S protein of the novel coronavirus strain (Genebank number: NC_045512.2). By optimizing the S protein gene, the expression level of the S protein is increased, thereby improving the immunogenicity of the vaccine.

[0046] First, use the Upgene software (Gao, W.Rzewski, A.Sun, H.Robbins, P.D. & Gambotto, A. UpGene: Application of a web-based DNA codon optimization algorithm. Biotechnol Prog, 2004.20 (2): p.443-8 .) Perform codon optimization to change most of the rare codons in the S protein gene to high frequency codons. Secondly, considering that software optimization may mechanically change codons to the most frequently used codons, protein translation efficiency may not be significantly improved due to the influence ...

Embodiment 2

[0107] Example 2. Immunological evaluation of different constructed recombinant adenoviruses on mouse models

[0108] 1. Vaccine humoral immune response detection

[0109] 100 SPF grade female BALB / c mice (6-8 weeks old) were randomly divided into 10 groups, 10 mice in each group. The mice were immunized with Ad5-nCoV according to the grouping conditions shown in Table 1. The way of intramuscular injection is to inject 100 μL into the inner side of the hind thigh, and the way of nasal drop immunization is to anesthetize the mice with isoflurane and instill 20 μL through the nasal cavity. The grouping situation is shown in Table 1.

[0110] Table 1. Grouping of mice for vaccine humoral immune response detection

[0111]

[0112]

[0113] Blood was collected from the mice at a specific time point after immunization, the serum was separated, and the IgG antibody titer against the new coronavirus S protein in the serum was detected by ELISA. Test results such as Figure...

Embodiment 3

[0127] Example 3. Immunological evaluation of Ad5-nCoV on guinea pig model

[0128] Fifty-six guinea pigs of SPF grade, weighing 200 to 250 grams, were randomly divided into 4 groups, 14 in each group, half male and half male. Guinea pigs were immunized with Ad5-nCoV according to the grouping conditions shown in Table 3. The way of immunization was intramuscular injection of 200 μL in the rear thigh.

[0129] Table 3. Grouping of Ad5-nCoV guinea pig immunogenicity detection

[0130]

[0131] Blood was collected from the guinea pigs at a specific time point after immunization, the serum was separated, and the IgG antibody titer against the novel coronavirus S protein in the serum was detected by ELISA. Test results such as Figure 14 Shown (ns, P≥0.05; *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001). The results showed that 14 days after Ad5-nCoV immunization in guinea pigs, high levels of serum IgG antibody titers were detected. There was no significant difference...

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Abstract

The invention provides a novel coronavirus vaccine using human type 5 replication-deficient adenovirus as a carrier. The vaccine uses a replication-defective human type 5 adenovirus with combined deletion of E1 and E3 as the carrier, HEK293 cells integrated with the adenovirus E1 gene as the packaging cell line, and the protective antigen gene carried is an optimized 2019 novel coronavirus (SARS‑CoV‑2) S protein gene (Ad5‑nCoV). After the S protein gene was optimized, the expression level in the transfected cells was significantly increased. The vaccine has good immunogenicity in both mouse and guinea pig models, and can induce strong cellular and humoral immune responses in the body in a short time. The protective effect study on hACE2 transgenic mice showed that a single immunization with Ad5-nCoV can significantly reduce the viral load in the lung tissue after 14 days, indicating that the vaccine has a good immune protective effect against 2019-nCoV. In addition, the preparation of the vaccine is quick and easy, and large-scale production can be realized in a short period of time for emergency outbreaks.

Description

technical field [0001] The invention relates to a recombinant novel coronavirus vaccine, aimed at preventing the epidemic of novel coronavirus. The invention belongs to the technical field of bioengineering. Background technique [0002] The 2019 novel coronavirus, SARS-CoV-2 (also known as 2019-nCoV), was named by the World Health Organization on January 12, 2020. It is a new strain of coronavirus that has never been found in humans before. This virus is the seventh coronavirus (CoV) that can infect humans. The incubation period of human infection with 2019-nCoV is generally 1 to 14 days. Common signs after infection with 2019-nCoV include respiratory symptoms, fever, cough, shortness of breath, and dyspnea. In more severe cases, infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and even death. As of March 17, 2020, my country has reported 81,116 confirmed cases and 3,231 deaths; other countries have reported 98,486 cases and a total of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/50C07K14/165C12N15/861A61K39/215A61P31/14
CPCC07K14/005C12N15/86A61K39/12A61P31/14C12N2770/20022C12N2710/10043C12N2770/20034A61K2039/575A61K2039/543A61K2039/545A61K2039/57A61K2039/572A61K39/215
Inventor 陈薇吴诗坡侯利华张哲王步森郭强张金龙宋小红付玲张军陈旖赵拯浩朱涛李荩莘春林
Owner ACADEMY OF MILITARY MEDICAL SCI
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