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Varicella-zoster virus mRNA vaccine as well as preparation method and application thereof

A technology of herpes zoster virus and chickenpox, which is applied in the field of varicella-zoster virus mRNA vaccine and its preparation, and can solve problems such as unstable glycosylation products, difficult quality control, and no herpes zoster vaccine

Pending Publication Date: 2022-08-05
浙江君怡生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In 2006, the US FDA approved the first live attenuated herpes zoster vaccine (Oka / Merck strain, Merck & Co), the vaccine was developed by Merck, based on its marketed live attenuated varicella vaccine (Varivax, Oka / Merck), which was concentrated and prepared, and the virus content was 14 times that of the live attenuated varicella vaccine, but the vaccine Only 60%-70% effective in preventing shingles
However, QS-21 is hemolytic and needs to be used in combination with cholesterol and liposomes to eliminate side effects
In addition, the use of CHO cell lines to produce glycoprotein E has disadvantages such as high cost, complicated fermentation and purification processes, unstable glycosylated products, and difficult quality control.
[0006] In recent years, some researchers have developed the The inactivated varicella-zoster vaccine prepared by γ-ray irradiation, the clinical trial results show that the vaccine is well tolerated, but due to its low immunogenicity, a four-dose immunization schedule is required, and it is effective for Ineffective in patients with hematologic malignancies
Although the herpes zoster vaccine has been promoted and applied in most countries around the world, unfortunately, there is still no herpes zoster vaccine on the market in China
There is no relevant report on the mRNA vaccine for the prevention of herpes zoster

Method used

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  • Varicella-zoster virus mRNA vaccine as well as preparation method and application thereof
  • Varicella-zoster virus mRNA vaccine as well as preparation method and application thereof
  • Varicella-zoster virus mRNA vaccine as well as preparation method and application thereof

Examples

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preparation example Construction

[0109] In some embodiments, the preparation method of the varicella-zoster virus nucleic acid vaccine comprises:

[0110] (a) Dissolving the optimized varicella-zoster virus glycoprotein E mRNA in an aqueous solution of auxiliary materials to obtain an aqueous phase; the aqueous solutions of the auxiliary materials include but are not limited to citrate buffer, sodium acetate solution or phosphate buffer, preferably is sodium acetate solution;

[0111] (b) dissolving protonatable cationic lipids, structured lipids, auxiliary lipids and surfactants in an organic solution according to the formula ratio to obtain an organic phase; the organic solution includes absolute ethanol, isopropanol or acetone, preferably is anhydrous ethanol;

[0112] (c) the aqueous phase obtained by (a) and the organic phase obtained by (b) are mixed to obtain a mixed solution, and the solvent in the replacement mixed solution is a buffer to obtain a varicella-zoster virus nucleic acid vaccine; The aq...

Embodiment 1

[0119] Example 1 UTR sequence optimization

[0120] The UTR sequence optimization scheme in the present embodiment is shown in Table 1, and the details are as follows:

[0121] The key elements of the F-1 mRNA sequence are ARCA, X-GLOBIN 5'-UTR, varicella-zoster virus glycoprotein E Genbank reference sequence (NCBI Reference Sequence: NC_001348.1), X-GLOBIN 3'-UTR and polyA sequences and subsequent components.

[0122] The key elements of the F-2 mRNA sequence are ARCA, A-GLOBIN 5'-UTR, varicella-zoster virus glycoprotein E Genbank reference sequence (NCBI Reference Sequence: NC_001348.1), A-GLOBIN 3'-UTR and polyA sequences and subsequent components.

[0123] The key elements of the F-3 mRNA sequence are ARCA, X-GLOBIN 5'-UTR, varicella-zoster virus glycoprotein E Genbank reference sequence (NCBI Reference Sequence: NC_001348.1), lipoxygenase gene 3'-UTR and polyA sequences and subsequent elements.

[0124] The key elements of the F-4 mRNA sequence are ARCA, NCA-7d 5'-UTR...

Embodiment 2

[0129] Example 2 T7 promoter sequence optimization

[0130] The T7 promoter sequence optimization scheme in the present embodiment is shown in Table 2, and the details are as follows:

[0131] pcDNA3.1(+)(Ampicillin) was used as the expression vector, and the T7 promoter sequence in the vector was transformed to construct T1, T2, and T3 expression vectors respectively. The T7 promoter sequence in the T1 expression vector was 5'-taatacgactcactataggg- 3' (SEQ ID No: 13); the T7 promoter sequence in the T2 expression vector is 5'-taatacgactcactataagg-3' (SEQ ID No: 14); the T7 promoter sequence in the T3 expression vector is 5'-taatacgactcactataag-3' (SEQ ID No: 15).

[0132] The key elements of the glycoprotein E DNA sequence are X-GLOBIN 5'-UTR, varicella-zoster virus glycoprotein EGenbank reference sequence (NCBI Reference Sequence: NC_001348.1), X-GLOBIN 3'-UTR and polyA (80bp) Sequence and subsequent elements (F-1 in Example 1). The above genes were cloned into modified T...

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Abstract

The invention relates to the field of nucleic acid vaccines, in particular to a varicella-herpes zoster mRNA vaccine and a preparation method and application thereof. The mRNA for coding the varicella-zoster virus antigen provided by the invention contains a coding region for coding glycoprotein E of the varicella-zoster virus, and the mRNA can induce specific antibody response and CD4 + T cell response aiming at the glycoprotein E when being delivered into a body.

Description

Technical field [0001] The present invention relates to the field of nucleic acid vaccines, and in particular to the varicella-zoster virus mRNA vaccine and its preparation method and application. Background technique [0002] Herpes zoster (HZ) is a neurocutaneous disease caused by the reactivation of varicella-zoster virus (VZV) infection. Its most common and serious complication is postherpetic neuralgia. neuralgia, PHN). Chickenpox is caused by VZV infection for the first time in childhood. After recovery, the virus lurks in the sensory ganglia in the body. A small number of patients have reduced immunity due to some reasons in adulthood, and the virus is reactivated to cause herpes zoster. Herpes zoster can occur in adults at any age, but is more common in the elderly and immunocompromised people. [0003] Vaccination is the most effective means of preventing and controlling shingles and its complications. There are currently two vaccines approved for marketing, name...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/38C12N15/85A61K39/25A61P31/22
CPCC07K14/005C12N15/85A61K39/00A61P31/22C12N2710/16722C12N2710/16734A61K2039/53C12N2800/107C12N2800/22C12N2830/50C12N2830/34Y02A50/30
Inventor 申镇维孙瑶胡晓李光勇
Owner 浙江君怡生物科技有限公司
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