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Varicella-herpes zoster mRNA vaccine composition, and preparation method and application thereof

A vaccine composition, herpes zoster technology, applied in the field of vaccines, can solve the problems of difficult quality control in the preparation process, stability needs to be further improved, active ingredients are sensitive to temperature, etc., to save time and economic costs, and avoid genome integration risk, the effect of enhancing antigen production

Pending Publication Date: 2022-02-25
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The key component of AS01B, QS21, is a saponin tree that grows only from the temperate regions of South America Quillaja saponaria The polysaccharide mixture extracted by reverse high-performance liquid chromatography in the bark cannot be artificially synthesized at present, and the source is limited, and the quality control of the preparation process is difficult (the purified components are non-monomers with complex components, and the active ingredients are sensitive to temperature), Limitations such as the need to add detoxification agents due to hemolytic activity
In addition, according to the instructions of Shingrix, the adjuvant component AS01B needs to be temporarily mixed with the antigen before use (Bedside mix), which indirectly suggests that the stability of the liposome system needs to be further improved when it is applied to vaccines
The above background makes Shingrix expensive (150-200 US dollars per injection, 2 injections in the whole process), but the supply is still in short supply (84.2% of vaccines will be sold in the United States in 2020, and there are basically no markets outside the United States at present)

Method used

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  • Varicella-herpes zoster mRNA vaccine composition, and preparation method and application thereof
  • Varicella-herpes zoster mRNA vaccine composition, and preparation method and application thereof
  • Varicella-herpes zoster mRNA vaccine composition, and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Embodiment 1——mRNA preparation:

[0070]Encoding the full-length amino acid sequence of varicella-zoster virus Oka strain glycoprotein E (UniProtKB / Swiss-Prot: Q9J3M8.1, see SEQ ID No.1), the amino acid sequence of mutations introduced into the carboxy-terminal intracellular region (Y569A, S593A, S595A, T596A, T598A, see SEQ ID No.2), the gene containing only the amino acid sequence of the extracellular region (see SEQ ID No.3) is optimized according to mammalian codon bias, together with the 5' untranslated region sequence, 3 The 'untranslated region sequence and the 3' end polyadenylation sequence were synthesized by Shanghai Sangong, and constructed between the XhoI and BamHI restriction sites on the plasmid pBlueScript II SK(+). After linearization, the mRNA synthesis kit was used (purchased from Shanghai Lanque Biology) was transcribed in vitro by co-transcription chemical substrate capping, and further purified using RNA purification kit (purchased from NEB). Qua...

Embodiment 2

[0071] Embodiment 2——Preparation of varicella-zoster mRNA vaccine composition

[0072] According to MC3: DSPC: cholesterol: DMG-PEG2000 molar ratio of 50:10:37.5:2.5, weigh the lipid (purchased from Shanghai Aiweituo) and dissolve it in absolute ethanol to form solution A; prepare 0.1 mg of Example 1 The mRNA was dissolved in 100mM, pH 4.0 citrate buffer to form solution B; use a microfluidic nanomedicine manufacturing system (NanoAssemblr Ignite from Precision Nanosystems, Canada) to mix solution A: solution B at a volume ratio of 1:3, and use 40 times the volume The varicella-zoster mRNA vaccine composition is obtained after dialysis with PBS buffer solution and sterilization through a 0.22 micron filter membrane.

[0073] For each mRNA vaccine composition prepared in the above embodiment 2, the following measurements were carried out:

[0074] 1. Particle size and polydispersity index

[0075] The particle size and polydispersity index of LNP were measured using Zetasizer...

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Abstract

The invention provides a varicella-herpes zoster mRNA vaccine composition, and a preparation method and application thereof. The vaccine composition comprises a messenger ribonucleic acid (mRNA) sequence for coding varicella-herpes zoster virus glycoprotein E, a derivative sequence of the messenger ribonucleic acid (mRNA) sequence and lipid nanoparticles (LNP), and the messenger ribonucleic acid (mRNA) sequence is prepared into particles with the diameter of 20-400 nanometers through microfluidic equipment. The vaccine composition can specifically enhance humoral immune response and cellular immune response against varicella-herpes zoster glycoprotein E, can be used as a varicella vaccine which does not cause latent infection of vaccine strains, and can also be used as a herpes zoster vaccine with unlimited productivity. All the components in the vaccine composition can be widely obtained, so that the vaccine cost is effectively reduced, and the vaccine yield is increased.

Description

technical field [0001] The invention belongs to the field of vaccines, and in particular relates to a varicella-zoster mRNA vaccine composition and a preparation method and application thereof. Background technique [0002] Almost all children have been infected with varicella-zoster virus (Varicella-Zoster Virus, VZV) before adulthood. The initial infection may produce chickenpox, and the virus will remain latent in the ganglion after the chickenpox self-heals. With the weakening of cellular immune response due to aging or other reasons (such as immunodeficiency virus infection or clinical need for immunosuppressive drugs), the latent virus will be reactivated in the body, leading to the occurrence of herpes zoster. [0003] The Oka strain live attenuated vaccine developed by Japanese Michiaki Takahashi was approved by FDA in 1995 for inoculating children and adults against chickenpox (inoculation volume 1 000-5 000 PFU <plaque forming unit, plaqueforming unit> ), w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/25A61P31/22A61P25/00
CPCA61K39/12A61P31/22A61P25/00A61K2039/53A61K2039/54A61K2039/575A61K2039/57C12N2710/16734
Inventor 刘存宝曹晗王云飞栾宁
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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