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Vaccines and compositions based on S antigen protein of SARS-CoV-2

A sars-cov-2, protein technology, applied in the direction of DNA / RNA vaccination, vaccines, virus antigen components, etc.

Active Publication Date: 2021-03-12
GUANGZHOU ARGORNA BIOPHARMACEUTICALS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no specific drug for the treatment of novel coronavirus pneumonia, and an effective vaccine is urgently needed

Method used

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  • Vaccines and compositions based on S antigen protein of SARS-CoV-2
  • Vaccines and compositions based on S antigen protein of SARS-CoV-2
  • Vaccines and compositions based on S antigen protein of SARS-CoV-2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1. Optimization of mRNA sequence

[0068] Sequence optimization of the coding sequence of SARS-CoV-2S protein (SEQ ID NO.1) was carried out through DNAWorks (v3.2.4) online software, including: adjusting codon bias for expression in the human body, Use the frequency to adjust, increase the GC content of the sequence, etc. The optimized nucleotide sequence is shown as SEQ ID NO.2. This nucleotide sequence makes the mRNA structure more stable after transcription, and the translation efficiency of the target protein in mammals and humans is higher.

Embodiment 2

[0069] Example 2. Expression of the SARS-CoV-2S trimeric protein in a pre-fusion stable form

[0070] The S protein consists of an extracellular domain (ECD), a transmembrane domain (TM) and a C-terminal cytoplasmic domain (CP). Among them, the extracellular domain can be further divided into signal peptide region (SP), N-terminal region (NTD), receptor binding domain (RBD), an internal membrane fusion peptide (FP) and two 7-peptide repeats (HR ), belonging to the first class of viral membrane fusion proteins. Schematic diagram of the domain structure of the S protein figure 1 As shown, the pre-fusion structure is the functional conformation of S protein, and a large number of sensitive neutralizing epitopes that only exist in the pre-fusion form are masked after fusion. Expression of a prefusion-stabilized form of the SARS-CoV-2S trimeric protein is key to the development of a safe and effective SARS-CoV-2 vaccine.

[0071] In this example, the following strategy was used ...

Embodiment 3

[0081] Embodiment 3.mRNA preparation

[0082] The mRNA encoding the prefusion-stabilized form of the SARS-CoV-2 viral S protein was prepared by in vitro transcription. The nucleic acid sequence of the coding region of the S protein is obtained by the whole gene synthesis method, and the nucleic acid sequence of the coding region is connected with a plasmid vector by a conventional molecular cloning method to obtain a DNA plasmid template for in vitro transcription.

[0083] Table 1. Transcription start sites

[0084]

[0085] The elements contained in the plasmid vector include a T7 promoter sequence (TAATACGACTCACTATA), and the starting nucleic acid sequence can be any of AGG / GGG / CGG, as shown in Table 1; 5'-UTR sequence (SEQ ID NO.41), 3'-UTR sequence (SEQID NO.43), 120nt 3'-terminal polyadenylic acid (polyA) sequence, including Xbal restriction site (TCTAGA) after the 3'-terminal polyadenylic acid sequence.

[0086] The mRNA sequence obtained by transcribing is shown i...

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Abstract

The present disclosure provides vaccines and compositions based on S antigen proteins of SARS-CoV-2, and specifically relates to recombinant SARS-CoV-2 spike proteins (S proteins) or antigenic fragments thereof, and mRNAs encoding the same. The present disclosure also relates to mRNA comprising a sequence selected from any one of SEQ ID NO. 44 to 47. The present disclosure further relates to mRNA-vector particles, such as lipid nanoparticles (LNPs), comprising the aforementioned mRNAs and compositions, such as vaccine compositions.

Description

technical field [0001] The disclosure belongs to the technical field of biomedicine and vaccines, and in particular relates to vaccines and compositions against novel coronavirus SARS-CoV-2. Background technique [0002] Coronaviridae virus is an enveloped single-stranded positive-sense RNA virus with the largest genome among RNA viruses, up to 27-32kb. Coronaviridae is divided into 4 genera α, β, γ and δ, among which α and β genera are susceptible to mammals, while γ and δ genera mainly infect poultry. So far, seven human coronaviruses have been discovered, such as the novel coronavirus (Severe acute respiratory syndrome coronavirus 2, SARS-CoV-2). SARS-CoV-2 is highly pathogenic and poses a serious threat to public health. Since the beginning of 2020, the novel coronavirus pneumonia epidemic caused by SARS-CoV-2 has swept the world. As of September 2020, the cumulative number of confirmed patients worldwide has exceeded 30 million, posing a huge threat to global public h...

Claims

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Application Information

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IPC IPC(8): C07K14/165C07K19/00C12N15/50C12N15/62A61K39/215A61P31/14A61K9/51A61K47/10A61K47/28A61K47/24
CPCC07K14/005A61K39/12A61P31/14A61K9/5146A61K9/5123C07K2319/02C07K2319/00C12N2770/20022C12N2770/20034A61K2039/53
Inventor 张必良李曼马林温剑张虹钟惠玲
Owner GUANGZHOU ARGORNA BIOPHARMACEUTICALS CO LTD
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