CN116808191A - Novel coronavirus vaccine, preparation method and application thereof - Google Patents

Abstract

The present disclosure provides a novel coronavirus vaccine, a preparation method and applications thereof, and relates to the technical field of vaccines. The novel coronavirus vaccine comprises a nucleic acid molecule encoding a novel coronavirus omnix Rong Zi variant ba.5S protein.

Description

Novel coronavirus vaccine, preparation method and application thereof

Cross Reference to Related Applications

The present application claims priority from chinese patent application (application number: 2022112076839, title: novel coronavirus vaccine, and methods for preparing and using the same), of which application date is 2022, 09, 30, and part of the contents are incorporated herein by reference in their entirety.

Technical Field

The disclosure relates to the technical field of vaccines, and in particular relates to a novel coronavirus vaccine, a preparation method and application thereof.

Background

The novel coronavirus is extremely easy to mutate, and the strains such as a novel coronavirus original strain, a novel coronavirus Alpha strain, a novel coronavirus Beta strain, a novel coronavirus Gamma mutant strain, a novel coronavirus Kappa strain, a novel coronavirus Delta strain, a novel coronavirus Omicron strain and the like appear successively from the discovery to date.

The current market and most of the new coronavirus vaccines in clinical trial stage are designed against the antigen of the original strain of the new coronavirus. The omickon (omacron) variant is at least 60 more mutations compared to the original new coronavirus original strain sequence, with more than 35 mutations in the spike protein (S protein) and 15 mutations in the most critical receptor binding domain within the spike protein (S protein), as opposed to only 2 mutations in this region. The omnikom variant can be divided into at least 5 sub variants, ba.1, ba.2.12.1, ba.2, ba.4, ba.5, etc., according to the difference in mutation sites.

It was found that the protective effect of the marketed vaccine against variants was reduced to varying degrees, in particular against the novel coronavirus omnikom strain. Therefore, new crown vaccines with better protection effect against variant strains are urgently needed to be developed.

Disclosure of Invention

According to various embodiments of the present disclosure, a novel coronavirus vaccine is provided, comprising a nucleic acid molecule encoding a novel coronavirus S protein, the amino acid sequence of which is selected from the group consisting of novel coronavirus omucotton ba.5 seed variant, novel coronavirus omucotton ba.2 seed variant, novel coronavirus omucotton ba.3 seed variant S protein or novel coronavirus omucon ba.5 seed variant S protein.

In one or more embodiments, the amino acid sequence of the novel coronavirus omucotton variant ba.1 subvariant S protein is preferably as shown in SEQ id No.6 or comprises an amino acid sequence at least 80% identical to SEQ id No.6, for example, but not limited to, an amino acid sequence comprising at least 80%, 85%, 90%, 95% or 98% identical to SEQ id No. 6.

In one or more embodiments, the amino acid sequence of the novel coronavirus omucotton variant ba.2 subvariant S protein is preferably as shown in SEQ id No.4 or comprises an amino acid sequence at least 80% identical to SEQ id No.4, for example, but not limited to, an amino acid sequence comprising at least 80%, 85%, 90%, 95% or 98% identical to SEQ id No. 4.

In one or more embodiments, the amino acid sequence of the novel coronavirus omucotton variant ba.3 subvariant S protein is preferably as shown in SEQ id No.8 or comprises an amino acid sequence at least 80% identical to SEQ id No.8, for example, but not limited to, an amino acid sequence comprising at least 80%, 85%, 90%, 95% or 98% identical to SEQ id No. 8.

In one or more embodiments, the amino acid sequence of the novel coronavirus omucotton variant ba.5 subvariant S protein is preferably as shown in or comprises an amino acid sequence at least 80% identical to SEQ id No.16, for example, but not limited to, an amino acid sequence comprising at least 80%, 85%, 90%, 95% or 98% identical to SEQ id No. 16.

In one or more embodiments, the amino acid sequence of the novel coronavirus S protein is selected from the S protein of the novel coronavirus Omikovia variant BA.5.

In one or more embodiments, the nucleic acid molecules include DNA molecules and/or RNA molecules.

In one or more embodiments, the DNA molecules include a chain DNA molecule and/or a circular DNA molecule.

In one or more embodiments, the RNA molecule comprises mRNA or circular RNA.

In one or more embodiments, the nucleic acid molecule is mRNA.

In one or more embodiments, the total GC% content of the open reading frame portion of the mRNA is 30-70%, and the GC% content of any one of the open reading frame fragments having a length of 60bp is not less than 40%.

In one or more embodiments, the portion of the open reading frame in the mRNA has a total gc% content of 50% to 60%, more preferably 54% to 60%.

In one or more embodiments, the mRNA further includes one or more of a 5' cap, a 5' utr, a 3' utr, a polyA tail, a start region, a termination region, a signal sequence region, and a linker sequence.

In one or more embodiments, the nucleotide sequence of the mRNA open reading frame is shown as SEQ ID No.17, SEQ ID No. 19-22, SEQ ID No.10, SEQ ID No.11, SEQ ID No.12, SEQ ID No.13, SEQ ID No.14, SEQ ID No.15, SEQ ID No. 18.

In one or more embodiments, the nucleotide sequence of the mRNA open reading frame is shown as SEQ ID.NO. 10-15, SEQ ID.NO. 17-18.

In one or more embodiments, the nucleotide sequence of the mRNA open reading frame is set forth in SEQ id No. 17.

In alternative embodiments, based on the provided RNA sequences, one of ordinary skill in the art will be able to obtain the corresponding DNA sequences (e.g., uracil to thymine). Likewise, based on the DNA sequences provided, one of ordinary skill in the art will obtain the corresponding RNA sequences (e.g., thymine to uracil conversion). In alternative embodiments, based on the provided RNA or DNA sequences, the person of ordinary skill in the art will be able to obtain the corresponding amino acid sequences.

In one or more embodiments, the 5' cap is selected from ARCA, m7G (5 "") ppp (5 "") (2 "" "ome) pG, m7 (3" "" ome) (5 "") ppp (5 "") (2 "" "ome) pG, mCAP, dmCAP, tmCAP, or dmapp.

In one or more embodiments, the poly (A) length is 50 to 200, preferably 80 to 200.

In one or more embodiments, the 5' UTR is preferably 10 to 200 nucleotides, preferably 15 to 100 nucleotides, in length.

In one or more embodiments, the 5' utr nucleotide sequence is set forth in SEQ id No. 1.

In one or more embodiments, the 3' utr sequence is set forth in SEQ id No. 2.

In one or more embodiments, one or more uridine in the mRNA molecule is replaced with a modified nucleoside; preferably, the modified nucleoside replacing uridine is pseudouridine (ψ), N1-methyl-pseudouridine (m 1 ψ) or 5-methyl-uridine (m 5U).

In one or more embodiments, the novel coronavirus vaccine contains nucleic acid lipid nanoparticles composed of the nucleic acid molecules and lipid components.

In one or more embodiments, the lipid component comprises 20 to 50% protonatable cationic lipid, 20 to 50% structural lipid, 5 to 20% helper lipid, and 1 to 5% surfactant, in mole percent, wherein the molar content of protonatable cationic lipid, structural lipid, helper lipid, and surfactant add up to 100%.

In one or more embodiments, the protonatable cationic lipid includes at least one of dlimc 3-DMA, DODMA, C-200 and DlinDMA.

In one or more embodiments, the helper lipid includes at least one of DSPC, DOPE, DOPC, DOPG and DOPS.

In one or more embodiments, the structural lipid includes cholesterol and/or cholesterol derivatives.

In one or more embodiments, the surfactant includes at least one of PEG-DMG, PEG-DSPE, and TPGS.

In one or more embodiments, the lipid component comprises Dlin-MC3-DMA 50%, DOPG 10%, cholesterol 38.5% and PEG-DMG 1.5% in mole percent.

In one or more embodiments, the lipid component comprises Dlin-MC3-DMA 50%, DOPG 20%, cholesterol 29% and PEG-DMG 1% in mole percent.

In one or more embodiments, the nucleic acid lipid nanoparticle is prepared according to the following method:

uniformly mixing an aqueous phase containing nucleic acid molecules and an organic phase containing the lipid component to obtain a mixed solution, and removing the organic phase to ensure that the concentration of the nucleic acid molecules in the system is 1-100 mug/ml to obtain the nucleic acid lipid nanoparticle;

the aqueous phase is an aqueous phase buffer solution containing 0.08-1.2 mg/L nucleic acid molecules, and the aqueous phase buffer solution is a citrate buffer solution or a sodium acetate buffer solution;

the organic phase is anhydrous C1-C4 low-carbon alcohol containing 5-7 mg/L lipid component; the volume ratio of the aqueous phase to the organic phase is 1:2-4.

In one or more embodiments, the novel coronavirus vaccine of any preceding claim is configured for use in a product for preventing or treating a novel coronavirus-induced disease.

In one or more embodiments, a product configured to prevent or treat a novel coronavirus-induced disease, comprising a novel coronavirus vaccine of any one of the preceding claims.

The details of one or more embodiments of the disclosure are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the disclosure will be apparent from the description and drawings, and from the claims.

Drawings

For a clearer description of embodiments of the disclosure or of the prior art, reference may be made to one or more of the accompanying drawings. The appended claims or examples used to describe the drawings should not be construed to limit the scope of the disclosed application, the presently described embodiments and or examples, and any of the best modes of carrying out the application presently understood. The drawings that are needed in the detailed description and prior art description are briefly presented below, and it will be apparent to those of ordinary skill in the art that other drawings may be derived from these drawings without undue burden as well as from some embodiments of the present disclosure.

FIG. 1 shows the results of test for pseudovirus neutralization activity of novel coronaviruses by serum produced after immunization of C57 mice with each vaccine formulation of example 4.

FIG. 2 shows the results of test of pseudovirus neutralization activity of novel coronaviruses by serum produced after immunization of C57 mice with each vaccine formulation of example 6.

Detailed Description

The technical solutions of the present disclosure will be clearly and completely described below in connection with embodiments, and it is apparent that the described embodiments are some embodiments of the present disclosure, but not all embodiments. Based on the embodiments in this disclosure, all other embodiments that a person of ordinary skill in the art would obtain without making any inventive effort are within the scope of protection of this disclosure.

It should be noted that: in the present disclosure, all embodiments and preferred methods of implementation mentioned herein may be combined with each other to form new solutions, if not specifically stated; all technical features and preferred features mentioned herein may be combined with each other to form new solutions; the components involved or their preferred components can be combined with one another to form new technical solutions.

The "range" disclosed in the present disclosure may be in the form of a lower limit and an upper limit, respectively, of one or more lower limits, and of one or more upper limits; unless otherwise indicated, the steps may or may not be performed in sequential order.

Unless otherwise defined, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any method or material similar or equivalent to those described may also be used in the present disclosure.

According to various embodiments of the present disclosure, the present disclosure provides a novel coronavirus vaccine comprising a nucleic acid molecule encoding a novel coronavirus ommicon (omicon) child variant ba.5S protein, predominantly with the nucleic acid molecule as an immunogenic substance. After the novel coronavirus vaccine is used for immunizing an organism, the novel coronavirus vaccine can express the variable strain BA.5S protein of the Omica Rong Zi in the organism.

"nucleic acid molecule" as used in this disclosure refers to a polymeric form of nucleotides of any length, including ribonucleotides and/or deoxyribonucleotides. Examples of nucleic acids include, but are not limited to, single-stranded, double-stranded or multi-stranded DNA or RNA, genomic DNA, cDNA; integrating vector DNA from an exogenous gene, such as an expression cassette or plasmid; DNA-RNA hybrids or polymers comprising purine and pyrimidine bases or other natural, chemically or biochemically modified, non-natural or derivatized nucleotide bases.

In the novel coronavirus vaccine provided by the disclosure, the S protein of the novel coronavirus encoded by the open reading frame is optionally the S protein obtained by mutating an omnikow variant under natural conditions; alternatively, the mutant and modified S protein can be an amino acid sequence of S protein which is obtained by wild type mutation and modification and accords with the Omikou variant; alternatively, the amino acid sequence of the S protein may be obtained by further mutating and modifying the amino acid sequence of the S protein of the Omikovia mutant strain.

In one or more embodiments, the amino acid sequence of the open reading frame encoded novel coronavirus S protein is selected from the group consisting of the amino acid sequence of the novel coronavirus omucon variant S protein derived from the ba.5 subvariant, the ba.2 subvariant, the ba.3 subvariant S protein, and the ba.5 subvariant S protein; or, the amino acid sequence of the S protein of the omnikom variant is optionally derived from the amino acid sequence obtained by mutating a wild-type S protein.

"sequence identity" between two nucleotide sequences indicates the percentage of identical nucleotides between the sequences. "sequence identity" between two amino acid sequences indicates the percentage of identical amino acids between the sequences.

The term "% identity" or similar terms refer to the percentage of identical nucleotides or amino acids between sequences to be compared under optimal alignment. The percentages are purely statistical and the differences between the two sequences may (but need not) be randomly distributed over the length of the sequences to be compared. Comparison of two sequences is typically performed by comparing the sequences with respect to a fragment or "comparison window" after optimal alignment to identify local regions of the corresponding sequences.

The S protein of the omacron ba.5 strain, SEQ id No.16, has a higher expression level by SN mutation (two proline residues substituted at positions 981 and 982 of the amino acid sequence of the full-length S protein).

The amino acid sequence positions are positioned by the full-length amino acid sequence of the wild-type S protein of the novel coronavirus original strain.

The novel coronavirus vaccine provided by the disclosure takes nucleic acid molecules as main efficacy components, and the novel coronavirus vaccine expresses S protein for producing virus Omikovia strains in vivo after being applied to an organism. The present disclosure also optimizes nucleic acid molecules and vaccine formulations in order to further enhance the immune effect of the vaccine.

The open reading frame (open reading frame, ORF) described in this disclosure is the normal nucleotide sequence of a structural gene, and the reading frame from the start codon to the stop codon can encode a complete polypeptide chain, with no stop codon present that interrupts translation.

In one or more embodiments, the nucleic acid molecule can optimize mRNA sequence by means of sequence optimization, improving characteristics related to expression efficacy after in vivo administration: for example, mRNA with improved properties can be obtained by increasing mRNA stability, increasing translational efficiency in the target tissue, reducing the number of truncated proteins expressed, improving folding or preventing misfolding of the expressed protein, reducing toxicity of the expressed product, reducing cell death caused by the expressed product, increasing and/or reducing protein aggregation. The sequence optimization purposes also include: optimizing the characteristics of formulation and delivery of nucleic acid-based therapeutics while maintaining structural and functional integrity; overcoming the threshold of expression; the expression rate is improved; half-life and/or protein concentration; optimizing protein localization; and avoiding adverse biological responses such as immune responses and/or degradation pathways. The sequence optimizing means includes: (1) Codon optimization to ensure proper folding and proper expression is performed according to codon frequency in a particular organ and/or host organism; (2) Regulating the G/C content to increase mRNA stability or decrease secondary structure; (3) Minimizing tandem repeat codons or base strings (base run) that may impair gene construction or expression; (4) custom transcription and translation control regions; (5) Reducing or eliminating problematic secondary structures within the polynucleotide.

The "original strain" identified on the graphs of FIGS. 1 and 2 is the EC50 value of the pseudovirus strain of the original strain of the new coronavirus; "Alpha" is the EC50 value of the pseudovirus strain of the novel coronavirus Alpha strain; "Beta" is the EC50 value of the pseudovirus strain of the new coronavirus Beta strain; "Gamma" is the EC50 value of the pseudovirus strain of the novel coronavirus Gamma strain; "Delta" is the EC50 value of the pseudovirus strain of the novel coronavirus Delta strain; "BA.1" is the EC50 value of the pseudovirus strain of the novel coronavirus Omikovia BA.1 subvariant strain; "BA.2" is the EC50 value of the pseudovirus strain of the novel coronavirus Omikovia BA.2 subvariant strain; "BA.4& BA.5" is the EC50 value of the pseudovirus strain of the novel coronavirus Omikovine BA.4& BA.5 subvariant (the S protein of the novel coronavirus Omikovine BA.4 subvariant is the same as that of the novel coronavirus Omikovine BA.5 subvariant).

The technical solution and advantageous effects of the present disclosure are further described below in conjunction with the preferred embodiments.

Example 1

The present example provides a method for preparing lipid nanoparticles comprising RNA, wherein the lipid nanoparticles comprise in mole percent: dlin-MC3-DMA 50%, DOPG 20%, cholesterol 29% and PEG-DMG 1%, and the preparation method is as follows:

(a) RNA was dissolved in citrate buffer at pH4 to adjust the concentration to 0.1. 0.1 mg/ml, to give an aqueous phase. (b) Dlin-MC3-DMA, DOPG, cholesterol and PEG-DMG were dissolved in absolute ethanol in the amounts of the formulation, and the concentration of the lipid component in the organic phase was adjusted to 6 mg/mL to give an organic phase.

(c) Mixing the aqueous phase of step (a) and the organic phase of step (b) at a volume ratio of 1:3 using a microfluidic device at a flow rate of 12 mL/min, immediately diluting the mixture 100-fold with PBS solution at pH7.4, removing ethanol component from the solution using Tangential Flow Filtration (TFF), and concentrating to a mRNA concentration of 55 μg/ml in the system to obtain lipid nanoparticles comprising RNA encoding SARS-CoV-2 virus antigen.

Example 2

The formulation of different vaccine vectors was studied by in vivo fluorescence imaging techniques using luciferase as a reporter gene (as shown in the following table, "MC3" means Dlin-MC3-DMA, "+" is the efficiency of delivering mRNA encoding luciferase gene in mice by in vivo fluorescence imaging system of small animals after administration) and the physicochemical index of different complex formulations (preparation method see example 1) was examined, and the results are shown in the table.

TABLE 1

。

It is found that increasing the mass ratio of lipid to mRNA is beneficial to increasing the encapsulation rate of mRNA in lipid nanoparticles, so that the lipid nanoparticles have higher stability, and moreover, moderately increasing the content of polyethylene glycol (PEG) in the formula is beneficial to increasing the expression efficiency of mRNA in vivo. Therefore, the factors such as the mRNA encapsulation efficiency, the in-vivo delivery efficiency of mRNA and the like are comprehensively considered, and formulas 3 and 4 are selected for the subsequent research of mRNA vaccines.

Example 3

The capacity of the cationic lipid nanoparticles of different formulations to encapsulate the mRNA encoding the full length of the S protein and the particle size data of the formed nanoparticles are shown in Table 2, and several formulations can compress the S protein mRNA into nanoparticles with a particle size below 100nm and a net neutral surface potential, and can encapsulate at least 50% of the mRNA, so that the cationic lipid nanoparticles have a certain in vivo delivery effect. "MC3" refers to Dlin-MC3-DMA.

TABLE 2

。

Example 4

Screening of Omicron subtype strain S protein:

this example designed a series of mRNA sequence information in which the sequence of the open reading frames is shown in table 3; in addition to the open reading frame sequence, the series of mRNA sequence features also include a 5 'cap (m 7G (5') (2 '-OMeA) pG), a 5' UTR (as shown in SEQ ID. NO. 1), a 3'UTR (as shown in SEQ ID. NO. 2), and the 3' tail of 100 polyAs (as shown in SEQ ID. NO. 3).

TABLE 3 mRNA sequence design scheme

。

Preparing four different antigens in samples 1-4 into vaccines according to the method of the embodiment 1, respectively preparing different mRNA mixtures into LNP preparations, wherein the LNP preparations have the detection encapsulation rate of more than 90% and the particle size of about 70 nm; sample 5 is shown as a blank (placebo).

The prepared mRNA LNP formulation was used in C57 mice immunization experiments, each mouse being injected 5 micrograms (in mRNA) via hind limb lateral thigh muscle, and two immunizations were performed after 7 days apart, 3 mice per group. Mice were serum withdrawn 14 days after the first immunization and sent to third party laboratories for the neutralization activity test of SARS-CoV-2 pseudovirus. Diluting mouse serum according to different proportions (initial dilution multiple is 30) in a 96-well plate, adding SARS-CoV-2 pseudovirus which can be infected, setting cell contrast and virus contrast, incubating for 1 hour, adding cells prepared in advance, culturing for 20-28 hours in a cell incubator, absorbing a part of supernatant, adding a luciferase detection reagent, repeatedly blowing and sucking cells in the well after light-shielding reaction at room temperature, and placing the cells in a chemiluminescent detector for reading a luminous value after the cells are fully lysed. And on the premise of ensuring that the virus control and the cell control are established, calculating an EC50 value by adopting a Reed-Muench method. EC50 values for pseudovirus strains of different antigens prepared from samples 1-4 are shown in figure 1.

As can be seen from the results of fig. 1: in the vaccine preparation prepared by the mRNA vaccine open reading frame encoding different S proteins (the amino acid sequences are shown as SEQ ID.NO.4, SEQ ID.NO.6, SEQ ID.NO.8 and SEQ ID.NO. 16), the mRNA vaccine encoding the S protein with the amino acid sequence shown as SEQ ID.NO.16 (the encoding amino acid is the S protein of the novel coronavirus Omicron BA.5 strain) stimulates mice to generate less neutralizing antibody activity reduction on pseudoviruses of various child variants of the novel coronavirus Omicron.

Example 5

Optimization of mRNA sequence encoding ba.5 child variant strain S protein:

this example shows a series of mRNA sequences designed based on the novel coronavirus Omacron strain S protein (amino acid sequence shown in SEQ ID. NO. 16) and mRNA sequence optimization principle, wherein the open reading frame information is shown in Table 4 below.

In addition to the open reading frames of Table 4, the series of mRNA sequence features included a 5 'cap (m 7G (5') (2 '-OMeA) pG), a 5' UTR (as shown in SEQ ID. NO. 1), a 3'UTR (as shown in SEQ ID. NO. 2) and the 3' tail of 100 polyAs (as shown in SEQ ID. NO. 3).

TABLE 4 mRNA sequence design and relative expression levels

。

"local GC% content" in Table 4: GC% content in the partial sequence was 60bp as window size from the 3 'end to the 5' end of the ORF.

The mRNA shown in Table 4 was transfected into cells, and the expression of the S full-length protein in the cells was examined, and the results are shown in Table 4. The detailed method is as follows: HEK293 cells transfected with each mRNA for 24 hours were lysed, and the target protein was specifically detected by SDS-PAGE immunoblotting at a loading of 10. Mu.g of total protein, and in this example, incubation was performed using an anti-SARS-S1 protein antibody as primary antibody and a goat anti-mouse-HRP antibody as secondary antibody, respectively, followed by color development. When the protein expression amount detection result is analyzed, the internal reference beta-actin is used for standardization and quantification, meanwhile, cells which are not transfected with mRNA are set as negative controls, and the difference of the expressed protein amount of transfected cells of different mRNA is compared. The detection results show that the expression of the whole S protein and the S1 subunit can be detected. The expression levels of the respective sequences were measured in terms of relative OD values as shown in Table 4; the method for calculating the relative OD value comprises the following steps: sample OD/OD of sample 1.

As can be seen from table 4: when the GC% content of the total mRNA open reading frame sequence is 54-60%, the relative expression amount of S protein is higher when the local GC% content is not lower than 40% (see SEQ ID. NO.17 and 19); in addition, the expression level of the S protein with the mRNA open reading frame sequence of SEQ ID No.17 is higher than that of the mRNA open reading frame sequence of SEQ ID No. 19.

Two different antigens of samples 1 and 2 in example 5 are respectively prepared into vaccines according to the method in example 1, different mRNA mixtures are respectively prepared into LNP preparations, the LNP preparations have encapsulation rates of more than 90% after detection, and the particle size is about 70nm. The two groups of vaccine formulations obtained were used in C57 mouse immunization experiments, the experimental procedure being shown in example 4. The serum corresponds to the group number and the results are shown in figure 2.

As can be seen from fig. 2: since the expression level of the mRNA sequence SEQ ID.NO.17 is high, the neutralizing capacity of the antibody produced by the mRNA vaccine preparation prepared by the mRNA sequence SEQ ID.NO.17 is optimal.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present disclosure, and not for limiting the same; although the present disclosure has been described in detail with reference to the foregoing embodiments, it should be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present disclosure.

Claims (10)

1. A novel coronavirus vaccine comprising a nucleic acid molecule encoding a novel coronavirus S protein selected from the group consisting of an S protein of a novel coronavirus ompick ba.1 variant shown in SEQ id No.6, an S protein of a novel coronavirus ompick ba.2 variant shown in SEQ id No.4, an S protein of a novel coronavirus ompick ba.3 variant shown in SEQ id No.8, and an S protein of a novel coronavirus ompick ba.5 variant shown in SEQ id No. 16.

2. The novel coronavirus vaccine of claim 1, wherein the amino acid sequence of the novel coronavirus S protein is selected from the group consisting of the S protein of the novel coronavirus omucon ba.5 variant.

3. The novel coronavirus vaccine of claim 1, wherein the nucleic acid molecules comprise DNA molecules and/or RNA molecules; the DNA molecules include a chain DNA molecule and/or a circular DNA molecule; the RNA molecule comprises mRNA or circular RNA.

4. The novel coronavirus vaccine of claim 3, wherein the nucleic acid molecule is mRNA; the total GC% content of the open reading frame part in the mRNA is 30-70%, and the GC% content in any fragment with the length of 60bp in the fragment of the open reading frame is not less than 40%.

5. The novel coronavirus vaccine of claim 4, wherein the nucleotide sequence of the mRNA open reading frame is shown in SEQ id.no. 10-15, SEQ id.no. 17-18.

6. The novel coronavirus vaccine of claim 3, wherein the mRNA further comprises one or more of a 5' cap, a 5' utr, a 3' utr, a polyA tail, a initiation region, a termination region, a signal sequence region, and a linker sequence;

the 5' cap of the mRNA molecule is selected from ARCA, m7G (5 "") ppp (5 "") (2 "" "ome) pG, m7 (3" "(5" ") ppp (5" ") (2" "" ome) pG, mCAP, dmCAP, tmCAP, m7 (3 "" (5 "") ppp (5 "") (2 "" "ome) pG or dmapp; the length of the poly (A) is 50-200; the nucleotide sequence of the 5' UTR is shown as SEQ ID.NO. 1; the 3' UTR sequence is shown as SEQ ID.NO. 2;

replacing one or more uridine in said mRNA molecule with a modified nucleoside; the modified nucleoside is pseudouridine (ψ), N1-methyl-pseudouridine (m 1 ψ) or 5-methyl-uridine (m 5U).

7. The novel coronavirus vaccine of claim 1, wherein the novel coronavirus vaccine comprises nucleic acid lipid nanoparticles comprised of the nucleic acid molecules and lipid components;

the lipid component comprises 20-50% of protonatable cationic lipid, 20-50% of structural lipid, 5-20% of auxiliary lipid and 1-5% of surfactant in terms of mole percent, wherein the mole content of the protonatable cationic lipid, the structural lipid, the auxiliary lipid and the surfactant is 100% in total;

the protonatable cationic lipid comprises at least one of dlimc 3-DMA, DODMA, C12-200 and DlinDMA;

the helper lipid comprises at least one of DSPC, DOPE, DOPC, DOPG and DOPS;

the structural lipid comprises cholesterol and/or cholesterol derivatives;

the surfactant includes at least one of PEG-DMG, PEG-DSPE and TPGS.

8. The novel coronavirus vaccine of claim 7, wherein the nucleic acid lipid nanoparticle is prepared according to the following method:

uniformly mixing an aqueous phase containing nucleic acid molecules and an organic phase containing the lipid component to obtain a mixed solution, and removing the organic phase to ensure that the concentration of the nucleic acid molecules in the system is 1-100 mug/ml to obtain the nucleic acid lipid nanoparticle;

the aqueous phase is an aqueous phase buffer solution containing 0.08-1.2 mg/L nucleic acid molecules, and the aqueous phase buffer solution is a citrate buffer solution or a sodium acetate buffer solution;

the organic phase is anhydrous C1-C4 low-carbon alcohol containing 5-7 mg/L lipid component; the volume ratio of the aqueous phase to the organic phase is 1:2-4.

9. Use of the novel coronavirus vaccine of any one of claims 1-8 in the preparation of a product configured to prevent or treat a novel coronavirus-induced disease.

10. A product configured for preventing or treating a novel coronavirus-induced disease, comprising the novel coronavirus vaccine of any one of claims 1-8.