mRNA vaccine for preventing varicella zoster, preparation method therefor, and use thereof

Abstract

Disclosed are an mRNA vaccine encoding a varicella zoster virus (VZV) protein, a preparation method therefor, and use thereof. The mRNA vaccine is characterized by encoding at least the varicella zoster virus (VZV) gE protein, gH protein, and gL protein.

Description

A varicella zoster vaccine, and a preparation method and application thereof TECHNICAL FIELD The present application relates to the technical field of biopharmaceuticals, and more particularly to a varicella zoster mRNA vaccine, and a preparation method and application thereof. BACKGROUND Varicella zoster virus (VZV) belongs to the alpha herpesvirus subfamily of the herpesviridae family, and only one serotype has been found so far. The primary infection caused by VZV manifests as chickenpox, and after self-healing, the virus can be latent in host sensory neurons. When the immune system is damaged or the immunity is reduced, VZV can be reactivated and cause herpes zoster (HZ). Up to 30% of patients with herpes zoster will develop post-herpetic neuralgia (PHN), and the pain rating can reach more than 7, which is severe pain and seriously affects the quality of life of patients. Vaccination is the most effective means of preventing and controlling varicella and herpes zoster and their complications. Currently, there are two types of varicella-zoster vaccines approved for marketing worldwide: attenuated live vaccines and recombinant subunit protein vaccines. Among them, the attenuated live vaccine Zostavax of Merck is mainly based on the Oka strain, which has limited protective efficacy and a low protection period. The effective rate of this vaccine for PHN in adults over 60 years old is only 39%. The recombinant subunit protein vaccine Shingrix of GSK is composed of VZV gE glycoprotein and AS01B adjuvant. Although Shingrix is significantly better than the VZV attenuated live vaccine Zostavax, the vaccine formulation is complex, the antigen needs to be freeze-dried and separated from the adjuvant, and the two are mixed when used, which is not convenient to operate. Moreover, AS01B is composed of monophosphoryl lipid A (MPL) and QS-21, and QS-21 cannot be chemically synthesized, which is relatively scarce in raw materials, has a relatively high production cost, is expensive, and has a serious shortage of domestic supply. The biggest disadvantage of this adjuvant is that the degree and incidence of side reactions are high. In addition, the glycoprotein E is produced by CHO cell lines, which has the disadvantages of high cost, complex fermentation and purification process, unstable glycosylation products, and great difficulty in quality control. Therefore, it is necessary to further explore and develop a VZV vaccine with small side effects, high effectiveness, and a simple formulation and low cost, which has far-reaching significance. In recent years, the key technology of mRNA vaccines has made a breakthrough, and has become a research hotspot due to its many advantages, such as no risk of genome integration, direct and rapid translation into protein, high safety, natural metabolic products, and small toxic and side effects. At the same time, the production of mRNA is convenient and fast, and the production cost is relatively low. At present, most of the reported VZV mRNA vaccines express VZV gE protein, such as CN117731769A, CN114081943A, CN114272369A, CN117247954A, WO2023062556A1, WO2024120490A1, WO2023214082A1, WO2024102813A1 and the like, and only a few are related to multiple proteins, such as WO2018170270A1, WO2023048760A1, WO2024156053A1, CN118340875A, WO2023056912A1 and the like. Although there are many reported VZV mRNA vaccines, most of them are still in the development stage at present, and the effects are different. The mRNA vaccine with more immune effect advantage is developed, and its application prospect is broader. SUMMARY In view of the above problems, the purpose of the present application is to provide an mRNA vaccine for preventing varicella zoster virus (VZV). The vaccine takes VZV envelope glycoprotein gE (including full-length gE, truncated secreted gE, truncated secreted gE-Fc), gH (including full-length gH, truncated secreted gH) and / or gL as a target. Another purpose of the present application is to provide a vaccine composition containing mRNA encoding VZV envelope glycoprotein gE (including full-length gE, truncated secreted gE, truncated secreted gE-Fc), gH (including full-length gH, truncated secreted gH) and / or gL protein. Still another purpose of the present application is to provide the application of the mRNA vaccine containing mRNA encoding VZV envelope glycoprotein gE (including full-length gE, truncated secreted gE, truncated secreted gE-Fc), gH (including full-length gH, truncated secreted gH) and / or gL protein or the vaccine composition thereof. Still another purpose of the present application is to provide a method for preventing varicella, zoster and postherpetic neuralgia (PHN), that is, administering an effective amount of mRNA vaccine encoding VZV envelope glycoprotein gE (including full-length gE, truncated secreted gE, truncated secreted gE-Fc), gH (including full-length gH, truncated secreted gH) and / or gL protein or the vaccine composition thereof to a subject in need thereof. The above purposes of the present application are achieved by the following scheme: An mRNA vaccine encoding at least varicella zoster virus (VZV) gE protein, gH protein and gL protein; The amino acid sequence of the gE protein is any one of SEQ ID NO: 5001-5003 or at least 95% similar to any one of SEQ ID NO: 5001-5003. The amino acid sequence of the gH protein is SEQ ID NO: 5004-5005 or any one of the sequences having at least 95% similarity to one of SEQ ID NO: 5004-5005. The amino acid sequence of the gL protein is SEQ ID NO: 5006 or a sequence having at least 95% similarity to SEQ ID NO: 5006. The gE protein has been proved to be an effective target for preventing herpes zoster. To further improve the immune effect of the vaccine, the influence of the protein variant or fusion protein on the immune effect is usually studied. The present application finds that the immunogenicity of the gE protein can be effectively improved by constructing a fusion protein of the extracellular part of the gE protein and Fc (Fc fragment of antibody IgG). In addition, gH and gL are also very important surface glycoproteins of VZV. The gH protein is fixed on the capsid membrane of the virion through the transmembrane region, and the gL protein is secreted to form a heterodimer (gH / gL) with the extracellular part of the gH protein. The present application finds that, compared with the mRNA vaccine group inoculated with gH / gL protein or gE protein alone, the mRNA vaccine group inoculated with gH / gL and gE protein has better specific cellular immune response and humoral immune response. It is also unexpectedly found that gH / gL can significantly improve the specific cellular immune response of gE protein. In one embodiment, the amino acid sequence of the gE protein is any one of SEQ ID NO: 5001-5003 or a sequence having at least 98% or 99% similarity to one of SEQ ID NO: 5001-5003. In one embodiment, the amino acid sequence of the gH protein is any one of SEQ ID NO: 5004-5005 or a sequence having at least 98% or 99% similarity to one of SEQ ID NO: 5004-5005. In one embodiment, the amino acid sequence of the gL protein is SEQ ID NO: 5006 or a sequence having at least 98% or 99% similarity to SEQ ID NO: 5006. In one embodiment, the amino acid sequence of the gE protein encoded by the mRNA vaccine is SEQ ID NO: 5001. In one embodiment, the amino acid sequence of the gE protein encoded by the mRNA vaccine is SEQ ID NO: 5002. In one embodiment, the amino acid sequence of the gE protein encoded by the mRNA vaccine is SEQ ID NO: 5003. In one embodiment, the amino acid sequence of the gH protein encoded by the mRNA vaccine is SEQ ID NO: 5004. In an embodiment, the amino acid sequence of the gH protein encoded by the mRNA vaccine is SEQ ID NO: 5005. In an embodiment, the amino acid sequence of the gL protein encoded by the mRNA vaccine is SEQ ID NO: 5006. In an embodiment, the mRNA comprises a coding region; the coding region is selected from the group consisting of SEQ ID NO: 57-68 or SEQ ID NO: 103. In an embodiment, the ORF is selected from the group consisting of SEQ ID NO: 57-68. Preferably, the coding region of the mRNA is SEQ ID NO: 60. Preferably, the coding region of the mRNA is SEQ ID NO: 63. Preferably, the coding region of the mRNA is SEQ ID NO: 103. In an embodiment, the mRNA further comprises a 5’UTR, the sequence of which is selected from any one of SEQ ID NO: 1, 3, 10, 12, 197, 201. In an embodiment, the mRNA further comprises a 5’UTR, the sequence of which is SEQ ID NO: 3 or SEQ ID NO: 197. In an embodiment, the mRNA further comprises a 5’UTR, the sequence of which is SEQ ID NO: 197. In an embodiment, the mRNA further comprises a 3’UTR, the sequence of which is selected from any one of SEQ ID NO: 16, 18, 205, 207, 209. In an embodiment, the mRNA further comprises a 3’UTR, the sequence of which is SEQ ID NO: 16 or SEQ ID NO: 205. In an embodiment, the mRNA further comprises a 3’UTR, the sequence of which is SEQ ID NO: 205. In an embodiment, the mRNA further comprises a poly A, which comprises at least 70 adenosines. In an embodiment, the poly A comprises 70-140 adenosines, or 70-120 adenosines, or 80-120 adenosines, or 90-120 adenosines, or 90-110 adenosines, or 90-100 adenosines, or 100-140 adenosines, or 100-120 adenosines, or 100-110 adenosines. In an embodiment, the poly A comprises 90-110 adenosines. In an embodiment, the mRNA comprises: (ii) 5'UTR; the 5'UTR is selected from any one of SEQ ID NO: 1, 3, 10, 12, 197, 201; (iii) coding region; the coding region is selected from SEQ ID NO: 60, SEQ ID NO: 63 or SEQ ID NO: 103; (iv) 3'UTR; the 3'UTR is selected from any one of SEQ ID NO: 16, 18, 205, 207, 209; (v) poly A; the poly A comprises at least 70 adenosines. In one embodiment, the coding region of the mRNA is SEQ ID NO: 60-62. In one embodiment, the coding region of the mRNA is SEQ ID NO: 60. In one embodiment, the coding region of the mRNA is SEQ ID NO: 63. In one embodiment, the coding region of the mRNA is SEQ ID NO: 103. In one embodiment, the mRNA further comprises a 5'UTR, and the sequence of the 5'UTR is SEQ ID NO: 3 or SEQ ID NO: 197. In one embodiment, the mRNA further comprises a 3'UTR, and the sequence of the 3'UTR is SEQ ID NO: 16 or SEQ ID NO: 205. In one embodiment, the sequence of the mRNA is SEQ ID NO: 9961, SEQ ID NO: 9974 or SEQ ID NO: 9975. In one embodiment, the sequence of the mRNA is SEQ ID NO: 9961, wherein the poly A comprises 90-110 adenosines. In one embodiment, the sequence of the mRNA is SEQ ID NO: 9974, wherein the poly A comprises 90-110 adenosines. In one embodiment, the sequence of the mRNA is SEQ ID NO: 9975, wherein the poly A comprises 90-110 adenosines. In one embodiment, the mRNA further comprises a 5' cap structure. In one embodiment, the 5' cap structure is selected from Cap0, Cap1, Cap2. In some embodiments, the 5' cap structure can be, but is not limited to, the following compounds: In one of the embodiments, the mRNA comprises at least one chemically modified nucleoside. In one of the embodiments, the chemically modified nucleoside is selected from pseudouracil, N1-methyl-pseudouracil, 1-ethylpseudouracil, 2-thiouracil, 4'-thiouracil, 5-methyluracil, or 5-methoxyuracil, or any combination thereof. In one of the embodiments, the chemically modified nucleoside is selected from pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from N1-methyl-pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from pseudouracil and / or N1-methyl-pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 1-ethylpseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 1-ethylpseudouracil and / or N1-methyl-pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 1-ethylpseudouracil and / or pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 4'-thiouracil. In one of the embodiments, the chemically modified nucleoside is selected from 4'-thiouracil and / or N1-methyl-pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 4'-thiouracil and / or pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 5-methyluracil. In one of the embodiments, the chemically modified nucleoside is selected from 5-methyluracil and / or N1-methyl-pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 5-methyluracil and / or pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 5-methoxyuracil. In one of the embodiments, the chemically modified nucleoside is selected from 5-methoxyuracil and / or N1-methyl-pseudouracil. In one of the embodiments, the chemically modified nucleoside is selected from 5-methoxyuracil and / or pseudouracil. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 10-100%, or 20-100%, or 30-100%, or 40-100%, or 50-100%, or 60-100%, or 70-100%, or 80-100%, or 90-100%, or 50-98%, or 50-95%, or 60-95%, or 70-95%, or 80-95%, or 90-95%, or 70-90%, or 80-90%, or 100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 10-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 20-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 30-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 40-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 50-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 60-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 70-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 80-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 90-100%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 50-98%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 50-95%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 60-95%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 70-95%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 80-95%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 90-95%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 70-90%. In one embodiment, the percentage of chemically modified nucleosides in the mRNA is 80-90%. In one embodiment, the ratio of the chemically modified nucleosides in the mRNA is 100%. The present application also protects a vaccine composition comprising the aforementioned mRNA vaccine and the delivery agent. In one embodiment, the delivery agent comprises a lipid nanoparticle; the lipid nanoparticle comprises: a cationic lipid, a phospholipid, a cholesterol, and a PEG lipid. In one embodiment, the molar ratio of the cationic lipid, the phospholipid, the cholesterol, and the PEG lipid in the lipid nanoparticle is 30-60:0-30:18.5-48.5:0-10. In one embodiment, the molar ratio of the cationic lipid, the phospholipid, the cholesterol, and the PEG lipid in the lipid nanoparticle is 35-55:0-25:20-45:0.5-8. In one embodiment, the molar ratio of the cationic lipid, the phospholipid, the cholesterol, and the PEG lipid in the lipid nanoparticle is 40-52:5-20:25-40:0.5-5. In one embodiment, the molar ratio of the cationic lipid, the phospholipid, the cholesterol, and the PEG lipid in the lipid nanoparticle is 45-52:8-15:30-40:1-5. In one embodiment, the molar ratio of the cationic lipid, the phospholipid, the cholesterol, and the PEG lipid in the lipid nanoparticle is 50:10:38.5:1.5. In one embodiment, the molar ratio of the cationic lipid, the phospholipid, the cholesterol, and the PEG lipid in the lipid nanoparticle is 48.5:11.1:38.9:1.5. In one embodiment, the molar ratio of the cationic lipid, the phospholipid, the cholesterol, and the PEG lipid in the lipid nanoparticle is 40:8.0:50.5:1.5. In one embodiment, the cationic lipid is one or more combinations of SM102, MC3, DOTAP, ALC-0315, H1, H2, H3, H4, H5, H6, H7, TM6. The structure is as follows: In one embodiment, the cationic lipid is shown in formula I: In one embodiment, the cationic lipid is shown in formula I: T1, L1 are each independently C 1-6 alkyl or absent; T2, L2are each independently -O-C(O)-, -C(O)-O-, -O-C(O)-O-, -NH-C(O)-, -C(O)-NH-, -NH-C(O)-O-, or -O-C(O)-NH-; T3, T4, L3, L4are each independently H, C 4-20 straight-chain or branched alkyl, C 4-20 straight-chain or branched alkenyl, or C 4-20 straight-chain or branched alkynyl; wherein T3and T4are not simultaneously H; L3and L4are not simultaneously H; X1, X2are each independently -NH-, C 1-4 alkyl, -O-, -C(O)-, -O-C(O)-, -C(O)-O-, -O-C(O)-O-, -NH-C(O)-, -C(O)-NH-, -NH-C(O)-O-, -O-C(O)-NH-, or nothing; X3is -NR2R3or wherein R2, R3are each independently C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 hydroxyalkyl, C 4-7 cycloalkyl, -C 1-4 alkyl-N C 1-4 alkyl(C 1-4 alkyl), -C 1-4 alkyl-C 4-7 cycloalkyl, -C 4-7 heterocycloalkyl, or -C 1-4 alkyl-C 4-7 heterocycloalkyl; Z1, Z2are each independently C or N; R4is H, C 1-4 alkyl, C 1-4 cycloalkyl, or C 1-4 hydroxyalkyl; n1, n2are each independently an integer from 1 to 3. In one embodiment, X0is H or F. In one embodiment, X0is H. In one embodiment, T1, L1are each independently ethyl, propyl, isopropyl, butyl, pentyl, hexyl, or nothing. In one embodiment, T1, L1are each independently ethyl, propyl, or nothing. In one embodiment, T1, L1are each independently ethyl or nothing. In one embodiment, T1, L1are each independently ethyl or propyl. In one embodiment, T2 and L2 are each independently -OC(O)-, -C(O)-O-, -OC(O)-O-, -NH-C(O)-, or -C(O)-NH-. In one embodiment, T2 and L2 are each independently -OC(O)-, -C(O)-O-, -NH-C(O)-, or -C(O)-NH-. In one implementation, T2 and L2 are each independently -OC(O)- or -NH-C(O)-. In one implementation, T2 and L2 are each independently -OC(O)- or -C(O)-O-. In one implementation, T3, T4, L3, and L4 are each independently H and C. 4-9 Alkyl, C 14-20 Alkyl, C 9-20 alkenyl or C 9-20 Alkyne group. In one embodiment, the structure of formula (I) Independently In one embodiment, the structure of formula (I) Independently In one embodiment, the structure of formula (I) Independently In one implementation, X1 and X2 are each independently C 1-4 alkyl. In one implementation, R2 and R3 are each independently C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Hydroxyalkyl, C 4-7 cycloalkyl, or -C 1-4 Alkyl-C 4-7 Heterocyclic alkyl groups. In one implementation, R2 and R3 are each independently C 1-4 alkyl. In one embodiment, the cationic lipid is selected from one of the following structures: In one embodiment, the phospholipid is DSPC or DOPE; the PEG lipid has a molecular weight of 1000 Da to 20 kDa PEG, PEG-modified compounds or mixtures thereof. In one embodiment, the mRNA is encapsulated in a lipid nanoparticle in the vaccine composition. In one embodiment, the vaccine composition further comprises a pharmaceutically acceptable excipient. The present application also protects the use of the aforementioned mRNA encoding VZV proteins or the aforementioned vaccine composition in the preparation of a vaccine for preventing varicella, herpes zoster and post-herpetic neuralgia (PHN). The present application also protects a method for preventing varicella, herpes zoster and post-herpetic neuralgia (PHN), administering an effective amount of the aforementioned mRNA encoding VZV proteins or the aforementioned vaccine composition to a subject in need thereof. Compared with the prior art, the present application has the following beneficial effects: The mRNA vaccine encoding gE, gH and / or gL proteins has better specific cellular immune and humoral immune responses, which is not only significantly superior to the marketed vaccine Shingrix, but also significantly superior to the mRNA vaccine encoding gH / gL proteins or gE proteins alone; and it is unexpectedly found that gH / gL proteins can significantly improve the specific cellular immune response of gE proteins. BRIEF DESCRIPTION OF DRAWINGS Figure 1 shows the number of gE and / or gH / gL specific IFN-γ+ cells in the spleen lymphocytes of each immunized group. Figure 2 shows the gE and / or gH / gL specific IgG titers in the serum of mice in each immunized group. Figure 3 shows the neutralizing antibody titers in the serum of mice in each immunized group. Figure 4 shows the number of gE and / or gH / gL specific IFN-γ+ cells in the spleen lymphocytes of each immunized group in Example 4. Figure 5 shows the gE and / or gH / gL specific IgG titers in the serum of mice in each immunized group in Example 4. Figure 6 shows the neutralizing antibody titers in the serum of mice in each immunized group in Example 4. Figure 7 shows the number of gE and / or gH / gL specific IFN-γ+ cells in the spleen lymphocytes of each immunized group in Example 5. Figure 8 shows the gE and / or gH / gL specific IgG titers in the serum of mice in each immunized group in Example 5. Figure 9 shows the neutralizing antibody titers in the serum of mice in each immunized group in Example 5. DETAILED DESCRIPTION TERMS As used herein and unless otherwise stated, the term "alkyl" refers to a saturated carbon alkyl group consisting only of carbon and hydrogen atoms.

[0089] A straight-chain or branched hydrocarbon chain group. In one embodiment, the alkyl group has, for example, 1 to 24 carbon atoms (C,-C, alkyl), 4 to 20 carbon atoms (C,-C, alkyl), 6 to 16 carbon atoms (C,C, alkyl), 6 to 9 carbon atoms (C,-C, alkyl), 1 to 15 carbon atoms (C,-C, alkyl), 1 to 12 carbon atoms (C,-C, alkyl), 1 to 8 carbon atoms (C,-C, alkyl), or 1 to 6 carbon atoms (C,-C, alkyl) and is linked to the remainder of the molecule by a single bond. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, etc. Unless otherwise stated, the alkyl group is optionally substituted. The term "lipid nanoparticle" or "LNP" refers to a particle having at least one nanometer (nm) scale size (e.g., 1 to 1000 nm) containing one or more types of lipid molecules. The LNPs described herein may further contain at least one non-lipid payload molecule (e.g., one or more nucleic acid molecules). In some embodiments, the LNP comprises a non-lipid payload molecule partially or completely encapsulated within a lipid shell. Specifically, in some embodiments, the payload is a negatively charged molecule (e.g., mRNA encoding a therapeutic protein), and the lipid component of the LNP comprises at least one cationic lipid. Without being bound by theory, it is anticipated that cationic lipids can interact with the negatively charged payload molecule and promote payload incorporation and / or encapsulation into the LNP during LNF formation. Other lipids that can form a portion of the LNPs described herein include, but are not limited to, neutral and charged lipids, such as steroids, polymer-bound lipids, and various zwitterionic lipids. The term "cationic lipid" refers to a lipid that carries a positive charge at any pH or hydrogen ion activity in its environment, or is capable of carrying a positive charge in response to the pH or hydrogen ion activity of its environment (e.g., the environment in which it is intended to be used). Therefore, the term "cationic" encompasses both "permanent cation" and "cationizable." In some embodiments, the positive charge in the cationic lipid is caused by the presence of a quaternary nitrogen atom. In some embodiments, the cationic lipid comprises a zwitterionic lipid that carries a positive charge in its intended environment (e.g., at physiological pH). As used herein and unless otherwise stated, the term "pharmaceutically acceptable salt" includes both acid addition salts and base addition salts. As used herein and unless otherwise indicated, the term "pharmaceutically acceptable carrier, diluent or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetening, diluting, preservative, dye / colorant, flavor-enhancing, surfactant, wetting, dispersing, suspending, stabilizing, isotonic, solvent, or emulsor agent approved by a regulatory agency of the Federal or a state government of the United States of America or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in mammals, such as, for example, humans and domestic animals. The term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts optionally selected, e.g., an mRNA molecule as provided herein. In certain embodiments, the mRNA is a monocistronic mRNA comprising only one ORF. In certain embodiments, the monocistronic mRNA encodes a polypeptide or protein comprising at least one epitope of a selected antigen, e.g., a pathogenic antigen or a tumor-associated antigen. In other embodiments, the mRNA is a multicistronic mRNA comprising two or more ORFs. In certain embodiments, the multicistronic mRNA encodes two or more polypeptides or proteins that can be the same or different from each other. In certain embodiments, each polypeptide or protein encoded by the multicistronic mRNA comprises at least one epitope of a selected antigen. In certain embodiments, different polypeptides or proteins encoded by the multicistronic mRNA each comprise at least one epitope of a different antigen. In any of the embodiments described herein, the at least one epitope can be at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 epitopes of an antigen. The term "administer" or "administration" refers to the act of injecting or otherwise physically delivering a substance (e.g., a lipid nanoparticle composition described herein) that is present outside of the body into the body of a patient, such as transmucosally, intradermally, intravenously, intramuscularly, and / or any other physical delivery method described herein or known in the art. When treating a disease, disorder, illness, or symptom thereof, administration of the substance is typically performed after the onset of the disease, disorder, illness, or symptom thereof. When preventing a disease, disorder, illness, or symptom thereof, administration of the substance is typically performed before the onset of the disease, disorder, illness, or symptom thereof. An "effective amount" is generally an amount sufficient to reduce the severity and / or frequency of symptoms; eliminate symptoms and / or their underlying cause; prevent an occurrence of symptoms and / or their underlying cause; and / or improve or remediate damage caused by or associated with a disease, disorder, or illness, including, for example, damage caused by or associated with infection and neoplastic formation. In some embodiments, an effective amount is a therapeutically effective amount or a prophylactically effective amount. As used herein, the term "therapeutically effective amount" refers to the amount of an agent (e.g., a lipid nanoparticle composition as described herein) sufficient to reduce and / or ameliorate the severity and / or duration of a given disease, disorder, or condition (e.g., an infectious disease, such as an infectious disease caused by a viral infection, or a neoplastic disease, such as a cancer). A "therapeutically effective amount" of a substance / molecule / agent (e.g., a lipid nanoparticle composition as described herein) of the present disclosure can vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance / molecule / agent to elicit a desired response in the individual. A therapeutically effective amount encompasses an amount of the substance / molecule / agent in which any toxic or detrimental effects of the substance / molecule / agent are outweighed by the therapeutically beneficial effects. In certain embodiments, the term "therapeutically effective amount" refers to the amount of a lipid nanoparticle composition as described herein, or a therapeutic or prophylactic agent (e.g., a therapeutic mRNA) contained therein, effective to treat a disease, disorder, or condition in a subject or mammal. The terms "treat," "treating," and "treatment" refer to all of the following: alleviating, in whole or in part, a disorder, disease, or condition, or one or more symptoms associated with the disorder, disease, or condition, or slowing or stopping further progression or worsening of those symptoms, or reducing or eliminating the cause of the disorder, disease, or condition itself. The terms "prevent," "preventing," and "prevention" refer to reducing the likelihood of onset (or recurrence) of a disease, disorder, condition, or associated symptoms. The term "side effects" encompasses unwanted and / or adverse effects of a therapy (e.g., a prophylactic or therapeutic agent). Unwanted effects are not necessarily adverse effects. Adverse effects of a therapy (e.g., a prophylactic or therapeutic agent) can be harmful, unpleasant, or risky. Examples of side effects include diarrhea, cough, gastroenteritis, wheezing, nausea, vomiting, anorexia, abdominal cramping, fever, pain, weight loss, dehydration, hair loss, dyspnea, insomnia, dizziness, mucositis, neurological and muscular effects, fatigue, mouth sores, loss of appetite, skin rash or swelling at the site of administration, flu-like symptoms such as fever, chills, and fatigue, digestive issues, and allergic reactions. Other unwanted effects experienced by patients are numerous and known in the art. The application will be further described in greater detail by way of specific Examples which are meant to explain the application but not to limit its scope. The following examples use the test methods described below unless otherwise specified. The materials and reagents used in the examples are commercially available unless otherwise specified. The application will be further described in conjunction with specific examples, which are used to explain the application and are not intended to limit the scope of the application. The test methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used are commercially available unless otherwise specified. Table 1 is part of the sequences involved in the application; Table 2 is the amino acid sequence; and Table 3 is part of the mRNA sequence. Table 1 Sequence The application also relates to SEQ ID NO: 21, 24, 27, 30, 33, 36, 39, 42, 45, 61, 74, 80, 95, 101, 122, 167, 201, 207, 209, and the specific sequences are shown in the computer readable form (CRF) sequence listing. Table 2 Amino acid sequence Part of the mRNA sequence formed by different ORFs combined with different 5'UTR sequences and 3'UTR sequences is shown in Table 3. When the ORF is any one of SEQ ID NO: 57, 58, 60, 61, 63, 64 or 66, the mRNA sequence that can be prepared is SEQ ID NO: 6001-6600. The ORF can also be replaced by any one of SEQ ID NO: 101-196 to prepare the mRNA sequence. The mRNA sequence prepared can be SEQ ID NO: 9961, wherein the ORF is SEQ ID NO: 60, the 5' UTR is SEQ ID NO: 197, the 3' UTR is SEQ ID NO: 205, and the poly A contains 70-140 adenosines. Any one of SEQ ID NO: 197 to 204 can be further substituted for SEQ ID NO: 197; any one of SEQ ID NO: 206 to 212 can be further substituted for SEQ ID NO: 205. The ORF in the sequence of SEQ ID NO: 6001 to 6600 or SEQ ID NO: 9961 can be further substituted with any one of 101 to 196. When there are two or more ORFs, each ORF is linked by a linker sequence, and the linker sequence is a P2A or an IRES sequence. The mRNA sequence prepared can be SEQ ID NO: 6721 to 6735, wherein the ORF is SEQ ID NO: 20, and the ORF is SEQ ID NO: 51. Any one of SEQ ID NO: 21 to 29 can be further substituted for SEQ ID NO: 20; or any one of SEQ ID NO: 52 to 56 or SEQ ID NO: 89 to 94 can be further substituted for SEQ ID NO: 51; or any one of SEQ ID NO: 197 to 204 can be further substituted for the 5' UTR; or any one of SEQ ID NO: 205 to 212 can be further substituted for the 3' UTR. Any one of SEQ ID NO: 39 to 50, SEQ ID NO: 71 to 88, or SEQ ID NO: 95 to 100 can be further substituted for SEQ ID NO: 20; or any one of SEQ ID NO: 36 to 38 can be further substituted for SEQ ID NO: 51. The mRNA sequence prepared can be SEQ ID NO: 8881 to 8895, wherein the ORF is SEQ ID NO: 20, the ORF is SEQ ID NO: 30, and the ORF is SEQ ID NO: 36. Any one of SEQ ID NO: 21 to 29 can be further substituted for SEQ ID NO: 20; or any one of SEQ ID NO: 31 to 35 can be further substituted for SEQ ID NO: 30; or any one of SEQ ID NO: 197 to 204 can be further substituted for the 5' UTR; or any one of SEQ ID NO: 205 to 212 can be further substituted for the 3' UTR. Table 3 Partial mRNA sequences Target point selection The following examples prepare corresponding mRNA vaccine molecules with envelope glycoprotein gE (including full-length gE, truncated secreted gE, truncated secreted gE-Fc), gH (including full-length gH, truncated secreted gH) and / or gL of VZV as the target point. Example 1 sequence screening 1.1 Screening of UTR sequences The selected 5' UTR sequences are SEQ ID NO: 1, 3, 10, 12, 197, 201; the selected 3' UTR sequences are SEQ ID NO: 16, 18, 205, 207, 209; one of the sequences of SEQ ID NO: 60, 61, 63 is an ORF, which is combined with different 5' UTR sequences and different 3' UTR sequences to obtain multiple groups of sequences. Taking SEQ ID NO: 60 as an example, SEQ ID NO: 6181, 6183, 6190, 6192, 6211, 6213, 6220, 6222, 9961, 9963, 9965 in Table 3 can be obtained. The sequences SEQ ID NO: 6181, 6183, 6190, 6192, 6211, 6213, 6220, 9961 are selected for testing, wherein the ORF is sequence SEQ ID NO: 60, and the 5' UTR sequence and the 3' UTR sequence are different. The effects of different 5' UTR sequences and different 3' UTR sequences are tested. The DNA sequences corresponding to the mRNA sequences of SEQ ID NO: 6181, 6183, 6190, 6192, 6211, 6213, 6220, 9961 are synthesized by GenScript, and the specific process is as follows: (1) Synthesize the DNA sequence corresponding to the mRNA; (2) Construct the DNA sequence on the plasmid vector by homologous recombination; wherein the number of adenosine in poly A is 100±10. The plasmid vector containing the target gene is amplified and purified by E. coli, and is further linearized. After linearization of the plasmid, the capped mRNA is prepared by "one-pot method", and the specific steps can be referred to the patent 202210973168.5. Among them, the cap structure is any one of compounds 1-54 listed in the application. Specifically, the cap structure used in the following example mRNA is compound 19. 1.2 mRNA expression in cells Transfection and expression: 293T cells at 0.6×10 6The cells were cultured at a density of 1 x 105 / mL in 6-well cell culture plates at 37°C in a 5% CO2 environment. The next day, the mRNA was transfected into the cells, and the mass / volume ratio of mRNA to transfection reagent jetMESSENGER was 1:2, as follows: 200 μL jetMESSENGER Buffer, 4 μg mRNA was added and mixed, 8 μL transfection reagent was added and mixed, and the mixture was allowed to stand at room temperature for 15 min, and then it was added to the cells, which were cultured at 37°C in a 5% CO2 environment. The medium was changed after 6 h of transfection, and the cells were collected after 24 h of transfection. Flow cytometry analysis of the positive cell rate and fluorescence value of cells expressing gE protein: the cells were collected, centrifuged at 400 g for 5 min at room temperature to remove the supernatant, 100 uL of PBS was added for washing, and the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant; the cells were resuspended with 100 uL of Cyto-Fast TM Fix / Perm solution, and the cells were incubated at room temperature for 20 min; 100 uL of 1X CytoFast TM Perm Wash Solution, and the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant; 200 uL of 1X CytoFast TM Perm Wash Solution, and the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant; the cells were resuspended with 1X CytoFast TM Perm Wash Solution, and the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant, 100 uL of 1X CytoFast TM Perm Wash Solution, and the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant; the cells were resuspended with 1X CytoFast TM Perm Wash Solution, and the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant; the cells were resuspended with 1X CytoFast TM Perm Wash Solution (diluted to 1X with deionized water), and the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant; the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant, 200 uL of PBS was added, the cells were resuspended, and the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant; the cells were centrifuged at 400 g for 5 min at room temperature to remove the supernatant, 100 uL of PBS was added to resuspend the cells; and the cells were detected on the machine: the cells were vortexed before being detected on the machine. The positive efficiency (the proportion of cells expressing gE protein) and fluorescence value were detected by mRNA transfection of cells to screen UTR sequences and ORF sequences. The detection results show that when the ORF sequence is SEQ ID NO. 60, the number of positive cells expressing the target protein of the mRNA composed of different 5'UTR sequences and different 3'UTR sequences is all more than 90%, and there is only a certain difference in the expression amount; it is shown that the 5'UTR sequence and the 3'UTR sequence of the application can be used for preparing the VZV mRNA vaccine. From the expression results, it can be known that when the 5'UTR sequence is SEQ ID NO. 3 and the 3'UTR sequence is SEQ ID NO. 16, and the 5'UTR sequence is SEQ ID NO. 197 and the 3'UTR sequence is SEQ ID NO. 205, the mRNA has the best expression, and therefore in the subsequent research, the 5'UTR sequence of SEQ ID NO. 3 or SEQ ID NO. 197 and the 3'UTR sequence of SEQ ID NO. 16 or SEQ ID NO. 205 are used to prepare the corresponding mRNA. 1.3 Screening of ORF sequence The mRNA is prepared, transfected and expressed according to 1.2. The sequence of the mRNA is the sequence listed in Table 3 or the sequence in Table 3, and the sequence obtained by replacing part of the ORF sequence. For example, the ORF sequence is SEQ ID NO: 60, 61, 63; the 5'UTR sequence is SEQ ID NO. 3, and the 3'UTR sequence is SEQ ID NO. 16, that is, the mRNA sequence is SEQ ID NO: 6183, 6243, 6363; or the 5'UTR sequence is SEQ ID NO. 197, the 3'UTR sequence is SEQ ID NO. 205, and the ORF sequence is SEQ ID NO: 60, that is, the mRNA sequence is SEQ ID NO: 9961 (wherein the poly A is 100±10 adenosines), the cell positive rate is all more than 95%, and the average fluorescence value is all more than 3.45E+05; it is shown that the prepared mRNA can well express the target protein in the cell, and the expression amount is high. Example 2 Encapsulation of mRNA The main components of the LNP include: cationic lipids, cholesterol, DSPC and DMG-PEG2000, and the molar ratio is 48.5%, 38.9%, 11.1% and 1.5% respectively. Among them, the cationic lipids are selected from one or more combinations of SM102, MC3, DOTAP, ALC-0315, TM6, H1-H41. 2.1.1 Lipid-ethanol solution: the required cationic lipids, cholesterol, DSPC and DMG-PEG2000 are dissolved in anhydrous ethanol according to the molar ratio, and a lipid-ethanol solution with a total lipid concentration of 8mM is prepared for standby. 2.1.2 mRNA-acetate buffer solution: mRNA stock solution was diluted to an appropriate concentration with acetate buffer solution (200 mM, pH 5.0) for use. 2.1.3 Encapsulation: mRNA-buffer solution was mixed with lipid-ethanol solution at a nitrogen to phosphorus ratio of 4.8 or other, with a flow rate ratio of 3:1 (buffer phase / ethanol phase), and a total flow rate of 3.6 mL / min, using a microfluidic instrument (syringe pump: SPM, Deka Industry; mixing chip: LNP-B0, FluidicLab). The mixed sample was dialyzed (25 or 30 kDa MwCO) overnight at 4°C against 34 volumes of Tris salt buffer solution (20.5 mM, pH 7.5, containing 8.95% sucrose) to obtain encapsulated mRNA LNPs, denoted as mRNA-LNP. The mass ratio of LNP to mRNA was about 5:1, 10:1, 15:1, and 20:1. To ensure accurate measurement of the amount of each component, the amount of each component was scaled up and prepared in proportion. The cationic lipid used in the following examples was H41, and the encapsulation was performed at a nitrogen to phosphorus ratio of 4.8. Example 3 Immunization of mRNA-LNP Referring to the methods in Example 1 and Example 2, test sample mRNA-LNP with different ORF sequences was prepared. 3.1 Immunization and sample collection mRNA-LNP immunization of mice (6-8 weeks): two intramuscular immunizations (day 0 and day 14), and the mice were sacrificed on day 42 to isolate splenic lymphocytes for cell immune response detection, and to collect whole blood to isolate serum for antigen-specific IgG detection by ELISA and evaluation of serum neutralizing antibody levels by in vitro VZV neutralization experiment. A negative control group and a positive control group were also set up. The negative control group was injected with Tris salt buffer solution (50 μL, 20.5 mM, pH 7.5, containing 8.95% sucrose); the positive control group was injected with Shingrix (50 μL, 5 μg per mouse); and the test group had four groups, denoted as Sample 1, Sample 2, Sample 3, and Sample 4. In Sample 1, the mRNA sequence was SEQ ID NO: 9969 (50 μL, 5 μg per mouse); In Sample 2, the mRNA sequences were SEQ ID NO: 9969 and SEQ ID NO: 9972, which were mixed in equal amounts after encapsulation (total volume 50 μL, 5 μg per mouse for each mRNA); The mRNA sequences in Sample 3 were SEQ ID NO: 9970 and SEQ ID NO: 9972, and the same amount of each was administered after encapsulation (total volume 50 μL, 5 μg of each mRNA per mouse); The mRNA sequence in Sample 4 was SEQ ID NO: 9961 (50 μL, 10 μg per mouse). The encapsulation of the mRNA was performed according to Example 2. 3.2 Isolation of mouse spleen lymphocytes After blood sampling on day 42, the mice were sacrificed by cervical dislocation and immersed in 75% ethanol for 1-5 min; the mice were dissected in a clean bench, and the spleens were removed with aseptic operation; the spleens were ground in a 35 mm culture dish containing 5 mL lymphocyte separation medium (at room temperature); the spleen cell suspension was transferred to a 15 mL centrifuge tube, and 1000 uL of RPMI 1640 medium was added (slowly added along the wall); Note: (1) After grinding one spleen, transfer it to the centrifuge tube in time to prevent the evaporation of the lymphocyte separation medium; (2) After grinding all the tissues or batch centrifugation of the tissues, add the medium liquid seal uniformly to prevent the medium and the lymphocyte separation medium from penetrating each other, affecting the separation effect; centrifuge at room temperature for 30 min at 800 g with a slow lifting speed (if there are ten gears and the tenth gear is the highest gear, the lifting speed should be adjusted to the second or third gear); aspirate the lymphocyte layer, and add 10 mL of RPMI 1640 medium and invert to wash. Centrifuge at room temperature for 10 min at 250 g to collect the cells; pour the supernatant, resuspend with 1 mL of RPMI 1640 medium containing 10% serum and count. 3.3 Detection of gE and / or gH / gL specific IFN-γ + Cells The detection kit was purchased from Mabtech (item number: FSP-41B-2), and the operation was carried out according to the instructions. The detection results are shown in Figure 1. Compared with the positive vaccine Shingrix, after two immunizations, on day 42, the gE specific IFN-γ + cells of Sample 1, Sample 2 and Sample 4 were significantly higher than those of the positive control group; the gH / gL specific IFN-γ + cells of Sample 2 and Sample 4 were significantly higher than those of the positive control group; the gE / gH / gL specific IFN-γ + cells of Sample 2 and Sample 4 were significantly higher than those of the positive control group. From the above results, it can be seen that the mRNA-LNP of Sample 2 and Sample 4 groups can obviously induce strong gE-specific cellular immune response and certain gH / gL-specific cellular immune response. 3.4 ELISA detection of gE, gH / gL-specific IgG titers in mouse serum Take out the overnight coated ELISA plate, remove the liquid in the plate, add 250-300 μL of 1 × PBST to wash the ELISA plate, a total of 3 times, each time shaking for 1 min, and the last time trying to remove the remaining liquid in the plate; add 100 μL of 5% BSA to each well, seal the plate with a sealing film, and incubate in a 37°C constant temperature incubator for 2 h; after incubation, remove the liquid in the plate, wash 3 times with 1 × PBST, and the last time try to remove the remaining liquid in the plate. According to the sample plate arrangement, add 100 μL of each serum dilution sample (30-fold starting, gradient dilution with 3 as the coefficient) to each well, set 3 replicate wells for each dilution serum, and incubate at 37°C for 1-1.5 h; remove the liquid in the plate, wash 5 times with 1 × PBST, and the last time try to remove the remaining liquid in the plate; add 100 μL of secondary antibody to each well, and incubate at 37°C for 30-50 min; remove the liquid in the plate, wash 5 times with 1 × PBST, and the last time try to remove the remaining liquid in the plate; prepare the color developing solution, mix the two bottles of liquid in the ELISA Color solution kit at a ratio of 1:1, i.e. 10 mL of H2O2 Chromogenic solution + 10 mL of TMB Chromogenic solution, and mix well; add 100 μL of color developing solution to each well, and incubate at 37°C in the dark for 10-15 min; after incubation, add 50 μL of ELISA Stopping Solution to each well, and mix well by tapping the plate. Read the values on the microplate reader at OD 450 nm-630 nm, and the detection should be performed immediately after adding the Stopping Solution, and the detection should be completed within 15 min at the latest. The detection results are shown in FIG. 2, and the gE-specific IgG titers in the serum of Sample 1, Sample 2, Sample 3, and Sample 4 are comparable to those of the positive control group. The serum of Sample 2, Sample 3, and Sample 4 can detect a higher content of gH / gL-specific IgG. 3.5 In vitro neutralization experiment to detect the serum anti-VZV ability Cell preparation: Adjust the density of MRC-5 cells, inoculate into 96-well plates, and place in a cell incubator (37°C, 5% CO2) overnight to ensure that the cell confluence is about 90% the next day; serum inactivation: inactivate in water bath at 56°C for 30 minutes; gradient dilution: dilute the serum by 30 times, dilute by 3 times by gradient, a total of 8 dilution degrees, 3 replicate wells; virus dilution: according to the PFU value of the virus, input the appropriate virus amount; neutralization reaction: add the diluted virus to the sample well and the virus control well, respectively, and sequentially dilute by 2 times by gradient, a total of 3 dilution degrees, 37°C, 5% CO2 incubator for about 1 hour; virus adsorption: add the virus and serum neutralization product and positive and back-dropping well virus to the pre-prepared cells, 50 μL per well, 2 replicate wells, 37°C, 5% CO2 incubator for about 2 hours, then change the liquid, add 100 μL of medium per well, and continue to culture for about 48 h; plate measurement: discard the supernatant, fix the cells, add fluorescently labeled detection antibodies, and use CTL instrument to read the plate. The results are shown in Figure 3. For the serum collected at day 42, higher levels of neutralizing antibodies were detected for Sample 2, Sample 3, and Sample 4 groups compared to the positive vaccine Shingrix, with Sample 3 and Sample 4 being comparable, and the neutralizing antibody titers being more than 10 times higher than the positive control group. Example 4 Immunization of mRNA-LNP Referring to the methods in Example 1 and Example 2, test sample mRNA-LNPs with different ORF sequences were prepared. The experimental scheme is the same as in Example 3, except that the test groups are different, and there are 9 test groups, denoted as Sample 1, Sample 2, Sample 3, Sample 5, Sample 6, Sample 7, Sample 8, Sample 9, and Sample 10. The mRNA in Sample 1, Sample 2, and Sample 3 is the same as in Example 3. The mRNA sequence in Sample 5 is SEQ ID NO: 9970 (50 μL, 5 μg per mouse). The mRNA sequence in Sample 6 is SEQ ID NO: 9973 (50 μL, 5 μg per mouse). The mRNA sequences in Sample 7 are SEQ ID NO: 9969 and SEQ ID NO: 9971, which are mixed in equal amounts after encapsulation (total volume 50 μL, 5 μg of each mRNA per mouse). The mRNA sequences in Sample 8 were SEQ ID NO:9970 and SEQ ID NO:9971, and the two were mixed in equal amounts after encapsulation (total volume 50 μL, 5 μg of each mRNA per mouse) for administration. The mRNA sequences in Sample 9 were SEQ ID NO:9973 and SEQ ID NO:9971, and the two were mixed in equal amounts after encapsulation (total volume 50 μL, 5 μg of each mRNA per mouse) for administration. The mRNA sequences in Sample 10 were SEQ ID NO:9973 and SEQ ID NO:9972, and the two were mixed in equal amounts after encapsulation (total volume 50 μL, 5 μg of each mRNA per mouse) for administration. The encapsulation of the mRNA was performed according to Embodiment 2. Detection of IFN-γ specific to gE and / or gH / gL + Cells The kit was purchased from Mabtech (item number: FSP-41B-2) and the operation was performed according to the instructions. The detection results are shown in FIG. 4. After two immunizations, at day 42, the gE-specific IFN-γ + The number of cells was significantly higher than that of the positive control group, and the number of gE-specific cells was the highest in Sample 2, followed by Sample 1; and the gH / gL-specific IFN-γ + cells were detected in the remaining groups, and the number of gH / gL-specific cells was the highest in Sample 9, followed by Sample 8. From the above results, it can be seen that the mRNA-LNP of Sample 1, Sample 5 and Sample 6 can only encode gE protein, and thus can induce stronger gE-specific cellular immunity, but cannot induce gH / gL-specific IFN-γ + cells. The mRNA-LNP of Sample 2, Sample 3, Sample 7, Sample 8, Sample 9 and Sample 10 can encode gE protein and gH / gL protein, and thus can induce both gE-specific cellular immune response and gH / gL-specific cellular immune response; among them, the gE-specific cellular immune response induced by Sample 2 was the strongest, followed by Sample 7; and the gH / gL-specific cellular immune response induced by Sample 9 was the strongest, followed by Sample 8. ELISA detection of gE, gH / gL specific IgG titers in mouse sera The detection method is the same as Example 3. The detection results are shown in Figure 5. The gE specific IgG titers in the sera of Sample 1, Sample 2, Sample 7 are comparable to the positive control group, and the rest are slightly lower. The sera of Sample 2, 7-10 groups can detect higher levels of gH / gL specific IgG, among which Sample 8 and Sample 9 groups are higher. Detection of serum anti-VZV ability by in vitro neutralization experiment The detection method is the same as Example 3. The detection results are shown in Figure 6. For the sera collected on day 42, compared with the positive vaccine Shingrix, higher neutralizing antibody titers were detected in the remaining groups except Sample 5 and Sample 6, among which the neutralizing antibody titers in Sample 8 and Sample 9 groups were significantly higher than those in other groups, followed by Sample 2 and Sample 10 groups. Example 5 Immunization of mRNA-LNP Referring to the methods in Example 1 and Example 2, test sample mRNA-LNP with different ORF sequences were prepared. The experimental scheme is the same as Example 3, except that the test groups are different. There are 6 groups in this test, which are Sample 2, Sample 4, Sample 8, Sample 9, Sample 11 and Sample 12. Among them, the mRNA of Sample 2 and Sample 4 groups is the same as Example 3; the mRNA of Sample 8 and Sample 9 groups is the same as Example 4; The mRNA sequence in Sample 11 is SEQ ID NO: 9974 (50 μL, 10 μg per mouse); The mRNA sequence in Sample 12 is SEQ ID NO: 9975 (50 μL, 10 μg per mouse). gE specific IFN-γ + Cell and gH / gL specific IFN-γ +The detection method of the cells is the same as that in Example 3, and the detection results are shown in Figure 7. It can be known from Figure 7 that Sample 2, Sample 4, Sample 8, Sample 9, Sample 11 and Sample 12 can simultaneously induce gE-specific cellular immune response and gH / gL-specific cellular immune response; Sample 4 induces the gE-specific cellular immune response most intensively, followed by Sample 2; Sample 9 induces the gH / gL-specific cellular immune response most intensively, followed by Sample 8. The detection method of the gE and gH / gL-specific IgG titers in the mouse serum is the same as that in Example 3, and the detection results are shown in Figure 8. It can be known from Figure 8 that the gE-specific IgG titers in the serum of the six test groups are comparable to those of the positive control group, and Sample 4 has a slight advantage; the gH / gL-specific IgG titers in the serum of the six test groups are comparable, and are significantly better than those of the positive control group. The detection method of the serum anti-VZV ability in the in vitro neutralization experiment is the same as that in Example 3, and the detection results are shown in Figure 9. It can be known from Figure 9 that the neutralizing antibody titers in the serum of the six test groups are significantly better than those of the positive control group, and the neutralizing antibody titer in Sample 12 is the highest, followed by Sample 11, Sample 4 and Sample 9. Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application and not to limit the protection scope of the present application. For those skilled in the art, based on the above description and ideas, other different forms of changes or modifications can also be made, which do not need to be or cannot be exhaustively listed here. Any modification, equivalent replacement and improvement made within the spirit and principle of the present application shall be included in the protection scope of the claims of the present application.

Claims

1. An mRNA vaccine, characterized in that, at least encodes a varicella zoster virus (VZV) gE protein, a gH protein, and a gL protein; wherein the amino acid sequence of the gE protein is any one of SEQ ID NOs: 5001-5003 or a sequence having at least 95% similarity to any one of SEQ ID NOs: 5001-5003; the amino acid sequence of the gH protein is any one of SEQ ID NOs: 5004-5005 or a sequence having at least 95% similarity to any one of SEQ ID NOs: 5004-5005; the amino acid sequence of the gL protein is SEQ ID NO: 5006 or a sequence having at least 95% similarity to SEQ ID NO: 5006.

2. The mRNA vaccine of claim 1, wherein, the amino acid sequence of the gE protein is any one of SEQ ID NOs: 5001-5003 or a sequence having at least 98% or 99% similarity to any one of SEQ ID NOs: 5001-5003; the amino acid sequence of the gH protein is any one of SEQ ID NOs: 5004-5005 or a sequence having at least 98% or 99% similarity to any one of SEQ ID NOs: 5004-5005; the amino acid sequence of the gL protein is SEQ ID NO: 5006 or a sequence having at least 98% or 99% similarity to SEQ ID NO: 5006.

3. The mRNA vaccine of claim 2, wherein, the mRNA comprises a coding region; the coding region is selected from SEQ ID NO: 60, SEQ ID NO: 63, or SEQ ID NO:

103.

4. The mRNA vaccine according to any one of claims 1 to 3, characterized in that, the mRNA further comprises a 5’ UTR, the sequence of which is selected from any one of SEQ ID NOs: 1, 3, 10, 12, 197, 201; Preferably, the mRNA further comprises a 5’ UTR, the sequence of which is SEQ ID NO: 3 or SEQ ID NO:

197.

5. The mRNA vaccine according to any one of claims 1 to 4, characterized in that, the mRNA further comprises a 3’ UTR, the sequence of which is selected from any one of SEQ ID NOs: 16, 18, 205, 207, 209; Preferably, the mRNA further comprises a 3’ UTR, the sequence of which is SEQ ID NO: 16 or SEQ ID NO:

205.

6. The mRNA vaccine according to any one of claims 1 to 5, characterized in that the mRNA further comprises a poly A, which comprises at least 70 adenosines.

7. An mRNA vaccine, characterized in that the mRNA comprises: (ii) a 5’ UTR; the 5’ UTR is selected from any one of SEQ ID NOs: 1, 3, 10, 12, 197, 201; (iii) a coding region; the coding region is selected from SEQ ID NO: 60, SEQ ID NO: 63, or SEQ ID NO: 103; (iv) a 3’ UTR; the 3’ UTR is selected from any one of SEQ ID NOs: 16, 18, 205, 207, 209; (v) a poly A; the poly A comprises at least 70 adenosines.

8. The mRNA vaccine of claim 7, wherein, the 5’ UTR sequence is SEQ ID NO: 3 or SEQ ID NO:

197. the 3' UTR sequence is SEQ ID NO: 16 or SEQ ID NO: 205; the ORF sequence is SEQ ID NO:

60.

9. The mRNA vaccine according to any one of claims 1 to 8, characterized in that, the poly A comprises 70-140 adenosines, or 70-120 adenosines, or 80-120 adenosines, or 90-120 adenosines, or 90-110 adenosines, or 90-100 adenosines, or 100-140 adenosines, or 100-120 adenosines, or 100-110 adenosines.

10. The mRNA vaccine according to any one of claims 1 to 9, characterized in that, the sequence of the mRNA is SEQ ID NO: 9961, SEQ ID NO: 9974 or SEQ ID NO: 9975.

11. The mRNA vaccine according to any one of claims 1 to 10, characterized in that, the mRNA further comprises a 5' cap structure.

12. The mRNA vaccine of claim 11, wherein, the 5' cap structure is selected from CapO, Capl, Cap2, or any one of the following compounds:

13. The mRNA vaccine according to any one of claims 1 to 12, characterized in that the mRNA comprises at least one chemically modified nucleoside.

14. The mRNA vaccine of claim 13, wherein, the chemically modified nucleoside is selected from pseudouracil, N1-methyl-pseudouracil, 1-ethylpseudouracil, 2-thiouracil, 4'-thiouracil, 5-methyluracil or 5-methoxyuracil, or any combination thereof.

15. The mRNA vaccine of claim 14, wherein the mRNA vaccine is characterized by, the proportion of chemically modified nucleosides in the mRNA is 10-100%, or 20-100%, or 30-100%, or 40-100%, or 50-100%, or 60-100%, or 70-100%, or 80-100%, or 90-100%, or 50-98%, or 50-95%, or 60-95%, or 70-95%, or 80-95%, or 90-95%, or 70-90%, or 80-90%, or 100%.

16. A vaccine composition comprising, the mRNA vaccine comprises the mRNA of any one of claims 1 to 15 and a delivery agent. the delivery agent comprises a lipid nanoparticle; the lipid nanoparticle comprises: a cationic lipid, a phospholipid, a cholesterol and a PEG lipid.

17. The vaccine composition of claim 16, wherein, the molar ratio of the cationic lipid, the phospholipid, the cholesterol and the PEG lipid in the lipid nanoparticle is any one of c1-c4: c1. the molar ratio of the cationic lipid, the phospholipid, the cholesterol and the PEG lipid is 30-60:0-30:18.5-48.5:0-10; c2. the molar ratio of the cationic lipid, the phospholipid, the cholesterol and the PEG lipid is 35-55:0-25:20-45:0.5-8; c3. the molar ratio of the cationic lipid, the phospholipid, the cholesterol and the PEG lipid is 40-52:5-20:25-40:0.5-5; c4. the molar ratio of the cationic lipid, the phospholipid, the cholesterol and the PEG lipid is 45-52:8-15:30-40:1-5; c5. the molar ratio of the cationic lipid, the phospholipid, the cholesterol and the PEG lipid is 50:10:38.5:1.5, 48.5:11.1:38.9:1.5 or 40:8.0:50.5:1.

5.

18. The vaccine composition according to claim 16 or 17, characterized in that, the cationic lipid is one or more of SM102, MC3, DOTAP, ALC-0315, TM6.

19. The vaccine composition according to any one of claims 16 to 18, wherein, The cationic lipid is of formula I: wherein X0is H; T1, L1are each independently C 1-6 alkyl or is absent; each of T2, L2is independently -O-C(O)-, -C(O)-O-, -O-C(O)-O-, -NH-C(O)-, -C(O)-NH-, -NH-C(O)-O-, or -O-C(O)-NH-; T3, T4, L3, L4 are each independently H, C 4-20 linear or branched alkyl, C 4-20 linear or branched alkenyl or C 4-20 linear or branched alkynyl; wherein T3and T4are not simultaneously H; L3and L4are not simultaneously H; X1, X2are each independently C 1-4 alkyl or nothing; X3is -NR2R3; wherein R2, R3are each independently C 1-4 alkyl or C 1-4 haloalkyl.

20. The vaccine composition of claim 19, wherein, each of said T1, L1is independently ethyl, propyl, isopropyl, butyl, pentyl, hexyl, or absent; each of said T1, L1is independently ethyl, propyl, or absent; each of said T1, L1is independently ethyl, propyl, or absent; each of said T1, L1is independently ethyl, propyl, or absent.

21. The vaccine composition according to claim 19 or 20, characterized in that, each of said T2, L2is independently -O-C(O)- or -C(O)-O-.

22. The vaccine composition according to any one of claims 19 to 21, wherein, The from independently being 23. The vaccine composition according to any one of claims 19 to 22, wherein, The cationic lipid is selected from one of the following structures:

24. The vaccine composition according to any one of claims 16 to 23, wherein, said phospholipid is DSPC or DOPE; said PEG lipid is a PEG with a molecular weight of 1000 Da to 20 kDa, a PEG modified compound, or a mixture thereof.

25. The vaccine composition according to any one of claims 16 to 24, wherein, said composition further comprises a pharmaceutically acceptable excipient.

26. Use of the mRNA vaccine of any one of claims 1 to 15 or the vaccine composition of any one of claims 16 to 25 in the manufacture of a vaccine for the prevention of varicella, herpes zoster, and post-herpetic neuralgia.

27. A method of preventing varicella, herpes zoster and / or postherpetic neuralgia (PHN), characterized in that, administering to a subject in need thereof an effective amount of the mRNA vaccine of any one of claims 1 to 15 or the vaccine composition of any one of claims 16 to 25.

Images