mRNA encoding nerve growth factor (NGF) and use thereof

Abstract

Provided are an mRNA encoding a nerve growth factor, and a composition thereof and the use thereof. The mRNA is obtained by means of obtaining an ORF sequence with a high protein expression level via codon optimization and screening, and by means of combinatorial 5' UTR and 3' UTR screening. The mRNA is delivered to animals, enabling successful expression of an NGF protein in animals, and can thereby be used for preventing and / or treating keratopathy, optic neuropathy, conjunctiva lesions, limbal stem cell deficiency, xerophthalmia, peripheral neuropathy or idiopathic facial nerve palsy, retinopathy, and disorders associated with central nervous system injuries and / or degeneration.

Description

Encoding Nerve Growth Factor (NGF) mRNA and its Uses This application claims priority to Chinese patent application 2025100791805, filed on January 17, 2025. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field This invention relates to the field of biopharmaceutical technology, and more specifically, to the encoding of nerve growth factor (NGF) mRNA and its uses. Background Technology Nerve growth factor (NGF) is one of the most important bioactive molecules in the nervous system, playing a crucial theoretical and clinical role in regulating neuronal growth, development, differentiation, survival, and the regeneration and repair of damaged nerves. Current basic research indicates that NGF can be applied to neurological diseases such as neurotoxicity, peripheral nerve injury, diabetic peripheral neuropathy, Alzheimer's disease, Parkinson's disease, facial neuritis, and nerve damage. Reported clinical trial indications include acute or degenerative neurological diseases such as neurotrophic keratitis, retinitis pigmentosa, and Alzheimer's disease. In addition to promoting the development, differentiation, growth, and maturation of central and peripheral neurons, maintaining normal nervous system function, and accelerating the repair function after nervous system injury, NGF can also promote the repair of corneal epithelial cells. The cornea is richly supplied with the trigeminal nerve, which expresses various epithelial neurotrophic mediators, such as substance P and calcitonin gene-related peptide, promoting DNA synthesis and stimulating corneal epithelial cell proliferation, adhesion, and migration. When the nerve is damaged, the secretion of these substances decreases, leading to impaired corneal epithelial physiological turnover and tear film function. Neurotrophic keratitis (NK) is a corneal regression disease caused by trigeminal nerve injury, characterized by decreased or absent corneal sensation, dry eye, corneal epithelial defects, and corneal ulcers, ultimately leading to corneal stromal melting and perforation. Cenegermin eye drops (rhNGF) received orphan drug designation from the FDA and are the only FDA-approved drug for the treatment of neurotrophic keratitis. It was approved for marketing in China on August 12, 2020, as a drug in short supply domestically. Clinical trial results showed that rhNGF can effectively restore corneal integrity, corneal sensitivity, and transparency. Topical application has no significant toxicity; in case of overdose, the eyes can be rinsed with warm water. It typically has negligible systemic absorption and is not distributed throughout the body, posing no risk of systemic toxicity. However, this drug has been withdrawn from the Chinese market, so there is currently a shortage of such drugs on the market. In recent years, with the advancement of science and technology, mRNA drugs have become a major research focus due to their numerous advantages, such as the absence of risk of genome integration, relatively low immunogenicity, direct and rapid translation into proteins, high safety, natural metabolites, and minimal toxicity. Furthermore, mRNA production is convenient and quick, with lower production costs, allowing for reduced dosing frequency. Therefore, compared to protein drugs, mRNA drugs offer significantly lower costs while achieving the same therapeutic effect; however, there are currently no reports of mRNA drugs that regulate nerve growth factor (NGF) expression. Summary of the Invention One of the objectives of this invention is to provide mRNA encoding nerve growth factor (NGF). Another object of the present invention is to provide a composition comprising mRNA encoding nerve growth factor (NGF). Another object of the present invention is to provide the application of mRNA encoding nerve growth factor (NGF). The above-mentioned objective of the present invention is achieved by the following solution: An mRNA encoding nerve growth factor, the mRNA comprising a coding region, a 5'UTR and a 3'UTR; the coding region is selected from SEQ ID NO:21 or a sequence having at least 98% similarity to SEQ ID NO:21; the 5'UTR is selected from SEQ ID NO:1001; the 3'UTR is selected from SEQ ID NO:1002. An mRNA encoding nerve growth factor, the mRNA comprising a coding region, a 5'UTR and a 3'UTR; the coding region is selected from SEQ ID NO:1134 or a sequence having at least 98% similarity to SEQ ID NO:1134. In one embodiment, the mRNA further includes a 5'UTR, the sequence of which is selected from SEQ ID NO:1001. In one embodiment, the mRNA further includes a 3'UTR, the sequence of which is selected from SEQ ID NO:1002. In one embodiment, the mRNA also includes polyA, which contains at least 70 adenosine molecules. In one embodiment, the mRNA includes: (ii) 5'UTR; 5'UTR is selected from SEQ ID NO:1001; (iii) Coding region; the coding region is selected from SEQ ID NO:21 or SEQ ID NO:1134, or a sequence that has at least 98% similarity to one of SEQ ID NO:21 or SEQ ID NO:1134; (iv) 3'UTR; 3'UTR is selected from SEQ ID NO:1002; (v)poly A; poly A contains at least 70 adenosine molecules. In one embodiment, poly A contains 70-140 adenosine, or 70-120 adenosine, or 80-120 adenosine, or 90-120 adenosine, or 100-140 adenosine, or 90-110 adenosine, or 90-100 adenosine, or 100-110 adenosine. In one embodiment, poly A contains 90-120 adenosine or 90-110 adenosine. In one embodiment, poly A contains 90-110 adenosine molecules. In one embodiment, poly A contains 90-100 adenosine or 100-110 adenosine. In one embodiment, the nucleotide sequence of the mRNA is shown in SEQ ID NO:880 or SEQ ID NO:881. An mRNA encoding nerve growth factor, the mRNA comprising a coding region; the coding region comprising one or more ORFs; the ORFs being selected from SEQ ID NO:20-27 or sequences having at least 98% similarity to one of SEQ ID NO:20-27. In this application, an ORF represents a nucleic acid sequence that encodes a sequence such as SEQ ID NO:2001 or 2002; or a nucleic acid sequence that encodes a C-terminus containing an HSA, ABD, or IgG4-Fc fragment, such as any one of SEQ ID NO:1041-1043 or SEQ ID NO:1091-1093, or any nucleic acid sequence formed by combining any one of SEQ ID NO:20-27 with SEQ ID NO:28 and SEQ ID NO:32. In the implementation, the ORF is one of the following: (1) ORF is SEQ ID NO:20; (2) ORF is SEQ ID NO:21; (3) ORF is SEQ ID NO:22; (4) ORF is SEQ ID NO:23; (5) ORF is SEQ ID NO:24; (6) ORF is SEQ ID NO:25; (7) ORF is SEQ ID NO:26; (8) ORF is SEQ ID NO:27. In one embodiment, the ORF is selected from SEQ ID NO:1041-1043, SEQ ID NO:1091-1093, SEQ ID NO:1131-1133, or a sequence that has at least 98% similarity to one of SEQ ID NO:1041-1043, SEQ ID NO:1091-1093, SEQ ID NO:1131-1133. In one implementation, the ORF is one of the following: (1) ORF is SEQ ID NO:1041; (2) ORF is SEQ ID NO:1042; (3) ORF is SEQ ID NO:1043; (4) ORF is SEQ ID NO:1091; (5) ORF is SEQ ID NO:1092; (6) ORF is SEQ ID NO:1093; (7) ORF is SEQ ID NO:1131; (8) ORF is SEQ ID NO:1132; (9) ORF is SEQ ID NO:1133. In one embodiment, when the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by any one of the sequences in SEQ ID NO: 28, 29 or 33. In one embodiment, when the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by a SEQ ID NO:28 or 29 as a spacer sequence. In one embodiment, when the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by a SEQ ID NO:28 or 33 as a spacer sequence. In one embodiment, when the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by a SEQ ID NO:29 or 33 as a spacer sequence. In one embodiment, the spacer sequence can be the Linker of SEQ ID NO:28, or it can be P2A or IRES. In one implementation, when the coding region contains at least three ORFs, the interval sequences may be the same or different. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO: 1011-1013, 1021-1023, 1031-1033, 1051-1053, 1061-1063, 1071-1073, 1081-1083, 1101-1103, 1111-1113, 1121-1123, and 1131-1133. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO: 1011-1013, 1021-1023 or 1031-1033. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1021-1023. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1031-1033. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO: 1051-1053, 1061-1063, 1071-1073 or 1081-1083. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1061-1063. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1071-1073. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1081-1083. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1101-1103. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1111-1113. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1121-1123. In one embodiment, the mRNA further includes a 5'UTR, the sequence of which is selected from any one of SEQ ID NO:1-15. In this implementation, the mRNA also includes a 5' UTR, the sequence of which is one of the following: (1) The 5'UTR sequence is SEQ ID NO:1; (2) The 5'UTR sequence is SEQ ID NO:2; (3) The 5'UTR sequence is SEQ ID NO:3; (4) The 5'UTR sequence is SEQ ID NO:4; (5) The 5'UTR sequence is SEQ ID NO:5; (6) The 5'UTR sequence is SEQ ID NO:6; (7) The 5'UTR sequence is SEQ ID NO:7; (8) The 5'UTR sequence is SEQ ID NO:8; (9) The 5'UTR sequence is SEQ ID NO:9; (10) The 5'UTR sequence is SEQ ID NO:10; (11) The 5'UTR sequence is SEQ ID NO:11; (12) The 5'UTR sequence is SEQ ID NO:12; (13) The 5'UTR sequence is SEQ ID NO:13; (14) The 5'UTR sequence is SEQ ID NO:14; (15) The 5'UTR sequence is SEQ ID NO:15. In one embodiment, the mRNA further includes a 5'UTR, the sequence of which is SEQ ID NO:3. In one embodiment, the mRNA further includes a 3'UTR, the sequence of which is selected from any one of SEQ ID NO:16-19. In this implementation, the mRNA also includes a 3'UTR, the sequence of which is one of the following: (1) The 3'UTR sequence is SEQ ID NO:16; (2) The 3'UTR sequence is SEQ ID NO:17; (3) The 3'UTR sequence is SEQ ID NO:18; (4) The 3'UTR sequence is SEQ ID NO:19. In one embodiment, the mRNA further includes a 3'UTR, the sequence of which is SEQ ID NO:16. In one embodiment, the mRNA also includes polyA, which contains at least 70 adenosine molecules. In one embodiment, poly A contains 70-140 adenosine, or 70-120 adenosine, or 80-120 adenosine, or 90-120 adenosine, or 100-140 adenosine, or 100-120 adenosine. In one embodiment, the mRNA includes: (ii) 5'UTR; the 5'UTR is selected from any one of SEQ ID NO:1-15; (iii) A coding region; the coding region contains one or more ORFs; the ORFs are selected from SEQ ID NO:20-27 or sequences that are at least 98% similar to one of SEQ ID NO:20-27; (iv) 3'UTR; the 3'UTR is selected from any one of SEQ ID NO:16-19; (v)poly A; the poly A contains at least 70 adenosine molecules. In the implementation, the ORF is one of the following: (1) ORF is SEQ ID NO:20; (2) ORF is SEQ ID NO:21; (3) ORF is SEQ ID NO:22; (4) ORF is SEQ ID NO:23; (5) ORF is SEQ ID NO:24; (6) ORF is SEQ ID NO:25; (7) ORF is SEQ ID NO:26; (8) ORF is SEQ ID NO:27. In the implementation, the 5'UTR sequence is one of the following: (1) The 5'UTR sequence is SEQ ID NO:1; (2) The 5'UTR sequence is SEQ ID NO:2; (3) The 5'UTR sequence is SEQ ID NO:3; (4) The 5'UTR sequence is SEQ ID NO:4; (5) The 5'UTR sequence is SEQ ID NO:5; (6) The 5'UTR sequence is SEQ ID NO:6; (7) The 5'UTR sequence is SEQ ID NO:7; (8) The 5'UTR sequence is SEQ ID NO:8; (9) The 5'UTR sequence is SEQ ID NO:9; (10) The 5'UTR sequence is SEQ ID NO:10; (11) The 5'UTR sequence is SEQ ID NO:11; (12) The 5'UTR sequence is SEQ ID NO:12; (13) The 5'UTR sequence is SEQ ID NO:13; (14) The 5'UTR sequence is SEQ ID NO:14; (15) The 5'UTR sequence is SEQ ID NO:15. In one embodiment, the mRNA further includes a 5'UTR, the sequence of which is SEQ ID NO:3. In the implementation, the sequence of 3'UTR is one of the following: (1) The 3'UTR sequence is SEQ ID NO:16; (2) The 3'UTR sequence is SEQ ID NO:17; (3) The 3'UTR sequence is SEQ ID NO:18; (4) The 3'UTR sequence is SEQ ID NO:19. In one embodiment, the mRNA further includes a 3'UTR, the sequence of which is SEQ ID NO:16. In one embodiment, when the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by any one of the sequences in SEQ ID NO:28, 29 or 33. In one embodiment, when the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by a SEQ ID NO:28 or 29 as a spacer sequence. In one embodiment, when the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by a SEQ ID NO:28 or 33 as a spacer sequence. In one embodiment, when the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by a SEQ ID NO:29 or 33 as a spacer sequence. In one implementation, when the coding region contains at least three ORFs, the interval sequences may be the same or different; When the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO: 1011-1013, 1021-1023, 1031-1033, 1051-1053, 1061-1063, 1071-1073, 1081-1083, 1101-1103, 1111-1113, 1121-1123. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO: 1011-1013, 1021-1023 or 1031-1033. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1021-1023. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1031-1033. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO: 1051-1053, 1061-1063, 1071-1073 or 1081-1083. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1061-1063. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1071-1073. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1081-1083. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1101-1103. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1111-1113. In one embodiment, when the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO:1121-1123. In one embodiment, the mRNA includes: (ii) 5'UTR; the 5'UTR is selected from SEQ ID NO:3; (iii) The coding region; the coding region is SEQ ID NO:21 or a sequence with at least 98% similarity to SEQ ID NO:21; (iv) 3'UTR; the 3'UTR is selected from SEQ ID NO:16; (v)poly A; the poly A contains at least 70 adenosine molecules. In one embodiment, the mRNA includes: (ii) 5'UTR; the 5'UTR is selected from SEQ ID NO:3; (iii) The coding region; the coding region is SEQ ID NO:1131, or a sequence that has at least 98% similarity to SEQ ID NO:1131; (iv) 3'UTR; the 3'UTR is selected from SEQ ID NO:16; (v)poly A; the poly A contains at least 70 adenosine molecules. In one embodiment, the mRNA includes: (ii) 5'UTR; the 5'UTR is selected from SEQ ID NO:3; (iii) The coding region; the coding region is SEQ ID NO:1132, or a sequence that has at least 98% similarity to SEQ ID NO:1132; (iv) 3'UTR; the 3'UTR is selected from SEQ ID NO:16; (v)poly A; the poly A contains at least 70 adenosine molecules. In one embodiment, the mRNA includes: (ii) 5'UTR; the 5'UTR is selected from SEQ ID NO:3; (iii) The coding region; the coding region is SEQ ID NO:1133, or a sequence that has at least 98% similarity to SEQ ID NO:1133; (iv) 3'UTR; the 3'UTR is selected from SEQ ID NO:16; (v)poly A; the poly A contains at least 70 adenosine molecules. In one embodiment, the mRNA comprises at least one chemically modified nucleoside. In one embodiment, 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 embodiment, the chemically modified nucleoside is selected from pseudouracil. In one embodiment, the chemically modified nucleoside is selected from N1-methyl-pseuuridine. In one embodiment, the chemically modified nucleoside is selected from pseudouracil and / or N1-methyl-pseudouracil. In one embodiment, the chemically modified nucleoside is selected from 1-ethylpseuuridine. In one embodiment, the chemically modified nucleoside is selected from 1-ethylpseudouracil and / or N1-methylpseudouracil. In one embodiment, the chemically modified nucleoside is selected from 1-ethylpseudouracil and / or pseudouracil. In one embodiment, the chemically modified nucleoside is selected from 4'-thiouracil. In one embodiment, the chemically modified nucleoside is selected from 4'-thiouracil and / or N1-methyl-pseuuridine. In one embodiment, the chemically modified nucleoside is selected from 4'-thiouracil and / or pseudouracil. In one embodiment, the chemically modified nucleoside is selected from 5-methyluracil. In one embodiment, the chemically modified nucleoside is selected from 5-methyluracil and / or N1-methyl-pseuuridine. In one embodiment, the chemically modified nucleoside is selected from 5-methyluracil and / or pseudouracil. In one embodiment, the chemically modified nucleoside is selected from 5-methoxyuracil. In one embodiment, the chemically modified nucleoside is selected from 5-methoxyuracil and / or N1-methyl-pseuuridine. In one embodiment, 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%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 10-100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 20% to 100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 30% to 100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 40-100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 50-100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 60-100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 70-100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 80-100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 90-100%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 50-98%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 50-95%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 60-95%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 70-95%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 80-95%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 90-95%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 70-90%. In one embodiment, the proportion of chemically modified nucleosides in the mRNA is 80-90%. In one embodiment, the amino acid sequence encoded by the mRNA includes any one of SEQ ID NO:2001, 2002, or 2015. In one embodiment, the amino acid sequence encoded by the mRNA includes SEQ ID NO:2001 or SEQ ID NO:2002. In one embodiment, the amino acid sequence encoded by the mRNA contains SEQ ID NO:2001 or 2015. In one embodiment, the amino acid sequence encoded by the mRNA comprises SEQ ID NO:2002 or 2015. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2015. In one embodiment, the amino acid sequence encoded by the mRNA is any one of SEQ ID NO:2001-2013. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2001. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2002. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2003. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2004. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2005. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2006. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2007. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2008. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2009. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2010. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2011. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2012. In one embodiment, the amino acid sequence encoded by the mRNA is as shown in SEQ ID NO:2013. In one embodiment, the mRNA further includes (i) a 5' cap structure, the 5' cap structure being selected from Cap0, Cap1, Cap2 or the structure shown in Formula I: Where R1 represents H and C 1-4 Alkyl, C 1-4 Halogenated alkyl, phenyl, C 1-4 Alkyl-substituted phenyl, halophenyl, benzyl, C 1-4 Alkyl-substituted benzyl or halobenzyl; R2 is OH, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Alkoxy-substituted C 1-4 Alkyl, C 1-4 Alkylamine substituent C 1-4 Alkyl, C 1-4 Amide group substituted C 1-4 Alkyl, benzyloxy, halobenzyloxy, benzyloxy-substituted C 1-4 Alkyl or halobenzyloxy-substituted C 1- 4-alkyl; R3 and R4 are independently H and C. 1-4 Alkyl or C 1-4 Halogenated alkyl groups; R5 represents H and C. 1-4 Alkoxy, F, Cl, I, or N3; R6 represents H and C. 1-4 Alkyl, C 1-4 alkenyl, C 1-4 alkynyl, phenyl, C 1-4 Alkyl-substituted phenyl, halophenyl, benzyl, C 1-4 Alkyl-substituted benzyl or halobenzyl; B1 and B2 are each independently a base G, A, U or C, and substituted bases include, but are not limited to, 6-MeA, 5-MeU or 5-MeC. In one embodiment, R6 is H or C. 1-4 Alkyl, C 1-4 alkenyl, C 1-4 Alkyne, phenyl, halophenyl, benzyl, or halobenzyl. In one embodiment, R6 is H or C. 1-4 Alkyl, phenyl, halophenyl, benzyl or halobenzyl. In one embodiment, R6 is H, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, phenyl, benzyl, 3-fluorophenyl, 4-fluorophenyl, 3,5-difluorophenyl, 3-chlorophenyl, 4-chlorophenyl, 3,5-dichlorophenyl, 3-fluorobenzyl, 4-fluorobenzyl, 3,5-difluorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl or 3,5-dichlorobenzyl. In one embodiment, R6 is H, methyl, ethyl, propyl, isopropyl, phenyl, benzyl, 3-fluorophenyl, 4-fluorophenyl, 3-chlorophenyl, 4-chlorophenyl, 3-fluorobenzyl, 4-fluorobenzyl, 3-chlorobenzyl or 4-chlorobenzyl. In one embodiment, R6 is H, methyl, ethyl, isopropyl, phenyl, benzyl, 4-fluorophenyl, 4-chlorophenyl, 4-fluorobenzyl or 4-chlorobenzyl. In one embodiment, R6 is H, methyl, ethyl, isopropyl, phenyl, benzyl, 4-fluorophenyl, or 4-fluorobenzyl. In one embodiment, R6 is H or methyl. In one embodiment, R6 is phenyl or 4-fluorophenyl. In one embodiment, R6 is benzyl or 4-fluorobenzyl. In one embodiment, R5 is H, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, F, Cl, I, or N3. In one embodiment, R5 is H, methyl, ethyl, propyl, isopropyl, tert-butyl, F, or N3. In one embodiment, R5 is H, methyl, ethyl, isopropyl, F, or N3. In one embodiment, R5 is H, methyl, ethyl, F, or N3. In one implementation, R5 is H or F. In one embodiment, R5 is H or methyl. In one implementation, R5 is F or N3. In one embodiment, R3 and R4 are each independently H, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, or trifluoroisopropyl. In one embodiment, R3 and R4 are each independently H, methyl, ethyl, isopropyl, tert-butyl, trifluoromethyl, trifluoroethyl, or trifluoroisopropyl. In one embodiment, R3 and R4 are each independently H, methyl, ethyl, isopropyl, trifluoromethyl, or trifluoroethyl. In one embodiment, R3 and R4 are each independently H, methyl, ethyl, or trifluoromethyl. In one embodiment, R3 and R4 are each independently H, methyl, or trifluoromethyl. In one embodiment, R2 is OH or C. 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Alkoxy-substituted C 1- 4-alkyl, C 1-4 Alkylamine substituent C 1-4 Alkyl, C 1-4 Amide group substituted C 1-4 Alkyl, benzyloxy or benzyloxy-substituted C 1-4 alkyl. In one embodiment, R2 is OH or C. 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-3 Alkoxy-substituted C 1- 3-alkyl, C 1-3 Alkylamine substituent C 1-3 Alkyl, C 1-3 Amide group substituted C 1-3 Alkyl, benzyloxy or benzyloxy-substituted C 1-3 alkyl. In one embodiment, R2 is OH or C. 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-3 Alkoxy-substituted C 1- 3-alkyl, C 1-3 Alkylamine substituent C 1-3 Alkyl or C 1-3 Amide group substituted C 1-3 alkyl. In one embodiment, R2 is OH or C. 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl or C 1-3 Alkoxy-substituted C 1- 3-alkyl group. In one embodiment, R2 is OH, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, methyloxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoroisopropyl, methoxymethyl, methoxyethyl, methoxypropyl, methoxyisopropyl, ethoxymethyl, ethoxyethyl, ethoxypropyl, ethoxyisopropyl, propoxymethyl, propoxyethyl, isopropoxymethyl, or isopropoxyethyl. In one embodiment, R2 is OH, methyl, ethyl, propyl, isopropyl, tert-butyl, methyloxy, ethoxy, propoxy, isopropoxy, tert-butoxy, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoroisopropyl, methoxymethyl, methoxyethyl, methoxyisopropyl, ethoxymethyl, ethoxyethyl, ethoxyisopropyl, propoxymethyl, propoxyethyl, isopropoxymethyl, or isopropoxyethyl. In one embodiment, R2 is OH, methyl, ethyl, methyloxy, ethoxy, isopropoxy, tert-butoxy, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoroisopropyl, methoxymethyl, methoxyethyl, ethoxymethyl, ethoxyethyl, propoxymethyl, propoxyethyl, isopropoxymethyl, or isopropoxyethyl. In one embodiment, R2 is OH, methyl, ethyl, methyloxy, ethoxy, trifluoromethyl, trifluoroethyl, methoxymethyl, methoxyethyl, ethoxymethyl, ethoxyethyl, propoxymethyl, propoxyethyl, isopropoxymethyl, or isopropoxyethyl. In one embodiment, R2 is OH, methyloxy, ethoxy, trifluoromethyl, methoxymethyl, or ethoxymethyl. In one embodiment, R2 is OH, methyloxy, or methoxymethyl. In one embodiment, R2 is -CH2NHCH3, -CH2NHC2H5, -CH2NHC3H7, -CH2NHC4H9, -C2H4NHCH3, -C2H4NHC2H5, -C2H4NHC3H7, -C2H4NHC4H9, -C3H6NHCH3, -C3H6NHC2H5, -C3H6NHC3H7, -C3H6NHC4H9, -CH2NHCOCH3, -CH2NHCOC2H5, -CH2NHCOC3H7, -C2H4NHCOC2H5, or -C2H4NHCOC3H7. In one embodiment, R2 is -CH2NHCH3, -CH2NHC2H5, -CH2NHC3H7, -C2H4NHCH3, -C2H4NHC2H5, -C2H4NHC3H7, -C3H6NHCH3, -C3H6NHC2H5, -C3H6NHC3H7, -CH2NHCOCH3, -CH2NHCOC2H5, -CH2NHCOC3H7, or -C2H4NHCOC2H5. In one embodiment, R2 is -CH2NHCH3, -CH2NHC2H5, -CH2NHC3H7, -C2H4NHCH3, -C2H4NHC2H5, -C3H6NHCH3, -C3H6NHC2H5, -CH2NHCOCH3, -CH2NHCOC2H5, or -C2H4NHCOC2H5. In one embodiment, R2 is -CH2NHCH3, -CH2NHC2H5, -CH2NHC3H7, -C2H4NHCH3, -C2H4NHC2H5, -CH2NHCOCH3, or -CH2NHCOC2H5. In one embodiment, R2 is -CH2NHCH3, -CH2NHC2H5, -CH2NHC3H7, -CH2NHCOCH3, or -CH2NHCOC2H5. In one embodiment, R2 is CH2NHCOCH3 or -CH2NHCOC2H5. In one embodiment, R1 is H, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoroisopropyl, phenyl, 2-methyl-phenyl, 3-methyl-phenyl, 4-methyl-phenyl, 2,6-dimethyl-phenyl, 3,5-dimethyl-phenyl, 2-ethyl-phenyl, 3-ethyl-phenyl, 4-ethyl-phenyl, 2,6-diethyl-phenyl, 3,5-diethyl-phenyl, 2-chloro-phenyl, 3-chloro-phenyl, 4-chloro-phenyl, 2,6-chloro-phenyl, 3,5-chloro-phenyl, 2-fluoro-phenyl, 3-fluoro-phenyl -Phenyl, 4-fluoro-phenyl, 2,6-fluoro-phenyl, 3,5-fluoro-phenyl, benzyl, 2-methyl-benzyl, 3-methyl-benzyl, 4-methyl-benzyl, 2,6-dimethyl-benzyl, 3,5-dimethyl-benzyl, 2-ethyl-benzyl, 3-ethyl-benzyl, 4-ethyl-benzyl, 2,6-diethyl-benzyl, 3,5-diethyl-benzyl, 2-chloro-benzyl, 3-chloro-benzyl, 4-chloro-benzyl, 2,6-chloro-benzyl, 3,5-chloro-benzyl, 2-fluoro-benzyl, 3-fluoro-benzyl, 4-fluoro-benzyl, 2,6-fluoro-benzyl or 3,5-fluoro-benzyl. In one embodiment, R1 is H, methyl, ethyl, propyl, isopropyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoroisopropyl, phenyl, 3-methyl-phenyl, 4-methyl-phenyl, 3,5-dimethyl-phenyl, 3-ethyl-phenyl, 4-ethyl-phenyl, 3,5-diethyl-phenyl, 3-chloro-phenyl, 4-chloro-phenyl, 3,5-chloro-phenyl, 3-fluoro-phenyl, 4-fluoro-phenyl, 3,5-fluoro-phenyl, benzyl, 3-methyl-benzyl, 4-methyl-benzyl, 3,5-dimethyl-benzyl, 3-ethyl-benzyl, 4-ethyl-benzyl, 3,5-diethyl-benzyl, 3-chloro-benzyl, 4-chloro-benzyl, 3,5-chloro-benzyl, 3-fluoro-benzyl, 4-fluoro-benzyl, or 3,5-fluoro-benzyl. In one embodiment, R1 is H, methyl, ethyl, isopropyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoroisopropyl, phenyl, 3-methyl-phenyl, 4-methyl-phenyl, 3,5-dimethyl-phenyl, 4-ethyl-phenyl, 3-chloro-phenyl, 4-chloro-phenyl, 3-fluoro-phenyl, 4-fluoro-phenyl, benzyl, 3-methyl-benzyl, 4-methyl-benzyl, 3-ethyl-benzyl, 4-ethyl-benzyl, 3-chloro-benzyl, 4-chloro-benzyl, 3-fluoro-benzyl, or 4-fluoro-benzyl. In one embodiment, R1 is H, methyl, ethyl, isopropyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoroisopropyl, phenyl, 4-methyl-phenyl, 4-ethyl-phenyl, 4-chloro-phenyl, 4-fluoro-phenyl, benzyl, 4-methyl-benzyl, 4-ethyl-benzyl, 4-chloro-benzyl or 4-fluoro-benzyl. In one embodiment, R1 is H, methyl, ethyl, isopropyl, trifluoromethyl, trifluoroethyl, benzyl, 4-methyl-benzyl, 4-ethyl-benzyl, 4-chloro-benzyl, or 4-fluoro-benzyl. In one embodiment, R1 is H, methyl, ethyl, isopropyl, trifluoromethyl, trifluoroethyl, benzyl, 4-methyl-benzyl, or 4-fluoro-benzyl. In some implementations, the 5' cap structure can be, but is not limited to, the following compounds: The present invention also protects compositions comprising the aforementioned mRNA encoding nerve growth factor and a delivery agent. In one embodiment, the delivery agent comprises lipid nanoparticles; the lipid nanoparticles include ionizable lipids, phospholipids, cholesterol, and PEG lipids. In one embodiment, the molar ratio of ionizable lipids, phospholipids, cholesterol and PEG lipids in the lipid nanoparticles is 30-60:0-30:18.5-48.5:0-10. In one embodiment, the molar ratio of ionizable lipids, phospholipids, cholesterol and PEG lipids in the lipid nanoparticles is 35-55:0-25:20-45:0.5-8. In one embodiment, the molar ratio of ionizable lipids, phospholipids, cholesterol and PEG lipids in the lipid nanoparticles is 40-52:5-20:25-40:0.5-5. In one embodiment, the molar ratio of ionizable lipids, phospholipids, cholesterol and PEG lipids in the lipid nanoparticles is 45-52:8-15:30-40:1-5. In one embodiment, the molar ratio of ionizable lipids, phospholipids, cholesterol and PEG lipids in the lipid nanoparticles is 50:10:38.5:1.5. In one embodiment, the molar ratio of ionizable lipids, phospholipids, cholesterol and PEG lipids in the lipid nanoparticles is 48.5:11.1:38.9:1.5. In one embodiment, the ionizable lipid is as shown in Formula II: Where n0 is an integer from 0 to 4; n1 is an integer from 2 to 7; n2 is an integer from 4 to 10; n3 is an integer from 2 to 8; n4 is an integer from 2 to 8; and n5 is an integer from 2 to 8. In one embodiment, n0 is 0, 1, 2, 3 or 4; preferably, n0 is 1, 2 or 3; more preferably, n0 is 1 or 2. In one embodiment, n1 is 2, 3, 4, 5, 6 or 7; preferably, n1 is 2, 3, 4 or 5; more preferably, n1 is 2, 3 or 4. In one embodiment, n2 is 4, 5, 6, 7, 8, 9 or 10; preferably, n2 is 6, 7, 8, 9 or 10; more preferably, n2 is 8, 9 or 10. In one embodiment, n3 is 2, 3, 4, 5, 6, 7 or 8; preferably, n3 is 3, 4, 5 or 6; more preferably, n3 is 4, 5 or 6. In one embodiment, n4 is 2, 3, 4, 5, 6, 7 or 8; preferably, n4 is 3, 4, 5 or 6; more preferably, n4 is 4, 5 or 6. In one embodiment, n5 is 2, 3, 4, 5, 6, 7 or 8; preferably, n5 is 3, 4, 5 or 6; more preferably, n5 is 4, 5 or 6. In one embodiment, the ionizable lipid is one or more combinations of SM102, MC3, DOTAP, ALC-0315, H1, H2, H3, H4, H5, and H6. In one embodiment, the ionizable lipid is one of the following: (1) The ionizable lipid is SM102; (2) The ionizable lipid is MC3; (3) The ionizable lipid is DOTAP; (4) The ionizable lipid is ALC-0315; (5) The ionizable lipid is H1; (6) The ionizable lipid is H2; (7) The ionizable lipid is H3; (8) The ionizable lipid is H4; (9) The ionizable lipid is H5; (10) The ionizable lipid is H6. Their structures are shown below: In one embodiment, the phospholipid is DSPC or DOPE; the PEG lipid has a molecular weight of PEG, PEG-modified compounds or mixtures thereof of 1000 Da to 20 kDa. In one embodiment, the mRNA in the composition is encapsulated in lipid nanoparticles. In one embodiment, the composition further includes pharmaceutically acceptable excipients. The present invention also protects the use of the aforementioned mRNA encoding nerve growth factor or the aforementioned composition in the preparation of medicaments for the prevention and / or treatment of, but not limited to, corneal lesions, optic neuropathy, conjunctival lesions, limbal stem cell deficiency, dry eye syndrome, peripheral neuropathy or idiopathic facial nerve palsy, retinal lesions, central nervous system injury and / or degenerative conditions. Preferably, the aforementioned mRNA encoding nerve growth factor or the aforementioned composition is used in the preparation of a medicament for the prevention and / or treatment of, but not limited to, corneal lesions, optic neuropathy, conjunctival lesions, limbal stem cell deficiency, dry eye syndrome, peripheral neuropathy, or idiopathic facial nerve palsy. Preferably, the aforementioned mRNA encoding nerve growth factor or the aforementioned composition is used in the preparation of medicaments for the prevention and / or treatment of, but not limited to, corneal lesions, optic neuropathy, conjunctival lesions, limbal stem cell deficiency, and dry eye syndrome. Preferably, the aforementioned mRNA encoding nerve growth factor or the aforementioned composition is used in the preparation of medicaments for the prevention and / or treatment of, but not limited to, corneal lesions, limbal stem cell deficiency, and dry eye. Preferably, the aforementioned mRNA encoding nerve growth factor or the aforementioned composition is used in the preparation of medicaments for the prevention and / or treatment of, but not limited to, corneal lesions and dry eye syndrome. Preferably, the aforementioned mRNA encoding nerve growth factor or the aforementioned composition is used in the preparation of a treatment for the prevention and / or treatment of peripheral neuropathy or idiopathic facial nerve palsy. Preferably, the aforementioned mRNA encoding nerve growth factor or the aforementioned composition is used in the preparation of a medicament for the prevention and / or treatment of retinal diseases, central nervous system injury, and / or degenerative conditions. In one embodiment, the corneal disease is selected from keratoconus, phototoxic keratopathy, persistent corneal epithelial defects, corneal ulcers, corneal dystrophy and degeneration, dry keratoconjunctivitis, or neurotrophic keratitis. In one embodiment, the corneal lesion is selected from corneal dystrophy and degeneration, dry keratoconjunctivitis, or neurotrophic keratitis. In one embodiment, the corneal lesion is neurotrophic keratitis. In one embodiment, the optic neuropathy is selected from glaucoma, as well as ischemic optic neuropathy, degenerative, traumatic, Leber hereditary optic neuropathy, and congenital optic atrophy. In one embodiment, dry eye is selected from aqueous dry eye, lipid-deficient dry eye, mucin-deficient dry eye, tear dynamics-deficient dry eye, and mixed dry eye. In one embodiment, dry eye is selected from aqueous dry eye, lipid abnormality dry eye, and mixed dry eye. The present invention also protects the use of the aforementioned mRNA encoding nerve growth factor or the aforementioned composition in the preparation of drugs for the prevention and / or treatment of peripheral neuropathy and idiopathic facial nerve palsy. In one embodiment, peripheral neuropathy includes, but is not limited to, HIV-associated sensory neuropathy, diabetic polyneuropathy, toxic peripheral neuropathy, and traumatic peripheral nerve injury. In one embodiment, the retinopathy is selected from diabetic retinopathy, retinal detachment, age-related macular degeneration, macular degeneration, macular atrophy, macular hole, macular edema, retinopathy of prematurity, retinal vascular occlusion, phototoxic retinopathy, or epiretinal membrane disease. In one embodiment, the retinal disease is selected from diabetic retinopathy, retinal detachment, age-related macular degeneration, macular degeneration, macular atrophy, macular hole, or macular edema. In one embodiment, the retinopathy is diabetic retinopathy. In one embodiment, the retinal disease is age-related macular degeneration, macular degeneration, macular atrophy, macular hole, or macular edema. In one embodiment, central nervous system injury and / or degeneration-related conditions include, but are not limited to, Alzheimer's disease, Parkinson's syndrome, and frontotemporal dementia. In one embodiment, the drug is a topical drug. In one embodiment, the drug is an injection, a topical preparation, a nasal spray, or eye drops. The present invention also protects a method for preventing and / or treating corneal diseases, optic neuropathy, conjunctival diseases, limbal stem cell deficiency or dry eye, peripheral neuropathy or idiopathic facial nerve palsy, comprising administering to a subject in need a therapeutically effective amount of the aforementioned mRNA encoding nerve growth factor or the aforementioned composition. In one embodiment, the administration of other medications to the subject is also included. In one embodiment, the corneal lesion is selected from keratoconus, phototoxic keratopathy, persistent corneal epithelial defects, corneal ulcers, corneal dystrophy and degeneration, dry keratoconjunctivitis, or neurotrophic keratitis. In one embodiment, the optic neuropathy is selected from glaucoma, as well as ischemic optic neuropathy, degenerative, traumatic, Leber hereditary optic neuropathy, and congenital optic atrophy. In one embodiment, the retinopathy is selected from diabetic retinopathy, retinal detachment, age-related macular degeneration, macular degeneration, macular atrophy, macular hole, macular edema, retinopathy of prematurity, retinal vascular occlusion, phototoxic retinopathy, or epiretinal membrane disease. In one embodiment, the retinal disease is selected from diabetic retinopathy, retinal detachment, age-related macular degeneration, macular degeneration, macular atrophy, macular hole, or macular edema. In one embodiment, the retinopathy is diabetic retinopathy. In one embodiment, the retinal disease is age-related macular degeneration, macular degeneration, macular atrophy, macular hole, or macular edema. In one embodiment, central nervous system injury and / or degeneration-related conditions include, but are not limited to, Alzheimer's disease, Parkinson's syndrome, and frontotemporal dementia. Compared with the prior art, the present invention has the following beneficial effects: This invention, through codon optimization and screening, obtains ORF sequences with high protein expression levels. Further screening using a combination of 5'UTR and 3'UTR yields mRNAs with excellent protein expression levels in animals. Delivery of these mRNAs to animals successfully expresses NGF protein, which can be used to improve NGF deficiency in animals, thereby preventing and / or treating various diseases such as corneal lesions, optic nerve diseases, conjunctival lesions, limbal stem cell deficiency, dry eye syndrome, peripheral neuropathy or idiopathic facial nerve palsy, retinal diseases, and central nervous system injury and / or degeneration-related conditions. Attached Figure Description Figure 1 shows the effect of mRNA-expressed proteins on PC12 cells in Example 2. Figure 2 shows the effect of mRNA-expressed proteins on HCEC cells in Example 3. Figure 3 shows the results of mouse weight recording in each group in Example 5. Figure 4 shows the corneal sensation recovery effect of each group of mice in Example 5. Figure 5 shows the IOD value test results of the corneal staining area of ​​mice in each group in Example 5. Figure 6 shows the results of the corneal staining area test in each group of mice in Example 5. Figure 7 shows the results of the corneal neuropathy grading test in each group of mice in Example 5. Figure 8 shows the effect of mRNA-expressed proteins on PC12 cells in Example 2. Figure 9 shows the effect of mRNA-expressed proteins on HCEC cells in Example 3. Figure 10 shows the effect of mRNA-expressed proteins on PC12 cells in Example 2. Figure 11 shows the results of mouse weight recording in each group in Example 5. Figure 12 shows the corneal sensation recovery effect of each group of mice in Example 5. Figure 13 shows the results of the corneal staining area test in each group of mice in Example 5. Figure 14 shows the results of detecting the number of surviving retinal ganglion cells in each group of rats in Example 6. Detailed Implementation the term 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] Straight-chain or branched hydrocarbon chain groups. In one embodiment, the alkyl group has, for example, 1 to 24 carbon atoms (C1-C2). 24 Alkyl groups, 4 to 20 carbon atoms (C4-C5) 20 Alkyl groups, 6 to 16 carbon atoms (C6-C5) 16Alkyl groups, 6 to 9 carbon atoms (C6-C9 alkyl groups), 1 to 15 carbon atoms (C1-C9 alkyl groups) 15 Alkyl groups, 1 to 12 carbon atoms (C1-C2) 12 Alkyl groups (1 to 8 carbon atoms (C1-C8 alkyl) or 1 to 6 carbon atoms (C1-C6 alkyl) are attached to the remainder of the molecule by single bonds. 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, alkyl groups are 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 contains at least one ionizable lipid. Without being bound by theory, it is anticipated that the ionizable lipid can interact with the negatively charged payload molecule and promote the incorporation and / or encapsulation of the payload 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 "ionizable 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). In some embodiments, the positive charge in an ionizable lipid is caused by the presence of a quaternary nitrogen atom. In some embodiments, an ionizable 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 stated, the term "pharmaceutically acceptable carrier, diluent, or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, gliding agent, sweetener, diluent, preservative, dye / coloring agent, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier approved for acceptable use in humans or livestock. The term "composition" is intended to cover products containing a specified amount of a specified ingredient (e.g., the mRNA molecule provided herein) optionally selected. In some embodiments, the mRNA is a monocistronic mRNA containing only one ORF. In some embodiments, the monocistronic mRNA encodes a peptide or protein containing at least one epitope of a selected antigen (e.g., a pathogenic antigen or tumor-associated antigen). In other embodiments, the mRNA is a polycistronic mRNA containing two or more ORFs. In some embodiments, the polycistronic mRNA encodes two or more peptides or proteins that may be the same as or different from each other. In some embodiments, each peptide or protein encoded by the polycistronic mRNA contains at least one epitope of a selected antigen. In some embodiments, the different peptides or proteins encoded by the polycistronic mRNA each contain at least one epitope of a different antigen. In any of the embodiments described herein, the at least one epitope may be at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten epitopes of an antigen. The term "nucleobase" encompasses purines and pyrimidines, including the natural compounds adenine, thymine, guanine, cytosine, uracil, inosine, and their natural or synthetic analogues or derivatives. The term "administer / administration" refers to the act of injecting or otherwise physically delivering a substance present outside the body (such as the lipid nanoparticle composition described herein) into a patient's body, such as via mucosal, intradermal, intravenous, intramuscular delivery, and / or any other physical delivery method described herein or known in this art. When treating a disease, condition, illness, or its symptoms, the substance is usually administered after the onset of the disease, condition, illness, or its symptoms. When preventing a disease, condition, illness, or its symptoms, the substance is usually administered before the onset of the disease, condition, illness, or its symptoms. An "effective amount" is generally an amount sufficient to reduce the severity and / or frequency of symptoms; eliminate symptoms and / or underlying causes; prevent the occurrence of symptoms and / or their underlying causes; and / or improve or remedy damage caused by or associated with a disease, condition, or disorder, including, for example, infection and lesion formation. In some implementations, an effective amount is a therapeutic effective amount or a preventive effective amount. As used herein, the term "therapeuticly effective amount" refers to an amount of agent (e.g., a lipid nanoparticle composition as described herein) sufficient to reduce and / or improve the severity and / or duration of a given disease, condition, or disorder and / or its associated symptoms (e.g., infectious diseases such as those caused by viral infections, or proliferative disorders such as cancer). The "therapeuticly effective amount" of the substances / molecules / agents disclosed herein (e.g., lipid nanoparticle compositions as described herein) can vary depending on numerous factors, such as an individual's disease state, age, sex, and weight, and the ability of the substance / molecule / agent to elicit a desired response in the individual. A therapeutically effective amount includes the amount in which the therapeutically beneficial effect of the substance / molecule / agent outweighs any of its toxic or harmful effects. In some embodiments, the term "therapeuticly effective amount" refers to an amount of a lipid nanoparticle composition as described herein, or a therapeutic or preventative agent (e.g., therapeutic mRNA) contained therein, that is effective in treating a disease, condition, or disorder in a subject or mammal. The term "treatment" refers to the complete or partial relief of a symptom, disease, or ailment, or one or more symptoms associated with a symptom, disease, or ailment, or the slowing or stopping of the further progression or worsening of those symptoms, or the reduction or eradication of the cause of the symptom, disease, or ailment itself. The term "preventing" refers to reducing the likelihood of the onset (or recurrence) of a disease, condition, illness, or related symptoms. The term "side effect" encompasses undesirable and / or adverse effects of a therapy (such as a preventative or therapeutic agent). An undesirable effect is not necessarily an adverse effect. Adverse effects of a therapy (such as a preventative or therapeutic agent) may be harmful, unpleasant, or risky. Examples of side effects include diarrhea, cough, gastroenteritis, wheezing, nausea, vomiting, anorexia, abdominal cramps, fever, pain, weight loss, dehydration, hair loss, difficulty breathing, insomnia, dizziness, mucositis, neuromuscular effects, fatigue, dry mouth, loss of appetite, rash or swelling at the application site, flu-like symptoms such as fever, chills, and fatigue, digestive problems, and allergic reactions. Numerous other undesirable effects experienced by patients are known in this technology. The present invention will be further described in detail below with reference to specific embodiments. These embodiments are only used to explain the present invention and are not intended to limit the scope of the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods; the materials and reagents used are commercially available unless otherwise specified. Table 1 shows some of the sequences involved in this invention; Table 2 shows some of the recombinant fusion sequences; Table 3 shows some of the amino acid sequences; Table 4 shows some of the mRNA sequences. Table 1 Sequence Table 2 Recombination and Fusion Sequences Table 3 Amino acid sequences Table 4 mRNA sequences The sequence of SEQ ID NO: 96, omitting poly A, is as follows: The sequence of SEQ ID NO: 880 for poly A is omitted as follows: The sequence of SEQ ID NO: 881, omitting poly A, is as follows: Example 1: Screening Sequence The amino acid sequence of the mature NGF protein is shown in SEQ ID NO:2001; the amino acid sequence of the ProNGF protein is shown in SEQ ID NO:2002; the ORF sequence of the wild-type mRNA of the NGF protein is shown in SEQ ID NO:27; the ORF sequence of the wild-type mRNA of the ProNGF protein is shown in SEQ ID NO:20. After codon optimization, the sequences SEQ ID NO:21-26 were designed. 1.1 Filtering UTR sequences The selected 5' UTR sequences are shown in SEQ ID NO:1-15; the selected 3' UTR sequences are shown in SEQ ID NO:16-19; using SEQ ID NO:20 as the ORF, it is combined with different 5' UTR sequences and different 3' UTR sequences to obtain sequences SEQ ID NO:34-93. The ORF sequences in SEQ ID NO:34-93 can be replaced with any one of SEQ ID NO:21-27, SEQ ID NO:1041-1043, SEQ ID NO:1091-1093, or SEQ ID NO:1131-1134 to obtain the mRNA sequences. Sequences SEQ ID NO:34-93 were selected for testing, where the ORF was all sequence SEQ ID NO:20, and the 5'UTR and 3'UTR sequences were different. The effects of different 5'UTR sequences and different 3'UTR sequences were tested. The DNA sequence corresponding to the mRNA of sequence SEQ ID NO:34-93 was synthesized by GenScript. The specific process is as follows: (1) Synthesize the DNA sequence corresponding to the mRNA; (2) Construct the DNA sequence on the plasmid vector through homologous recombination; wherein the number of adenosine in poly A is 100±10. The plasmid vector containing the target gene was amplified and purified by E. coli and further linearized. After linearization, the plasmid was used to prepare capped mRNA by a one-pot method. For specific steps, please refer to patent 202210973168.5. The cap structure can be any one of compounds 1-50 listed in this invention. Specifically, the cap structure used in the mRNA of the following examples is compound 19. Detection of mRNA expression levels in cells Transfection and expression: 293T cells were transfected with 0.6 × 10⁻⁶ cells. 6 Cells were seeded at a density of [number] cells / mL in 6-well cell culture plates and cultured at 37°C with 5% CO2. The next day, mRNA was transfected into cells at a mass-to-volume ratio of 1:2 for mRNA to transfection reagent jetMESSENGER, as follows: 200 μL jetMESSENGER Buffer was mixed with 4 μg mRNA, followed by 8 μL transfection reagent. The mixture was incubated at room temperature for 15 min, then added to the cells and cultured at 37°C with 5% CO2. The medium was changed 6 h after transfection, and the supernatant was collected 24 h after transfection. The expression level of NGF in the supernatant was measured by ELISA. The measurement results are shown in Table 5. Table 5. mRNA expression results in cells. As shown in Table 5, the 5'UTR and 3'UTR sequences used in this invention, after being used to prepare mRNA, can both express proteins normally in cells. When the ORF sequence is SEQ ID NO:20, mRNAs composed of different 5'UTR sequences and different 3'UTR sequences show significant differences in expression levels. From the above results, it can be seen that the mRNA expression is optimal when the 5'UTR sequence is SEQ ID NO:3 and the 3'UTR sequence is SEQ ID NO:16. Following the steps outlined above, sequence SEQ ID NO:880-904 was selected for testing, and the results are shown in Table 5. When the ORF sequences were SEQ ID NO:21 and SEQ ID NO:1131-1134, the mRNAs composed of different 5'UTR sequences and different 3'UTR sequences showed essentially equivalent expression levels. 1.2 Screening ORF sequences The preparation, transfection, and expression of mRNA are described in 1.1. The ORF sequences of the mRNA are SEQ ID NO: 20-26; the 5' UTR sequence is SEQ ID NO: 3 or SEQ ID NO: 1001; and the 3' UTR sequence is SEQ ID NO: 16 or SEQ ID NO: 1002, which are the sequences SEQ ID NO: 36, 96, 156, 216, 276, 336, 396, 880. The measured expression results are shown in Table 6. Table 6. mRNA expression results in cells. As shown in Table 6, the multiple mRNAs prepared by this invention can all express the target protein in cells, and the expression levels are good, all superior to the wild-type sequence SEQ ID NO:20; especially sequences SEQ ID NO:96, SEQ ID NO:156, SEQ ID NO:216, and SEQ ID NO:880, among which SEQ ID NO:880 has a higher expression level. 1.3 Screening for recombination and fusion sequences To further improve mRNA stability and expression efficiency, multiple recombinant fusion sequences were designed to screen for mRNAs with better performance. The synthesis, transfection, and expression detection of the recombinant fusion sequences are described in section 1.1. The measurement results are shown in Table 7. Table 7. mRNA expression results in cells. As shown in Table 7, the recombinant and fused sequences all showed a certain degree of increase in protein expression compared with the non-recombinant and fused sequences, indicating that recombinant and fused ORF can increase mRNA expression and protein expression by more than 1.5 times. Example 2: Effects of mRNA-expressed proteins on PC12 cells The selected mRNA was used to express the protein in cells, and a cell supernatant containing the target protein was obtained. Its effect on PC12 cells was then tested. The test method is as follows: (1) Preparation of cell supernatant containing NGF protein mRNA was prepared using the sequence obtained after screening, and transfected into 293T cells for expression according to the method in Example 1. After 24 hours of expression, the cell supernatant containing NGF protein was collected for later use. (2) Testing the effect on PC12 cells PC12 cells were used at a rate of 1×10 5 Cells were seeded at a density of [number] cells / mL in 6-well plates coated with rat tail collagenase and cultured at 37°C under 5% CO2. After 24 hours of culture, the cell culture medium was removed, the cells were washed twice with PBS, and the cell supernatant was replaced with either NGF protein or non-NGF protein. Cells were then cultured for 5 days and photographed. The final concentration of NGF protein used was 50 ng / μL. The growth of PC12 cells in different culture media was observed. The length of a neurite is determined by comparing the neurite length with the average diameter of the cell body, and is reported as neurite length / cell body. As shown in Figures 1, 8, and 10, compared with the control group, the treatment group with cell supernatant containing mRNA-expressing proteins showed a significant increase in both neurite length and cell body length. Example 3: mRNA-expressed proteins promote the proliferation and migration of HCECs. (1) Preparation of cell supernatant containing NGF protein Refer to (1) in Example 2. (2) Testing the effect on HCEC Corneal epithelial cells HCEC at 3.5 × 10 5 Cells were seeded at a density of 10 cells / mL in 6-well cell culture plates and cultured at 37°C under 5% CO2. After confluence, the cells were replaced with serum-free medium and starved for 8 hours. Cells were then scratched with a sterile 200 μL pipette tip, washed three times with PBS, and cultured in cell supernatant with or without NGF protein. The final NGF concentration used was 50 ng / μL. Cells were photographed at 24 and 48 hours, and migration rate was calculated. The results are shown in Figures 2 and 9. Compared with the control, the wound healing rate was significantly increased in the group treated with cell supernatant containing mRNA-expressing proteins. Example 4: mRNA encapsulation and in vivo expression 4.1 mRNA encapsulation The main components of LNP include: ionizable lipids, cholesterol, DSPC, and DMG-PEG2000. Among them, the ionizable lipids are selected from one or more combinations of SM102, MC3, DOTAP, ALC-0315, H1, H2, H3, H4, H5, H6, H7, and H8. When the ionizable lipid is one of SM102, MC3, DOTAP, ALC-0315, H1, H2, H3, H4, H5, H6 or H8, the molar proportions of ionizable lipid, cholesterol, DSPC and DMG-PEG2000 in LNP are 48.5%, 38.9%, 11.1% and 1.5%, respectively, and the nitrogen-phosphorus ratio is 4.8 (i.e., the mass ratio of LNP to mRNA is 20:1). 4.1.1 Lipid-ethanol solution: Mix the required ionizable lipids, cholesterol, DSPC and DMG-PEG2000 in anhydrous ethanol at the desired molar ratio to prepare a lipid-ethanol solution with a total lipid concentration of 8 mM for later use. 4.1.2 mRNA-acetic acid buffer: Dilute the mRNA stock solution with acetate buffer (200mM, pH 5.0) to a suitable concentration and set aside. 4.1.3 Encapsulation: The mRNA-acetic acid buffer and lipid-ethanol solution were mixed at a flow rate ratio of 3:1 (buffer phase / ethanol phase) and a total flow rate of 3.6 mL / min using a microfluidic system (syringe pump: SPM, Duko Industrial; mixing chip: LNP-B0, FluidicLab). The mixed sample was dialyzed overnight at 4°C with 34 volumes of Tris salt buffer (20.5 mM, pH 7.5, containing 8.95% sucrose) (25 or 30 kDa MwCO) to obtain LNPs encapsulating mRNA. The ratio of LNP to mRNA is shown in Tables 8.1 and 8.2. To ensure accurate weighing of each component, the amount can be scaled up and prepared proportionally. Table 8.1 Components and Dosage Table 8.2 Components and Dosage 4.2 In vivo expression in rats mRNA-LNP (0.1 mg / kg) was injected into the tail vein of rats (where the cationic lipid in LNP is H7). After administration, blood samples were collected from mice at 0.5, 1, 2, 4, 6, 8, 10, 12, 24, 32, 48, 96, and 120 h to detect the concentration of NGF in the serum. The pharmacokinetic parameters are shown in Table 9. Table 9 The half-life of the recombinant fusion mRNA is significantly prolonged, and a single dose can maintain its effect for a longer period of time, which is beneficial for preparing it into a long-acting drug, reducing the frequency of use, and improving convenience and patient compliance. The cationic lipids in LNP were replaced with H8, and the above experiment was repeated. The results showed that the expression data of mRNA-LNP prepared by the three groups of SEQ ID NO:96, 880, and 881 are as shown above. The results indicate that after one administration, the expression levels of SEQ ID NO:880 and SEQ ID NO:881 in rats were significantly better than those of SEQ ID NO:96. Example 5: Pharmacodynamic experiment of a mouse model of benzalkonium chloride-induced neurotrophic keratitis. I. Pharmacological Experiments in Mouse Models Forty healthy female C57BL / 6J mice were randomly selected and a neurotrophic keratitis model was induced in mice with benzalkonium chloride (0.1%, bilateral modeling, twice a day, 5 μL / eye). Day 1 of modeling was designated as D1. On D7, 30 animals with a significant decrease in corneal sensation were selected based on corneal perception and randomly divided into 5 groups: model control group, positive control group, high-dose administration group (Qid), medium-dose administration group (Bid), and low-dose administration group (Qd). From day 8 to day 21, all animals were maintained in the model (method as before). The model control group received eye drops of diluted mRNA-LNP solution (20.5 mM Tris buffer containing 8.95% sucrose, pH 7.5) four times a day, 5 μL per eye each time. The positive control group received eye drops of Recombinant human beta NGF protein at a concentration of 40 ng / μL four times a day, 5 μL per eye each time. The Qid, Bid, and Qd groups all received eye drops of mRNA-LNP (SEQ ID NO: 96, SM102) at a concentration of 500 ng / μL, 5 μL per eye each time. The high-dose administration group (Qid) consisted of administering mRNA-LNP eye drops four times a day. The medium-dose administration group (Bid) consisted of two eye drops of mRNA-LNP per day. The low-dose group (Qd) consisted of one eye drop of mRNA-LNP once a day. During the experiment, animals underwent routine clinical observation daily and were weighed twice per week. On days 1, 7, 14, and 21, slit-lamp examination, corneal sensation testing, and corneal fluorescein staining were performed on each group of animals. All experimental data are expressed as mean ± standard error for each group of mice. 5.1 General Clinical Indicators During the trial, observations were made at least once a day, including but not limited to physical appearance, coat, general behavior, mental state, glandular secretions, skin and mucous membrane color, respiratory status, fecal characteristics, genitals, and death. The results showed that no abnormal clinical reactions were observed in any of the groups of animals. 5.2 Weight During the experiment, there were no statistically significant differences in body weight among the groups at each time point (P>0.05), as shown in Figure 3. 5.3 Corneal Sensation Before modeling (D1), there was no statistically significant difference in corneal perception among the groups (P>0.05). Before drug administration (D7), corneal perception in all groups was significantly reduced compared to D1, but there was no statistically significant difference between groups (P>0.05), indicating that eye drops of benzalkonium chloride solution can reduce corneal perception in mice, and there was no difference in the degree of lesions among the groups before drug administration. Data were collected from both eyes of each mouse, and the data for each group are the mean ± standard error of the data collected from all mice in that group. At D14 and D21, corneal perception in the mRNA-LNP administration group (Qd, Bid, Qid) and the positive control group was significantly higher than that in the model control group (P≤0.05). There was no significant difference in corneal perception between the Qd and Bid administration groups and the positive control group. The corneal perception recovery effect was more obvious in the Qid administration group, as shown in Figure 4. 5.4 Corneal fluorescein sodium staining indicators Before modeling (D1), the IOD value and area of ​​the stained corneal region (evaluation index of corneal fluorescein staining) of all groups were 0. Before drug administration (D7), the corneal fluorescein staining evaluation index increased significantly among all groups, but there was no statistically significant difference between groups (P>0.05), indicating that benzalkonium chloride solution can induce corneal epithelial damage in mice by bilateral eye drops, and the degree of lesions was similar among the groups before drug administration. Data were collected from both eyes of each mouse, and the data for each group are the mean ± standard error of the data collected from all mice in the group. At D14 and D21, the corneal fluorescein sodium staining evaluation index of the mRNA-LNP administration group (Qd, Bid, Qid, and positive control group) was significantly lower than that of the model control group (P≤0.05). At D14, there was no significant difference in corneal staining area and IOD value of corneal staining area between the Qd administration group and the positive control group. The corneal sensation recovery effect of the Qid and Bid administration groups was better, indicating that the corneal sensation recovery speed of mice in the Qid and Bid administration groups was faster during D7-D14, as shown in Figure 5-6. 5.5 Corneal neuropathy At day 22, mice were sacrificed and their left eyes were examined. The corneal neuropathy evaluation index of the mRNA-LNP treatment group (Qd, Bid, Qid) and the positive control group was significantly lower than that of the model control group (P≤0.05), indicating that the mRNA-LNP treatment group (Qd, Bid, Qid) showed a good therapeutic effect on corneal neuropathy in mice, and the effect was comparable to that of the positive control group, as shown in Figure 7. In summary, benzalkonium chloride solution eye drops successfully induced a mouse model of neurotrophic keratitis. The mRNA-LNP administration groups (Qd, Bid, and Qid) showed significant improvement in corneal sensory impairment and corneal epithelial damage in the model mice after continuous eye drops for 7 and 14 days, respectively. Among them, the Bid and Qid administration groups showed a faster recovery rate in corneal sensory perception during the first week of administration. II. Mouse Model Drug Efficacy Experiment 2 Fifty healthy female C57BL / 6J mice were randomly selected and a neurotrophic keratitis model was induced in mice with benzalkonium chloride (0.1%, bilateral modeling, twice a day, 5 μL / eye). Day 1 of modeling was designated as D1. On D7, 40 animals with a significant decrease in corneal sensation were selected based on corneal perception and randomly divided into 5 groups: model control group, positive control group, high-dose group (SEQ ID NO:96-Qid), medium-dose group (SEQ ID NO:880-Bid), and medium-dose group (SEQ ID NO:881-Bid). From day 8 to day 21, all animals were maintained in the model (using the same method as before). The model control group received eye drops of diluted mRNA-LNP solution (20.5 mM Tris buffer containing 8.95% sucrose, pH 7.5) in both eyes, 5 μL per eye each time, 4 times a day. The positive control group received eye drops of Recombinant human beta NGF protein at a concentration of 40 ng / μL in both eyes, 5 μL per eye each time, 4 times a day. SEQ96-Qid is administered as 500 ng / μL eye drops four times a day to both eyes, with each drop being 5 μL per eye. SEQ880-Bid is a group that administers eye drops twice a day to both eyes at a concentration of 500 ng / μL, with 5 μL administered to each eye each time. SEQ881-Bid is a group that administers eye drops twice a day to both eyes at a concentration of 250 ng / μL, with 5 μL administered to each eye each time. The cationic lipid in the LNP in the above samples is H8. During the experiment, animals underwent routine clinical observation daily and were weighed twice per week. On days 1, 7, 14, and 21, slit-lamp examination, corneal sensation testing, and corneal fluorescein staining were performed on each group of animals. All experimental data are expressed as mean ± standard error for each group of mice. 5.1 General Clinical Indicators During the trial, observations were made at least once a day, including but not limited to physical appearance, coat, general behavior, mental state, glandular secretions, skin and mucous membrane color, respiratory status, fecal characteristics, genitals, and death. The results showed that no abnormal clinical reactions were observed in any of the groups of animals. 5.2 Weight During the experiment, there were no statistically significant differences in body weight among the groups at any time point (P>0.05). As shown in Figure 11, there were no significant differences in body weight among the groups of mice. 5.3 Corneal Sensation Before modeling (D1), there was no statistically significant difference in corneal perception among the groups (P>0.05). Before drug administration (D7), corneal perception in all groups was significantly reduced compared to D1, but there was no statistically significant difference between groups (P>0.05), indicating that eye drops of benzalkonium chloride solution can reduce corneal perception in mice, and there was no difference in the degree of lesions among the groups before drug administration. Data were collected from both eyes of each mouse, and the data for each group are the mean ± standard error of the data collected from all mice in that group. As shown in Figure 12, at D14 and D21, corneal sensation was significantly higher in all mRNA-treated groups and the positive control group than in the model control group (P≤0.05). The effects of the three treatment groups, SEQ880-Bid, SEQ881-Bid, and SEQ96-Qid, were comparable. 5.4 Corneal fluorescein sodium staining indicators Before modeling (D1), the corneal staining area (corneal fluorescein sodium staining evaluation index) of all groups was 0. Before drug administration (D7), the corneal fluorescein sodium staining evaluation index increased significantly among all groups, but there was no statistically significant difference between groups (P>0.05), indicating that benzalkonium chloride solution can induce corneal epithelial damage in mice by bilateral eye drops, and the degree of lesions was similar in all groups before drug administration. Data were collected from both eyes of each mouse, and the data for each group are the mean ± standard error of the data collected from all mice in the group. As shown in Figure 13, at D14 and D21, the corneal fluorescein staining area in all mRNA treatment groups and the positive control group was significantly lower than that in the model control group (P≤0.05). The effects of the three treatment groups, SEQ880-Bid, SEQ881-Bid, and SEQ96-Qid, were comparable. The cationic lipid in the LNP of the mRNA-LNP sample used in the following experiments is H8. Example 6: Pharmacological Experiment of a Ganglion Cell Injury Model Induced by Anterior Chamber Perfusion with Physiological Saline in SD Rats Rats were anesthetized by intraperitoneal injection of ketamine (5 mL / kg), and physiological saline was instilled into the anterior chamber at a height of 220 cm for 90 minutes. The model animals were randomly divided into four groups of six: a model control group, an 880 eye drop group, an 881 eye drop group, an 880 subconjunctival injection group, and an 881 subconjunctival injection group; normal rats served as the blank control group. The sample diluent was 20.5 mM Tris buffer containing 8.95% sucrose at pH 7.5. (1) The model control group 1 was injected subconjunctivally with sample dilution once a week, 50 μL per eye each time, for 6 consecutive weeks. (2) 880 eye drop group: The mRNA-LNP sequence of the drug is SEQ ID NO:880. The drug is administered 4 times a day, 5 μL per eye each time, for 6 consecutive weeks. (3) 881 eye drop group: The mRNA-LNP sequence of the drug is SEQ ID NO:881. The drug is administered 4 times a day, 5 μL per eye each time, for 6 consecutive weeks. (4) 880 subconjunctival injection group: The mRNA-LNP sequence was SEQ ID NO:880. It was injected once a week, 50 μL per eye each time, for 6 consecutive weeks. (5) 881 subconjunctival injection group: The mRNA-LNP sequence of the drug is SEQ ID NO:881. It is injected once a week, 50 μL per eye each time, for 6 consecutive weeks. The mRNA concentration in the four treatment groups was 500 ng / μL. The results are shown in Figure 14: (1) Compared with the blank control group, the number of surviving retinal ganglion cells in the model control group was significantly reduced; (2) Compared with the model control group, the number of surviving retinal ganglion cells in the SEQ ID NO:880 and SEQ ID NO:881 drug administration groups was significantly increased, with the subconjunctival injection drug administration group showing a greater increase in the number of surviving retinal ganglion cells. Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. For those skilled in the art, other variations or modifications can be made based on the above description and ideas, and it is neither necessary nor possible to exhaustively describe all implementation methods here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. An mRNA encoding nerve growth factor, characterized in that, The mRNA comprises a coding region, a 5'UTR, and a 3'UTR; the coding region is selected from SEQ ID NO:21 or a sequence having at least 98% similarity to SEQ ID NO:21; the 5'UTR is selected from SEQ ID NO:1001; and the 3'UTR is selected from SEQ ID NO:1002.

2. An mRNA encoding nerve growth factor, characterized in that, The mRNA contains a coding region; the coding region is selected from SEQ ID NO:1134 or a sequence that has at least 98% similarity to SEQ ID NO:1134.

3. The mRNA according to claim 2, characterized in that, The mRNA also includes a 5'UTR, the sequence of which is selected from SEQ ID NO:1001.

4. The mRNA encoding nerve growth factor according to claim 2 or 3, characterized in that, The mRNA also includes a 3'UTR, the sequence of which is selected from SEQ ID NO:1002.

5. The mRNA encoding nerve growth factor according to any one of claims 1 to 4, characterized in that, The mRNA also includes poly A, which contains at least 70 adenosine molecules.

6. The mRNA encoding nerve growth factor according to any one of claims 1 to 5, characterized in that, The mRNA includes: (ii) 5'UTR; the 5'UTR is selected from SEQ ID NO:1001; (iii) The coding region; said coding region is selected from SEQ ID NO:21, SEQ ID NO:1134 or a sequence having a similarity of at least 98% to one of SEQ ID NO:21, SEQ ID NO:1134; (iv) 3'UTR; the 3'UTR is selected from SEQ ID NO:1002; (v)poly A; the poly A contains at least 70 adenosine molecules.

7. The mRNA encoding nerve growth factor according to any one of claims 1 to 6, characterized in that, The poly A contains 70-140 adenosine, or 70-120 adenosine, or 80-120 adenosine, or 90-120 adenosine, or 100-140 adenosine, or 90-110 adenosine, or 90-100 adenosine, or 100-110 adenosine.

8. The mRNA encoding nerve growth factor according to any one of claims 1 to 7, characterized in that, The nucleotide sequence of the mRNA is shown in SEQ ID NO:96, SEQ ID NO:880 or SEQ ID NO:

881.

9. An mRNA encoding nerve growth factor, characterized in that, The mRNA contains a coding region; the coding region contains one or more ORFs; the ORFs are selected from SEQ ID NO:20-27 or sequences that are at least 98% similar to one of SEQ ID NO:20-27.

10. An mRNA encoding nerve growth factor, characterized in that, The mRNA contains a coding region; the coding region contains one or more ORFs; the ORFs are selected from SEQ ID NO:1041-1043, SEQ ID NO:1091-1093, SEQ ID NO:1131-1133, or sequences that have at least 98% similarity to one of SEQ ID NO:1041-1043, SEQ ID NO:1091-1093, SEQ ID NO:1131-1133.

11. The mRNA encoding nerve growth factor according to claim 9 or 10, characterized in that, When the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by any one of the sequences in SEQ ID NO:28, 29 or 33; Alternatively, when the coding region contains at least three ORFs, the interval sequences may be the same or different.

12. The mRNA encoding nerve growth factor according to claim 11, characterized in that, When the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO: 1011-1013, 1021-1023, 1031-1033, 1051-1053, 1061-1063, 1071-1073, 1081-1083, 1101-1103, 1111-1113, 1121-1123, and 1131-1133.

13. The mRNA encoding nerve growth factor according to any one of claims 9 to 12, characterized in that, The mRNA also includes a 5'UTR, the sequence of which is selected from any one of SEQ ID NO:1-15.

14. The mRNA encoding nerve growth factor according to any one of claims 9 to 13, characterized in that, The mRNA also includes a 3'UTR, the sequence of which is selected from any one of SEQ ID NO:16-19.

15. The mRNA encoding nerve growth factor according to any one of claims 9 to 14, characterized in that, The mRNA also includes poly A, which contains at least 70 adenosine molecules.

16. The mRNA encoding nerve growth factor according to claim 15, characterized in that, The poly A contains 70-140 adenosine, or 70-120 adenosine, or 80-120 adenosine, or 90-120 adenosine, or 100-140 adenosine, or 100-120 adenosine.

17. The mRNA encoding nerve growth factor according to any one of claims 9 to 16, characterized in that, The mRNA includes: (ii) 5'UTR; the 5'UTR is selected from any one of SEQ ID NO:1-15; (iii) A coding region; the coding region contains one or more ORFs; the ORFs are selected from SEQ ID NO:20-27 or sequences that are at least 98% similar to one of SEQ ID NO:20-27; (iv) 3'UTR; the 3'UTR is selected from any one of SEQ ID NO:16-19; (v)poly A; the poly A contains at least 70 adenosine molecules.

18. The mRNA encoding nerve growth factor according to claim 17, characterized in that, The ORF is one of the following: (1) The ORF is SEQ ID NO:20; (2) The ORF is SEQ ID NO:21; (3) The ORF is SEQ ID NO:22; (4) The ORF is SEQ ID NO:23; (5) The ORF is SEQ ID NO:24; (6) The ORF is SEQ ID NO:25; (7) The ORF is SEQ ID NO:26; (8) The ORF is SEQ ID NO:

27.

19. The mRNA encoding nerve growth factor according to claim 17 or 18, characterized in that, The 5'UTR sequence is one of the following: (1) The 5'UTR sequence is SEQ ID NO:1; (2) The 5'UTR sequence is SEQ ID NO:2; (3) The 5'UTR sequence is SEQ ID NO:3; (4) The 5'UTR sequence is SEQ ID NO:4; (5) The 5'UTR sequence is SEQ ID NO:5; (6) The 5'UTR sequence is SEQ ID NO:6; (7) The 5'UTR sequence is SEQ ID NO:7; (8) The 5'UTR sequence is SEQ ID NO:8; (9) The 5'UTR sequence is SEQ ID NO:9; (10) The 5'UTR sequence is SEQ ID NO:10; (11) The 5'UTR sequence is SEQ ID NO:11; (12) The 5'UTR sequence is SEQ ID NO:12; (13) The 5'UTR sequence is SEQ ID NO:13; (14) The 5'UTR sequence is SEQ ID NO:14; (15) The 5'UTR sequence is SEQ ID NO:

14. ID NO:

15.

20. The mRNA encoding nerve growth factor according to any one of claims 17 to 19, characterized in that, The sequence of the 3'UTR is one of the following: (1) The 3'UTR sequence is SEQ ID NO:16; (2) The 3'UTR sequence is SEQ ID NO:17; (3) The 3'UTR sequence is SEQ ID NO:18; (4) The 3'UTR sequence is SEQ ID NO:

19.

21. The mRNA encoding nerve growth factor according to any one of claims 17 to 20, characterized in that, When the coding region contains at least two ORFs, the sequences of the ORFs may be the same or different; each ORF is separated by any one of the sequences in SEQ ID NO:28, 29 or 33.

22. The mRNA encoding nerve growth factor according to claim 21, characterized in that, When the coding region contains at least three ORFs, the interval sequences may be the same or different; When the coding region contains at least two ORFs, the coding region is selected from any one of SEQ ID NO: 1011-1013, 1021-1023, 1031-1033, 1041-1043, 1051-1053, 1061-1063, 1071-1073, 1081-1083, 1091-1093, 1101-1103, 1111-1113, 1121-1123, and 1131-1133.

23. The mRNA encoding nerve growth factor according to any one of claims 1 to 22, characterized in that, The mRNA contains at least one chemically modified nucleoside.

24. The mRNA encoding nerve growth factor according to claim 23, characterized in that, The chemically modified nucleoside is selected from pseudouracil, N1-methyl-pseudorazine, 1-ethylpseudorazine, 2-thiouracil, 4'-thiouracil, 5-methyluracil, or 5-methoxyuracil, or any combination thereof.

25. The mRNA encoding nerve growth factor according to claim 23 or 24, characterized in that, The chemically modified nucleosides are selected from one of the following: (1) The chemically modified nucleosides are selected from pseudouracil and / or N1-methyl-pseudorazine; (2) The chemically modified nucleoside is selected from 1-ethylpseuuridine; (3) The chemically modified nucleoside is selected from 2-thiouracil; (4) The chemically modified nucleoside is selected from 4'-thiouracil; (5) The chemically modified nucleoside is selected from 5-methyluracil; (6) The chemically modified nucleoside is selected from 5-methoxyuracil.

26. The mRNA encoding nerve growth factor according to any one of claims 23 to 25, characterized in that, 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%.

27. The mRNA encoding nerve growth factor according to any one of claims 1 to 27, characterized in that, The amino acid sequence encoded by the mRNA includes any one of SEQ ID NO:2001, 2002 or 2015.

28. The mRNA encoding nerve growth factor according to any one of claims 1 to 27, characterized in that, The mRNA also includes a 5' cap structure.

29. The mRNA encoding nerve growth factor according to claim 28, characterized in that, The 5' cap structure is selected from Cap0, Cap1, Cap2, or the structure shown in Formula I: Where R1 represents H and C 1-4 Alkyl, C 1-4 Halogenated alkyl, phenyl, C 1-4 Alkyl-substituted phenyl, halophenyl, benzyl, C 1-4 Alkyl-substituted benzyl or halobenzyl; R2 represents OH, halogen, or OC. 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Alkoxy-substituted C 1-4 Alkyl, C 1-4 Alkylamine substituent C 1-4 Alkyl, C 1-4 Amide group substituted C 1-4 Alkyl, benzyloxy, halobenzyloxy, benzyloxy-substituted C 1-4 Alkyl or halobenzyloxy-substituted C 1-4 alkyl; R3 and R4 are independently H and C. 1-4 Alkyl or C 1-4 Halogenated alkyl groups; R5 represents H and C. 1-4 Alkoxy, F, Cl, I, or N3; R6 represents H and C. 1-4 Alkyl, C 1-4 alkenyl, C 1-4 alkynyl, phenyl, C 1-4 Alkyl-substituted phenyl, halophenyl, benzyl, C 1-4 Alkyl-substituted benzyl or halobenzyl; B1 and B2 are each independently a base G, A, U or C, and substituted bases include, but are not limited to, 6-MeA, 5-MeU or 5-MeC.

30. The mRNA encoding nerve growth factor according to claim 29, characterized in that, R6 is any one of the following a1-a6: a1. R6 is H, C 1-4 Alkyl, C 1-4 alkenyl, C 1-4 Alkyne, phenyl, halophenyl, benzyl, or halobenzyl; a2. R6 is H, C 1-4 Alkyl, phenyl, halophenyl, benzyl or halobenzyl; a3. The R6 is H, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, phenyl, benzyl, 3-fluorophenyl, 4-fluorophenyl, 3,5-difluorophenyl, 3-chlorophenyl, 4-chlorophenyl, 3,5-dichlorophenyl, 3-fluorobenzyl, 4-fluorobenzyl, 3,5-difluorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl or 3,5-dichlorobenzyl; a4. The R6 is H, methyl, ethyl, propyl, isopropyl, phenyl, benzyl, 3-fluorophenyl, 4-fluorophenyl, 3-chlorophenyl, 4-chlorophenyl, 3-fluorobenzyl, 4-fluorobenzyl, 3-chlorobenzyl or 4-chlorobenzyl; a5. The R6 is H, methyl, ethyl, isopropyl, phenyl, benzyl, 4-fluorophenyl, 4-chlorophenyl, 4-fluorobenzyl or 4-chlorobenzyl; a6. The R6 is H, methyl, ethyl, isopropyl, phenyl, benzyl, 4-fluorophenyl or 4-fluorobenzyl.

31. The mRNA encoding nerve growth factor according to claim 29 or 30, characterized in that, R3 and R4 are any one of the following b1-b4: b1. R3 and R4 are each independently H, methyl, ethyl, isopropyl, tert-butyl, trifluoromethyl, trifluoroethyl or trifluoroisopropyl; b2. R3 and R4 are each independently H, methyl, ethyl, isopropyl, trifluoromethyl, or trifluoroethyl; b3. R3 and R4 are each independently H, methyl, ethyl, or trifluoromethyl; b4. R3 and R4 are each independently H, methyl or trifluoromethyl.

32. The mRNA encoding nerve growth factor according to any one of claims 29 to 31, characterized in that, R2 is any one of the following c1-c4: c1. R2 is OH, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Alkoxy-substituted C 1-4 Alkyl, C 1-4 Alkylamine substituent C 1-4 Alkyl, C 1-4 Amide group substituted C 1-4 Alkyl, benzyloxy or benzyloxy-substituted C 1-4 alkyl; c2. The R2 is OH, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-3 Alkoxy-substituted C 1-3 Alkyl, C 1-3 Alkylamine substituent C 1-3 Alkyl, C 1-3 Amide group substituted C 1-3 Alkyl, benzyloxy or benzyloxy-substituted C 1-3 alkyl; c3. The R2 is OH, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-3 Alkoxy-substituted C 1-3 Alkyl, C 1-3 Alkylamine substituent C 1-3 Alkyl or C 1-3 Amide group substituted C 1-3 alkyl; c4. The R2 is OH, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl or C 1-3 Alkoxy-substituted C 1-3 alkyl.

33. The mRNA encoding nerve growth factor according to any one of claims 29 to 32, characterized in that, R1 is any one of the following d1-d5: d1: R1 is H, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoroisopropyl, phenyl, 2-methyl-phenyl, 3-methyl-phenyl, 4-methyl-phenyl, 2,6-dimethyl-phenyl, 3,5-dimethyl-phenyl, 2-ethyl-phenyl, 3-ethyl-phenyl, 4-ethyl-phenyl, 2,6-diethyl-phenyl, 3,5-diethyl-phenyl, 2-chloro-phenyl, 3-chloro-phenyl, 4-chloro-phenyl, 2,6-chloro-phenyl, 3,5-chloro-phenyl, 2-fluoro-phenyl, 3-fluoro-phenyl , 4-fluoro-phenyl, 2,6-fluoro-phenyl, 3,5-fluoro-phenyl, benzyl, 2-methyl-benzyl, 3-methyl-benzyl, 4-methyl-benzyl, 2,6-dimethyl-benzyl, 3,5-dimethyl-benzyl, 2-ethyl-benzyl, 3-ethyl-benzyl, 4-ethyl-benzyl, 2,6-diethyl-benzyl, 3,5-diethyl-benzyl, 2-chloro-benzyl, 3-chloro-benzyl, 4-chloro-benzyl, 2,6-chloro-benzyl, 3,5-chloro-benzyl, 2-fluoro-benzyl, 3-fluoro-benzyl, 4-fluoro-benzyl, 2,6-fluoro-benzyl or 3,5-fluoro-benzyl; d2: R1 is H, methyl, ethyl, propyl, isopropyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoropropyl, trifluoroisopropyl, phenyl, 3-methyl-phenyl, 4-methyl-phenyl, 3,5-dimethyl-phenyl, 3-ethyl-phenyl, 4-ethyl-phenyl, 3,5-diethyl-phenyl, 3-chloro-phenyl, 4-chloro-phenyl, 3,5-chloro-phenyl, 3-fluoro-phenyl, 4-fluoro-phenyl, 3,5-fluoro-phenyl, benzyl, 3-methyl-benzyl, 4-methyl-benzyl, 3,5-dimethyl-benzyl, 3-ethyl-benzyl, 4-ethyl-benzyl, 3,5-diethyl-benzyl, 3-chloro-benzyl, 4-chloro-benzyl, 3,5-chloro-benzyl, 3-fluoro-benzyl, 4-fluoro-benzyl or 3,5-fluoro-benzyl; d3: R1 is H, methyl, ethyl, isopropyl, tert-butyl, trifluoromethyl, trifluoroethyl, trifluoroisopropyl, phenyl, 4-methyl-phenyl, 4-ethyl-phenyl, 4-chloro-phenyl, 4-fluoro-phenyl, benzyl, 4-methyl-benzyl, 4-ethyl-benzyl, 4-chloro-benzyl or 4-fluoro-benzyl; d4: R1 is H, methyl, ethyl, isopropyl, trifluoromethyl, trifluoroethyl, benzyl, 4-methyl-benzyl, 4-ethyl-benzyl, 4-chloro-benzyl, or 4-fluoro-benzyl; d5: R1 is H, methyl, ethyl, isopropyl, trifluoromethyl, trifluoroethyl, benzyl, 4-methyl-benzyl or 4-fluoro-benzyl.

34. The mRNA encoding nerve growth factor according to any one of claims 29 to 33, characterized in that, R5 is any one of the following e1-e6: e1: R5 is H, methyl, ethyl, propyl, isopropyl, tert-butyl, F, or N3; e2: R5 is H, methyl, ethyl, isopropyl, F or N3; e3: R5 is H, methyl, ethyl, F or N3; e4: R5 is H or F; e5: R5 is H or methyl; e6: R5 is F or N3.

35. A composition, characterized in that, It comprises the mRNA encoding nerve growth factor as described in any one of claims 1 to 34 and a delivery agent.

36. The composition according to claim 35, characterized in that, The delivery agent comprises lipid nanoparticles; the lipid nanoparticles contain ionizable lipids, phospholipids, cholesterol, and PEG lipids.

37. The composition according to claim 36, characterized in that, The molar ratio of ionizable lipids, phospholipids, cholesterol, and PEG lipids in the lipid nanoparticles is any one of the following c1-c4: c1. The molar ratio of ionizable lipids, phospholipids, cholesterol and PEG lipids is 30–60:0–30:18.5–48.5:0–10; The molar ratio of c2 ionizable lipids, phospholipids, cholesterol and PEG lipids is 35-55:0-25:20-45:0.5-8; The molar ratio of c3 ionizable lipids, phospholipids, cholesterol and PEG lipids is 40-52:5-20:25-40:0.5-5; The molar ratio of C4 ionizable lipids, phospholipids, cholesterol and PEG lipids is 45-52:8-15:30-40:1-5; The molar ratio of c5 ionizable lipids, phospholipids, cholesterol and PEG lipids is 50:10:38.5:1.5 or 48.5:11.1:38.9:1.

5.

38. The composition according to any one of claims 35 to 37, characterized in that, The ionizable lipid is shown in Formula II: Where n0 is an integer from 0 to 4; n1 is an integer from 2 to 7; n2 is an integer from 4 to 10; n3 is an integer from 2 to 8; n4 is an integer from 2 to 8; n5 is an integer from 2 to 8. Alternatively, the ionizable lipid may be one or more combinations of SM102, MC3, DOTAP, ALC-0315, H1, H2, H3, H4, H5, H6, H7, or H8.

39. The composition according to any one of claims 35 to 38, characterized in that, The phospholipid is DSPC or DOPE; the PEG lipid has a molecular weight of PEG, PEG-modified compounds or mixtures thereof of 1000 Da to 20 kDa.

40. The composition according to any one of claims 35 to 39, characterized in that, The composition also includes pharmaceutically acceptable excipients.

41. The use of the mRNA encoding nerve growth factor according to any one of claims 1 to 34 or the composition according to any one of claims 35 to 40 in the preparation of a medicament for the prevention and / or treatment of, but not limited to, corneal diseases, optic neuropathy, conjunctival diseases, limbal stem cell deficiency, dry eye, peripheral neuropathy or idiopathic facial nerve palsy, retinal diseases, central nervous system injury and / or degeneration-related conditions.

42. The application according to claim 41, characterized in that, The corneal lesions are selected from keratoconus, phototoxic keratopathy, persistent corneal epithelial defects, corneal ulcers, corneal dystrophy and degeneration, dry keratoconjunctivitis, or neurotrophic keratitis. The optic neuropathy is selected from glaucoma, ischemic optic neuropathy, degenerative, traumatic, Leber hereditary optic neuropathy, and congenital optic atrophy; The dry eye syndrome is selected from aqueous dry eye, lipid abnormality dry eye, mucin abnormality dry eye, tear dynamics abnormality dry eye and mixed dry eye.

43. The application according to claim 41, characterized in that, The peripheral neuropathy includes, but is not limited to, HIV-associated sensory neuropathy, diabetic polyneuropathy, toxic peripheral neuropathy, and traumatic peripheral nerve injury.

44. The application according to claim 41, characterized in that, The retinal lesions are selected from diabetic retinopathy, retinal detachment, age-related macular degeneration, macular degeneration, macular atrophy, macular hole, macular edema, retinopathy of prematurity, retinal vascular occlusion, phototoxic retinopathy, or epiretinal membrane disease.

45. The application according to claims 41, characterized in that, The central nervous system injury and / or degeneration-related conditions include, but are not limited to, Alzheimer's disease, Parkinson's syndrome, and frontotemporal dementia.

46. ​​The application according to any one of claims 41 or 45, characterized in that, The drug is a topical medication.

47. The application according to claim 46, characterized in that, The drug is an injection, a topical preparation, a nasal spray, or an eye drop.

48. A method for preventing and / or treating corneal pathological conditions, optic neuropathy, conjunctival pathological conditions, limbal stem cell deficiency or dry eye, peripheral neuropathy, or idiopathic facial nerve palsy, characterized in that, Administer to a subject in need a therapeutically effective amount of the mRNA encoding nerve growth factor as described in any one of claims 1 to 34 or the composition as described in any one of claims 35 to 40.

49. The method according to claim 48, characterized in that, It also includes administering other medications to the subject.

50. The method according to claim 48, characterized in that, The corneal pathological conditions are selected from keratoconus, phototoxic corneal lesions, persistent epithelial defects, corneal ulcers, corneal dystrophy and degeneration, dry keratoconjunctivitis, or neurotrophic keratitis. The optic nerve lesions mentioned are selected from glaucoma and ischemic, degenerative, traumatic, hereditary and congenital optic nerve lesions; The dry eye syndrome is selected from aqueous-deficient dry eye, lipid-dysplastic dry eye, mucin-dysplastic dry eye, tear dynamics-dysplastic dry eye, and mixed dry eye. The peripheral neuropathy includes, but is not limited to, HIV-associated sensory neuropathy, diabetic polyneuropathy, toxic peripheral neuropathy, and traumatic peripheral nerve injury.

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