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EP-4737571-A1 - VARICELLA ZOSTER VIRUS (VZV) VACCINE

EP4737571A1EP 4737571 A1EP4737571 A1EP 4737571A1EP-4737571-A1

Abstract

The present invention relates to a non-natural nucleic acid, a genetic engineering vector, a host cell, a delivery carrier, a pharmaceutical composition and use thereof, and a varicella zoster virus (VZV) vaccine, wherein the non-natural nucleic acid comprises a polynucleotide encoding a VZV gE protein or a fragment thereof.

Inventors

  • Li, Linxian

Assignees

  • Shenzhen Shenxin Biotechnology Co., Ltd.

Dates

Publication Date
20260506
Application Date
20240628

Claims (20)

  1. A non-natural nucleic acid comprising a polynucleotide encoding a VZV gE protein or a fragment thereof.
  2. The nucleic acid according to claim 1, wherein the amino acid sequence of the VZV gE protein comprises or is an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence set forth in SEQ ID NO: 1.
  3. The nucleic acid according to claim 1, wherein the VZV gE protein is a full-length gE protein.
  4. The nucleic acid according to claim 1, wherein the fragment of the VZV gE protein is one of the follows: (1) a truncated polypeptide of positions 23 to 623, positions 31 to 623, positions 1 to 561, positions 1 to 573, positions 1 to 543, positions 31 to 573 or positions 31 to 543 of the VZVgE protein; and (2) a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence of the truncated polypeptide of (1).
  5. The nucleic acid according to any one of claims 1-4, wherein the VZV gE protein or a fragment thereof further has a mutation; preferably, the mutation is one or both of the mutations: Y569A and Y582G.
  6. The nucleic acid according to any one of claims 1-5, wherein the VZV gE protein or a fragment thereof comprises or is one of the following polypeptides or proteins: (1) a polypeptide or protein having an amino acid sequence as set forth in one of SEQ ID NOs: 1-8; and (2) a polypeptide or protein comprising an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to one of the sequences SEQ ID NOs: 1-8.
  7. The nucleic acid according to any one of claims 1-6, wherein the VZV gE protein or a fragment thereof is further linked to a heterologous signal peptide and/or ferritin.
  8. The nucleic acid according to claim 7, wherein the heterologous signal peptide comprises one or more of: an IgE signal peptide and an IgGκ signal peptide; preferably, the amino acid sequence of the IgGκ signal peptide is set forth in SEQ ID NO: 9; and/or, the amino acid sequence of ferritin is set forth in SEQ ID NO: 10.
  9. The nucleic acid according to any one of claims 1-6, wherein the nucleic acid is RNA; preferably, the polynucleotide encoding the VZV gE protein or a fragment thereof is codon optimized; preferably, the polynucleotide encoding the VZV gE protein or a fragment thereof comprises or is one of the follows: (1) an RNA corresponding to a polynucleotide having a nucleotide sequence set forth in one of SEQ ID NOs: 11-18; and (2) an RNA corresponding to a polynucleotide comprising a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the nucleotide sequence set forth in one of SEQ ID NOs: 11-18.
  10. The nucleic acid according to any one of claims 1-9, wherein the nucleic acid is RNA, and the nucleic acid comprises or is one of the following RNAs: (1) an RNA corresponding to a polynucleotide having a nucleotide sequence set forth in one of SEQ ID NOs: 19-24; and (2) an RNA corresponding to a polynucleotide comprising a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the nucleotide sequence set forth in one of SEQ ID NOs: 19-24.
  11. The nucleic acid according to claim 8-10, wherein the nucleic acid is mRNA; preferably, the mRNA comprises at least one of a 5'-cap structure, a 5'-UTR, a 3'-UTR and a poly(A) tail.
  12. The nucleic acid according to claim 11, wherein the DNA sequence corresponding to the 5'-UTR is set forth in SEQ ID NO: 25; and/or, the DNA sequence corresponding to the 3'-UTR is set forth in SEQ ID NO: 26; and/or, the nucleotides constituting the poly(A) tail comprises at least 20, at least 40, at least 80, at least 100 or at least 120 A nucleotides; preferably, the nucleotides constituting the poly(A) tail comprises at least 20, at least 40, at least 80, at least 100 or at least 120 continuous A nucleotides; preferably, the nucleotides constituting the poly(A) tail comprises one or more nucleotides other than the A nucleotide; more preferably, the DNA sequence corresponding to the poly(A) tail is set forth in SEQ ID NO: 27.
  13. The nucleic acid according to any one of claims 1-8, wherein the nucleic acid is DNA; preferably, the DNA can be transcribed into RNA, preferably, the polynucleotide encoding the VZV gE protein or a fragment thereof is codon optimized; preferably, the polynucleotide encoding the VZV gE protein or a fragment thereof comprises or is one of the follows: (1) a polynucleotide having a nucleotide sequence set forth in one of SEQ ID NOs: 11-18; and (2) a polynucleotide comprising a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the nucleotide sequence set forth in one of SEQ ID NOs: 11-18.
  14. The nucleic acid according to claim 13, comprising or being one of the following polynucleotides: (1) a polynucleotide having a nucleotide sequence set forth in one of SEQ ID NOs: 19-24; and (2) a polynucleotide comprising a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the nucleotide sequence set forth in one of SEQ ID NOs: 19-24.
  15. The nucleic acid according to any one of claims 1-14, comprising a modified nucleotide; preferably, the nucleic acid comprises a modified nucleoside; preferably, the modified nucleoside comprises at least one of a modified uridine, a modified cytidine, a modified adenosine, and a modified guanosine.
  16. A genetic engineering vector, comprising the nucleic acid according to any one of claims 1-15, or comprising a polynucleotide that can be transcribed into the nucleic acid according to any one of claims 1-15.
  17. A host cell comprising the genetic engineering vector according to claim 16.
  18. A protein or polypeptide encoded by the nucleic acid according to any one of claims 1-15.
  19. A delivery carrier comprising the nucleic acid according to any one of claims 1-15, or the genetic engineering vector according to claim 16.
  20. The delivery carrier according to claim 19, wherein the delivery carrier is a lipid nanoparticle (LNP), a cationic liposome, a cationic protein or a lipopolyplex (LPP); preferably, the delivery carrier is a lipid nanoparticle comprising a cationic lipid comprising the following compound (IV), or a pharmaceutically acceptable salt or stereoisomer thereof: where: L 3 and L 4 are the same or different, and are each independently C1-C12 alkylene, C2-C12 alkenylene or C2-C12 alkynylene; preferably L 3 and L 4 are the same or different, and are each independently C3-C10 alkylene, C3-C10 alkenylene or C3-C10 alkynylene; further preferably, L 3 and L 4 are the same or different, and are each independently C3-C10 alkylene; most preferably, L 3 and L 4 are the same or different, and are each independently C5-C8 alkylene; G 4 and G 5 are the same or different, and are each independently -O-(C=O)-, -(C=O)-O-, -C(=O)-, -O-, -C(=O)-S- or -S-C(=O)-; preferably, G 4 and G 5 are the same or different, and are each independently -O-(C=O)-, -(C=O)-O-, -C(=O)- or -O-; most preferably, G 4 and G 5 are the same or different, and are each independently selected from -O-(C=O)- or -(C=O)-O-; R 18 and R 19 are the same or different, and are each independently C5-C27 alkyl or C5-C27 alkenyl containing one or more double bonds; preferably, R 18 and R 19 are the same or different, and are each independently C8-C20 alkyl or C8-C20 alkenyl containing one or more double bonds; further preferably, R 18 and R 19 are the same or different, and are each independently C9-C17 alkyl or C9-C18 alkenyl containing one or two double bonds; most preferably, R 18 and R 19 are the same or different, and are each independently or R 20 is halogen, hydroxyl, cyano, C1-C6 alkyl, nitro, C1-C6 alkoxy, C1-C6 alkylcarbonyloxy, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C1-C6 alkylcarbonylamino; preferably, R 20 is halogen, hydroxyl, cyano, C1-C6 alkoxy, C1-C6 alkylcarbonyloxy, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C1-C6 alkylcarbonylamino; further preferably, R 20 is halogen, hydroxyl, cyano, C1-C4 alkoxy, C1-C4 alkylcarbonyloxy, C1-C4 alkoxycarbonyl, C1-C4 alkylaminocarbonyl or C1-C4 alkylcarbonylamino; most preferably, R 20 is fluorine, hydroxyl, cyano, methoxy, acetoxy, methoxycarbonyl, butylaminocarbonyl or acetamido; z is 1, 2 or 3.

Description

Technical Field The present invention belongs to the field of biotechnology, and relates to a varicella-zoster virus (VSV) vaccine. Background Herpes zoster (HZ) is caused by the reactivation of previously infected varicella-zoster virus (VZV). The varicella-zoster virus is highly infectious, and the person infected by the virus is the main infectious source. The ways of infection mainly include direct contact and droplet transmission. The virus would be excreted through the lacrimal gland or saliva two days before the onset of skin rashes in patients. Viruses in blister fluid from varicella-zoster patients are infectious and are infected by contact with skin lesions or inhalation. The onset of herpes zoster is usually manifested as a skin rash that occurs on one side of the body and is accompanied by pain and itching, which can last two to four weeks, and papulovesicle and blisters may occur sysmatically, accompanied by severe pain. A common complication for herpes zoster is post-herpes zoster neuralgia, which can last months to years and seriously affect the patients' normal work and life. With the increase of age, the immune function of the human body decreases, and the incidence of herpes zoster increases sharply. The global incidence of people over 60-year-old can reach 8-12/1000 per year, while at least half or more of the elderly people over 85-year-old have suffered from herpes zoster at least once. Summary The present disclosure provides a non-natural nucleic acid that can induce a significant immune response in the body. In addition, the present disclosure further provides a polypeptide or protein encoded by the non-natural nucleic acid, a genetic engineering vector comprising the non-natural nucleic acid, a delivery carrier, a pharmaceutical composition and a vaccine comprising the non-natural nucleic acid, the polypeptide or protein encoded by the non-natural nucleic acid, or the genetic engineering vector, and use thereof. A non-natural nucleic acid comprising a polynucleotide encoding a VZV gE protein or a fragment thereof. In some embodiments, the amino acid sequence of the VZV gE protein comprises or is an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the VZV gE protein is a full-length gE protein. In some embodiments, the fragment of the VZVgE protein is one of the follows: (1) a truncated polypeptide of positions 23 to 623, 31 to 623, 1 to 561, 1 to 573, 1 to 543, 31 to 573 or 31 to 543 of the VZVgE protein; and(2) a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence of the truncated polypeptide of (1). In one embodiment, the VZV gE protein or fragment thereof further has a mutation; preferably, the mutation is one or both of mutations Y569A and Y582G. In one embodiment, the VZV gE protein or fragment thereof comprises or is one of the following polypeptides or proteins: (1) a polypeptide or protein having an amino acid sequence as set forth in one of SEQ ID NOs: 1-8; and(2) a polypeptide or protein comprising an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to one of the sequences SEQ ID NOs: 1-8. In some embodiments, the VZV gE protein or fragment thereof is further linked to a heterologous signal peptide and/or a ferritin. In some embodiments, the heterologous signal peptide comprises one or more of an IgE signal peptide and an IgGκ signal peptide. In some embodiments, the amino acid sequence of the IgGκ signal peptide is set forth in SEQ ID NO: 9. In some embodiments, the amino acid sequence of the ferritin is set forth in SEQ ID NO: 10. In some embodiments, the nucleic acid is RNA. In some embodiments, the polynucleotide encoding the VZV gE protein or fragment thereof is codon optimized. In some embodiments, the polynucleotide encoding the VZV gE protein or fragment thereof comprises or is one of the follows: (1) an RNA corresponding to a polynucleotide having a nucleotide sequence set forth in one of SEQ ID NOs: 11-18; and(2) an RNA corresponding to a polynucleotide comprising a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the nucleotide sequence set forth in one of SEQ ID NOs: 11-18. In some embodiments, the nucleic acid is RNA, and the nucleic acid comprises or is one of the following RNAs: (1) an RNA corresponding to a polynucleotide having a nucleotide sequence set forth in one of SEQ ID NOs: 19-24; and(2) an RNA corresponding to a polynucleotide comprising a nucleotide sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the nucleotide sequence set forth in one of SEQ ID NOs: 19-2