Search

BR-102025014234-A2 - Respiratory Syncytial Virus mRNA Vaccine, Method of Preparation and Use.

BR102025014234A2BR 102025014234 A2BR102025014234 A2BR 102025014234A2BR-102025014234-A2

Abstract

This disclosure pertains to the technical field of mRNA vaccines and relates particularly to a respiratory syncytial virus (RSV) vaccine and a method for its preparation and use. The vaccine provided in this disclosure comprises RNA encoding an RSV F protein or a variant thereof. The vaccine can prevent RSV infection and its complications.

Inventors

  • Gengshen Song
  • Limin Liang
  • Huanyu Wang
  • KAI DONG
  • Chen PAN
  • Wang Wang
  • Yuting Zhou
  • Xin CHAI
  • Jing Li
  • Xiaowei LANG
  • Jinyu Zhang

Assignees

  • Hangzhou Tianlong Pharmaceutical Co., Ltd

Dates

Publication Date
20260317
Application Date
20250709
Priority Date
20240709

Claims (20)

  1. 1. Immunogenic composition, characterized in that it comprises a ribonucleic acid (RNA) of a respiratory syncytial virus (RSV), wherein the RNA encodes a wild-type RSV F protein or a variant thereof; wherein a sequence of the wild-type RSV F protein is SEQ ID NO: 1; the variant of the RSV F protein comprises the mutations P102A, I379V and M447V; the variant of the RSV F protein is a truncated protein in which the amino acids at positions 550 to 574 of SEQ ID NO: 1 are truncated and the amino acids at positions 104 to 144 are replaced by a GS linker; and the variant of the RSV F protein further comprises a combination of site mutations selected from any of the following groups from Group 1 to Group 5, thus obtaining the RSV F protein variant selected from YK-RSV-061, YK-RSV-003 and/or YK-RSV-062:
  2. 2. Composition according to claim 1, characterized in that an amino acid sequence of the RSV F protein variant is: SEQ ID NO: 204, SEQ ID NO: 12 or SEQ ID NO: 208; or, a nucleotide sequence of an open reading frame (ORF) encoding the RSV F protein variant is: SEQ ID NO: 203, SEQ ID NO: 11 or SEQ ID NO: 207; or, an RSV RNA sequence is: SEQ ID NO: 205, SEQ ID NO: 13 or SEQ ID NO: 209; or, a DNA sequence encoding the RSV F protein variant is: SEQ ID NO: 202, SEQ ID NO: 10 or SEQ ID NO: 206.
  3. 3. Composition according to claim 1, characterized in that the RSV RNA further comprises a 5' untranslated region (UTR).
  4. 4. Composition according to claim 3, characterized in that an untranslated 5' region sequence is: SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220 or SEQ ID NO: 221.
  5. 5. Composition according to claim 1, characterized in that the RSV RNA further comprises a 3' untranslated region (UTR).
  6. 6. Composition according to claim 5, characterized in that an untranslated 3' region sequence is: SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224 or SEQ ID NO: 225.
  7. 7. Composition according to claim 1, characterized in that the RSV RNA further comprises a poly(A) tail.
  8. 8. Composition according to claim 1, characterized in that the RSV RNA further comprises a 5' cap structure.
  9. 9. Composition according to claim 8, characterized in that the cap 5' structure is: 7mG(5')ppp(5')NlmpNp.
  10. 10. Composition according to claim 1, characterized in that a sequence of an open reading frame (ORF) encoding the RSV F protein or a variant thereof in RSV RNA is codon-optimized.
  11. 11. Composition according to claim 10, characterized in that the ORF sequence comprises at least one base modification.
  12. 12. Composition according to claim 11, characterized in that the base modification comprises any one or more selected from the following: pseudouridine, N1-methylpseudouridine, N1-ethylpseudouridine, 2-thiouridine, 4'-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine and 2'-O-methyluridine.
  13. 13. Composition according to claim 1, characterized in that the composition is a vaccine and further comprises a pharmaceutically acceptable excipient.
  14. 14. Composition according to claim 13, characterized in that the pharmaceutically acceptable excipient comprises a lipid mixture and the lipid mixture is a lipid nanoparticle (LNP).
  15. 15. Composition, according to claim 13, characterized in that the vaccine is an mRNA vaccine; or, an effective dose of RSV RNA is 25 to 200 μg; or, an effective dose of RSV RNA is 50 to 100 μg.
  16. 16. Composition according to claim 14, characterized in that the lipid nanoparticle comprises a cationic lipid, a neutral lipid, a structural lipid, and a lipid conjugated with a polymer.
  17. 17. Composition according to claim 16, characterized in that the cationic lipid is a compound with a structure of formula I, or an N-oxide, a solvate, a pharmaceutically acceptable salt or a stereoisomer thereof, wherein G1 is C1-6 alkylene; G2 is C2-8 alkylene; G3 is C1-3 alkylene; L1 is C6-15 linear alkyl; L2 is C12-25 branched alkyl A cationic lipid is a compound with a structure of formula II, or an N-oxide, a solvate, a pharmaceutically acceptable salt, or a stereoisomer thereof, wherein Gi is C2-8 alkylene; G2 is C2-8 alkylene; Li is -C(O)O- or -OC(O)-; L2 is -C(O)O- or -OC(O)-; Ri is linear or branched C6-25 alkyl; R2 is linear or branched C6-25 alkyl; G3 is HO(CH2)2- or HO(CH2)3-; G4 is HO(CH2)2- or HO(CH2)4-; L is -(CH2)2- or -(CH2)3- or -(CH2)4- The cationic lipid is a compound with a formula III structure, or an N-oxide, a solvate, a pharmaceutically acceptable salt, or a stereoisomer thereof, wherein G1 is C1-6 alkylene; G2 is C2-8 alkylene; R1 is C6-20 linear or branched alkyl; R2 is C12-25 branched alkyl; G3 is: HO(CH2)2N(CH3)(CH2)2-, HO(CH2)2N(CH2CH3)(CH2)2-, (HO(CH2)2)2N(CH2)2-, CH3O(CH2)2N(CH3)(CH2)2-, (CH3)2N(CH2)3SC(O)O(CH2)2-, (CH3)2N(CH2)3SC(O)-, CH3NH(CH2)2N(CH3)(CH2)2-, or CH3CH2NH(CH2)2 The cationic lipid is a compound having a structure of formula IV, or an N-oxide, a solvate, a pharmaceutically acceptable salt, or a stereoisomer thereof, wherein G1 is C1-8 alkylene; G2 is C2-8 alkylene; R1 is C6-25 linear or branched alkyl; R2 is C12-25 linear or branched alkyl; G3 is HO(CH2)2N(R3)CH2CH(OH)CH2-, wherein R3 is -CH3, or -CH2CH3, or -CH2CH2OH; The cationic lipid is a compound with a structure of formula V, or an N-oxide, a solvate, a pharmaceutically acceptable salt, or a stereoisomer thereof, wherein G1 and G2 are each independently unsubstituted C6-C10 alkylene; G3 is unsubstituted C1-C12 alkylene; R1 and R2 are each independently C6-C24 alkyl or C6-C24 alkenyl; R3 is OR5, N, -C(=O)OR4, -OC(=O)R4 or -NR5C(=O)R4; R4 is C1-C12 alkyl; and R5 is H or C1-C6 alkyl. The cationic lipid is a compound with a formula VI structure, or an N-oxide, a solvate, a pharmaceutically acceptable salt, or a stereoisomer thereof, wherein R4 is selected from -(CH2)nQ and -(CH2)nCHQR; Q is selected from the group consisting of: -OR, -OH, -O(CH2)nN(R)2, -OC(O)R, -CX3, -CN, -N(R)C(O)R, -N(H)C(O)R, -N(R)S(O)2R, -N(H)S(O)2R, -N(R)C(O)N(R)2, -N(H)C(O)N(R)2, -N(H)C(O)N(H)(R), -N(R)C(S)N(R)2, -N(H)C(S)N(H)(R), -N(R)S(O)2R8 and a heterocyclic ring; n is 1, 2 or 3; where R is hydrogen, C1-3 alkyl, C2-3 alkenyl or (CH2) qOR*, where q is 1, 2 or 3, R* is C1-12 alkyl or C2-12 alkenyl; X is fluoro, chlorine, bromine or iodine; R8 is C3-6 cycloalkyl or a heterocyclic ring A cationic lipid is a compound with a formula VII structure, or an N-oxide, a solvate, a pharmaceutically acceptable salt, or a stereoisomer thereof. The cationic lipid is selected from the following compounds: YK-009, YK-401, YK-305, ALC0315, SM102, DLIN-MC3
  18. 18. Composition according to claim 16, characterized in that the cationic lipid and the neutral lipid are in a molar ratio of (1-10):1; or, the cationic lipid and the structural lipid are in a molar ratio of (1-5):1.
  19. 19. Composition according to claim 16, characterized in that the cationic lipid, the neutral lipid, the structural lipid and the polymer-conjugated lipid are in a molar ratio of (25-75):(5-25):(15-65):(0.5-10); or the cationic lipid, the neutral lipid, the structural lipid and the polymer-conjugated lipid are in a molar ratio of (35-49):(7.5-15):(35-55):(1-5).
  20. 20. Composition according to claim 16, characterized in that the neutral lipid is selected from one or more of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, ceramide, sterol and derivatives thereof; The neutral lipid is selected from one or more of the following: 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphorylcholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecadienyl-sn-glycero-3-phosphocholine (18:0 Diether PC). 1-oleoyl-2-cholesterol-hemisuccinyl-sn-glycero-3-phosphocholine(OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonyl-sn-glycero-3-phosphorylcholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphanyl-sn-glycero-3-phosphoethanolamine (ME 16:0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, sodium salt of 1,2-dioleoyl-sn-glycero-3-phosphorac-(1-glycerol) (DOPG), dipalmitoyl phosphatidylglycerol (DPPG), palmitoyl oleoyl phosphatidylethanolamine (POPE), distearoyl phosphatidylethanolamine (DSPE), dipalmitoyl phosphatidylethanolamine (DPPE), dimyristoyl phosphatidylethanolamine (DMPE), 1-stearyl-2-oleoyl-stearoylethanolamine (SOPE), 1-stearoyl-2-oleoyl phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyl oleoyl phosphatidylcholine, lysophosphatidylcholine and lysophosphatidylethanolamine (LPE); or the neutral lipid is DOPE and/or DSPC; or the structural lipid is selected from one or more of the following: cholesterol, non-sterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol and corticosteroid.

Description

FIELD OF TECHNIQUE [0001] This disclosure pertains to the technical field of nucleic acid vaccines and, in particular, relates to a respiratory syncytial virus mRNA vaccine and a method for its preparation and use. BACKGROUND [0002] Respiratory syncytial virus (RSV) is a negative-sense RNA virus of the genus Pneumovirus in the family Paramyxoviridae. In 1956, scientists isolated RSV from respiratory specimens of chimpanzees, which was named respiratory syncytial virus because it caused adjacent cells to fuse during cell culture and the cells to transform into a syncytium-like structure. [0003] RSV virus is an important pathogen that causes lower respiratory tract infections in infants, the elderly, and patients with impaired immune function. RSV is airborne, enters the trachea and lungs by inhalation through the mouth and nose, and invades epithelial cells to cause airway trauma, so that respiratory mucus is secreted from the airway surface to block the lumen, causing dyspnea. Common symptoms of an RSV infection include runny nose, fever, cough, and asthma, which can cause shortness of breath and even airway obstruction in severe cases, leading to respiratory failure and even death. [0004] RSV is a common respiratory virus. It usually causes symptoms similar to those of a common cold. RSV infection occurs mainly in the fall, winter, and spring, and is a leading cause of severe respiratory illness in infants under 5 years of age and the elderly over 65 years of age. For the vast majority of young people, RSV usually causes mild cold symptoms, but for infants, young children, and the elderly, RSV infection is a potentially fatal illness. [0005] RSV infection has become a global public health problem. RSV is the most common viral pathogen causing acute lower respiratory tract infections (RRTIs) in children under 5 years of age worldwide and is the leading cause of hospitalization for viral respiratory tract infections in infants and young children. Data show that RSV infection accounts for 28% of all RRTIs, with RSV-related hospital deaths in children accounting for 13% to 22% of RRTI deaths. [0006] RSV infection is distinguished by a widespread global epidemic, and the epidemic is affected by factors such as geographic location, temperature, and humidity. For example, the RSV epidemic season begins from mid-October to mid-May of the following year in northern China and frequently occurs in winter and spring in southern China, which is closely related to temperature. RSV has only one serotype, which is divided into subtypes A and B. These two subtypes are alternately epidemic in the northern part of the country. [0007] Vaccines are the most economical and effective means of preventing RSV infection. The WHO has listed the development of RSV vaccines as one of the urgent problems to be solved. However, since the beginning of RSV vaccine development in the 1960s, no effective vaccine certified by regulatory agencies has been approved for commercialization. Recently, major advances in RSV vaccines have been made one after another. On May 3, 2023, a respiratory syncytial virus (RSV) vaccine developed by GSK, Arexvy, received FDA marketing approval, becoming the world's first RSV vaccine to be approved for commercialization. The vaccine is primarily used in patients over 60 years of age to prevent lower respiratory tract diseases (bronchi and lungs) caused by RSV infection. This was followed on June 1, 2023, by the US FDA's approval for marketing of Pfizer's RSV protein vaccine, Abrysvo™, for the prevention of acute respiratory tract diseases and lower respiratory tract diseases caused by RSV in adults over 60 years of age. [0008] Currently, the technical routes of RSV vaccines under development include live attenuated vaccines, subunit vaccines, recombinant vector vaccines, mRNA vaccines, etc. Traditional vaccines are generally based on inactivated or attenuated live virus vaccines, or subunit proteins derived from pathogens. Although these vaccines produce an effective immune response, they have several associated safety problems. For example, instead of having a protective effect on those vaccinated, early-developed inactivated RSV vaccines develop enhanced respiratory disease (ERD), i.e., they not only fail to prevent those vaccinated from becoming infected with the RSV virus, but also lead to a worsening of the condition of those vaccinated after becoming infected with the RSV virus. In a clinical trial of an inactivated RSV vaccine, 16 of the 20 infants in the experimental group were severely ill and required hospitalization, with a hospitalization rate of up to 80%, and two of them died. [0009] mRNA vaccines work by introducing mRNA encoding a viral antigen into the body and producing an antigen through expression in the body, which in turn stimulates the body to produce an immune response. They have relatively high safety because mRNA exists only in the cytoplasm, is not integrated into the host genome, and