Nucleic acid vaccines

Abstract

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use ribonucleic acid vaccines (NAVs) comprising polynucleotide molecules encoding one or more antigens.

1 .- 23 . (canceled) 
     
     
         24 . A nucleic acid vaccine, comprising:
 one or more RNA polynucleotides having an open reading frame encoding a picornavirus polypeptide, formulated in a cationic lipid nanoparticle having a molar ratio of about 20-60% cationic lipid:about 5-25% non-cationic lipid:about 25-55% sterol; and about 0.5-15% PEG-modified lipid.   
     
     
         25 . The vaccine of  claim 24 , wherein the picornavirus polypeptide is an enterovirus polypeptide derived from a virus selected from the group consisting of: enteroviruses, rhinoviruses, polioviruses, coxsackieviruses, and echoviruses. 
     
     
         26 . The vaccine of  claim 25 , wherein the enterovirus polypeptide is a human enterovirus 68 (HEV68) polypeptide or a human enterovirus 71 (HEV71) polypeptide. 
     
     
         27 . The vaccine of  claim 24 , wherein the RNA polynucleotide comprises a polynucleotide encoding an HEV71 P1 polyprotein amino acid sequence having at least 90% identity to SEQ ID NO: 2233. 
     
     
         28 . The vaccine of  claim 24 , wherein the HEV71 RNA polynucleotide comprises a polynucleotide having 90% sequence identity to SEQ ID NO: 2113. 
     
     
         29 . The vaccine of  claim 25 , wherein the enterovirus polypeptide is a human rhinovirus (HRV) polypeptide. 
     
     
         30 . The vaccine of  claim 24 , wherein the cationic lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid. 
     
     
         31 . The vaccine of  claim 24 , wherein the cationic lipid is an ionizable cationic lipid and the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol. 
     
     
         32 . The vaccine of  claim 24 , wherein the open reading frame is codon-optimized. 
     
     
         33 . The vaccine of  claim 24 , wherein the one or more RNA polynucleotides comprise at least one chemical modification. 
     
     
         34 . The vaccine of  claim 24 , wherein the nucleic acid vaccine is multivalent. 
     
     
         35 . The vaccine of  claim 33 , wherein the chemical modification is selected from the group consisting of pseudouridine, N1-methylpseudouridine, 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-methyl uridine. 
     
     
         36 . The vaccine of  claim 24 , wherein the nanoparticle has a polydispersity value of less than 0.4. 
     
     
         37 . A nucleic acid vaccine, comprising:
 one or more RNA polynucleotides having an open reading frame encoding a picornavirus polypeptide, wherein at least 80% of the uracil in the open reading frame have a chemical modification.   
     
     
         38 . The vaccine of  claim 37 , wherein 100% of the uracil in the open reading frame have a chemical modification. 
     
     
         39 . The vaccine of  claim 37 , wherein the chemical modification is in the 5-position of the uracil. 
     
     
         40 . The vaccine of  claim 37 , wherein the chemical modification is a N1-methyl pseudouridine. 
     
     
         41 . A nucleic acid vaccine, comprising:
 one or more RNA polynucleotides having an open reading frame encoding a picornavirus polypeptide, at least one 5′ terminal cap and at least one chemical modification, formulated within a cationic lipid nanoparticle.   
     
     
         42 . The vaccine of  claim 41 , wherein the 5′ terminal cap is 7mG(5′)ppp(5′)NlmpNp. 
     
     
         43 . The vaccine of  claim 41 , wherein the chemical modification is selected from the group consisting of pseudouridine, N1-methylpseudouridine, 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-methyl uridine.