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 . A nucleic acid vaccine, comprising:
 one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide that is derived from an infectious agent, at least one 5′ terminal cap and at least one chemical modification, formulated within a cationic lipid nanoparticle.   
     
     
         2 . The nucleic acid vaccine of  claim 1 , wherein the vaccine is one RNA polynucleotide. 
     
     
         3 . The nucleic acid vaccine of  claim 1 , wherein the cationic lipid nanoparticle has a molar ratio of about 20-60% ionizable cationic lipid:about 5-25% non-cationic lipid:about 25-55% sterol; and about 0.5-15% PEG-modified lipid. 
     
     
         4 . The nucleic acid vaccine of  claim 1 , wherein the RNA polynucleotide has at least two chemical modifications. 
     
     
         5 . The nucleic acid vaccine of  claim 1 , wherein the open reading frame is codon-optimized. 
     
     
         6 . The nucleic acid vaccine of  claim 3 , wherein the cationic lipid nanoparticle has a polydispersity value of less than 0.4. 
     
     
         7 . The nucleic acid vaccine of  claim 1 , wherein the polynucleotide has a poly-A tail of 80-250 nucleotides in length. 
     
     
         8 . The nucleic acid vaccine of  claim 1 , wherein the nucleic acid vaccine is multivalent. 
     
     
         9 . The nucleic acid vaccine of  claim 1 , wherein the open reading frame of the one or more RNA polynucleotides encode at least 2, 3, or 4 antigenic polypeptides. 
     
     
         10 . The nucleic acid vaccine of  claim 1 , wherein the open reading frame of the one or more RNA polynucleotides encode 1 antigenic polypeptide. 
     
     
         11 . The nucleic acid vaccine of  claim 1 , wherein the cationic lipid nanoparticle comprises a molar ratio of about 55% ionizable cationic lipid, about 2.5% PEG lipid, about 32.5% cholesterol and about 10% non-cationic lipid. 
     
     
         12 . The nucleic acid vaccine of  claim 1 , wherein the nanoparticle has a mean diameter of 50-200 nm. 
     
     
         13 . The nucleic acid vaccine of  claim 1 , wherein the nanoparticle has a mean diameter of 80-100 nm. 
     
     
         14 . The nucleic acid vaccine of  claim 1 , wherein at least 80% of the uracil in the open reading frame have a chemical modification. 
     
     
         15 . The nucleic acid vaccine of  claim 1 , wherein 100% of the uracil in the open reading frame have a chemical modification. 
     
     
         16 . The nucleic acid vaccine of  claim 1 , wherein the chemical modification is in the 5-position of the uracil. 
     
     
         17 . The nucleic acid vaccine of  claim 1 , 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. 
     
     
         18 . The nucleic acid vaccine of  claim 17 , wherein the chemical modification is a N1-methyl pseudouridine. 
     
     
         19 . The nucleic acid vaccine of  claim 1 , wherein the RNA polynucleotide is formulated to produce neutralizing antibodies following a single administration of the nucleic acid vaccine. 
     
     
         20 . The nucleic acid vaccine of  claim 1 , wherein the vaccine includes 1.5 mg/mL of RNA polynucleotide and 35-45 mg/mL lipids in the cationic lipid nanoparticle. 
     
     
         21 . The nucleic acid vaccine of  claim 1 , wherein the vaccine includes about 2 mg/mL of RNA polynucleotide and about 40 mg/mL lipids in the cationic lipid nanoparticle. 
     
     
         22 . The nucleic acid vaccine of  claim 1 , wherein the RNA polynucleotide does not include a histone stem-loop stabilization element. 
     
     
         23 . The nucleic acid vaccine of  claim 1 , wherein the RNA polynucleotide is formulated with an adjuvant, wherein the adjuvant is selected from a cationic peptide and an immunostimulatory nucleic acid.