LIPID NANOPARTICLE mRNA VACCINES

A method for stimulating an anti-influenza immune response in a mammalian subject comprising administering to the subject an effective amount of a pharmaceutical composition comprising:

• (a) at least a first mRNA comprising a first coding sequence encoding an HA3 antigen from an influenza A virus at least 90% identical to the amino acid sequence of SEQ ID NO:
• 13853 (H3N2), said first coding sequence comprising a RNA sequence at least 85% identical to the HA3 antigen coding sequence of SEQ ID NO:
• 224181,
• wherein the first mRNA comprises, from 5′ to 3′:
• (i) a 5′-CAP structure;
• (ii) a 5′-untranslated region (UTR) element;
• (iii) the first coding sequence;
• (iv) a 3′-UTR element;
• and (v) a poly(A) sequence comprising 60 to 250 consecutive adenosines;
• wherein the mRNA is formulated with:
• (b) lipid nanoparticles (LNPs) comprising:
• (i) a cationic lipid with the formula III:
• or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:
• L1 and L2 are each independently —O(C═O)—, —(C═O)O—;
• G1 and G2 are each independently unsubstituted C1-C12 alkylene;
• G3 is C1-C24 alkylene;
• R1 and R2 are each independently C6-C24 alkyl;
• R3 is OR5;
• and
• R5 is H or C1-C6 alkyl;
• and
• (ii) a PEGylated lipid,
• wherein the pharmaceutical composition is administered by intradermal, subcutaneous or intramuscular injection.

A method for producing a pharmaceutical composition comprising:

• (I) obtaining at least a first mRNA comprising a first coding sequence encoding an HA3 antigen from an influenza A virus at least 90% identical to the amino acid sequence of SEQ ID NO:
• 13853 (H3N2), said first coding sequence comprising a RNA sequence at least 85% identical to the HA3 antigen coding sequence of SEQ ID NO:
• 224181,
• wherein the first mRNA comprises, from 5′ to 3′:
• (i) a 5′-CAP structure;
• (ii) a 5′-untranslated region (UTR) element;
• (iii) the first coding sequence;
• (iv) a 3′-UTR element;
• and (v) a poly(A) sequence comprising 60 to 250 consecutive adenosines;
• and
• (II) formulating said first mRNA in LNPs comprising:
• (i) a cationic lipid with the formula III:
• or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:
• L1 and L2 are each independently —O(C═O)—, —(C═O)O—, —C(═O)—, —O—, —S(O)x—, —S—S—, —C(═O)S—, —SC(═O)—, —NRaC(═O)—, —C(═O)NRa—, —NRaC(═O)NRa—, —OC(═O)NRa— or —NRaC(═O)O—;
• G1 and G2 are each independently unsubstituted C1-C12 alkylene or C1-C12 alkenylene;
• G3 is C1-C24 alkylene, C1-C24 alkenylene, C3-C8 cycloalkylene, or C3-C8 cycloalkenylene;
• Ra is, at each occurrence, independently H or C1-C12 alkyl;
• R1 and R2 are each independently C6-C24 alkyl or C6-C24 alkenyl;
• R3 is OR5, CN, —C(═O)OR4, —OC(═O)R4 or —NR5C(═O)R4;
• R4 is C1-C12 alkyl;
• R5 is H or C1-C6 alkyl;
• and
• x is 0, 1 or 2;
• and
• (ii) a PEGylated lipid.