RSV RNA molecules and compositions for vaccination
The present invention is directed to an artificial nucleic acid, particularly to an artificial RNA suitable for use in treatment and/or prophylaxis of an infection with Respiratory syncytial virus (RSV) or a disorder related to such an infection. The invention further concerns a method of treating or preventing a disorder or a disease, first and second medical uses of the artificial RNA, compositions, and vaccines. Further, the invention is directed to a kit, particularly to a kit of parts, comprising the artificial RNA, compositions and vaccines.
1. A purified RNA molecule, said RNA molecule comprising:
a) a 5′-cap structure;
b) a heterologous 5′ untranslated region (UTR);
c) at least one coding sequence encoding an antigenic RSV F protein having an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 4957, wherein said at least one coding sequence is at least 90% identical to the RNA sequence of SEQ ID NO: 4962; and
d) at least one poly(A) sequence comprising 30 to 150 consecutive adenosine nucleotides.
2. The purified RNA molecule of claim 1 , further comprising a heterologous 3′ UTR.
3. The purified RNA molecule of claim 2 , wherein the heterologous 3′-UTR is derived from a 3′-UTR of an alpha-globin gene.
4. The purified RNA molecule of claim 1 , wherein the 5′-cap structure is a Cap1.
5. The purified RNA molecule of claim 2 , wherein the antigenic RSV F protein has an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 4957.
6. The purified RNA molecule of claim 1 , wherein the RNA molecule comprises at least one chemically modified nucleotide.
7. The purified RNA molecule of claim 1 , wherein all uracil nucleotides of the RNA molecule are replaced by N1-methylpseudouridine (m1ψ) nucleotides.
8. The purified RNA molecule of claim 7 , wherein the RNA molecule is a mRNA molecule.
9. The purified RNA molecule of claim 1 , wherein the RNA molecule is a self-replicating RNA molecule.
10. The purified RNA molecule of claim 1 , wherein the at least one poly(A) sequence is located at the 3′ terminus of the RNA molecule.
11. The purified RNA molecule of claim 1 , wherein the amino acid sequence of the antigenic RSV F protein comprises the following amino acid substitutions relative to the amino acid sequence of a native RSV F protein: S155C, S290C, S190F, V207L, A149C and Y458C.
12. The purified RNA molecule of claim 11 , wherein the amino acid sequence of the antigenic RSV F protein comprises a GS linker in place of amino acids 104-144 of the amino acid sequence of the native RSV F protein.
13. The purified RNA molecule of claim 12 , wherein the amino acid sequence of the antigenic RSV F protein lacks amino acids 554-574 of the amino acid sequence of the native RSV F protein.
14. The purified RNA molecule of claim 12 , wherein the antigenic RSV F protein comprises the native RSV F protein transmembrane domain.
15. A pharmaceutical composition comprising purified RNA molecules of claim 8 formulated in a pharmaceutically acceptable carrier.
16. The pharmaceutical composition of claim 15 , wherein the RNA molecules are formulated with lipid nanoparticles (LNPs).
17. The pharmaceutical composition of claim 16 , wherein the LNPs comprise a cationic lipid.
18. The pharmaceutical composition of claim 17 , wherein the LNPs comprise a cationic lipid, a neutral lipid, a sterol, and a polymer conjugated lipid.
19. The pharmaceutical composition of claim 18 , wherein the LNPs comprise a cationic lipid, a neutral lipid, a sterol, and a PEG-lipid.
20. The pharmaceutical composition of claim 19 , wherein the LNPs comprise a molar ratio of about 20-60% cationic lipid, 5-25% neutral lipid, 25-55% sterol, and 0.5-15% PEG-lipid.
21. The pharmaceutical composition of claim 18 , wherein the amino acid sequence of the antigenic RSV F protein comprises the following amino acid substitutions relative to the amino acid sequence of a native RSV F protein: S155C, S290C, S190F, V207L, A149C and Y458C.
22. The pharmaceutical composition of claim 21 , wherein the amino acid sequence of the antigenic RSV F protein comprises a GS linker in place of amino acids 104-144 of the amino acid sequence of the native RSV F protein.
23. The pharmaceutical composition of claim 22 , wherein the amino acid sequence of the antigenic RSV F protein lacks amino acids 554-574 of the amino acid sequence of the native RSV F protein.
24. The pharmaceutical composition of claim 23 , wherein the antigenic RSV F protein comprises the native RSV F protein transmembrane domain.
25. A method of stimulating an anti-RSV immune response in a subject comprising administering to the subject the pharmaceutical composition of claim 20 .
26. The method of claim 25 , wherein the pharmaceutical composition is administered by intramuscular injection.
27. The method of claim 26 , wherein the amino acid sequence of the antigenic RSV F protein is at least 98% identical to the amino acid sequence of SEQ ID NO: 4957 and comprises the following amino acid substitutions relative to the amino acid sequence of a native RSV F protein: S155C, S290C, S190F, V207L, A149C and Y458C.
28. The method of claim 27 , wherein the amino acid sequence of the antigenic RSV F protein comprises the native RSV F protein transmembrane domain but lacks amino acids 554-574 of the amino acid sequence of the native RSV F protein.
29. The method of claim 27 , wherein the pharmaceutical composition is administered in an amount effective to reduce nasal RSV production following RSV infection of the subject.
30. The purified RNA molecule of claim 1 , wherein:
the RNA molecule is an mRNA molecule;
the amino acid sequence of the antigenic RSV F protein is at least 98% identical to the amino acid sequence of SEQ ID NO: 4957;
the amino acid sequence of the antigenic RSV F protein comprises the following amino acid substitutions relative to the amino acid sequence of a native RSV F protein: S155C, S290C, S190F, V207L, A149C and Y458C; and
wherein said at least one coding sequence is at least 92% identical to the RNA sequence of SEQ ID NO: 4962.
31. The purified RNA molecule of claim 30 , wherein the antigenic RSV F protein comprises the native RSV F protein transmembrane domain.
32. The purified RNA molecule of claim 31 , wherein the amino acid sequence of the antigenic RSV F protein lacks amino acids 554-574 of the amino acid sequence of the native RSV F protein.
33. The purified RNA molecule of claim 32 , wherein the amino acid sequence of the antigenic RSV F protein comprises a GS linker in place of amino acids 104-144 of the amino acid sequence of the native RSV F protein.
34. The purified RNA molecule of claim 30 , wherein the amino acid sequence of the antigenic RSV F protein is at least 99% identical to the amino acid sequence of SEQ ID NO: 4957, and wherein said at least one coding sequence is at least 93% identical to the RNA sequence of SEQ ID NO: 4962.
35. The purified RNA molecule of claim 31 , wherein the amino acid sequence of the antigenic RSV F protein is at least 99% identical to the amino acid sequence of SEQ ID NO: 4957, and wherein said at least one coding sequence is at least 93% identical to the RNA sequence of SEQ ID NO: 4962.
36. The purified RNA molecule of claim 32 , wherein the amino acid sequence of the antigenic RSV F protein is at least 99% identical to the amino acid sequence of SEQ ID NO: 4957, and wherein said at least one coding sequence is at least 93% identical to the RNA sequence of SEQ ID NO: 4962.
37. The purified RNA molecule of claim 33 , wherein the amino acid sequence of the antigenic RSV F protein is at least 99% identical to the amino acid sequence of SEQ ID NO: 4957, and wherein said at least one coding sequence is at least 93% identical to the RNA sequence of SEQ ID NO: 4962.
38. The purified RNA molecule of claim 33 , wherein said at least one coding sequence is adapted to human codon usage with most frequently used codons such that the at least one coding sequence has a codon adaptation index (CAI) of at least 0.9.
39. The purified RNA molecule of claim 38 , wherein said at least one coding sequence has a CAI of at least 0.95.
40. A pharmaceutical composition comprising purified mRNA molecules of claim 39 , wherein said mRNA molecules are formulated with lipid nanoparticles (LNPs), said LNPs comprising a molar ratio of about 20-60% cationic lipid, 5-25% neutral lipid, 25-55% sterol, and 0.5-15% PEG-lipid.
41. The pharmaceutical composition of claim 40 , wherein the neutral lipid is distearoylphosphatidylcholine (DSPC), the sterol is cholesterol, and the PEG-lipid is 1-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG-DMG).
42. The pharmaceutical composition of claim 41 , wherein all uracil nucleotides of the mRNA molecule are replaced by N1-methylpseudouridine (ml) nucleotides and wherein the 5′-cap structure is a cap1 or modified cap1 structure.
43. The purified RNA molecule of claim 30 , wherein said at least one coding sequence has a human CAI of at least 0.95.
44. The purified RNA molecule of claim 31 , wherein said at least one coding sequence has a human CAI of at least 0.95.
45. The purified RNA molecule of claim 32 , wherein said at least one coding sequence has a human CAI of at least 0.95.