Rotavirus vaccines
The present invention provides mRNA sequences comprising at least one coding region, encoding for at least one epitope of a protein, or of a fragment, variant or derivative thereof, of a virus of the genus rotavirus. Particularly preferred is the protein respectively the protein cleavage product VP8* of rotavirus. The mRNA sequence may be used as a vaccine or generally as a pharmaceutical composition for prophylaxis or treatment of rotavirus infections.
1. A method for stimulating an anti-rotavirus response in a patient comprising administering an effective amount of an immune stimulating composition comprising a lipid nanoparticle (LNP) comprising an mRNA molecule comprising at least one region encoding a polypeptide comprising at least 100 amino acids of a VP8* cleavage product of a rotavirus VP4 protein, wherein the mRNA comprises a 5′ Cap and a poly(A) tail of 60 to about 250 adenosine at the 3′ end of the mRNA,
wherein the composition is administered by intramuscular injection.
2. The method of claim 1 , wherein the VP8* cleavage product is derived from the P[4], P[6], or P[8] serotype.
3. The method of claim 1 , wherein the mRNA molecule further comprises a sequence encoding a helper peptide and a sequence encoding a peptide linker, wherein the sequence encoding the helper peptide is located at the 5′ end of the at least one region encoding at least one epitope.
4. The method of claim 3 , wherein the helper peptide is derived from tetanus toxin.
5. The method of claim 1 , wherein the mRNA molecule further comprises a sequence encoding a signal peptide derived from tissue plasminogen activator, albumin, CD5, HLA-A2, luciferase, immunoglobulin, or IL-2.
6. The method of claim 1 , wherein the at least one region encoding at least one epitope is mutated to delete at least one predicted glycosylation site.
7. The method of claim 1 , wherein the mRNA molecule further comprises a sequence encoding a transmembrane domain, wherein the sequence encoding the transmembrane domain is located at the 3′ end of the at least one region encoding at least one epitope.
8. The method of claim 1 , wherein the mRNA molecule further comprises a sequence encoding at least one peptide enabling virus like particle (VLP) formation, wherein the peptide is derived from Woodchuck hepatitis virus core antigen (WHcAg) or from Alfalfa mosaic virus coat protein.
9. The method of claim 1 , wherein the mRNA molecule further comprises at least a second region encoding at least one epitope of a structural or non-structural protein of a different serotype of a rotavirus.
10. The method of claim 9 , wherein the second region encodes a further VP8* epitope.
11. The method of claim 1 , wherein the at least one region encoding the polypeptide has a G/C content of the region encoding the polypeptide that is increased compared to the G/C content of a wild type mRNA encoding the polypeptide.
12. The method of claim 1 , wherein the mRNA molecule is at least partially complexed with a cationic or polycationic compound or polymeric carrier.
13. The method of claim 1 , wherein the immune stimulating composition further comprises a pharmaceutical carrier.
14. The method of claim 1 , wherein the immune stimulating composition further comprises at least one adjuvant component.
15. The method of claim 1 , wherein the method induces a cross-reactive immune response to multiple serotypes of rotavirus.
16. The method of claim 1 , wherein the 5′ cap is a Cap structure is a CAP1.
17. The method of claim 4 , wherein the helper peptide is the P2 helper peptide of tetanus toxin.
18. The method of claim 1 , wherein the mRNA comprises a nucleotide analogue.
19. The method of claim 18 , wherein the nucleotide analogue is pseudouridine or 1-methyl-pseudouridine.
20. The method of claim 1 , wherein the LNP comprises an ionizable amino lipid, phospholipid, cholesterol and a PEGylated lipid.