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 pharmaceutical composition comprising a purified RNA molecule comprising a region encoding a polypeptide comprising at least 100 amino acids of a VP8* cleavage product of a rotavirus VP4 protein, said RNA comprising a 5′ Cap structure and a poly(A) region of 60 to 250 adenosine nucleotides, wherein said RNA is formulated with a lipid nanoparticle (LNP).
2. The pharmaceutical composition of claim 1 , wherein the 5′ Cap structure is a CAP1.
3. The pharmaceutical composition of claim 1 , wherein the poly(A) region is located at the 3′ terminus of the RNA.
4. The pharmaceutical composition of claim 1 , wherein the VP8* cleavage product is derived from the P[4], P[6], or P[8] serotype.
5. The pharmaceutical composition of claim 1 , wherein the polypeptide comprises an amino acid substitution to remove at least one predicted glycosylation site.
6. The pharmaceutical composition of claim 1 , wherein the RNA is a mRNA that comprises, from 5′ to 3′:
(i) a 5′ Cap structure;
(ii) a 5′ untranslated region (UTR);
(iii) the region encoding the polypeptide;
(iv) a 3′ UTR; and
(v) the poly(A) region.
7. The pharmaceutical composition of claim 4 , wherein the composition comprises at least a second RNA encoding a VP8* cleavage product of a rotavirus VP4 from a different rotavirus strain as the first RNA.
8. The pharmaceutical composition of claim 4 , wherein the LNP comprises an ionizable amino lipid, phospholipid, cholesterol and a PEGylated lipid.
9. The pharmaceutical composition of claim 4 , wherein the RNA further encodes a sequence encoding a VLP forming protein.
10. The pharmaceutical composition of claim 9 , wherein the VLP forming protein is derived from an enveloped virus.
11. The pharmaceutical composition of claim 4 , wherein the wherein the G/C content of the region encoding the polypeptide is increased compared to the G/C content of the coding region of a corresponding original polypeptide.
12. The pharmaceutical composition of claim 4 , said RNA further comprising a sequence encoding a helper peptide, wherein the sequence encoding the helper peptide is located at the 5′ end of the polypeptide.
13. The pharmaceutical composition of claim 12 , wherein the helper peptide is derived from tetanus toxin.
14. The pharmaceutical composition of claim 4 , wherein the RNA molecule further comprises a sequence encoding a signal peptide derived from tissue plasminogen activator, albumin, CD5, HLA-A2, luciferase, immunoglobulin or IL-2.
15. The pharmaceutical composition of claim 4 , wherein the RNA comprises 1-methyl-pseudouridine substitutions.
16. A kit comprising a pharmaceutical composition of claim 1 and instructions for administering said composition.