Flavivirus vaccine
The present invention is directed to an artificial nucleic acid and to a polypeptide suitable for use in the treatment or prophylaxis of an infection with a flavivirus, in particular an infection with yellow fever virus or with dengue virus, or of a disorder related to such an infection. The present invention is also directed to a composition, preferably an immunogenic composition, comprising the artificial nucleic acid or the inventive polypeptide. In particular, the present invention concerns an immunogenic composition against a flavivirus, such as yellow fever virus or dengue virus. Further, the invention concerns a kit, particularly a kit of parts, comprising the artificial nucleic acid, polypeptide or (immunogenic) composition. The invention is further directed to a method of treating or preventing a disorder or a disease, first and second medical uses of the artificial nucleic acid, polypeptide, composition, in particular the first and second medical uses of the immunogenic composition according to the invention.
1. A pharmaceutical formulation comprising RNA formulated in a lipid nanoparticle (LNP), said RNA comprising
a) at least one coding region encoding at least one polypeptide comprising a flavivirus premembrane protein (prM) and a flavivirus envelope protein (E), wherein the at least one coding region comprises a nucleic acid sequence at least 95% identical to the coding sequence of SEQ ID NO: 2630, and
b) an untranslated region (UTR) comprising at least one heterologous UTR element,
wherein the flavivirus a dengue virus,
wherein said LNP comprises: (i) at least one cationic lipid; (ii) a neutral lipid; (iii) a sterol; and (iv) a PEG-lipid.
2. The pharmaceutical formulation according to claim 1 , wherein the at least one encoded polypeptide further comprises a flavivirus non-structural protein or a flavivirus capsid protein (C).
3. The pharmaceutical formulation according to claim 1 , wherein the RNA further comprises a heterologous nucleic acid sequence.
4. The pharmaceutical formulation according to claim 1 , wherein the at least one encoded polypeptide comprises at least one signal sequence, wherein the at least one signal sequence is a signal sequence of a secretory protein or a signal sequence of a membrane protein.
5. The pharmaceutical formulation according to claim 1 , wherein the at least one encoded polypeptide further comprises at least one amino acid sequence; promotes virus-like particle (VLP) formation.
6. The pharmaceutical formulation according to claim 1 , wherein the at least one encoded polypeptide further comprises at least one amino acid sequence that promotes antigen clustering and/or formation of nanoparticles.
7. The pharmaceutical formulation according to claim 2 , wherein the at least one encoded polypeptide further comprises at least one amino acid sequence that promotes self-cleavage of the encoded polypeptide.
8. The pharmaceutical formulation according to claim 1 , wherein the at least one encoded polypeptide comprises a mutated furin cleavage site.
9. The pharmaceutical formulation according to claim 1 , wherein the RNA is an mRNA, which comprises a histone stem-loop, a 3′-UTR element, a 5′-UTR element, a poly(A) sequence, and/or a poly(C) sequence.
10. A kit or kit of parts comprising the pharmaceutical formulation according to claim 1 , and comprising technical instructions providing information on administration and dosage of the components.
11. A method for treating or inhibiting a dengue virus infection comprising administering the pharmaceutical formulation of claim 1 to a patient in need thereof.
12. The pharmaceutical formulation according to claim 5 , wherein the amino acid sequence promoting virus-like particle (VLP) formation is from a hepatitis B virus core antigen.
13. The pharmaceutical formulation according to claim 1 , wherein the coding region comprises a nucleic acid sequence having the coding sequence of SEQ ID NO: 2630.
14. The pharmaceutical formulation according to claim 1 , wherein the coding region encodes a protein that is at least 95% identical to SEQ ID NO: 1101.
15. The pharmaceutical formulation according to claim 14 , wherein the coding region encodes a protein having the sequence of any one of SEQ ID NOs: 1070 and 1097-1103.
16. The pharmaceutical formulation according to claim 14 , wherein the coding region encodes a protein having the sequence of SEQ ID NO: 1101.
17. The pharmaceutical formulation according to claim 1 , wherein the coding region comprises the nucleic acid sequence of any one of SEQ ID NOs: 1375, 1402-1408, 1531, and 1558-1563.
18. The pharmaceutical formulation according to claim 1 , wherein the coding region comprises a nucleic acid sequence having the sequence of SEQ ID NO: 1406 or 1562.
19. The pharmaceutical formulation according to claim 1 , wherein the LNP comprises (i) the cationic lipid; (ii) the neutral lipid; (iii) the sterol; and (iv) the PEG-lipid, in a molar ratio of about 20-60% cationic lipid: 5-25% neutral lipid: 25-55% sterol; and 0.5-15% PEG-lipid.
20. The pharmaceutical formulation according to claim 19 , wherein the PEG-lipid is PEG-DMG or PEG-cDMA.