Application of mRNA for use as a therapeutic against tumour diseases
The present invention relates to a pharmaceutical composition comprising at least one mRNA comprising at least one coding region for at least one antigen from a tumor, in combination with an aqueous solvent and preferably a cytokine, e.g. GM-CSF, and a process for the preparation of the pharmaceutical composition. The pharmaceutical composition according to the invention is used in particular for therapy and/or prophylaxis against cancer.
1. A method of stimulating an antitumour immune response in a subject comprising administering an effective amount of a cell-free composition comprising a mRNA encoding a MUC-1 tumour antigen to a subject having a tumour, thereby stimulating a T-cell mediated cytotoxic antitumor immune response in the subject.
2. The method of claim 1 , wherein the composition comprises at least one RNase inhibitor.
3. The method of claim 1 , wherein the mRNA is complexed with at least one cationic or polycationic agent.
4. The method of claim 3 , wherein the cationic or polycationic agent is chosen from the group consisting of protamine, poly-L-lysine, poly-L-arginine and histones.
5. The method of claim 4 , wherein the mRNA is complexed with protamine.
6. The method of claim 1 , administering one or more adjuvant(s) to the subject.
7. The method of claim 6 , wherein the adjuvant is chosen from the Group consisting of lipopolysaccharide, TNF-α, CD40 ligand, GP96, oligonucleotides With aCpG motif, aluminium hydroxide, Freund's adjuvant, lipopeptides and cytokines.
8. The method of claim 7 , wherein the cytokine is GM-CSF.
9. The method of claim 1 , wherein the wherein the mRNA encoding MUC-1 is modified compared with the wild-type mRNA encoding MUC-1 such that it has no destabilizing sequence element.
10. The method of claim 1 , wherein the wherein the mRNA comprises a 5′ cap structure, at least one IRES and/or a poly(A + ) tail of at least about 25 nucleotides.
11. The method of claim 1 , wherein the wherein the mRNA comprises at least one 5′-stabilizing sequence and/or at least one 3′-stabilizing sequence.
12. The method of claim 11 , wherein the 5′- and/or the 3′-stabilizing sequence(s) is/are chosen from the group consisting of untranslated sequences (UTR) of the β-globin gene and a stabilizing sequence of the general formula (C/U)CCAN x CCC(U/A)Py x UC(C/U)CC.
13. The method of claim 1 , wherein the wherein the mRNA comprises least one analogue of naturally occurring nucleotide selected from the group consisting of phosphorothioates, phosphoroamidates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine.
14. The method of claim 1 , further comprising administering one or more additional mRNA(s) encoding additional tumour antigen(s) to the subject.
15. The method of claim 14 , further comprising administering mRNA encoding a PSA tumour antigen and mRNA encoding a PSM tumour antigen.
16. The method of claim 14 , further comprising administering mRNA encoding a MAGE tumour antigen and mRNA encoding a NY-ESO-1 tumour antigen.
17. The method of claim 1 , wherein the mRNA is administered by injection of an aqueous solution comprising the mRNA.
18. The method of claim 17 , wherein the mRNA is administered intradermally.
19. The method of claim 1 , wherein the mRNA is administered two or more times.
20. The method of claim 1 , wherein the subject has been determined to have a MUC-1 positive tumour.