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 tumour, 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 pharmaceutical composition comprising at least one mRNA comprising at least one coding region for at least one antigen from a tumour, in combination with an aqueous solvent.
2 . The pharmaceutical composition according to claim 1 , wherein the coding region for the antigen(s) from a tumour and/or the 5′ and/or the 3′ untranslated region of the mRNA is modified compared with the wild-type mRNA such that it has no destabilizing sequence element.
3 . The pharmaceutical composition according to claim 1 , wherein the mRNA has a 5′ cap structure and/or a poly(A + ) tail of at least about 25 nucleotides and/or at least one internal ribosomal entry site (IRES) and/or at least one 5′-stabilizing sequence and/or at least one 3′-stabilizing sequence.
4 . The pharmaceutical composition according to claim 3 , 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.
5 . The pharmaceutical composition according to claim 1 , wherein the mRNA contains at least one analogue of naturally occurring nucleotides.
6 . The pharmaceutical composition according to claim 5 , wherein the analogue is chosen from the group consisting of phosphorothioates, phosphoroamidates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine.
7 . The pharmaceutical composition according to claim 1 , wherein the antigen(s) from a tumour is/are a polyepitope of antigens from a tumour.
8 . The pharmaceutical composition according to claim 7 , wherein the polyepitope is modified by deletion, addition and/or substitution of one or more amino acid radicals.
9 . The pharmaceutical composition according to claim 1 , wherein the mRNA additionally codes for at least one cytokine.
10 . The pharmaceutical composition according to one claim 1 , which also comprises one or more adjuvants.
11 . The pharmaceutical composition according to claim 10 , wherein the adjuvant is chosen from the group consisting of lipopolysaccharide, TNF-α, CD40 ligand, GP96, oligonucleotides with the CpG motif, aluminium hydroxide, Freund's adjuvant, lipopeptides and cytokines.
12 . The pharmaceutical composition according to claim 11 , wherein the cytokine is GM-CSF.
13 . The pharmaceutical composition according to claim 1 , wherein the mRNA is present in a form complexed or fused with at least one cationic or polycationic agent.
14 . The pharmaceutical composition according to claim 13 , wherein the cationic or polycationic agent is chosen from the group consisting of protamine, poly-L-lysine, poly-L-arginine and histones.
15 . The pharmaceutical composition according to claim 1 , which also comprises at least one RNase inhibitor.
16 . The pharmaceutical composition according to claim 15 , wherein the RNase inhibitor is RNasin.
17 . The pharmaceutical composition according to one claim 1 , which comprises a majority of mRNA molecules which represent a cDNA library, or a part thereof, of a tumour tissue.
18 . The pharmaceutical composition according to claim 17 , wherein the part of the cDNA library codes for the tumour-specific antigens.
19 . Pharmaceutical composition according to claim 1 , wherein the antigen(s) from a tumour is/are chosen from the group consisting of 707-AP, AFP, ART-4, BAGE, β-catenin/m, Bcr-abl, CAMEL, CAP-1, CASP-8, CDC27/m, CDK4/m, CEA, CT, Cyp-B, DAM, ELF2M, ETV6-AML1, G250, GAGE, GnT-V, Gp100, HAGE, HER-2/neu, HLA-A*0201-R170I, HPV-E7, HSP70-2M, HAST-2, hTERT (or hTRT), iCE, KIAA0205, LAGE, LDLR/FUT, MAGE, MART-1/melan-A, MC1R, myosin/m, MUC1, MUM-1, -2, -3, NA88-A, NY-ESO-1, p190 minor bcr-abl, Pml/RARα, PRAME, PSA, PSM, RAGE, RU1 or RU2, SAGE, SART-1 or SART-3, TEL/AML1, TPI/m, TRP-1, TRP-2, TRP-2/INT2 and WT1.
20 . The pharmaceutical composition according to claim 1 , wherein the mRNA contains a sequence region which serves to increase the translation rate.
21 . The pharmaceutical composition according to claim 1 , comprising at least one further pharmaceutically acceptable carrier and/or at least one further pharmaceutically acceptable vehicle.
22 . The pharmaceutical composition according to claim 1 , for therapy and/or prophylaxis of cancer.
23 . A process for the preparation of a pharmaceutical composition according to claim 1 , comprising the steps:
(a) preparation of a cDNA library, or a part thereof, from tumour tissue of a patient,
(b) preparation of a matrix for in vitro transcription of RNA with the aid of the cDNA library or a part thereof and
(c) in vitro transcribing of the matrix.
24 . The process according to claim 23 , wherein the part of the cDNA library of the tumour tissue codes for the tumour-specific antigens.
25 . The process according to claim 24 , in which the sequences of the tumour-specific antigens are ascertained before step (a).
26 . The process according to claim 25 , wherein the ascertaining of the sequences of the tumour-specific antigens comprises an alignment with a cDNA library from healthy tissue.
27 . The process according to claim 25 , wherein the ascertaining of the sequences of the tumour-specific antigens comprises a diagnosis by a microarray.