Nucleic acid molecules
The present invention provides novel artificial nucleic acid molecules encoding at least one antigenic peptide or protein and at least one additional sequence preferably targeting the antigenic peptides or proteins to cellular compartments of interest. Further, the invention provides (pharmaceutical) compositions or vaccines and kits comprising said nucleic acid molecules. The nucleic acid molecules, (pharmaceutical) compositions or vaccines and kits are useful for treating a variety of diseases such as cancer, infectious diseases, autoimmune diseases, allergies or graft-versus host disease.
1. A method of treating or preventing cancer, autoimmune diseases, or infectious diseases, wherein the method comprises administering, to a patient in need thereof, an effective amount of an artificial nucleic acid molecule comprising at least one coding region encoding:
(a) at least one antigenic peptide or protein; and
(b) at least one additional amino acid sequence derived from at least one immune response activing signal transduction protein located in the external plasma membrane (IRST epm ), wherein said at least one additional amino acid sequence derived from at least one IRST epm comprises at least one transmembrane domain from CTLA4 (Cytotoxic T-lymphocyte protein 4) and localizes the antigenic peptide or protein to the plasma membrane.
2. The method of claim 1 , wherein said at least one additional amino acid sequence further comprises at least one cytoplasmic domain of CTLA4.
3. The method of claim 2 , wherein the at least one antigenic peptide or protein is from KRAS.
4. The method of claim 2 , wherein the at least one antigenic peptide or protein is from a viral antigen.
5. The method of claim 1 , wherein said at least one coding region further encodes at least one signal peptide.
6. The method of claim 5 , wherein the at least one signal peptide comprises or consists of an amino acid sequence corresponding to any one of SEQ ID NOs: 1-156, 76948-76951, or a fragment, variant or derivative thereof, and is optionally encoded by a nucleic acid sequence according to any one of SEQ ID NOs: 209-364, 76952-76955, 625-780, 76960-76963, 833-988, 76964-76967, 417-572, 76956-76959, 1249-1404, 76972-76975, 1457-1612, 76976-76979, 1665-1820, 76980-76983, 1873-2028, 76984-76987, 2081-2236, 76988-76991, 2289-2444, 76992-76995, 2497-2652, 76996-76999, 2705-2860, 77000-77003, 1041-1196, or 76968-76971.
7. The method of claim 1 , wherein said at least one antigenic peptide or protein is selected from or derived from tumor antigens, viral, bacterial, protozoal, fungal or allogenic antigens.
8. The method of claim 7 , wherein said at least one antigenic peptide or protein comprises or consists of an amino acid sequence corresponding to any one of SEQ ID NOs: 3719-27945; 76420-76439, 76440-76474, and is optionally encoded by a nucleic acid sequence according to any one of SEQ ID NOs: 27946-52172; 76495-76514, 52173-76399; 76570-76589, 76515-76549, 76590-76624.
9. The method of claim 1 , wherein said antigenic peptide or protein is from a tumor antigen is selected from BRAF, PIK3CA, KRAS, IDH1, TP53, NRAS, AKTI, SF3B1, CDKN2A, RPSAP58, EGFR, NY-ESO1, MUC-1, 5T4, Her2, MAGE-A3, LY6K, CEACAM6, CEA, MCAK, KK-LC1, Gastrin, VEGFR2, MMP-7, MPHOSPH1, MAGE-A4, MAGE-A1, MAGE-C1, PRAME, Survivin, MAGE-A9, MAGE-C2, FGFR2, WT1, PSA, PSMA, Prostate-specific antigen precursor, Kita-kyushu lung cancer antigen 1, Trophoblast glycoprotein, Cyclin-dependent kinase inhibitor 2A, Cyclin-dependent kinase inhibitor 2A, isoforms 1/2/3, multiple tumor suppressor 1/cyclin-dependent kinase 4 inhibitor p16, GTPase NRas or any combination thereof.
10. The method of claim 1 , wherein said at least one IRST epm comprises or consists of an amino acid sequence corresponding to SEQ ID NO: 169, optionally encoded by a nucleic acid sequence corresponding to any one of SEQ ID NOs: 585, 793, 1001, 1209, 1417, 1625, 1833, 2041, 2249, 2457, 2665 or 2873.
11. The method of claim 1 , further encoding in its at least one coding region at least one linker.
12. The method of claim 11 , wherein said linker is a non-immunogenic linker, optionally comprising or consisting of an amino acid sequence according to any one of SEQ ID NOs: 2937, 76400-76418, 77018-77058 optionally encoded by a nucleic acid sequence according to any one of SEQ ID NOs: 2936, 76494, 76569, 76475-76493, 76550-76568, or 77059-77061.
13. The method of claim 1 , wherein said at least one coding region further encodes at least one T helper epitope.
14. The method of claim 13 , wherein said at least one helper epitope sequence comprises or consists of an amino acid sequence corresponding to any one of SEQ ID NOs: 3083-3294, optionally encoded by a nucleic acid sequence according to any one of SEQ ID NOs: 3295-3506, or 3507-3718.
15. The method of claim 1 , wherein said artificial nucleic acid molecule is an RNA.
16. The method of claim 15 , which comprises, preferably in 5′ to 3′ direction, the following elements:
a) a 5′-CAP structure,
b) optionally a 5′-UTR element,
c) said at least one coding region sequence,
d) optionally a 3′-UTR element,
e) optionally a poly(A) tail,
f) optionally a poly(C) tail, and
g) optionally a histone stem-loop.
17. The method of claim 1 , wherein the method is a method of treating or preventing cancer.
18. The method of claim 17 , wherein the at least one antigenic peptide or protein is a KRAS epitope having a mutation selected from the group consisting of A146T; G12A; G12C; G12D; G12F; G12R; G12S; G12V; G13C; G13D; K117N; Q61H; Q61K; Q61L; and Q61R.
19. The method of claim 18 , wherein the wherein the method is a method of treating or preventing cancer and wherein the artificial nucleic acid molecule is a mRNA.
20. The method of claim 19 , wherein the patient has a pancreatic cancer.
21. The method of claim 1 , wherein the method is a method of treating or preventing infectious diseases selected from the group consisting of viral, bacterial, fungal and protozoal infections.