COMPOSITION COMPRISING A COMPLEXED (m)RNA AND A NAKED mRNA FOR PROVIDING OR ENHANCING AN IMMUNOSTIMULATORY RESPONSE IN A MAMMAL AND USES THEREOF

Abstract

The present invention relates to an immunostimulatory composition comprising a) an adjuvant component, comprising or consisting of at least one (m)RNA, complexed with a cationic or polycationic compound, and b) at least one free mRNA, encoding at least one therapeutically active protein, antigen, allergen and/or antibody, wherein the immunostimulatory composition is capable to elicit or enhance an innate and optionally an adaptive immune response in a mammal. The inventive immunostimulatory composition may be a pharmaceutical composition or a vaccine. The invention furthermore relates to a method of preparation of the inventive immunostimulatory composition. The invention also relates to the use of the inventive immunostimulatory composition or its components (for the preparation of a pharmaceutical composition or a vaccine) for the treatment of various diseases. Finally, the invention relates to kits containing the inventive immunostimulatory composition, its components and/or the pharmaceutical composition or vaccine.

1 - 17 . (canceled) 
     
     
         18 . An immunostimulatory composition comprising:
 a) an adjuvant component, comprising at least one RNA, complexed with protamine wherein the weight ratio of the at least one mRNA to protamine in the adjuvant component is 2:1 to 3:1; and   b) at least one free mRNA, encoding at least one antigen, wherein the molar ratio of the RNA of the adjuvant component to the at least one free mRNA of the second component b) is 1:1 to 1:4.   
     
     
         19 . A pharmaceutical composition, comprising an immunostimulatory composition according to  claim 18  and optionally a pharmaceutically acceptable carrier, adjuvant, and/or vehicle. 
     
     
         20 . A method for preparing an immunostimulatory composition as defined according to  claim 18 , comprising following steps: (i) preparing the adjuvant component by mixing in a specific ratio the at least one mRNA and the protamine; and (ii) preparing the immunostimulatory composition by adding in a specific ratio of the at least one free mRNA to the adjuvant component prepared according to step (i). 
     
     
         21 . A kit comprising the immunostimulatory composition according to  claim 18 , and a pharmaceutically acceptable carrier and technical instructions with information on the administration and dosage of the immunostimulatory composition and/or the pharmaceutically acceptable carrier. 
     
     
         22 . A method for treating or preventing cancer in a subject comprising administering by injection an effect amount of at least a first immunostimulatory composition to the subject, said immunostimulatory composition comprising:
 a) an adjuvant component, comprising at least one RNA, complexed with protamine; and   b) at least one free mRNA, encoding at least a first tumor antigen, wherein the molar ratio of the RNA of the adjuvant component a) to the at least one free mRNA of the second component b) is in the range of 0.01:1 to 1:0.01.   
     
     
         23 . The method of  claim 22 , wherein the N/P ratio of the RNA to the protamine in the adjuvant component is in the range of 0.1-10. 
     
     
         24 . The method of  claim 22 , wherein the N/P ratio of the RNA to the protamine in the adjuvant component is in the range of 0.3-4. 
     
     
         25 . The method of  claim 22 , wherein the molar ratio of the RNA of the adjuvant component a) to the at least one free mRNA of the second component b) is in the range of 1:1 to 1:4. 
     
     
         26 . The method of  claim 25 , wherein the molar ratio of the RNA of the adjuvant component a) to the at least one free mRNA of the second component b) is in the range of 2:1 to 3:1. 
     
     
         27 . The method of  claim 22 , wherein the at least one free RNA is a mRNA. 
     
     
         28 . The method of  claim 27 , wherein the at least one free mRNA and the at least one mRNA of the adjuvant component are identical to each other. 
     
     
         29 . The method of  claim 27 , wherein the G/C content of the coding region of at least one free mRNA is increased compared with the G/C content of the coding region of a native RNA encoding the tumor antigen. 
     
     
         30 . The method of  claim 22 , wherein the at least one free mRNA encodes a tumor antigen, selected from 5T4, 707-AP, 9D7, AFP, AlbZIP HPG1, alpha5beta1-Integrin, alpha5beta6-Integrin, alpha-methylacyl-coenzyme A racemase, ART-4, B7H4, BAGE-1, BCL-2, BING-4, CA 15-3/CA 27-29, CA 19-9, CA 72-4, CA125, calreticulin, CAMEL, CASP-8, cathepsin B, cathepsin L, CD19, CD20, CD22, CD25, CD30, CD33, CD4, CD52, CD55, CD56, CD80, CEA, CLCA2, CML28, Coactosin-like protein, Collagen XXIII, COX-2, CT-9/BRD6, Cten, cyclin B1, cyclin D1, cyp-B, CYPB1, DAM-10/MAGE-B1, DAM-6/MAGE-B2, EGFR/Her1, EMMPRIN, EpCam, EphA2, EphA3, ErbB3, EZH2, FGF-5, FN, Fra-1, G250/CAIX, GAGE-1, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7b, GAGE-8, GDEP, GnT-V, gp100, GPC3, HAGE, HAST-2, hepsin, Her2/neu/ErbB2, HERV-K-MEL, HNE, homeobox NKX 3.1, HOM-TES-14/SCP-1, HOM-TES-85, HPV-E6, HPV-E7, HST-2, hTERT, iCE, IGF-1R, IL-13Ra2, IL-2R, IL-5, immature laminin receptor, kallikrein 2, kallikrein 4, Ki67, KIAA0205, KK-LC-1, KM-HN-1, LAGE-1, Livin, MAGE-A1, MAGE-A10, MAGE-A12, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A9, MAGE-B1, MAGE-B10, MAGE-B16, MAGE-B17, MAGE-B2, MAGE-B3, MAGE-B4, MAGE-B5, MAGE-B6, MAGE-C1, MAGE-C2, MAGE-C3, MAGE-D1, MAGE-D2, MAGE-D4, MAGE-E1, MAGE-E2, MAGE-F1, MAGE-H1, MAGEL2, mammaglobin A, MART-1/Melan-A, MART-2, matrix protein 22, MC1R, M-CSF, Mesothelin, MG50/PXDN, MMP 11, MN/CA IX-antigen, MRP-3, MUC1, MUC2, NA88-A, N-acetylglucos-aminyltransferase-V, Neo-PAP, NGEP, NMP22, NPM/ALK, NSE, NY-ESO-1, NY-ESO-B, OA1, OFA-iLRP, OGT, OS-9, osteocalcin, osteopontin, p15, p15, p190 minor bcr-abl, p53, PAGE-4, PAI-1, PAI-2, PAP, PART-1, PATE, PDEF, Pim-1-Kinase, Pin1, POTE, PRAME, prostein, proteinase-3, PSA, PSCA, PSGR, PSM, PSMA, RAGE-1, RHAMM/CD168, RU1, RU2, 8400, SAGE, SART-1, SART-2, SART-3, SCC, Sp7, SSX-1, SSX-2/HOM-MEL-40, SSX-4, STAMP-1, STEAP, survivin, survivin-213, TA-90, TAG-72, TARP, TGFb, TGFbR11, TGM-4, TRAG-3, TRG, TRP-1, TRP-2/6b, TRP-2/INT2, Trp-p8, Tyrosinase, UPA, VEGF, VEGFR-2/FLK-1, WT1; alpha-actinin-4/m, ARTC1/m, ber/abl, beta-Catenin/m, BRCA1/m, BRCA2/m, CASP-5/m, CASP-8/m, CDC27/m, CDK4/m, CDKN2A/m, CML66, COA-1/m, DEK-CAN, EFTUD2/m, ELF2/m, ETV6-AML1, FN1/m, GPNMB/m, HLA-A*0201-R170I, HLA-A11/m, HLA-A2/m, HSP70-2M, KIAA0205/m, K-Ras/m, LDLR-FUT, MART2/m, ME1/m, MUM-1/m, MUM-2/m, MUM-3/m, Myosin class 1/m, neo-PAP/m, NFYC/m, N-Ras/m, OGT/m, OS-9/m, p53/m, Pml/RARa, PRDX5/m, PTPRX/m, RBAF600/m, SIRT2/m, SYT-SSX-1, SYT-SSX-2, TEL-AML1, TGFbRII, and TPI/m. 
     
     
         31 . The method of  claim 22 , further comprising administering an effect amount of at least a second immunostimulatory composition to the subject, said immunostimulatory composition comprising:
 a) an adjuvant component, comprising at least one RNA, complexed with protamine; and   b) at least one free mRNA, encoding at least a second tumor antigen, wherein the molar ratio of the RNA of the adjuvant component a) to the at least one free mRNA of the second component b) is in the range of 0.01:1 to 1:0.01.   
     
     
         32 . The method of  claim 31 , further comprising administering an effect amount of at least a third, fourth, fifth, or sixth immunostimulatory composition to the subject, said immunostimulatory composition comprising:
 a) an adjuvant component, comprising at least one RNA, complexed with protamine; and   b) at least one free mRNA, encoding at least a third, fourth, fifth, or sixth tumor antigen, wherein the molar ratio of the RNA of the adjuvant component a) to the at least one free mRNA of the second component b) is in the range of 0.01:1 to 1:0.01.   
     
     
         33 . The method of  claim 31 , wherein the method comprises administration of mRNAs encoding the hTERT, WT1, MAGE-A2, 5T4, MAGE-A3, MUC1, Her-2/neu, NY-ESO-1, CEA, Survivin, MAGE-C1, and/or MAGE-C2 tumor antigens. 
     
     
         34 . The method of  claim 33 , comprising administering mRNAs encoding the NY-ESO-1, 5T4, Survivin, MAGE-C1, and MAGE-C2 tumor antigens. 
     
     
         35 . The method of  claim 22 , wherein the composition is administered by intradermal injection. 
     
     
         36 . The method of  claim 22 , wherein the subject has cancer. 
     
     
         37 . The method of  claim 36 , wherein the subject has non-small cell lung cancer (NSCLC). 
     
     
         38 . The method of  claim 23 , wherein the composition is administered by intradermal injection and wherein the subject has non-small cell lung cancer (NSCLC).