COMPOSITION FOR TREATING PROSTATE CANCER (PCa)

Abstract

The present invention relates to an active (immunostimulatory) composition comprising at least one RNA preferably an mRNA, encoding at least two (preferably different) antigens capable of eliciting an (adaptive) immune response in a mammal wherein the antigens are selected from the group consisting of PSA (Prostate-Specific Antigen), PSMA (Prostate-Specific Membrane Antigen), PSCA (Prostate Stem Cell Antigen), and STEAP (Six Transmembrane Epithelial Antigen of the Prostate). The invention furthermore relates to a vaccine comprising an active (immunostimulatory) composition, and to the use of the active (immunostimulatory) composition (for the preparation of a vaccine) and/or of the vaccine for eliciting an (adaptive) immune response for the treatment of prostate cancer (PCa), preferably of neoadjuvant and/or hormone-refractory prostate cancers, and diseases or disorders related thereto. Finally, the invention relates to kits, particularly to kits of parts, containing the active (immunostimulatory) composition and/or the vaccine.

1 . Active (immunostimulatory) composition comprising at least one RNA, encoding at least two, three or four different antigens,
 a) wherein at least one antigen is selected from:
 STEAP (Six Transmembrane Epithelial Antigen of the Prostate); and 
   b) wherein the further antigen(s) is (are) selected from at least one antigen of any of the following antigens or specific combinations thereof:
 PSA (Prostate-Specific Antigen), or 
 PSMA (Prostate-Specific Membrane Antigen), or 
 PSCA (Prostate Stem Cell Antigen); 
   or
 PSA and PSMA, or 
 PSA and PSCA, or 
 PSMA and PSCA; 
   or
 PSA, PSMA and PSCA. 
   
     
     
         2 . Active (immunostimulatory) composition according to  claim 1  comprising at least one RNA, encoding four different antigens selected from PSA, PSMA, PSCA and STEAP. 
     
     
         3 . Active (immunostimulatory) composition according to any of  claims 1  to  2 , wherein the at least one RNA comprises a length of 250 to 20000 nucleotides. 
     
     
         4 . Active (immunostimulatory) composition according to any of  claims 1  to  3 , wherein the at least one RNA is an mRNA. 
     
     
         5 . Active (immunostimulatory) composition according to any of  claims 1  to  4 , wherein the at least one RNA is a monocistronic, bicistronic or even multicistronic RNA. 
     
     
         6 . Active (immunostimulatory) composition according to  claim 5 , wherein the at least two antigens are each encoded by a monocistronic RNA. 
     
     
         7 . Active (immunostimulatory) composition according to  claim 5 , wherein the at least two antigens are encoded by a bi- or multicistronic RNA. 
     
     
         8 . Active (immunostimulatory) composition according to  claim 5 , wherein the at least two antigens are encoded by a mixture of monocistronic, bicistronic and/or even multicistronic RNAs. 
     
     
         9 . Active (immunostimulatory) composition according to any of  claims 1  to  8 , wherein the at least one RNA comprises a RNA sequence encoding a fragment, a variant or an epitope of an antigen as defined in any of  claims 1  or  2 . 
     
     
         10 . Active (immunostimulatory) composition according to any of  claims 1  to  9 , wherein at least one RNA comprises an RNA selected from RNAs being identical or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% identical to the RNA sequences of  FIG. 2, 5, 8 , or  11  (SEQ ID NOs: 2, 5, 8, or 11). 
     
     
         11 . Active (immunostimulatory) composition according to any of  claims 1  to  10 , wherein the at least one RNA is a modified RNA, in particular a stabilized mRNA. 
     
     
         12 . Active (immunostimulatory) composition according to  claim 11 , wherein the G/C content of the coding region of the at least one RNA is increased compared to the G/C content of the coding region of the wild-type RNA, the coded amino acid sequence of the at least one RNA preferably not being modified compared to the coded amino acid sequence of the wild-type RNA. 
     
     
         13 . Active (immunostimulatory) composition according to any of  claims 11  to  12 , wherein the A/U content in the environment of the ribosome binding site of the at least one RNA is increased compared with the A/U content in the environment of the ribosome binding site of the wild-type RNA. 
     
     
         14 . Active (immunostimulatory) composition according to any of  claims 11  to  12 , wherein the coding region and/or the 5′ and/or 3′ untranslated region of the modified mRNA is modified compared to the wild-type RNA such that it contains no destabilizing sequence elements, the coded amino acid sequence of the modified mRNA preferably not being modified compared to the wild-type RNA. 
     
     
         15 . Active (immunostimulatory) composition according to any of  claims 11  to  14 , wherein the modified mRNA has a 5′ cap structure and/or a poly(A) tail, preferably of 10 to 200 adenosine nucleotides, and/or a poly(C) tail, preferably of 10 to 200 cytosine nucleotides, and/or at least one IRES and/or at least one 5′ and/or 3′ stabilizing sequence. 
     
     
         16 . Active (immunostimulatory) composition according to any of  claims 1  to  15 , wherein at least one RNA comprises an RNA selected from RNAs being identical or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% identical to the RNA sequences of  FIG. 1, 3, 4, 6, 7, 9, 10 or 12  (SEQ ID NOs: 1, 3, 4, 6, 7, 9, 10 or 12). 
     
     
         17 . Active (immunostimulatory) composition according to any of  claims 1  to  16 , wherein the at least one RNA is complexed with one or more polycations, preferably with protamine or oligofectamine, most preferably with protamine. 
     
     
         18 . Active (immunostimulatory) composition according to any of  claims 1  to  17 , wherein the active composition additionally comprises at least one adjuvant. 
     
     
         19 . Active (immunostimulatory) composition according to  claim 18 , wherein the at least one adjuvant is selected from the group consisting of:
 cationic or polycationic compounds, comprising cationic or polycationic peptides or proteins, including protamine, nucleoline, spermin or spermidine, poly-L-lysine (PLL), poly-arginine, basic polypeptides, cell penetrating peptides (CPPs), including HIV-binding peptides, Tat, HIV-1 Tat (HIV), Tat-derived peptides, Penetratin, VP22 derived or analog peptides, HSV VP22 (Herpes simplex), MAP, KALA or protein transduction domains (PTDs, PpT620, prolin-rich peptides, arginine-rich peptides, lysine-rich peptides, MPG-peptide(s), Pep-1, L-oligomers, Calcitonin peptide(s), Antennapedia-derived peptides (particularly from  Drosophila antennapedia ), pAntp, plsl, FGF, Lactoferrin, Transportan, Buforin-2, Bac715-24, SynB, SynB(1), pVEC, hCT-derived peptides, SAP, protamine, spermine, spermidine, or histones, cationic polysaccharides, including chitosan, polybrene, cationic polymers, including polyethyleneimine (PEI), cationic lipids, including DOTMA: [1-(2,3-sioleyloxy)propyl)]-N,N,N-trimethylammonium chloride, DMRIE, di-C14-amidine, DOTIM, SAINT, DC-Chol, BGTC, CTAP, DOPC, DODAP, DOPE: Dioleyl phosphatidylethanol-amine, DOSPA, DODAB, DOIC, DMEPC, DOGS: Dioctadecylamidoglicylspermnin, DIMRI: Dimyristo-oxypropyl dimethyl hydroxyethyl ammonium bromide, DOTAP: dioleoyloxy-3-(trimethylammonio)propane, DC-6-14: O,O-ditetradecanoyl-N-(α-trimethylammonioacetyl)diethanolamine chloride, CLIP1: rac-[(2,3-dioctadecyloxypropyl)(2-hydroxyethyl)]-dimethylammonium chloride, CLIP6: rac-[2(2,3-dihexadecyloxypropyl-oxymethyloxy)ethyl]trimethylammonium, CLIP9: rac-[2(2,3-dihexadecylxypropy-oxysuccinyloxy)ethyl]-trimethylammonium, oligofectamine, or cationic or polycationic polymers, including modified polyaminoacids, including β-aminoacid-polymers or reversed polyamides, modified polyethylenes, including PVP (poly(N-ethyl-4-vinylpyridinium bromide)), modified acrylates, including pDMAEMA (poly(dimethylaminoethyl methylacrylate)), modified Amidoamines including pAMAM (poly(amidoamine)), modified polybetaaminoester (PBAE), including diamine end modified 1,4 butanediol diacrylate-co-5-amino-1-pentanol polymers, dendrimers, including polypropylamine dendrimers or pAMAM based dendrimers, polyimine(s), including PEI: poly(ethyleneimine), poly(propyleneimine), polyallylamine, sugar backbone based polymers, including cyclodextrin based polymers, dextran based polymers, Chitosan, etc., silan backbone based polymers, such as PMOXA-PDMS copolymers, etc., Blockpolymers consisting of a combination of one or more cationic blocks selected of a cationic polymer as mentioned before, and of one or more hydrophilic- or hydrophobic blocks (e.g polyethyleneglycole);   or   cationic or polycationic proteins or peptides, selected from following proteins or peptides having the following total formula (I): (Arg) l ; (Lys) m ; (His) n ; (Orn) o ; (Xaa) x , wherein l+m+n+o+x=8-15, and l, m, n or o independently of each other may be any number selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15, provided that the overall content of Arg, Lys, His and Orn represents at least 50% of all amino acids of the oligopeptide; and Xaa may be any amino acid selected from native (=naturally occurring) or non-native amino acids except of Arg, Lys, His or Orn; and x may be any number selected from 0, 1, 2, 3 or 4, provided, that the overall content of Xaa does not exceed 50% of all amino acids of the oligopeptide;   or   nucleic acids having the formula (II): G l X m G n , wherein: G is guanosine, uracil or an analogue of guanosine or uracil; X is guanosine, uracil, adenosine, thymidine, cytosine or an analogue of the above-mentioned nucleotides; l is an integer from 1 to 40, wherein when l=1 G is guanosine or an analogue thereof, when l>1 at least 50% of the nucleotides are guanosine or an analogue thereof; m is an integer and is at least 3; wherein when m=3 X is uracil or an analogue thereof, when m>3 at least 3 successive uracils or analogues of uracil occur; n is an integer from 1 to 40, wherein when n=1 G is guanosine or an analogue thereof, when n>1 at least 50% of the nucleotides are guanosine or an analogue thereof;   or   nucleic acids having the formula (III): C l X m C n , wherein: C is cytosine, uracil or an analogue of cytosine or uracil; X is guanosine, uracil, adenosine, thymidine, cytosine or an analogue of the above-mentioned nucleotides; l is an integer from 1 to 40, wherein when l=1 C is cytosine or an analogue thereof, when l>1 at least 50% of the nucleotides are cytosine or an analogue thereof; m is an integer and is at least 3; wherein when m=3 X is uracil or an analogue thereof, when m>3 at least 3 successive uracils or analogues of uracil occur; n is an integer from 1 to 40, wherein when n=1 C is cytosine or an analogue thereof, when n>1 at least 50% of the nucleotides are cytosine or an analogue thereof.   
     
     
         20 . Vaccine, comprising an active (immunostimulatory) composition according to any of  claims 1  to  19 . 
     
     
         21 . Vaccine according to  claim 20 , wherein the active (immunostimulatory) composition according to any of  claims 1  to  19  elicits an adaptive immune response. 
     
     
         22 . Vaccine according to any of  claims 20  and  21 , wherein the vaccine further comprises a pharmaceutically acceptable carrier. 
     
     
         23 . Use of an active (immunostimulatory) composition according to any of  claims 1  to  19  for preparing a vaccine for the treatment of prostate cancer (PCa), preferably of neoadjuvant and/or hormone-refractory prostate cancers, and diseases or disorders related thereto. 
     
     
         24 . Kit, preferably kit of parts, comprising the active (immunostimulatory) composition according to any of  claims 1  to  19 , and/or a vaccine according to any of  claims 20  to  22 , and optionally technical instructions with information on the administration and dosage of the active composition and/or the vaccine.