Fusion Comprising A Cell Penetrating Peptide, A Multi Epitope And A TLR Peptide Agonist For Treatment Of Cancer

Abstract

The present invention provides a complex for use in the prevention and/or treatment of cancer, the complex comprising a) a cell penetrating peptide, b) at least one antigen or antigenic epitope, and c) at least one TLR peptide agonist, wherein the components a)-c) are covalently linked. In particular, compositions for use in the prevention and/or treatment of cancer, such as a pharmaceutical compositions and vaccines are provided.

1 . A complex comprising:
 a) a cell penetrating peptide;   b) at least one antigen or antigenic epitope; and   c) at least one TLR peptide agonist,   wherein the components a)-c) are covalently linked,   for use in the prevention and/or treatment of colorectal cancer.   
     
     
         2 . The complex for use according to  claim 1 , wherein the complex is a recombinant polypeptide or a recombinant protein. 
     
     
         3 . The complex for use according to  claim 1  or  2 , wherein the cell penetrating peptide
 (i) has a length of the amino acid sequence of said peptide of 5 to 50 amino acids in total, preferably of 10 to 45 amino acids in total, more preferably of 15 to 45 amino acids in total; and/or 
 (ii) has an amino acid sequence comprising a fragment of the minimal domain of ZEBRA, said minimal domain extending from residue 170 to residue 220 of the ZEBRA amino acid sequence according to SEQ ID NO: 3, wherein, optionally, 1, 2, 3, 4, or 5 amino acids have been substituted, deleted, and/or added without abrogating said peptide's cell penetrating ability, or a variant thereof. 
 
     
     
         4 . The complex for use according to  claim 3 , wherein the cell penetrating peptide has an amino acid sequence comprising a Ser (S) at the equivalent of position 189 of the ZEBRA amino acid sequence according to SEQ ID NO: 3. 
     
     
         5 . The complex for use according to  claim 3  or  4 , wherein the cell penetrating peptide has an amino acid sequence comprising:
 (i) a sequence according to the following general formula (I):
   X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 SX 13 X 14 X 15 X 16 X 17    
 
 with 0, 1, 2, 3, 4, or 5 amino acids which are substituted, deleted, and/or added without abrogating said peptide's cell penetrating ability, wherein 
 X 1  is K, R, or H, preferably X 1  is K or R; 
 X 2  is R, K, or H, preferably X 2  is R or K; 
 X 3  is Y, W, or F, preferably X 3  is Y, W, or F; 
 X 4  is K, R, or H, preferably X 4  is K or R; 
 X 5  is N or Q; 
 X 6  is R, K, or H, preferably X 6  is R or K; 
 X 7  is V, I, M, L, F, or A, preferably X 7  is V, I, M or L; 
 X 8  is A, V, L, I, or G, preferably X 8  is A or G; 
 X 9  is S or T; 
 X 10  is R, K, or H, preferably X 10  is R or K; 
 X 11  is K, R, or H, preferably X 11  is K or R; 
 X 13  is R, K, or H, preferably X 13  is R or K; 
 X 14  is A, V, L, I, or G, preferably X 14  is A or G; 
 X 15  is K, R, or H, preferably X 15  is K or R; 
 X 16  is F, L, V, I, Y, W, or M, preferably X 16  is F, Y or W; and 
 X 17  is K, R, or H, preferably X 17  is K or R. 
 
     
     
         6 . The complex for use according to  claim 3  or  4 , wherein the cell penetrating peptide has an amino acid sequence comprising an amino acid sequence selected from the group consisting of amino acid sequences according to SEQ ID NOs: 4-13, or sequence variants thereof without abrogating said peptide's cell penetrating ability, in particular sequence variants thereof sharing at least 70% sequence identity, preferably at least 80% sequence identity and more preferably at least 90% sequence identity without abrogating said peptide's cell penetrating ability. 
     
     
         7 . The complex for use according to  claim 6 , wherein the cell penetrating peptide has an amino acid sequence comprising or consisting of an amino acid sequence according to SEQ ID NO: 6 (CPP3/Z13), SEQ ID NO: 7 (CPP4/Z14), SEQ ID NO: 8 (CPP5/Z15), or SEQ ID NO: 11 (CPP8/Z18), or sequence variants thereof without abrogating said peptide's cell penetrating ability, in particular sequence variants thereof sharing at least 70% sequence identity, preferably at least 80% sequence identity and more preferably at least 90% sequence identity without abrogating said peptide's cell penetrating ability. 
     
     
         8 . The complex for use according to  claim 7 , wherein the cell penetrating peptide comprises or consists of a peptide having an amino acid sequence according to SEQ ID NO: 6 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         9 . The complex for use according to any of  claims 1  to  8 , wherein the at least one antigen or antigenic epitope is selected from the group consisting of: (i) a peptide, a polypeptide, or a protein, (ii) a polysaccharide, (iii) a lipid, (iv) a lipoprotein, (v) a glycolipid, (vi) a nucleic acid, and (vii) a small molecule drug or a toxin. 
     
     
         10 . The complex for use according to any of  claims 1  to  9 , wherein the at least one antigen or antigenic epitope comprises or consists of at least one tumor epitope, preferably of at least one colorectal cancer epitope. 
     
     
         11 . The complex for use according to any of  claims 1  to  10 , wherein the at least one antigen or antigenic epitope is at least one CD4 +  epitope and/or at least one CD8 +  epitope. 
     
     
         12 . The complex for use according to any of  claims 1  to  11 , wherein the at least one antigen or antigenic epitope is a peptide, a polypeptide, or a protein. 
     
     
         13 . The complex for use according to any of  claims 1  to  12 , wherein the complex comprises more than one antigen or antigenic epitope, in particular 2, 3, 4, 5, 6, 7, 8, 9, 10 or more antigens or antigenic epitopes. 
     
     
         14 . The complex for use according to  claim 13 , wherein the more than one antigen or antigenic epitope, in particular 2, 3, 4, 5, 6, 7, 8, 9, 10 or more antigens or antigenic epitopes, are positioned consecutively in the complex. 
     
     
         15 . The complex for use according to any of  claims 10  to  14 , wherein the at least one tumor epitope is an epitope of an antigen selected from the group consisting of EpCAM, HER-2, MUC-1, TOMM34, RNF 43, KOC1, VEGFR, βhCG, survivin, CEA, TGFβR2, p53, KRas, OGT, CASP5, COA-1, MAGE, SART and IL13Ralpha2. 
     
     
         16 . The complex for use according to any of  claims 10  to  14 , wherein the at least one tumor epitope is an epitope of an antigen selected from the group consisting of ASCL2, EpCAM, HER-2, MUC-1, TOMM34, RNF 43, KOC1, VEGFR, βhCG, survivin, CEA, TGFβR2, p53, KRas, OGT, GASPS, COA-1, MAGE, SART and IL13Ralpha2. 
     
     
         17 . The complex for use according to  claim 15 , wherein the at least one tumor epitope is an epitope of an antigen selected from the group consisting of EpCAM, MUC-1, survivin, CEA, KRas, MAGE-A3 and IL13Ralpha2, preferably the at least one tumor epitope is an epitope of an antigen selected from the group consisting of EpCAM, MUC-1, survivin, CEA, KRas and MAGE-A3, more preferably the at least one tumor epitope is an epitope of an antigen selected from the group consisting of EpCAM, MUC-1, survivin and CEA. 
     
     
         18 . The complex for use according to  claim 15  or  16 , wherein the at least one tumor epitope is an epitope of an antigen selected from the group consisting of ASCL2, EpCAM, MUC-1, survivin, CEA, KRas, MAGE-A3 and IL13Ralpha2, preferably the at least one tumor epitope is an epitope of an antigen selected from the group consisting of ASCL2, EpCAM, MUC-1, survivin, CEA, KRas and MAGE-A3, more preferably the at least one tumor epitope is an epitope of an antigen selected from the group consisting of ASCL2, EpCAM, MUC-1, survivin and CEA and even more preferably the at least one tumor epitope is an epitope of an antigen selected from the group consisting of ASCL2, EpCAM, survivin and CEA. 
     
     
         19 . The complex for use according to  claim 18 , wherein the complex comprises an epitope of survivin. 
     
     
         20 . The complex for use according to  claim 19 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 52, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         21 . The complex for use according to  claim 20 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 53. 
     
     
         22 . The complex for use according to any one of  claims 19 - 21 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 95 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         23 . The complex for use according to any one of  claims 18 - 22 , wherein the complex comprises an epitope of CEA. 
     
     
         24 . The complex for use according to  claim 23 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 54, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         25 . The complex for use according to  claim 24 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 55 and/or a peptide having an amino acid sequence according to SEQ ID NO: 56. 
     
     
         26 . The complex for use according to any one of  claims 23 - 25 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 96 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         27 . The complex for use according to any one of  claims 18 - 26 , wherein the complex comprises an epitope of ASCL2. 
     
     
         28 . The complex for use according to  claim 27 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 92, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         29 . The complex for use according to  claim 28 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 93 and/or a peptide having an amino acid sequence according to SEQ ID NO: 94. 
     
     
         30 . The complex for use according to any one of  claims 27 - 29 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 97 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         31 . The complex for use according to any of  claims 12  to  30 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof; 
 b) one or more epitopes of MUC-1 or functional sequence variants thereof; 
 c) one or more epitopes of survivin or functional sequence variants thereof; 
 d) one or more epitopes of CEA or functional sequence variants thereof; 
 e) one or more epitopes of KRas or functional sequence variants thereof; and/or 
 f) one or more epitopes of MAGE-A3 or functional sequence variants thereof. 
 
     
     
         32 . The complex for use according to any of  claims 12  to  30 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof; 
 b) one or more epitopes of MUC-1 or functional sequence variants thereof; 
 c) one or more epitopes of survivin or functional sequence variants thereof; 
 d) one or more epitopes of CEA or functional sequence variants thereof; 
 e) one or more epitopes of KRas or functional sequence variants thereof; 
 f) one or more epitopes of MAGE-A3 or functional sequence variants thereof; and/or 
 g) one or more epitopes of ASCL2 or functional sequence variants thereof. 
 
     
     
         33 . The complex for use according to  claim 31  or  32 , wherein the complex comprises
 a) a fragment of EpCAM comprising one or more epitopes or a functional sequence variant thereof; 
 b) a fragment of MUC-1 comprising one or more epitopes or a functional sequence variant thereof; 
 c) a fragment of survivin comprising one or more epitopes or a functional sequence variant thereof; 
 d) a fragment of CEA comprising one or more epitopes or a functional sequence variant thereof; 
 e) a fragment of KRas comprising one or more epitopes or a functional sequence variant thereof; and/or 
 f) a fragment of MAGE-A3 comprising one or more epitopes or a functional sequence variant thereof. 
 
     
     
         34 . The complex for use according to  claim 31  or  32 , wherein the complex comprises
 a) a fragment of EpCAM comprising one or more epitopes or a functional sequence variant thereof; 
 b) a fragment of MUC-1 comprising one or more epitopes or a functional sequence variant thereof; 
 c) a fragment of survivin comprising one or more epitopes or a functional sequence variant thereof; 
 d) a fragment of CEA comprising one or more epitopes or a functional sequence variant thereof; 
 e) a fragment of KRas comprising one or more epitopes or a functional sequence variant thereof; 
 f) a fragment of MAGE-A3 comprising one or more epitopes or a functional sequence variant thereof; and/or 
 g) a fragment of ASCL2 comprising one or more epitopes or a functional sequence variant thereof. 
 
     
     
         35 . The complex for use according to any of  claims 31 - 34 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   d) one or more epitopes of CEA or functional sequence variants thereof; and   f) one or more epitopes of MAGE-A3 or functional sequence variants thereof.   
     
     
         36 . The complex for use according to any of  claims 31 - 34 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   b) one or more epitopes of MUC-1 or functional sequence variants thereof;   d) one or more epitopes of CEA or functional sequence variants thereof; and   f) one or more epitopes of MAGE-A3 or functional sequence variants thereof.   
     
     
         37 . The complex for use according to any of  claims 31 - 34 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   b) one or more epitopes of MUC-1 or functional sequence variants thereof;   d) one or more epitopes of CEA or functional sequence variants thereof; and   e) one or more epitopes of KRas or functional sequence variants thereof.   
     
     
         38 . The complex for use according to any of  claims 31 - 34 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   c) one or more epitopes of survivin or functional sequence variants thereof;   d) one or more epitopes of CEA or functional sequence variants thereof; and   f) one or more epitopes of MAGE-A3 or functional sequence variants thereof.   
     
     
         39 . The complex for use according to any of  claims 31 - 34 , wherein the complex comprises
 b) one or more epitopes of MUC-1 or functional sequence variants thereof;   c) one or more epitopes of survivin or functional sequence variants thereof; and/or   f) one or more epitopes of MAGE-A3 or functional sequence variants thereof.   
     
     
         40 . The complex for use according to any of  claims 31 - 34 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   b) one or more epitopes of MUC-1 or functional sequence variants thereof;   c) one or more epitopes of survivin or functional sequence variants thereof; and/or   d) one or more epitopes of CEA or functional sequence variants thereof.   
     
     
         41 . The complex for use according to  claim 40 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   b) one or more epitopes of MUC-1 or functional sequence variants thereof;   c) one or more epitopes of survivin or functional sequence variants thereof; and   d) one or more epitopes of CEA or functional sequence variants thereof.   
     
     
         42 . The complex for use according to  claim 40 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   b) one or more epitopes of MUC-1 or functional sequence variants thereof; and   d) one or more epitopes of CEA or functional sequence variants thereof.   
     
     
         43 . The complex for use according to  claim 40 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof; and   d) one or more epitopes of CEA or functional sequence variants thereof.   
     
     
         44 . The complex for use according to  claim 40 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof.   
     
     
         45 . The complex for use according to  claim 40 , wherein the complex comprises
 d) one or more epitopes of CEA or functional sequence variants thereof.   
     
     
         46 . The complex for use according to any of  claims 31 - 34 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   c) one or more epitopes of survivin or functional sequence variants thereof;   d) one or more epitopes of CEA or functional sequence variants thereof; and/or   g) one or more epitopes of ASCL2 or functional sequence variants thereof.   
     
     
         47 . The complex for use according to  claim 46 , wherein the complex comprises
 a) one or more epitopes of EpCAM or functional sequence variants thereof;   c) one or more epitopes of survivin or functional sequence variants thereof;   d) one or more epitopes of CEA or functional sequence variants thereof; and   g) one or more epitopes of ASCL2 or functional sequence variants thereof.   
     
     
         48 . The complex for use according to  claim 46 , wherein the complex comprises
 c) one or more epitopes of survivin or functional sequence variants thereof;   d) one or more epitopes of CEA or functional sequence variants thereof; and   g) one or more epitopes of ASCL2 or functional sequence variants thereof.   
     
     
         49 . The complex for use according to  claim 48 , wherein the complex comprises in N- to C-terminal direction:
 (i) one or more epitopes of CEA or functional sequence variants thereof;   (ii) one or more epitopes of survivin or functional sequence variants thereof; and   (iii) one or more epitopes of ASCL2 or functional sequence variants thereof.   
     
     
         50 . The complex for use according to  claim 49 , wherein the complex comprises in N- to C-terminal direction:
 (i) a peptide having an amino acid sequence according to SEQ ID NO: 54, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity;   (ii) a peptide having an amino acid sequence according to SEQ ID NO: 52, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity; and   (iii) a peptide having an amino acid sequence according to SEQ ID NO: 92, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity.   
     
     
         51 . The complex for use according to  claim 50 , wherein the C-terminus of (i) the peptide having an amino acid sequence according to SEQ ID NO: 54 or the fragment or variant thereof is directly linked to the N-terminus of (ii) the peptide having an amino acid sequence according to SEQ ID NO: 52 or the fragment or variant thereof; and the C-terminus of (ii) the peptide having an amino acid sequence according to SEQ ID NO: 52 or the fragment or variant thereof is directly linked to the N-terminus of (iii) the peptide having an amino acid sequence according to SEQ ID NO: 92 or the fragment or variant thereof. 
     
     
         52 . The complex for use according to any one of  claims 48 - 51 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 96 or a functional sequence variant thereof having at least 70% sequence identity; a peptide having an amino acid sequence according to SEQ ID NO: 95 or a functional sequence variant thereof having at least 70% sequence identity; and a peptide having an amino acid sequence according to SEQ ID NO: 97 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         53 . The complex for use according to any one of  claims 48 - 52 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 98 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         54 . The complex for use according to  claim 46 , wherein the complex comprises
 g) one or more epitopes of ASCL2 or functional sequence variants thereof.   
     
     
         55 . The complex for use according to any of  claims 10  to  14 , wherein the at least one tumor epitope is an epitope of a neoantigen, preferably an epitope of a colorectal cancer-specific neoantigen. 
     
     
         56 . The complex for use according to any of  claims 1  to  55 , wherein the at least one TLR peptide agonist is a TLR2, TLR4 and/or TLR5 peptide agonist, preferably a TLR2 peptide agonist and/or a TLR4 peptide agonist. 
     
     
         57 . The complex for use according to  claim 56 , wherein the at least one TLR peptide agonist comprises or consists of an amino acid sequence according to SEQ ID NO: 15 or 71; or of a sequence variant thereof, in particular a sequence variant thereof sharing at least 70% sequence identity, preferably at least 80% sequence identity and more preferably at least 90% sequence identity without abrogating said peptide's TLR agonist ability. 
     
     
         58 . The complex for use according to  claim 57 , wherein the at least one TLR peptide agonist comprises or consists of a peptide having an amino acid sequence according to SEQ ID NO: 71; or a functional sequence variant thereof having at least 70% sequence identity, preferably at least 80% sequence identity and more preferably at least 90% sequence identity (without abrogating said peptide's TLR agonist ability). 
     
     
         59 . The complex for use according to any of  claims 1  to  58 , wherein the complex comprises more than one TLR peptide agonists, in particular 2, 3, 4, 5, 6, 7, 8, 9, 10 or more TLR peptide agonists. 
     
     
         60 . The complex for use according to any of  claims 1  to  59 , wherein the at least one antigen or antigenic epitope is positioned C-terminally of the cell penetrating peptide, whereby the cell penetrating peptide and the at least one antigen or antigenic epitope are optionally linked by a further component, e.g. a linker, a spacer, or by the at least one TLR peptide agonist. 
     
     
         61 . The complex for use according to  claim 60 , wherein the at least one antigen or antigenic epitope is positioned C-terminally of the cell penetrating peptide, whereby the cell penetrating peptide and the at least one antigen or antigenic epitope are optionally linked by a further component, e.g. a linker, a spacer, but not by the at least one TLR peptide agonist. 
     
     
         62 . The complex for use according to any of  claims 2  to  61 , wherein said complex is a recombinant polypeptide or a recombinant protein and the components a) to c) are positioned in N-terminal→C-terminal direction of the main chain of said complex in the order:
 (α) component a)-component b)-component c); or 
 (β) component c)-component a)-component b), 
 wherein the components may be linked by a further component, in particular by a linker or a spacer. 
 
     
     
         63 . The complex for use according to any one of  claims 1 - 62  comprising, preferably in N- to C-terminal direction, the following components:
 (i) a peptide having an amino acid sequence according to SEQ ID NO: 6 or a functional sequence variant thereof having at least 70% sequence identity; 
 (ii) a peptide having an amino acid sequence according to SEQ ID NO: 96, or a functional sequence variant thereof having at least 70% sequence identity; 
 (iii) a peptide having an amino acid sequence according to SEQ ID NO: 95, or a functional sequence variant thereof having at least 70% sequence identity; 
 (iv) a peptide having an amino acid sequence according to SEQ ID NO: 97, or a functional sequence variant thereof having at least 70% sequence identity; and 
 (v) a peptide having an amino acid sequence according to SEQ ID NO: 71; or a functional sequence variant thereof having at least 70% sequence identity. 
 
     
     
         64 . The complex for use according to any of  claims 1  to  62 , wherein said complex comprises or consists of an amino acid sequence according to any of SEQ ID NOs: 26-28, 33-34, 37-41, 46, 69, or 72-87; or said complex comprises or consists of an amino acid sequence sharing at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with any of SEQ ID NOs: 26-28, 33-34, 37-41, 46, 69, or 72-87. 
     
     
         65 . The complex for use according to any of  claims 1  to  63 , wherein said complex comprises or consists of an amino acid sequence according to any of SEQ ID NOs: 26-28, 33-34, 37-41, 46, 69, or 72-91; or said complex comprises or consists of an amino acid sequence sharing at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with any of SEQ ID NOs: 26-28, 33-34, 37-41, 46, 69, or 72-91. 
     
     
         66 . The complex for use according to any of  claims 1  to  63  and  65 , wherein said complex comprises or consists of a polypeptide having an amino acid sequence according to SEQ ID NO: 89, or a functional sequence variant thereof having at least 50% sequence identity. 
     
     
         67 . A complex comprising:
 a) a cell penetrating peptide;   b) at least three antigenic epitopes; and   c) at least one TLR peptide agonist,   wherein the components a)-c) are covalently linked, and   wherein the at least three antigenic epitopes comprise (i) one or more epitopes of survivin or (a) functional sequence variant(s) thereof, (ii) one or more epitopes of CEA or (a) functional sequence variant(s) thereof, and (iii) one or more epitopes of ASCL2 or (a) functional sequence variant(s) thereof.   
     
     
         68 . The complex according to  claim 67 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 52, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         69 . The complex according to  claim 68 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 53. 
     
     
         70 . The complex according to any one of  claims 67 - 69 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 95 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         71 . The complex according to any one of  claims 67 - 70 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 54, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         72 . The complex according to  claim 71 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 55 and/or a peptide having an amino acid sequence according to SEQ ID NO: 56. 
     
     
         73 . The complex according to  claim 71  or  72 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 96 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         74 . The complex according to any one of  claims 67 - 73 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 92, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         75 . The complex according to  claim 74 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 93 and/or a peptide having an amino acid sequence according to SEQ ID NO: 94. 
     
     
         76 . The complex according to  claim 74  or  75 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 97 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         77 . The complex according to any one of  claims 67 - 76 , wherein the complex comprises in N- to C-terminal direction:
 (i) one or more epitopes of CEA or functional sequence variants thereof;   (ii) one or more epitopes of survivin or functional sequence variants thereof; and   (iii) one or more epitopes of ASCL2 or functional sequence variants thereof.   
     
     
         78 . The complex according to  claim 77 , wherein the complex comprises in N- to C-terminal direction:
 (i) a peptide having an amino acid sequence according to SEQ ID NO: 54, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity;   (ii) a peptide having an amino acid sequence according to SEQ ID NO: 52, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity; and   (iii) a peptide having an amino acid sequence according to SEQ ID NO: 92, or a fragment thereof having a length of at least 10 amino acids, or a functional sequence variant thereof having at least 70% sequence identity.   
     
     
         79 . The complex according to  claim 78 , wherein the C-terminus of (i) the peptide having an amino acid sequence according to SEQ ID NO: 54 or the fragment or variant thereof is directly linked to the N-terminus of (ii) the peptide having an amino acid sequence according to SEQ ID NO: 52 or the fragment or variant thereof; and the C-terminus of (ii) the peptide having an amino acid sequence according to SEQ ID NO: 52 or the fragment or variant thereof is directly linked to the N-terminus of (iii) the peptide having an amino acid sequence according to SEQ ID NO: 92 or the fragment or variant thereof. 
     
     
         80 . The complex according to any one of  claims 67 - 79 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 96 or a functional sequence variant thereof having at least 70% sequence identity; a peptide having an amino acid sequence according to SEQ ID NO: 95 or a functional sequence variant thereof having at least 70% sequence identity; and a peptide having an amino acid sequence according to SEQ ID NO: 97 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         81 . The complex according to any one of  claims 67 - 80 , wherein the complex comprises a peptide having an amino acid sequence according to SEQ ID NO: 98 or a functional sequence variant thereof having at least 70% sequence identity. 
     
     
         82 . The complex according to any one of  claims 67 - 81 , wherein the complex is a recombinant polypeptide or a recombinant protein. 
     
     
         83 . The complex according to any one of  claims 67 - 82 , wherein the cell penetrating peptide is as defined in any one of  claims 3 - 8 . 
     
     
         84 . The complex according to any one of  claims 67 - 83 , wherein the at least one TLR peptide agonist is as defined in any one of  claims 56 - 58 . 
     
     
         85 . The complex according to any one of  claims 67 - 84  comprising, preferably in N- to C-terminal direction, the following components:
 (i) a peptide having an amino acid sequence according to SEQ ID NO: 6 or a functional sequence variant thereof having at least 70% sequence identity; 
 (ii) a peptide having an amino acid sequence according to SEQ ID NO: 96, or a functional sequence variant thereof having at least 70% sequence identity; 
 (iii) a peptide having an amino acid sequence according to SEQ ID NO: 95, or a functional sequence variant thereof having at least 70% sequence identity; 
 (iv) a peptide having an amino acid sequence according to SEQ ID NO: 97, or a functional sequence variant thereof having at least 70% sequence identity; and 
 (v) a peptide having an amino acid sequence according to SEQ ID NO: 71; or a functional sequence variant thereof having at least 70% sequence identity, wherein components (i)-(v) are optionally linked by a linker or spacer. 
 
     
     
         86 . The complex according to any of  claims 67 - 85 , wherein said complex comprises or consists of a polypeptide having an amino acid sequence according to SEQ ID NO: 89, or a functional sequence variant thereof having at least 50% sequence identity. 
     
     
         87 . A nucleic acid encoding the complex as defined in any of  claims 1  to  66  for use in the prevention and/or treatment of colorectal cancer, wherein the complex is a polypeptide or a protein. 
     
     
         88 . A nucleic acid encoding the complex according to any of  claims 67  to  86 , wherein the complex is a polypeptide or a protein. 
     
     
         89 . The nucleic acid according to  claim 87  or  88 , wherein the nucleic acid comprises a polynucleotide encoding an amino acid sequence according to any of SEQ ID NOs: 26-28, 33-34, 37-41, 46, 69, or 72-87 or an amino acid sequence sharing at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with any of SEQ ID NOs: 26-28, 33-34,37-41,46,69, or 72-87. 
     
     
         90 . The nucleic acid according to  claim 87  or  88 , wherein the nucleic acid comprises a polynucleotide encoding an amino acid sequence according to any of SEQ ID NOs: 26-28, 33-34, 37-41, 46, 69, or 72-91 or an amino acid sequence sharing at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with any of SEQ ID NOs: 26-28, 33-34, 37-41, 46, 69, or 72-91. 
     
     
         91 . The nucleic acid according to  claim 90 , wherein the nucleic acid comprises a polynucleotide encoding an amino acid sequence according to SEQ ID NO: 89, or a functional sequence variant thereof having at least 50% sequence identity. 
     
     
         92 . The nucleic acid according to any of  claims 87 - 91 , wherein the nucleic acid is a DNA molecule or an RNA molecule; preferably selected from genomic DNA; cDNA; siRNA; rRNA; mRNA; antisense DNA; antisense RNA; ribozyme; complimentary RNA and/or DNA sequences; RNA and/or DNA sequences with or without expression elements, regulatory elements, and/or promoters; a vector; and combinations thereof. 
     
     
         93 . A vector comprising the nucleic acid as defined in any of  claims 87 - 92 , preferably for use in the prevention and/or treatment of colorectal cancer. 
     
     
         94 . A host cell comprising the vector as defined in  claim 93 , preferably for use in the prevention and/or treatment of colorectal cancer. 
     
     
         95 . A method of preparing the complex as defined in any of  claims 1  to  86 , the complex being a polypeptide or a protein, comprising cultivating a host cell as defined in  claim 72  in a culture medium and separating said complex from the culture medium or from the host cell lysate after host cell lysis. 
     
     
         96 . A cell loaded with a complex as defined in any one of  claims 1  to  86 , preferably for use in the prevention and/or treatment of colorectal cancer. 
     
     
         97 . A cell according to  claim 96 , wherein said cell is an antigen presenting cell, preferably a dendritic cell. 
     
     
         98 . A composition comprising at least one of:
 (i) a complex as defined in any one of  claims 6  to  86 ;   (ii) a nucleic acid as defined in any one of  claims 87 - 92 ;   (i) a vector as defined in  claim 93 ;   (iv) a host cell as defined in  claim 94 ; or   (v) a cell loaded with a complex as defined in  claim 96  or  97  preferably for use in the prevention and/or treatment of colorectal cancer.   
     
     
         99 . A vaccine comprising at least one of:
 (i) a complex as defined in any one of  claims 1  to  86 ;   (ii) a nucleic acid as defined in any one of  claims 87 - 92 ;   (ii) a vector as defined in  claim 93 ;   (iv) a host cell as defined in  claim 94 ; or   (v) a cell loaded with a complex as defined in  claim 96  or  97  preferably for use in the prevention and/or treatment of colorectal cancer.   
     
     
         100 . A pharmaceutical composition comprising at least one complex as defined in any one of  claims 1  to  86  or at least one cell as defined in  claim 96  or  97 , and a pharmaceutically acceptable carrier, preferably for use in the prevention and/or treatment of colorectal cancer. 
     
     
         101 . The pharmaceutical composition according to  claim 100 , wherein said composition comprises at least two different complexes. 
     
     
         102 . A diagnostic composition comprising a complex as defined in any one of  claims 1  to  86 . 
     
     
         103 . A combination of
 (i) a complex as defined in any of  claims 1  to  86 ; and   (ii) a chemotherapeutic agent, a targeted drug and/or an immunotherapeutic agent, such as an immune checkpoint modulator,   preferably for use in the prevention and/or treatment of colorectal cancer.   
     
     
         104 . The combination for use according to  claim 103 , wherein (i) the complex and (ii) the chemotherapeutic agent, the targeted drug and/or the immunotherapeutic agent, such as an immune checkpoint modulator, are administered at about the same time. 
     
     
         105 . The combination for use according to  claim 103  or  104 , wherein (i) the complex and (ii) the chemotherapeutic agent, the targeted drug and/or the immunotherapeutic agent, such as an immune checkpoint modulator, are administered consecutively. 
     
     
         106 . The combination for use according to any of  claims 103  to  105 , wherein (i) the complex and (ii) the chemotherapeutic agent, the targeted drug and/or the immunotherapeutic agent, such as an immune checkpoint modulator, are administered via the same route of administration. 
     
     
         107 . The combination for use according to any of  claims 103  to  105 , wherein (i) the complex and (ii) the chemotherapeutic agent, the targeted drug and/or the immunotherapeutic agent, such as an immune checkpoint modulator, are administered via distinct routes of administration. 
     
     
         108 . The combination for use according to any of  claims 103  to  107 , wherein (i) the complex and (ii) the chemotherapeutic agent, the targeted drug and/or the immunotherapeutic agent, such as an immune checkpoint modulator, are provided in distinct compositions. 
     
     
         109 . The combination for use according to any of  claims 103  to  106 , wherein (i) the complex and (ii) the chemotherapeutic agent, the targeted drug and/or the immunotherapeutic agent, such as an immune checkpoint modulator, are provided in the same composition. 
     
     
         110 . The combination for use according to any of  claims 103  to  109 , wherein the chemotherapeutic agent, the targeted drug and/or the immunotherapeutic agent is an immune checkpoint modulator. 
     
     
         111 . The combination for use according to  claim 110 , wherein the immune checkpoint modulator is a PD-1 inhibitor. 
     
     
         112 . A kit comprising the complex as defined in any of  claims 1  to  86 , the nucleic acid as defined in any one of  claims 87 - 92 , the vector as defined in  claim 93 , the cell as defined in  claim 94 ,  96  or  97 , the composition as defined in  claim 98 , the vaccine as defined in  claim 99 , and/or the pharmaceutical composition as defined in  claim 100  or  101 , preferably for use in the prevention and/or treatment of colorectal cancer. 
     
     
         113 . The kit according to  claim 112 , wherein the kit further comprises a package insert or instruction leaflet with directions to prevent or to treat colorectal cancer by using the complex as defined in any one of  claims 1  to  86 , the nucleic acid as defined in any one of  claims 87 - 92 , the vector as defined in  claim 93 , the cell as defined in  claim 94 ,  96  or  97 , the composition as defined in  claim 98 , the vaccine as defined in  claim 99 , and/or the pharmaceutical composition as defined in  claim 100  or  101 . 
     
     
         114 . The kit according to  claim 112  or  113 , wherein the kit further comprises a chemotherapeutic agent, a targeted drug and/or an immunotherapeutic agent, such as an immune checkpoint modulator. 
     
     
         115 . A method for preventing and/or treating colorectal cancer or initiating, enhancing or prolonging an anti-tumor-response in a subject in need thereof comprising administering to the subject a complex comprising:
 a) a cell penetrating peptide;   b) at least one antigen or antigenic epitope; and   c) at least one TSR peptide agonist,   wherein the components a)-c) are covalently linked.   
     
     
         116 . The method of  claim 115 , wherein the complex as defined in any one of  claims 1  to  86 , the nucleic acid as defined in any one of  claims 87 - 92 , the vector as defined in  claim 93 , the cell as defined in  claim 94 ,  96  or  97 , the composition as defined in  claim 98 , the vaccine as defined in  claim 99 , and/or the pharmaceutical composition as defined in  claim 100  or  101  is administered to the subject. 
     
     
         117 . The method of  claim 115  or  116 , wherein the subject has colorectal cancer.