A Novel Complex Comprising A Cell Penetrating Peptide, A Cargo And A TLR Peptide Agonist

Abstract

The present invention provides a novel 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. Moreover, the present invention also provides a nucleic acid encoding such a complex, wherein the complex is a peptide or a protein. Such a nucleic acid may be comprised by a vector, and such a vector may be comprised by a host cell. In particular, compositions, such as a pharmaceutical compositions and vaccines are provided, which may be useful for example in the prevention and/or treatment of a diseases and/or a disorder including cancer, hematological disorders, infectious diseases, autoimmunity disorders and transplant rejections.

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.   
     
     
         2 . The complex according to  claim 1 , wherein the complex is a recombinant polypeptide or a recombinant protein. 
     
     
         3 . The complex according to  claim 1 , 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 .- 6 . (canceled) 
     
     
         7 . The complex according to  claim 3 , 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 (CPPB/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 according to  claim 1 , 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. 
     
     
         9 . The complex according to  claim 1 , wherein the at least one antigen or antigenic epitope comprises or consists of at least one pathogen epitope and/or at least one tumor epitope, preferably the at least one antigen or antigenic epitope comprises or consists of at least one tumor epitope. 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . The complex according to  claim 1 , 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, wherein the more than one antigen or antigenic epitope, in particular 2, 3, 4, 5, 6, 7, 8, 9, 10 or more antigens or epitopes, are positioned consecutively in the complex. 
     
     
         13 . (canceled) 
     
     
         14 . The complex according to  claim 1 , 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. 
     
     
         15 . The complex according to  claim 14 , wherein the at least one TLR peptide agonist comprises or consists of an amino acid sequence according to SEQ ID NO: 15 or 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. 
     
     
         16 .- 18 . (canceled) 
     
     
         19 . The complex according to  claim 2 , wherein 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. 
 
     
     
         20 . A nucleic acid encoding the complex according to  claim 1 , wherein the complex is a polypeptide or a protein. 
     
     
         21 . A vector comprising the nucleic acid according to  claim 20 . 
     
     
         22 . A host cell comprising the vector according to  claim 21 . 
     
     
         23 .- 24 . (canceled) 
     
     
         25 . A cell loaded with a complex according to  claim 1 , wherein said cell is an antigen presenting cell, preferably a dendritic cell. 
     
     
         26 .- 27 . (canceled) 
     
     
         28 . A vaccine comprising at least one of:
 (i) a complex according to  claim 1 ;   (ii) a nucleic acid encoding the complex of (i);   (iii) a vector comprising the nucleic acid according to (ii);   (iv) a host cell comprising the vector according to (iii); or   (v) a cell loaded with a complex according to (i).   
     
     
         29 . A pharmaceutical composition comprising at least one complex according to  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         30 .- 40 . (canceled) 
     
     
         41 . A method for treating 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 TLR peptide agonist,
 wherein the components a)-c) are covalently linked. 
   
     
     
         42 . The method of  claim 41 , wherein the method comprises administering to the subject
 a cell loaded with the complex,   a pharmaceutical composition comprising the complex; and/or   a vaccine comprising
 i) the complex, 
 ii) a nucleic acid encoding the complex, 
 iii) a vector comprising the nucleic acid according to (ii), 
 iv) a host cell comprising the vector according to (iii), or 
 v)a cell loaded with the complex. 
   
     
     
         43 . (canceled) 
     
     
         44 . A method for treating cancer and/or a hematological disorder, preferably a malignant neoplasm of the brain or a malignant neoplasm of lymphoid, hematopoietic and related tissue in a subject in need thereof, most preferably glioblastoma, 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 TLR peptide agonist,
 wherein the components a)-c) are covalently linked. 
   
     
     
         45 . The method of  claim 44 , wherein the method comprises administering to the subject
 a cell loaded with the complex,   a pharmaceutical composition comprising the complex; and/or   a vaccine comprising
 i) the complex, 
 ii) a nucleic acid encoding the complex, 
 iii) a vector comprising the nucleic acid according to (ii), 
 iv) a host cell comprising the vector according to (iii), or 
 v) a cell loaded with the complex.