Complex comprising a cell penetrating peptide, a cargo and a TLR peptide agonist
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 toll-like receptor (TLR) peptide agonist, wherein the TLR peptide agonist is a TLR2 or TLR4 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 2 , wherein the components a) to c) of said complex are positioned in N-terminal→C-terminal direction in the order:
(α) component a)-component b)-component c); or
(β) component c)-component a)-component b).
4. The complex according to claim 3 , wherein the components a) to c) of said complex are positioned in N-terminal→C-terminal direction in the order:
component a)-component b)-component c).
5. The complex according to claim 3 , wherein the components a) to c) of said complex are positioned in N-terminal→C-terminal direction in the order:
component c)-component a)-component b).
6. The complex according to claim 3 , wherein the components are linked by a further component.
7. The complex according to claim 1 , wherein the cell penetrating peptide:
(i) has a length of 5 to 50 amino acids in total; and
(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, or a variant thereof wherein zero, 1, 2, 3, 4, or 5 amino acids have been substituted, deleted, and/or added without abrogating said peptide's cell penetrating ability.
8. The complex according to claim 7 , wherein the cell penetrating peptide has an amino acid sequence comprising 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 a variant thereof sharing at least 90% sequence identity with at least one of SEQ ID NOs: 6, 7, 8, or 11 without abrogating said peptide's cell penetrating ability.
9. The complex according to claim 7 , wherein the cell penetrating peptide has a length of 10 to 45 amino acids in total.
10. The complex according to claim 9 , wherein the cell penetrating peptide has a length of 15 to 45 amino acids in total.
11. The complex according to claim 7 , wherein the cell penetrating peptide has an amino acid sequence 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 a variant thereof sharing at least 90% sequence identity with at least one of SEQ ID NOs: 6, 7, 8, or 11 without abrogating said peptide's cell penetrating ability.
12. 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.
13. The complex according to claim 1 , wherein the at least one antigen or antigenic epitope comprises at least one pathogen epitope, at least one tumor epitope, or a combination thereof.
14. The complex according to claim 13 , wherein the at least one antigen or antigenic epitope comprises at least one tumor epitope.
15. The complex according to claim 1 , wherein the complex comprises more than one antigen or antigenic epitope positioned consecutively in the complex.
16. The complex according to claim 15 , wherein the complex comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, or more antigens or antigenic epitopes.
17. The complex according to claim 1 , wherein the at least one TLR peptide agonist comprises an amino acid sequence according to SEQ ID NO: 15 or a variant thereof sharing at least 90% sequence identity with SEQ ID NO:15 without abrogating said peptide's TLR agonist ability.
18. A pharmaceutical composition comprising at least one complex according to claim 1 and a pharmaceutically acceptable carrier.
19. The complex according to claim 1 , wherein the cell penetrating peptide has an amino acid sequence length of 5 to 50 amino acids in total.
20. The complex according to claim 1 , wherein the cell penetrating peptide 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, or a variant thereof wherein, zero, 1, 2, 3, 4, or 5 amino acids have been substituted, deleted, and/or added without abrogating said peptide's cell penetrating ability.
21. The complex according to claim 1 , wherein the at least one antigen or antigenic epitope consists of at least one pathogen epitope, at least one tumor epitope, or a combination thereof.
22. The complex according to claim 1 , wherein the at least one TLR peptide agonist consists of an amino acid sequence according to SEQ ID NO: 15 or a variant thereof sharing at least 90% sequence identity with SEQ ID NO:15 without abrogating said peptide's TLR agonist ability.
23. The complex according to claim 1 , wherein the components a) to c) of said complex are positioned in N-terminal→C-terminal direction.
24. 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).
25. The vaccine of claim 24 , wherein the components a) to c) of said complex are positioned in N-terminal→C-terminal direction in the order:
(α) component a)-component b)-component c); or
(β) component c)-component a)-component b).
26. A complex comprising:
a) a cell penetrating peptide;
b) at least one antigen or antigenic epitope; and
c) at least one toll-like receptor (TLR) peptide agonist, wherein the TLR peptide agonist is a TLR2 or TLR4 peptide agonist,
wherein the components a)-c) are covalently linked, and
wherein the complex is a recombinant polypeptide or a recombinant protein.
27. The complex according to claim 26 , wherein the cell penetrating peptide has an amino acid sequence length of 5 to 50 amino acids in total.
28. The complex according to claim 26 , wherein the cell penetrating peptide 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, or a variant thereof wherein, zero, 1, 2, 3, 4, or 5 amino acids have been substituted, deleted, and/or added without abrogating said peptide's cell penetrating ability.
29. The complex according to claim 28 , wherein the cell penetrating peptide has an amino acid sequence 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 a variant thereof sharing at least 90% sequence identity with at least one of SEQ ID NOs: 6, 7, 8, or 11 without abrogating said peptide's cell penetrating ability.
30. The complex according to claim 26 , wherein the at least one antigen or antigenic epitope comprises at least one pathogen epitope, at least one tumor epitope, or a combination thereof.
31. The complex according to claim 30 , wherein the at least one antigen or antigenic epitope comprises at least one tumor epitope.
32. The complex according to claim 26 , wherein the at least one TLR peptide agonist consists of an amino acid sequence according to SEQ ID NO: 15 or a variant thereof sharing at least 90% sequence identity with SEQ ID NO:15 without abrogating said peptide's TLR agonist ability.
33. The complex according to claim 26 , wherein the components a) to c) of said complex are positioned in N-terminal→C-terminal direction.
34. The complex according to claim 26 , wherein the complex comprises more than one antigen or antigenic epitope positioned consecutively in the complex.
35. The complex according to claim 34 , wherein the complex comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, or more antigens or antigenic epitopes.
36. A pharmaceutical composition comprising at least one complex according to claim 26 and a pharmaceutically acceptable carrier.
37. A vaccine comprising at least one of:
(i) a complex according to claim 26 ;
(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).
38. The vaccine of claim 37 , wherein the components a) to c) of said complex are positioned in N-terminal→C-terminal direction in the order:
(α) component a)-component b)-component c); or
(β) component c)-component a)-component b).