Anticancer fusion protein comprising TRAIL and interferon
A fusion protein comprising domain (a) which is a functional fragment of hTRAIL protein sequence, which fragment begins with an amino acid at a position not lower than hTRAIL95, or a homolog of said functional fragment having at least 70% sequence identity; and domain (b) which is a sequence of an immunostimulating effector peptide, wherein the sequence of domain (b) is attached at the C-terminus or N-terminus of domain (a). The fusion protein can be used for the treatment of cancer diseases.
1. A fusion protein comprising:
domain (a) which is a functional fragment of human Tumor Necrosis Factor-Related Apoptosis Inducing Ligand (hTRAIL) protein, wherein said fragment begins with an amino acid at a position not lower than hTRAIL95 and ends with the amino acid at position hTRAIL281, or a homolog of said functional fragment having at least 70% sequence identity; and
domain (b) which is an immunostimulating effector peptide selected from the group consisting of pseudodimer of interferon gamma of SEQ ID NO: 19 and the pseudodimer of interferon alpha 2b of SEQ ID NO: 46;
wherein domain (b) is attached at the C-terminus or N-terminus of domain (a)
and wherein hTRAIL has the sequence set forth as SEQ ID NO: 16.
2. The fusion protein according to claim 1 wherein domain (a) comprises the fragment of hTRAIL protein that begins with an amino acid from the range hTRAIL95 to hTRAIL122, inclusive, and ends with the amino acid hTRAIL281.
3. A pharmaceutical composition, comprising as an active ingredient the fusion protein as defined in claim 2 in combination with a pharmaceutically acceptable carrier.
4. The fusion protein according to claim 2 wherein domain (a) is selected from the group consisting of fragments of hTRAIL95-281 that begin with an amino acid in position 95, 116, 120, 121 or 122.
5. A pharmaceutical composition, comprising as an active ingredient the fusion protein as defined in claim 4 in combination with a pharmaceutically acceptable carrier.
6. The fusion protein according to claim 1 , which between domain (a) and domain (b) contains at least one domain (c) comprising a protease cleavage site selected from the group consisting of a sequence recognized by metalloprotease MMP, a sequence recognized by urokinase uPA, and combinations thereof.
7. A pharmaceutical composition, comprising as an active ingredient the fusion protein as defined in claim 6 in combination with a pharmaceutically acceptable carrier.
8. The fusion protein according to claim 6 , wherein the sequence recognized by metalloprotease MMP is SEQ ID NO: 20, and the sequence recognized by urokinase uPA is SEQ ID NO: 21.
9. A pharmaceutical composition, comprising as an active ingredient the fusion protein as defined in claim 8 in combination with a pharmaceutically acceptable carrier.
10. The fusion protein according to claim 6 , wherein domain (c) is a combination of the sequence recognized by metalloprotease MMP and the sequence recognized by urokinase uPA located next to each other.
11. A pharmaceutical composition, comprising as an active ingredient the fusion protein as defined in claim 10 in combination with a pharmaceutically acceptable carrier.
12. The fusion protein according to claim 6 , which contains two domains (c) and between two domains (c) contains a domain (d) of a linker for attachment of a PEG molecule, wherein domain (d) has a sequence selected from the group consisting of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, and SEQ ID NO: 25.
13. The fusion protein according to claim 12 which additionally comprises a glycine-serine flexible steric linker between domains (a), (b), (c) or (d).
14. A pharmaceutical composition, comprising as an active ingredient the fusion protein as defined in claim 12 in combination with a pharmaceutically acceptable carrier.
15. The fusion protein according to claim 6 which additionally comprises a glycine-serine flexible steric linker between domains (a), (b) or (c).
16. The fusion protein according to claim 1 the amino acid sequence of which is selected from the group consisting SEQ ID NO: 12; SEQ ID NO: 13; SEQ ID NO: 14; SEQ ID NO: 15; and SEQ ID NO: 44.
17. A pharmaceutical composition, comprising as an active ingredient the fusion protein as defined in claim 1 in combination with a pharmaceutically acceptable carrier.
18. A method of treatment of cancer in a mammal comprising administration to the mammal in need thereof of an antineoplastic-effective amount of the fusion protein as defined in claim 1 .
19. The method of claim 18 , wherein the mammal is a human.