Lytic domain fusion constructs and methods of making and using same
The invention relates to fusion constructs, methods of using fusion constructs and methods of treating undesirable or aberrant cell proliferation or hyperproliferative disorders, such as tumors, cancers, neoplasia and malignancies.
1. A fusion construct comprising a first lytic domain and a second domain, wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKK (SEQ. ID NO. 1) or peptide KFAKFAKKFAKFAKKFAK (SEQ. ID NO. 4); and said second domain comprises a luteinizing hormone releasing hormone (LHRH), an LHRH fragment, or an LHRH analog.
2. The fusion construct of claim 1 , wherein said LHRH comprises a sequence QHWSYGLRPG (SEQ. ID NO. 9).
3. The fusion construct of claim 1 , wherein said first lytic domain is positioned at the NH 2 -terminus relative to said second domain.
4. The fusion construct of claim 1 , wherein said second domain is positioned at the NH 2 -terminus relative to said first lytic domain.
5. The fusion construct of claim 1 , wherein said first lytic domain or said second domain has one or more D-amino acids.
6. The fusion construct of claim 1 , wherein said first lytic domain has a D-amino acid at any K, F or A residue.
7. The fusion construct of claim 1 , wherein said first lytic domain forms an amphipathic alpha-helix.
8. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by a covalent bond.
9. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by a peptide or a non-peptide linker.
10. The fusion construct of claim 9 , wherein said non-peptide linker comprises a linear carbon chain linker.
11. The fusion construct of claim 9 , wherein said non-peptide linker comprises a linear 6 carbon chain linker.
12. The fusion construct of claim 1 , wherein said peptide linker has a length from 1 to 25 amino acid residues.
13. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by peptide linker comprising one or more A, S or G amino acid residues.
14. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by peptide linker comprising or consisting of: GSGGS (SEQ. ID NO. 10), or ASAAS (SEQ. ID NO. 11).
15. The fusion construct of claim 1 , wherein said fusion construct is isolated or purified.
16. The fusion construct of claim 1 , wherein said fusion construct has greater anti-cell proliferative activity than Phor21-βCG-ala, Phor21-(SEQ. ID NO. 10)-βCG-ala, Phor21-(SEQ. ID NO. 11)-βCG-ala, or Phor 14-βCG-ala, as ascertained by a lower IC 50 value.
17. The fusion construct of claim 1 , wherein said fusion construct has a smaller IC 50 /HA 50 (hemolytic activity) ratio, than Phor21-βCG-ala, Phor21-(SEQ. ID NO. 10)-βCG-ala, Phor21-(SEQ. ID NO. 11)-βCG-ala, or Phor 14-βCG-ala.
18. The fusion construct of claim 1 , wherein said fusion construct has an IC 50 /HA 50 (hemolytic activity) ratio of less than about 0.02, 0.01, or 0.005.
19. The fusion construct of claim 1 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKK (SEQ. ID NO. 1).
20. The fusion construct of claim 1 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKKFAK (SEQ. ID NO. 4).
21. The fusion construct of claim 1 wherein the LHRH analog comprises Lupron (leuprolide).
22. The fusion construct of claim 1 , wherein the LHRH analog comprises zoladex (goserelin).
23. The fusion construct of claim 1 , wherein the LHRH analog comprises supprelin (histrelin).
24. The fusion construct of claim 1 , wherein the LHRH analog comprises triptorelin.
25. The fusion construct of claim 1 , wherein the LHRH analog comprises buserelin.
26. The fusion construct of claim 1 , wherein the LHRH analog comprises centrorelix.
27. The fusion construct of claim 3 , wherein the LHRH analog comprises ganirelix.
28. The fusion construct of claim 1 , wherein the LHRH analog comprises abarelix.
29. The fusion construct of claim 1 , wherein the LHRH analog comprises antide.
30. The fusion construct of claim 1 , wherein the LHRH analog comprises teverelix.
31. The fusion construct of claim 1 , wherein the LHRH analog comprises degarelix (Fe200486).
32. A composition comprising the fusion construct of claim 1 .
33. A pharmaceutical composition comprising the fusion construct of claim 1 .
34. A composition, comprising the fusion construct of claim 1 and an alkylating agent, anti-metabolite, plant extract, plant alkaloid, nitrosourea, hormone, nucleoside or a nucleotide analog.
35. A composition, comprising the fusion construct of claim 1 and cyclophosphamide, azathioprine, cyclosporin A, prednisolone, melphalan, chlorambucil, mechlorethamine, busulphan, methotrexate, 6-mercaptopurine, thioguanine, 5-fluorouracil, cytosine arabinoside, AZT, 5-azacytidine (5-AZC), bleomycin, actinomycin D, mithramycin, mitomycin C, carmustine, lomustine, semustine, streptozotocin, hydroxyurea, cisplatin, carboplatin, oxiplatin, mitotane, procarbazine, dacarbazine, taxol (paclitaxel), vinblastine, vincristine, doxorubicin, dibromomannitol, irinotecan, topotecan, etoposide, teniposide, gemcitabine, or pemetrexed.