Lytic domain fusion constructs and methods of making and using same

1. A fusion construct comprising a first lytic domain and a second domain, wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKKFA (SEQ ID NO:3), or KFAKFAKKFAKFAKKFAKF (SEQ ID NO.5) or KFAKFAKKFAKFAKKFAKFA (SEQ ID NO.6); and said second domain comprises a binding moiety, wherein the binding moiety comprises a luteinizing hormone releasing hormone (LHRH), a LHRH analogue, a fragment of a LHRH, or a fragment of a LHRH analogue that binds to a LHRH receptor, or wherein the binding moiety comprises a chorionic gonadotropin beta subunit (β-CG), a β-CG analogue, a fragment of a β-CG or a β-CG analogue that binds to a luteinizing hormone (LH)/chorionic gonadotropin (CG) receptor.

2. The fusion construct of claim 1 , wherein said binding moiety binds a cell.

3. The fusion construct of claim 2 , wherein said cell is a hyperproliferative cell.

4. The fusion construct of claim 2 , wherein said cell is a cancer cell.

5. The fusion construct of claim 1 , wherein said binding moiety comprises a luteinizing hormone releasing hormone (LHRH) or a LHRH analogue that binds to a LHRH receptor.

6. The fusion construct of claim 1 , wherein said binding moiety comprises a luteinizing hormone releasing hormone (LHRH), a LHRH analogue, a fragment of a LHRH, or a fragment of a LHRH analogue that binds to a LHRH receptor.

7. The fusion construct of claim 1 , wherein said LHRH is the amino acid sequence set forth as: QHWSYGLRPG (SEQ. ID NO. 9).

8. The fusion construct of claim 1 , wherein said LHRH analogue is zoladex, supprelin, triptorelin, buserelin, centrorelix, ganirelix, abarelix, antide, teverelix or degarelix (Fe200486).

9. The fusion construct of claim 1 , wherein said binding moiety comprises a chorionic gonadotropin beta subunit (β-CG) or a β-CG analogue that binds to a LH/CG receptor.

10. The fusion construct of claim 1 , wherein said binding moiety is a chorionic gonadotropin beta subunit (β-CG), a β-CG analogue, a fragment of a β-CG or a β-CG analogue that binds to a luteinizing hormone (LH)/chorionic gonadotropin (CG) receptor.

11. The fusion construct of claim 1 , wherein said fragment of β-CG is the amino acid sequence set forth as: SYAVALSAQAALARR (SEQ. ID NO. 8).

12. The fusion construct of claim 1 , wherein said binding moiety has a linear or cyclic structure.

13. The fusion construct of claim 1 , wherein said first lytic domain is positioned at the NH 2 -terminus relative to said second domain.

14. The fusion construct of claim 1 , wherein said second domain is positioned at the NH 2 -terminus relative to said first domain.

15. The fusion construct of claim 1 , wherein said first lytic domain or said second domain has one or more D-amino acids.

16. The fusion construct of claim 1 , wherein said first lytic domain has a D-amino acid at any K, F or A residue.

17. The fusion construct of claim 1 , wherein said first lytic domain forms an amphipathic alpha-helix.

18. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by a covalent bond.

19. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by a peptide or a non-peptide linker.

20. The fusion construct of claim 19 , wherein said non-peptide linker comprises a linear carbon chain linker.

21. The fusion construct of claim 19 , wherein said non-peptide linker comprises a linear 6 carbon chain linker.

22. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by peptide linker sequence having from 1 to 25 amino acid residues.

23. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by peptide linker sequence comprising one or more A, S or G amino acid residues.

24. The fusion construct of claim 1 , wherein said first lytic and second domains are joined by peptide sequence comprising or consisting of: GSGGS (SEQ ID NO:10), or ASAAS(SEQ ID NO.11).

25. The fusion construct of claim 1 , further comprising a third, fourth, fifth, sixth or seventh domain.

26. The fusion construct of claim 1 , wherein said fusion construct is isolated or purified.

27. The fusion construct of claim 1 , wherein said fusion construct comprises a mixture.

28. 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.

29. 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.

30. 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.

31. A composition comprising the fusion construct of claim 1 .

32. The composition of claim 31 , further comprising an alkylating agent, anti-metabolite, plant extract, plant alkaloid, nitrosourea, hormone, nucleoside or a nucleotide analog.

33. The composition of claim 31 , further comprising 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.

34. A composition comprising the fusion construct of claim 1 , and an anti-cell proliferative or immune stimulating agent.

35. A pharmaceutical composition comprising the fusion construct of claim 1 .

36. The pharmaceutical composition of claim 35 , further comprising an alkylating agent, anti-metabolite, plant extract, plant alkaloid, nitrosourea, hormone, nucleoside or a nucleotide analog.

37. The pharmaceutical composition of claim 35 , further comprising further comprising 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.

38. A unit dosage, comprising the fusion construct of claim 1 in a pharmaceutical composition.

39. A kit comprising the fusion construct of claim 1 and instructions for reducing or inhibiting proliferation of a cell, reducing or inhibiting proliferation of a hyperproliferating cell, reducing or inhibiting proliferation of a neoplastic, tumor or cancer cell, treating a subject having a hyperproliferative disorder, treating a subject having a neoplasia, tumor or cancer, or reducing fertility of an animal.

40. The fusion construct of claim 1 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKKFAKF (SEQ ID NO.5).

41. The fusion construct of claim 1 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKKFA (SEQ ID NO.3).

42. The fusion construct of claim 1 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKKFAKFA (SEQ ID NO.6).

43. An isolated or purified peptide consisting of KFAKFAKKFAKFAKKFA (SEQ ID NO:3), KFAKFAKKFAKFAKKFAKF (SEQ ID NO:5) or KFAKFAKKFAKFAKKFAKFA (SEQ ID NO:6).