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 composition comprising 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 composition of claim 1 , wherein said second domain consists of or comprises the amino acid sequence set forth as: QHWSYGLRPG (SEQ. ID NO. 9).
3. The composition of claim 1 , wherein said first lytic domain is positioned at the NH 2 -terminus relative to said second domain.
4. The composition of claim 1 , wherein said second domain is positioned at the NH 2 -terminus relative to said first lytic domain.
5. The composition of claim 1 , wherein said first lytic domain or said second domain has one or more D-amino acids.
6. The composition of claim 1 , wherein said first lytic domain has a D-amino acid at any K, F or A residue.
7. The composition of claim 1 , wherein said first lytic domain forms an amphipathic alpha-helix.
8. The composition of claim 1 , wherein said first lytic and second domains are joined by a covalent bond.
9. The composition of claim 1 , wherein said first lytic and second domains are joined by a peptide or a non-peptide linker.
10. The composition of claim 9 , wherein said peptide linker has a length from 1 to 25 amino acid residues.
11. The composition of claim 9 , wherein said non-peptide linker comprises a linear carbon chain linker.
12. The composition of claim 9 , wherein said non-peptide linker comprises a linear 6 carbon chain linker.
13. The composition 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 composition 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 composition of claim 1 , wherein said fusion construct is isolated or purified.
16. A pharmaceutical composition comprising the composition of claim 1 .
17. The composition 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.
18. The composition 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.
19. The composition 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.
20. The composition of claim 1 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKK (SEQ. ID NO. 1).
21. The composition of claim 1 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKKFAK (SEQ. ID NO. 4).
22. The composition of claim 1 , wherein the LHRH analog comprises Lupron (leuprolide).
23. The composition of claim 1 , wherein the LHRH analog comprises zoladex (goserelin).
24. The composition of claim 1 , wherein the LHRH analog comprises supprelin (histrelin).
25. The composition of claim 1 , wherein the LHRH analog comprises triptorelin.
26. The composition of claim 1 , wherein the LHRH analog comprises buserelin.
27. The composition of claim 1 , wherein the LHRH analog comprises cetrorelix.
28. The composition of claim 1 , wherein the LHRH analog comprises ganirelix.
29. The composition of claim 1 , wherein the LHRH analog comprises abarelix.
30. The composition of claim 1 , wherein the LHRH analog comprises antide.
31. The composition of claim 1 , wherein the LHRH analog comprises teverelix.
32. The composition of claim 1 , wherein the LHRH analog comprises degarelix (Fe200486).
33. The composition of claim 1 , further comprising an alkylating agent, anti-metabolite, plant extract, plant alkaloid, nitrosourea, hormone, nucleoside or a nucleotide analog.
34. The composition of claim 1 , 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.
35. A composition comprising 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 chorionic gonadotropin beta subunit (βCG), a βCG fragment or a βCG analog.
36. The composition of claim 35 , wherein said first lytic domain is positioned at the NH 2 -terminus relative to said second domain.
37. The composition of claim 35 , wherein said second domain is positioned at the NH 2 -terminus relative to said first lytic domain.
38. The composition of claim 35 , wherein said first lytic domain or said second domain has one or more D-amino acids.
39. The composition of claim 35 , wherein said first lytic domain has a D-amino acid at any K, F or A residue.
40. The composition of claim 35 , wherein said first lytic domain forms an amphipathic alpha-helix.
41. The composition of claim 35 , wherein said first and second domains are joined by a covalent bond.
42. The composition of claim 35 , wherein said first and second domains are joined by a peptide or a non-peptide linker.
43. The composition of claim 42 , wherein said peptide linker has a length from 1 to 25 amino acid residues.
44. The composition of claim 42 , wherein said non-peptide linker comprises a linear carbon chain linker.
45. The composition of claim 42 , wherein said non-peptide linker comprises a linear 6 carbon chain linker.
46. The composition of claim 35 , wherein said first lytic and second domains are joined by peptide linker comprising one or more A, S or G amino acid residues.
47. The composition of claim 35 , wherein said first lytic and second domains are joined by peptide linker sequence comprising or consisting of: GSGGS (SEQ ID NO. 10), or ASAAS (SEQ ID NO. 11).
48. The composition of claim 35 , wherein said fusion construct is isolated or purified.
49. A pharmaceutical composition comprising the-composition of claim 35 .
50. The composition of claim 35 , 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.
51. The composition of claim 35 , 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.
52. The composition of claim 35 , wherein said fusion construct has an IC 50 /HA 50 (hemolytic activity) ratio of less than about 0.02, 0.01, or 0.005.
53. The composition of claim 35 , wherein the KG comprises a sequence SYAVALSAQAALARR (SEQ. ID NO. 8).
54. The composition of claim 35 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKK (SEQ. ID NO. 1).
55. The composition of claim 35 , wherein said first lytic domain consists of peptide KFAKFAKKFAKFAKKFAK (SEQ. ID NO. 4).
56. The composition of claim 35 , further comprising an alkylating agent, anti-metabolite, plant extract, plant alkaloid, nitrosourea, hormone, nucleoside or a nucleotide analog.
57. The composition of claim 35 , 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.