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 domain and a second domain, wherein said first domain consists of a 12 to 28 amino acid sequence that includes a peptide KFAKFAKKFAKFAKKFAK; and said second domain comprises a sequence SYAVALSAQAALARR.
2 . A fusion construct comprising a first domain and a second domain, wherein said first domain consists of an amino acid sequence KFAKFAKKFAKFAKKFAK; and said second domain comprises a sequence SYAVALSAQAALARR.
3 .- 14 . (canceled)
15 . The fusion construct of claim 1 , wherein said first or second domain consists of or comprises an amino acid sequence of about 1 to 10, 10 to 20, 15 to 20, 20 to 30, 30 to 40, 40 to 50, 60 to 70, 70 to 80, 80 to 90, 90 to 100 or more amino acids.
16 .- 19 . (canceled)
20 . The fusion construct of claims 1 or 2 , wherein said first domain is positioned at the NH 2 -terminus relative to said second domain.
21 . The fusion construct of claims 1 or 2 , wherein said second domain is positioned at the NH 2 -terminus relative to said first domain.
22 . The fusion construct of claims 1 or 2 , wherein said first domain or said second domain has one or more D-amino acids.
23 . The fusion construct of claims 1 or 2 , wherein said first domain has a D-amino acid at any K, F or A residue.
24 . The fusion construct of claims 1 or 2 , wherein said first domain forms an amphipathic alpha-helix.
25 . The fusion construct of claims 1 or 2 , wherein said first and second domains are joined by a covalent bond.
26 . The fusion construct of claims 1 or 2 , wherein said first and second domains are joined by a peptide or a non-peptide linker.
27 . The fusion construct of claims 1 or 2 , wherein said first and second domains are joined by peptide sequence having from 1 to 25 amino acid residues, or a linear carbon chain.
28 . The fusion construct of claims 1 or 2 , wherein said first and second domains are joined by peptide sequence comprising one or more A, S or G amino acid residues.
29 . The fusion construct of claims 1 or 2 , wherein said first and second domains are joined by peptide sequence comprising or consisting of: GSGGS, ASAAS, or CCCCCC.
30 . (canceled)
31 . The fusion construct of claims 1 or 2 , wherein said fusion construct is isolated or purified.
32 . (canceled)
33 . A composition comprising the fusion construct of claims 1 or 2 .
34 . A pharmaceutical composition comprising the fusion construct of claims 1 or 2 .
35 .- 39 . (canceled)
40 . A nucleic acid molecule that encodes the fusion construct of claim 1 .
41 . A vector comprising the nucleic acid molecule of claim 40 .
42 . A host cell transformed with the vector of claim 41 .
43 . A cell that expresses the fusion construct of claims 1 or 2 .
44 . A method of reducing or inhibiting proliferation of a cell, comprising contacting a cell with the fusion construct of claim 1 in an amount sufficient to reduce or inhibit proliferation of the cell.
45 .- 46 . (canceled)
47 . A method of reducing or inhibiting proliferation of a neoplastic, tumor, cancer or malignant cell, comprising contacting the cell with the fusion construct of claim 1 in an amount sufficient to reduce or inhibit proliferation of the neoplastic, tumor, cancer or malignant cell.
48 .- 56 . (canceled)
57 . A method of selectively reducing or inhibiting proliferation of a neoplastic, tumor, cancer or malignant cell that expresses a receptor or antigen, comprising contacting the cell with the fusion construct of claim 1 in an amount sufficient to reduce or inhibit proliferation of the neoplastic, tumor, cancer or malignant cell, wherein the binding moiety of said fusion construct binds to the receptor, ligand, or antigen expressed by the cell.
58 .- 63 . (canceled)
64 . A method of treating a subject having a neoplasia, tumor, cancer or malignancy, comprising administering to the subject an amount of the fusion construct of any of claims 1 or 2 sufficient to reduce or inhibit proliferation of the neoplasia, tumor, cancer or malignancy.
65 . (canceled)
66 . The method of claim 64 , wherein the neoplasia, tumor, cancer or malignancy is metastatic, non-metastatic or benign.
67 . The method of claim 64 , wherein the neoplasia, tumor, cancer or malignancy comprises a solid cellular mass.
68 . The method of claim 64 , wherein the neoplasia, tumor, cancer or malignancy comprises hematopoietic cells.
69 . The method of claim 64 , wherein the neoplasia, tumor, cancer or malignancy comprises a carcinoma, sarcoma, lymphoma, leukemia, adenoma, adenocarcinoma, melanoma, glioma, glioblastoma, meningioma, neuroblastoma, retinoblastoma, astrocytoma, oligodendrocytoma, mesothelioma, reticuloendothelial, lymphatic or haematopoietic neoplasia, tumor, cancer or malignancy.
70 . The method of claim 64 , wherein the sarcoma comprises a lymphosarcoma, liposarcoma, osteosarcoma, chondrosarcoma, leiomyosarcoma, rhabdomyosarcoma or fibrosarcoma.
71 . (canceled)
72 . The method of claim 64 , wherein the neoplasia, tumor, cancer or malignancy comprises a lung, thyroid, head or neck, nasopharynx, throat, nose or sinuses, brain, spine, breast, adrenal gland, pituitary gland, thyroid, lymph, gastrointestinal (mouth, esophagus, stomach, duodenum, ileum, jejunum (small intestine), colon, rectum), genito-urinary tract (uterus, ovary, cervix, endometrial, bladder, testicle, penis, prostate), kidney, pancreas, liver, bone, bone marrow, lymph, blood, muscle, or skin neoplasia, tumor, or cancer.
73 .- 86 . (canceled)
87 . The method of claim 64 , wherein the treatment results in partial or complete destruction of the neoplastic, tumor, cancer or malignant cell mass, volume, size or numbers of cells, stimulating, inducing or increasing neoplastic, tumor, cancer or malignant cell necrosis, lysis or apoptosis, reducing neoplasia, tumor, cancer or malignancy volume size, cell mass, inhibiting or preventing progression or an increase in neoplasia, tumor, cancer or malignancy volume, mass, size or cell numbers, or prolonging lifespan.
88 . The method of claim 64 , wherein the treatment results in reducing or decreasing severity, duration or frequency of an adverse symptom or complication associated with or caused by the neoplasia, tumor, cancer or malignancy.
89 .- 90 . (canceled)
91 . The method of claim 64 , wherein the subject is a mammal.
92 . The method of claim 64 , wherein the subject is a human.
93 .- 95 . (canceled)
96 . A method of reducing or treating endometriosis of an animal, comprising administering to the animal an amount of a fusion construct of claim 1 sufficient to treat endometriosis.
97 . A method of reducing or treating benign prostate hyperplasia, comprising administering to the animal an amount of a fusion construct of claim 1 sufficient to treat benign prostate hyperplasia.
98 . A method of reducing or treating a fibroid or polyp, comprising administering to the animal an amount of a fusion construct of claim 1 sufficient to treat the fibroid or polyp.
99 . The method of claim 98 , wherein the fibroid or polyp is present in breast, uterus, vagina, cervix or fallopian tube.
100 . The fusion construct of claims 1 or 2 , wherein said fusion construct has greater anti-cell proliferative activity than Phor21-βCG-ala, Phor21-GSGGS-βCG-ala, Phor21-ASAAS-βCG-ala, or Phor 14-βCG-ala, as ascertained by a lower IC50 value.
101 . The fusion construct of claims 1 or 2 , wherein said fusion construct has a smaller IC50/(HA50, hemolytic activity) ratio, than Phor21-βCG-ala, Phor21-GSGGS-βCG-ala, Phor21-ASAAS-βCG-ala, or Phor 14-βCG-ala.
102 . The fusion construct of claims 1 or 2 , wherein said fusion construct has an IC50/(HA50, hemolytic activity) ratio of less than about 0.02, 0.01, or 0.005.
103 .- 108 . (canceled)