Lytic-peptide-Her2/neu (human epidermal growth factor receptor 2) ligand conjugates and methods of use
The invention relates to conjugates that bind to Her2/neu, methods of using conjugates that bind to Her2/neu and methods of treating undesirable or aberrant cell proliferation or hyperproliferative disorders, such as tumors, cancers, neoplasia and malignancies that express Her2/neu.
1. A conjugate comprising an antibody or an antibody fragment that binds to Her2/neu and a second domain, wherein said second domain consists of a 15 to 20 amino acid sequence that includes a peptide selected from the group consisting of KFAKFAKKFAKFAKK (SEQ. ID NO. 1), KFAKFAKKFAKFAKKF (SEQ. ID NO. 2), KFAKFAKKFAKFAKKFA (SEQ. ID NO. 3), KFAKFAKKFAKFAKKFAK (SEQ. ID NO. 4), KFAKFAKKFAKFAKKFAKF (SEQ. ID NO. 5) and KFAKFAKKFAKFAKKFAKFA (SEQ. ID NO. 6).
2. A conjugate comprising an antibody or an antibody fragment that binds to Her2/neu and a second domain, wherein said second domain consists of an amino acid sequence selected from the group consisting of KFAKFAKKFAKFAKK (SEQ. ID NO. 1), KFAKFAKKFAKFAKKF (SEQ. ID NO. 2), KFAKFAKKFAKFAKKFA (SEQ. ID NO. 3), KFAKFAKKFAKFAKKFAK (SEQ. ID NO. 4), KFAKFAKKFAKFAKKFAKF (SEQ. ID NO. 5) and KFAKFAKKFAKFAKKFAKFA (SEQ. ID NO. 6).
3. The conjugate of claim 1 or 2 , wherein said antibody fragment comprises an Fab, Fab′, F(ab) 2 , Fv, Fd, single-chain Fv (scFv), disulfide-linked Fvs (sdFv), V L , V H , Camel Ig, V-NAR, VHH, trispecific (Fab 3 ), bispecific (Fab 2 ), diabody ((V L -V H ) 2 or (V H -V L ) 2 ), triabody (trivalent), tetrabody (tetravalent), minibody ((scF v -C H 3) 2 ), bispecific single-chain Fv (Bis-scFv), IgGdeltaC H 2, scFv-Fc, (scFv) 2 -Fc, affibody, aptamer, avimer or nanobody.
4. The conjugate of claim 1 or 2 , wherein said antibody or antibody fragment comprises a monoclonal antibody.
5. The conjugate of claim 1 or 2 , wherein said antibody or antibody fragment comprises a mammalian antibody or antibody fragment.
6. The conjugate of claim 1 or 2 , wherein said antibody or antibody fragment comprises a human, humanized, primatized, or chimeric antibody or antibody fragment.
7. The conjugate of claim 1 or 2 , wherein said antibody comprises trastuzumab or pertuzumab.
8. The conjugate of claim 1 or 2 , wherein said Her2/neu is expressed on a cell.
9. The conjugate of claim 8 , wherein said cell is a hyperproliferative cell.
10. The conjugate of claim 8 , wherein said cell is a breast, ovarian, uterine, cervical, prostate, stomach, lung, gastric, colon, bladder, glial, hematologic or endometrial cell.
11. The conjugate of claim 1 or 2 , wherein said second domain is positioned at the NH 2 -terminus relative to said antibody or antibody fragment that binds to Her2/neu, or wherein said second domain is positioned at the C-terminus relative to said antibody or antibody fragment that binds to Her2/neu, or wherein said antibody or antibody fragment that binds to Her2/neu is positioned at the NH 2 -terminus relative to said second domain, or wherein said antibody or antibody fragment that binds to Her2/neu is positioned at the C-terminus relative to said second domain.
12. The conjugate of claim 1 or 2 , wherein said second domain has one or more D-amino acids.
13. The conjugate of claim 1 or 2 , wherein said second domain forms an amphipathic alpha-helix.
14. The conjugate of claim 1 or 2 , wherein said antibody or antibody fragment that binds to Her2/neu and said second domain are joined by a covalent bond.
15. The conjugate of claim 1 or 2 , wherein said antibody or antibody fragment that binds to Her2/neu and said second domain are joined by a peptide linker.
16. The conjugate of claim 1 or 2 , wherein said antibody or antibody fragment that binds to Her2/neu and said second domain are joined by a peptide sequence having from 1 to 25 amino acid residues.
17. The conjugate of claim 1 or 2 , further comprising a third, fourth, fifth, sixth or seventh domain.
18. The conjugate of claim 1 or 2 , wherein said conjugate is isolated or purified.
19. The conjugate of claim 1 or 2 , wherein said conjugate comprises a mixture.
20. A composition comprising the conjugate of claim 1 or 2 .
21. A pharmaceutical composition comprising the conjugate of claim 1 or 2 .
22. A composition comprising the conjugate of claim 1 or 2 , and an anti-cell proliferative or immune stimulating agent.
23. A unit dosage, comprising the conjugate of claim 1 or 2 in an amount effective to treat a subject having a neoplasia, tumor or cancer.
24. A kit comprising the conjugate of claim 1 or 2 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, or treating a subject having a neoplasia, tumor or cancer.
25. A nucleic acid molecule that encodes the conjugate of claim 1 or 2 .
26. A method of reducing or inhibiting proliferation of a cell, comprising contacting a cell with the conjugate of claim 1 or 2 in an amount sufficient to reduce or inhibit proliferation of the cell.
27. A method of reducing or inhibiting proliferation of a hyperproliferating cell, comprising contacting a cell with the conjugate of claim 1 or 2 in an amount sufficient to reduce or inhibit proliferation of the hyperproliferating cell.
28. A method of reducing or inhibiting proliferation of a neoplastic, tumor, cancer or malignant cell, comprising contacting the cell with the conjugate of claim 1 or 2 in an amount sufficient to reduce or inhibit proliferation of the neoplastic, tumor, cancer or malignant cell.
29. The method of any of claims 26 or 28 , wherein the cell expresses Her2/neu.
30. A method of selectively reducing or inhibiting proliferation of a cell that expresses Her2/neu, comprising contacting the cell with the conjugate of claim 1 or 2 in an amount sufficient to reduce or inhibit proliferation of the cell that expresses Her2/neu.
31. 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 conjugate of claim 1 or 2 in an amount sufficient to reduce or inhibit proliferation of the neoplastic, tumor, cancer or malignant cell that expresses Her2/neu.
32. A method of treating a subject having a hyperproliferative disorder, comprising administering to the subject an amount of the conjugate of claim 1 or 2 sufficient to treat the hyperproliferative disorder.
33. A method of treating a subject having a neoplasia, tumor, cancer or malignancy, comprising administering to the subject an amount of the conjugate of claim 1 or 2 sufficient to reduce or inhibit proliferation of the neoplasia, tumor, cancer or malignancy.
34. The method of claim 31 or 33 , wherein the neoplasia, tumor, cancer or malignancy is metastatic, non-metastatic or benign.
35. The method of claim 31 or 33 , wherein the neoplasia, tumor, cancer or malignancy comprises a solid cellular mass.
36. The method of claim 31 or 33 , wherein the neoplasia, tumor, cancer or malignancy comprises hematopoietic cells.
37. The method of claim 31 or 33 , 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.
38. The method of claim 37 , wherein the sarcoma comprises a lymphosarcoma, liposarcoma, osteosarcoma, chondrosarcoma, leiomyosarcoma, rhabdomyosarcoma or fibrosarcoma.
39. The method of claim 37 , wherein the haematopoietic neoplasia, tumor, cancer or malignancy comprises a myeloma, lymphoma or leukemia.
40. The method of claim 31 or 33 , 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, wilm's, biliary tract, B-ALL, or hematologic neoplasia, tumor, or cancer.
41. The method of claim 40 , wherein the lung neoplasia, tumor, cancer or malignancy comprises small cell lung or non-small cell lung cancer.
42. The method of claim 31 or 33 , further comprising administering an anti-cell proliferative, anti-neoplastic, anti-tumor, anti-cancer or immune-enhancing treatment or therapy.
43. The method of claim 31 or 33 , 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.
44. The method of claim 31 or 33 , wherein the subject is a mammal.
45. The method of claim 31 or 33 , wherein the subject is a human.
46. The conjugate of claim 15 , wherein the peptide linker comprises a GS linker.
47. The conjugate of claim 1 or 2 , wherein said ligand that binds to Her2/neu and said second domain are joined by a linear carbon chain.