Methods and compositions for inducing apoptosis

Abstract

The C-terminal domain of focal adhesion kinase (FAK-CD) was isolated using a Baculoviral system. Using phage display techniques, a phage encoding a 12 amino-acid peptide (peptide 35) and AV3 that binds to FAK-CD were identified. The peptides were also conjugated to TAT-FITC to produce a fluorescently labeled chimeric molecule capable of penetrating cell membranes. Contacting various breast cancer cell lines with these molecule caused detachment, rounding, apoptosis and cell death. These effects were not observed in normal (non-cancerous) breast cells.

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         4 . A method for inducing apoptosis in a cancer cell, the method comprising contacting the cancer cell with an agent that specifically binds focal adhesion kinase at a site that is specifically bound by a peptide comprising the amino acid sequences of SEQ ID NO: 1 and/or SEQ ID NO: 3. 
     
     
         5 . The method of  claim 4 , wherein the agent comprises the amino acid sequence of SEQ ID NO:1 and/or SEQ ID NO: 3 or variants thereof. 
     
     
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         12 . A method of treating cancer comprising:
 administering to a patient a composition comprising SEQ ID NO: 1 and/or SEQ ID NO: 3, derivatives, fragments and variants thereof;   contacting a cancer cell with the composition;   binding of the composition to focal adhesion kinase at a site that is specifically bound by a peptide comprising an amino acid sequence of SEQ ID NO:1 and/or SEQ ID NO: 3, derivatives, variants and fragments thereof; and,   treating cancer.   
     
     
         13 . The method of  claim 12 , wherein the composition enters a cell via a cellular membrane. 
     
     
         14 . The method of  claim 12 , wherein the composition induces apoptosis in an abnormal cell expressing focal adhesion kinase. 
     
     
         15 . The method of  claim 12 , wherein the composition inhibits cell motility. 
     
     
         16 . The method of  claim 12 , wherein the composition inhibits metastasis of a tumor cell. 
     
     
         17 . The method of  claim 12 , wherein contacting a cell with the composition induces apoptosis and/or inhibits cell motility and or metastasis. 
     
     
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         25 . A vector comprising a focal adhesion kinase binding chimeric molecule. 
     
     
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         27 . The vector of  claim 26 , wherein the focal adhesion kinase binding domain is identified by SEQ ID NO: 1 and/or SEQ ID NO: 3, derivatives, fragments and variants thereof. 
     
     
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         32 . A method of treating a cancer patient comprising:
 administering a chimeric fusion protein composition to a patient; and,   contacting a tumor cell with the chimeric fusion protein composition;   modulating the activity of the tumor cell; thereby, treating a cancer patient.   
     
     
         33 . The method of  claim 32 , wherein the chimeric fusion molecule comprises a first domain which binds to focal adhesion kinase molecules in or on a cell. 
     
     
         34 . The method of  claim 33 , wherein the focal adhesion kinase molecule binding first domain of the chimeric fusion protein is identified by SEQ ID NO: 1 and/or SEQ ID NO: 3, derivatives, fragments and variants thereof. 
     
     
         35 . The method of  claim 32 , wherein the chimeric fusion protein composition comprises a second domain comprising a cell permeabilization domain. 
     
     
         36 . The method of  claim 32 , wherein the activity of a tumor cell is apoptosis, motility and invasion. 
     
     
         37 . The method of  claim 32 , wherein the chimeric fusion protein composition induces apoptosis in a tumor cell. 
     
     
         38 . The method of  claim 32 , wherein the chimeric fusion protein inhibits cell motility and invasion. 
     
     
         39 . The method of  claim 32 , wherein the chimeric fusion protein inhibits metastasis of a tumor cell. 
     
     
         40 . The method of  claim 32 , wherein the chimeric fusion protein is co-administered with one or more chemotherapeutic agents. 
     
     
         41 . The method of  claim 40 , wherein the chemotherapeutic agent comprises cyclophosphamide (CTX, 25 mg/kg/day, p.o.), taxanes (paclitaxel or docetaxel), busulfan, cisplatin, cyclophosphamide, methotrexate, daunorubicin, doxorubicin, melphalan, cladribine, vincristine, vinblastine, and chlorambucil. 
     
     
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