TISSUE INHIBITOR OF METALLOPROTEINASES (Timp) LINKED TO GLYCOSYLPHOSPHATIDYLINOSITOL (Gpi) - ANCHORS FOR TREATMENT OF CANCER AND SKIN LESIONS

Abstract

The present invention relates to fusion constructs of glycosylphosphatidylinositol (GPI)-anchored tissue inhibitors of metalloproteinases (TIMPs) and their use for the treatment of cancer and in regenerative medicine. By this approach, the GPI-anchored TIMP proteins are incorporated into the surface membrane of tumor cells and render tumor cells sensitive to FAS-induced apoptosis. Furthermore, the fusion constructs of the present invention are effective agents useful in wound healing applications. In one embodiment, the TIMP is linked to mucin followed by GPI in order to enhance surface presentation. The use of GPI to link TIMP renders the resulting fusion protein particularly useful as an anti-cancer agent for the treatment of cancer, and, in particular, any residual cancer following an incomplete surgical resection of primary tumors in an individual.

1 - 14 . (canceled) 
     
     
         15 . A method of treating cancer in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising a fusion construct
 comprising an amino acid sequence of a tissue inhibitor of metalloproteinases (TIMP) or a functionally active portion thereof which retains the activity of TIMP, wherein the functionally active portion comprises a TIMP molecule truncated to the first 50 to 152 N-terminal amino acid residues, and wherein said TIMP or functionally active portion thereof is linked to a glycosylphosphatidylinositol (GPI)-anchor.   
     
     
         16 . The method of  claim 15 , wherein said cancer is selected from the group consisting of breast cancer, renal cancer, prostate cancer, leukemias, seminomas, melanomas, teratomas, lymphomas, neuroblastomas, gliomas, rectal cancer, endometrial cancer, kidney cancer, adrenal cancer, thyroid cancer, blood cancer, skin cancer, cancer of the brain, cervical cancer, intestinal cancer, liver cancer, colon cancer, stomach cancer, intestine cancer, gastrointestinal cancer, lymph node cancer, esophagus cancer, colorectal cancer, pancreas cancer, ear, nose and throat (ENT) cancer, cancer of the uterus, ovarian cancer and lung cancer and the metastases thereof. 
     
     
         17 . The method of  claim 15 , wherein said cancer is residual cancer after surgical removal. 
     
     
         18 . The method of  claim 17 , wherein said fusion construct is locally administered as anti-tumor adjuvant for the treatment of residual cancer in breast cancer patients and patients with glioblastoma (astrocytoma VI). 
     
     
         19 . The method of  claim 17 , wherein said fusion construct is administered at a concentration of 0.5 to 5 μg/ml, preferably 1 μg/ml. 
     
     
         20 . The method of  claim 15 , wherein said fusion construct is administered by spraying into the wound and/or injection into regions that are not available for surgery. 
     
     
         21 . The method of  claim 15 , wherein said fusion construct is a fusion construct (TIMP-GPI) that does not include the mucin domain. 
     
     
         22 . An in-vitro method for inhibition of cancer cell proliferation comprising the step of subjecting a cancer cell line to an effective amount of TIMP-GPI fusion construct
 comprising an amino acid sequence of a tissue inhibitor of metalloproteinases (TIMP) or a functionally active portion thereof which retains the activity of TIMP, wherein the functionally active portion comprises a TIMP molecule truncated to the first 50 to 152 N-terminal amino acid residues, and wherein said TIMP or functionally active portion thereof is linked to a glycosylphosphatidylinositol (GPI)-anchor.   
     
     
         23 . The method of  claim 22 , wherein said cell line is a renal cell carcinoma (RCC) cell line. 
     
     
         24 - 30 . (canceled) 
     
     
         31 . The method of  claim 17 , wherein said fusion construct is administered at a concentration of 1 μg/ml.