Membrane resident peptide in anti-cancer peptides causes tumor cell necrosis rather than apoptosis of cancer cells

Abstract

An aspect of the invention provides a method of selectively necrosing cells, comprising: providing a plurality cells, including at least one cancer cell and at least one non-cancerous cell; and administering to the cells a compound, including an HDM-2 targeting component and a cytotoxic component attached to the HDM-2 targeting component, wherein said compound comprises a membrane-active form.

What is claimed is: 
     
         1 . A method of selectively necrosing cells, comprising:
 providing a plurality cells, including at least one cancer cell and at least one non-cancerous cell;   administering to the cells a compound, including an HDM-2 targeting component and a cytotoxic component, said cytotoxic component attached to said HDM-2 targeting component, wherein said compound comprises a membrane-active form.   
     
     
         2 . The method of  claim 1 , wherein the cytotoxic component is selected from the group consisting of: a membrane resident peptide, a toxin, a drug, a radionuclide, a whole antibody, an antibody fragment, and combinations thereof. 
     
     
         3 . The method of  claim 1 , wherein the HDM-2 targeting component is selected from the group consisting of: a small molecule, a peptide, a protein, a glycoprotein, a whole antibody, an antibody fragment, and combinations thereof. 
     
     
         4 . The method of  claim 1 , further comprising the step of observing in a cell medium a level of LDH. 
     
     
         5 . The method of  claim 1 , further comprising the step of observing necrosis in the cancer cells. 
     
     
         6 . The method of  claim 1 , further comprising the step of observing a non-response in the non-cancerous cell, wherein the non-response indicates the non-cancerous cell is unaffected. 
     
     
         7 . The method of  claim 1 , wherein the administering step further comprises administering a PNC-27, a PNC-28 peptide, or combinations thereof. 
     
     
         8 . A method of causing membranolysis in at least one cancer cell, comprising:
 administering to at least one cancer cell a compound comprising an HDM-2 targeting component and a pore-forming component attached to said HDM-2 targeting component, wherein said administering step results in at least one transmembrane pore in a cancer cell membrane.   
     
     
         9 . The method of  claim 8 , further wherein the administering step further comprises administering said compound at a dosage. 
     
     
         10 . The method of  claim 8 , further comprising the step of observing membranolysis in the cancer cell by: detecting an LDH amount, performing electron microscopy, observing cell morphology, and combinations thereof. 
     
     
         11 . The method of  claim 8 , further comprising the step of observing necrosis of the cancer cell. 
     
     
         12 . A method of treating cancer in a subject in need thereof, comprising:
 administering to the subject in need thereof a therapeutically effective amount of a compound having an HDM-2 targeting component and a cytotoxic component, said HDM-2 targeting component and said cytotoxic component having a membrane-active form.   
     
     
         13 . The method of  claim 12 , wherein the cytotoxic component is selected from the group consisting of: a membrane resident peptide, a toxin, a drug, a radionuclide, a whole antibody, an antibody fragment, and combinations thereof. 
     
     
         14 . The method of  claim 12 , wherein the HDM-2 targeting component is selected from the group consisting of: a small molecule, a peptide, a protein, a glycoprotein, a whole antibody, an antibody fragment, and combinations thereof. 
     
     
         15 . The method of  claim 12  further comprising the step of determining whether a plurality of cancerous cells have undergone membranolysis. 
     
     
         16 . The method of  claim 12 , further comprising the step of, after the administering step, determining whether a plurality subsequent cancer cells exists. 
     
     
         17 . A method of screening cancer treatments, comprising:
 providing a plurality of cancerous cells; each of said cells having HDM-2 in said cellular membranes;   administering a candidate cancer treatment to the plurality of cancer cells; and   measuring the level of LDH present in a cellular medium.   
     
     
         18 . The method of  claim 17 , further comprising the step of determining whether the candidate cancer treatment has membrane active conformation. 
     
     
         19 . The method of  claim 17 , further comprising the step of determining whether the candidate cancer treatment includes an HDM-2 targeting component. 
     
     
         20 . The method of  claim 17 , further comprising the step of observing the cancer cell membranes for an area of pore formation. 
     
     
         21 . A method of selectively necrosing cancer cells, comprising:
 providing a plurality of cells, including at least one cancer cell and at least one non-cancerous cell; and   contacting the plurality of cells with an HDM-2 targeting compound which includes a membrane resident peptide (MRP), wherein the HDM-2 targeting compound colocalizes to HDM-2 present in at least one cancer cell membrane, binding to a cell membrane of the at least one cancer cell and adopting a membrane-resident conformation within said cancer cell membrane.   
     
     
         22 . The method of  claim 21 , wherein adopting the membrane-resident conformation further includes forming a pore in said cancer cell membrane. 
     
     
         23 . The method of  claim 21 , comprising observing necrosis in the plurality of cancer cells. 
     
     
         24 . The method of  claim 21 , comprising observing a non-result in the non-cancerous cells, indicative of non-targeting of said non-cancerous cells. 
     
     
         25 . A method of identifying cancer cells, comprising:
 providing a plurality of cells, wherein at least one of said cells is a candidate cancer cell; and   administering to the plurality of cells an HDM-2 recognition agent.   
     
     
         26 . The method of  claim 25 , further comprising of the step of observing the plurality of cells for the HDM-2 recognition agent colocalization with at least one cell membrane. 
     
     
         27 . The method of  claim 25 , wherein said HDM-2 recognition agent further comprises an HDM-2 targeting component tagged with an observation aid, said observation aid selected from the group consisting of: a dye, an enzyme, a fluorescing agent, a radiopaque material, a radioactive isotope and combinations thereof. 
     
     
         28 . The method of  claim 25 , further comprising identifying at least one cancer cell. 
     
     
         29 . The method of  claim 25 , further comprising mapping a plurality of cancer cells.