Consumable Cryopreserved Cells Transiently Overexpressing Gene(s) Encoding Drug Transporter Protein(s) and/or Drug Metabolizing Enzyme(s)

Abstract

The present invention discloses cryopreserved recombinant cells for screening drug candidates that transiently overexpress one or more drug transporter proteins and/or drug metabolizing enzymes. Advantageously, such cells provide a cost-efficient consumable product that streamlines the process of screening whether drug candidates are substrates or inhibitors of drug transporter proteins and/or drug metabolizing enzymes.

What is claimed is: 
     
         1 . A cryopreserved recombinant cell comprising one or more transiently overexpressed genes encoding a protein selected from the group consisting of a drug transporter protein and a drug metabolizing enzyme, or a combination thereof wherein activity of the drug transporter protein or the drug metabolizing enzyme or combinations is detectable in a population of the cryopreserved recombinant cell following thaw from cryopreservation. 
     
     
         2 . The recombinant cell of  claim 1 , wherein said one or more genes encodes a drug metabolizing enzyme. 
     
     
         3 . The recombinant cell of  claim 2 , wherein the drug metabolizing enzyme is selected from the group consisting of cytochrome P450, UDP-glucouronyl transferase, alcohol dehydrogenase, monoamine oxidase and aldehyde oxidase. 
     
     
         4 . The recombinant cell of  claim 1 , which transiently overexpresses one or more genes encoding a protein selected from the group consisting of an ATP binding cassette transporter and a solute carrier transporter protein. 
     
     
         5 . The recombinant cell of  claim 4 , wherein said one or more genes is selected from the group consisting of MDR1/Mdr1a/Mdr1b, MRP1/Mrp1, MRP2/Mrp2, MRP3/Mrp3, MRP4/Mrp4, MRP5/Mrp5, MRP6/Mrp6, MRP7/Mrp7, MRP 8/Mrp8, BCRP/Bcrp, BSEP/Bsep, OATP2/Oatp2, OATP1B3/Oatp1b3, OAT1/Oat1, OAT2/Oat2, OAT3/Oat3, OAT4/Oat4, OCT1/Oct1, OCT2/Oct2, OATP1/Oatp1, PEPT1/Pept1, PEPT2/Pept2, OCTN1/Octn1, OCTN2/Octn2, MATE1/Mate1, MATE2K/Mate2, URAT1/Urat1, ASBT/Asbt, and NTCP/Ntcp or a combination. 
     
     
         6 . The recombinant cell of  claim 4 , wherein said one or more genes is selected from OATP1B1*1a, OATP1B1*1b, OATP1B3, OAT1, OAT3, MATE1 and MATE2K. 
     
     
         7 . The recombinant cell of  claim 1 , wherein the one or more genes is derived individually from human or an animal species selected from mouse, rat, guinea pig, dog, and monkey. 
     
     
         8 . The recombinant cell of  claim 1 , wherein said cell is selected from the group consisting of HEK293, CHO, MDCK, LLC-PK1, Caco-2 and V79 cells. 
     
     
         9 . The recombinant cell of  claim 1 , wherein said cell comprises a hepatocyte. 
     
     
         10 . The recombinant cell of  claim 1 , wherein said cell comprises an endothelial cell. 
     
     
         11 . The recombinant cell of  claim 1 , wherein activity of the protein(s) is detectable in a population of said cell at least 24 hours post plating following thaw from cryopreservation. 
     
     
         12 . The recombinant cell of  claim 1 , wherein activity of the protein(s) is detectable in a population of said cell at least 48 hours post plating following thaw from cryopreservation. 
     
     
         13 . The recombinant cell of  claim 1 , wherein activity of the protein(s) is detectable in a population of said cell at least 72 hours post plating following thaw from cryopreservation. 
     
     
         14 . A process of preparing transiently transfected recombinant cells which transiently overexpress one or more genes encoding a protein selected from a drug transporter protein and a drug metabolizing enzyme, comprising transiently transfecting cells with one or more genes encoding a drug transporter protein or a drug metabolizing enzyme and cryopreserving the transiently transfected recombinant cells within 48 hrs of transfection. 
     
     
         15 . The process of  claim 14 , wherein the transient transfection step comprises electroporation. 
     
     
         16 . The process of  claim 14 , wherein the transiently transfected recombinant cells are cryopreserved at 30 minutes to 24 hours post transfection.