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

Patent metadata

Field	Value
Publication number	US-2016102131-A1
Application number	US-201514972012-A
Country	US
Kind code	A1
Filing date	Dec 16, 2015
Priority date	Sep 11, 2012
Publication date	Apr 14, 2016
Grant date	—

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.

First claim

What is claimed is:

1 . A cryopreserved recombinant cell comprising one or more transiently overexpressed genes encoding a drug transporter protein, wherein activity of the drug transporter protein 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 encode a drug transporter protein selected from the group consisting of an ATP binding cassette transporter and a solute carrier transporter protein.

3 . The recombinant cell of claim 2 , 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, OATP1A2/Oatp1a2, OATP2B1/Oatp2b1, PEPT1/Pept1, PEPT2/Pept2, OCTN1/Octn1, OCTN2/Octn2, MATE1/Mate1, MATE2K/Mate2, URAT1/Urat1, ASBT/Asbt, NTCP/Ntcp and combinations thereof.

4 . The recombinant cell of claim 2 , wherein said one or more genes is selected from the group consisting of OATP2/Oatp2, OATP1B3/Oatp1b3, OAT1/Oat1, OAT2/Oat2, OAT3/Oat3, OAT4/Oat4, OCT1/Oct1, OCT2/Oct2, OATP1A2/Oatp1a2, OATP2B1/Oatp2b1, PEPT1/Pept1, PEPT2/Pept2, OCTN1/Octn1, OCTN2/Octn2, MATE1/Mate1, MATE2K/Mate2, URAT1/Urat1, ASBT/Asbt, NTCP/Ntcp and combinations thereof.

5 . The recombinant cell of claim 2 , wherein said one or more genes is selected from the group consisting of OATP1B1*1a, OATP1B1*1b, OATP1B3, OAT1, OAT3, MATE1, MATE2K and combinations thereof.

6 . The recombinant cell of claim 1 , wherein said one or more genes is derived individually from human or an animal species selected from mouse, rat, guinea pig, dog, and monkey.

7 . 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.

8 . The recombinant cell of claim 1 , wherein said cell comprises a hepatocyte.

9 . The recombinant cell of claim 1 , wherein said cell comprises an endothelial cell.

10 . The recombinant cell of claim 1 , wherein activity of the drug transporter protein is detectable in a population of said cell at least 24 hours post plating following thaw from cryopreservation.

11 . The recombinant cell of claim 1 , wherein activity of the drug transporter protein is detectable in a population of said cell at least 48 hours post plating following thaw from cryopreservation.

12 . The recombinant cell of claim 1 , wherein activity of the drug transporter protein is detectable in a population of said cell at least 72 hours post plating following thaw from cryopreservation.

13 . A process of preparing transiently transfected recombinant cells which transiently overexpress one or more genes encoding a drug transporter protein, comprising transiently transfecting cells with one or more genes encoding a drug transporter protein and cryopreserving the transiently transfected recombinant cells within 48 hrs of transfection.

14 . The process of claim 13 , wherein the transient transfection step comprises electroporation.

15 . The process of claim 13 , wherein the transiently transfected recombinant cells are cryopreserved at 30 minutes to 24 hours post transfection.

16 . The process of claim 13 , wherein said one or more genes encode a drug transporter protein selected from the group consisting of an ATP binding cassette transporter and a solute carrier transporter protein.

17 . The process of claim 16 , 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, OATP1A2/Oatp1a2, OATP2B1/Oatp2b1, PEPT1/Pept1, PEPT2/Pept2, OCTN1/Octn1, OCTN2/Octn2, MATE1/Mate1, MATE2K/Mate2, URAT1/Urat1, ASBT/Asbt, NTCP/Ntcp and combinations thereof.

18 . The process of claim 16 , wherein said one or more genes is selected from the group consisting of OATP2/Oatp2, OATP1B3/Oatp1b3, OAT1/Oat1, OAT2/Oat2, OAT3/Oat3, OAT4/Oat4, OCT1/Oct1, OCT2/Oct2, OATP1A2/Oatp1a2, OATP2B1/Oatp2b1, PEPT1/Pept1, PEPT2/Pept2, OCTN1/Octn1, OCTN2/Octn2, MATE1/Mate1, MATE2K/Mate2, URAT1/Urat1, ASBT/Asbt, NTCP/Ntcp, and combinations thereof.

19 . The process of claim 16 , wherein said one or more genes is selected from the group consisting of OATP1B1*1a, OATP1B1*1b, OATP1B3, OAT1, OAT3, MATE1, MATE2K and combinations thereof.

20 . A cryopreserved transiently transfected recombinant cell prepared according to the process of claim 13 .

Assignees

• Corning Inc

Inventors

• Li Na
• Wang Jie
• Patten Christopher J

Classifications

• G08B29/185

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• G08B25/08

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• G08B13/00

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• G08B13/196

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• G05B2219/2642

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