Green Tea (Camellia sinensis) 19124972
Inhibitory Effects of Green Tea and (-)-Epigallocatechin Gallate on Transport by OATP1B1, OATP1B3, OCT1, OCT2, MATE1, MATE2-K and P-Glycoprotein CSV JSON

Green tea catechins inhibit the function of organic anion transporting polypeptides (OATPs) that mediate the uptake of a diverse group of drugs and endogenous compounds into cells. The present study was aimed at investigating the effect of green tea and its most abundant catechin epigallocatechin gallate (EGCG) on the transport activity of several drug transporters expressed in enterocytes, hepatocytes and renal proximal tubular cells such as OATPs, organic cation transporters (OCTs), multidrug and toxin extrusion proteins (MATEs), and P-glycoprotein (P-gp). Uptake of the typical substrates metformin for OCTs and MATEs and bromosulphophthalein (BSP) and atorvastatin for OATPs was measured in the absence and presence of a commercially available green tea and EGCG. Transcellular transport of digoxin, a typical substrate of P-gp, was measured over 4 hours in the absence and presence of green tea or EGCG in Caco-2 cell monolayers. OCT1-, OCT2-, MATE1- and MATE2-K-mediated metformin uptake was significantly reduced in the presence of green tea and EGCG (P < 0.05). BSP net uptake by OATP1B1 and OATP1B3 was inhibited by green tea [IC50 2.6% (v/v) and 0.39% (v/v), respectively]. Green tea also inhibited OATP1B1- and OATP1B3-mediated atorvastatin net uptake with IC50 values of 1.9% (v/v) and 1.0% (v/v), respectively. Basolateral to apical transport of digoxin was significantly decreased in the presence of green tea and EGCG. These findings indicate that green tea and EGCG inhibit multiple drug transporters in vitro. Further studies are necessary to investigate the effects of green tea on prototoypical substrates of these transporters in humans, in particular on substrates of hepatic uptake transporters (e.g. statins) as well as on P-glycoprotein substrates.

1 . Inhibition of MATE1 by (-)-epigallocatechingallate (id=NPDI-GaLUxw)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

Metformin

  • MATE1 (SLC47A1)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

8

10 µM

100 µM

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

2 . Inhibition of OCT2 by (-)-epigallocatechin gallate (id=NPDI-bVuJYQ)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

Metformin

  • OCT2 (SLC22A2)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

10 µM

100 µM

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

3 . Inhibition of OCT1 by (-)-epigallocatechin gallate (id=NPDI-27Q7-A)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

Metformin

  • OCT1 (SLC22A1)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

10 µM

100 µM

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

4 . Inhibition of MATE2-K by (-)-epigallocatechingallate (id=NPDI-DGEDfw)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

Metformin

  • MATE2-K (SLC47A2)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

5.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

72 h

Assay Conditions

10 min

37 deg C

8

10 µM

100 µM

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

5 . Inhibition of MATE2-K by Green Tea (id=NPDI-GdcFMg)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — >20% inhibition vs. control

Metformin

green tea leaf

  • MATE2-K (SLC47A2)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Percent inhibition in MATE2-K cells was estimated from Fig. 1. The IC50 value for inhibition of metformin by green tea (0.1-30% v/v) could not be calculated.

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Sodium butyrate (10 mM) was added 24 h prior to uptake experiments to nonspecifically induce transporter expression.

Cell Culture Conditions

5.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

72 h

Assay Conditions

10 min

37 deg C

8

10 µM

30% v/v

Healthya green tea (Kao Corp., Tokyo, Japan)

Control Experiment Information

6 . Inhibition of OCT2 by Green Tea (id=NPDI-nm2fYg)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

Metformin

green tea leaf

  • OCT2 (SLC22A2)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

10 µM

0.1-30 v/v

Healthya green tea (Kao Corp., Tokyo, Japan)

Control Experiment Information

7 . Inhibition of MATE1 by Green Tea (id=NPDI-cpRc8Q)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — N/A

Metformin

green tea leaf

  • MATE1 (SLC47A1)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

8

10 µM

0.1-30% v/v

Healthya green tea (Kao Corp., Tokyo, Japan)

Control Experiment Information

8 . Inhibition of OCT1 by Green Tea (id=NPDI-8cPLyw)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

Metformin

green tea leaf

  • OCT1 (SLC22A1)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

10 µM

0.1-30 v/v

Healthya green tea (Kao Corp., Tokyo, Japan)

Control Experiment Information

9 . Inhibition of OATP1B1 by (-)-epigallocatechin gallate (id=NPDI-8SDYnQ)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

atorvastatin

  • OATP1B1 (SLCO1B1)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

2.5 µM

100 µM

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

10 . Inhibition of OATP1B1 by Green Tea (id=NPDI-KsBTjg)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

atorvastatin

green tea leaf

  • OATP1B1 (SLCO1B1)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

2.5 µM

0.01-10 % v/v

Healthya green tea (Kao Corp., Tokyo, Japan)

Control Experiment Information

11 . Inhibition of OATP1B3 by Green Tea (id=NPDI-OBEe_w)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

atorvastatin

green tea leaf

  • OATP1B3 (SLCO1B3)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

2.5 µM

0.01-10 % v/v

Healthya green tea (Kao Corp., Tokyo, Japan)

Control Experiment Information

12 . Inhibition of OATP1B3 by (-)-epigallocatechin gallate (id=NPDI-sKz_0w)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

  • OATP1B3 (SLCO1B3)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

5 min

37 deg C

1 µM

100 µM (percent inhibition); 0-200 µM (IC50 determination)

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

13 . Inhibition of OATP1B1 by Green Tea Leaf (id=NPDI-VWdSJg)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

bromosulfophthalein

green tea leaf

  • OATP1B1 (SLCO1B1)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

5 min

37 deg C

0.05 µM

0.01-10 % v/v

Healthya green tea (Kao Corp., Tokyo, Japan)

Control Experiment Information

14 . Inhibition of OATP1B3 by Green Tea (id=NPDI-wmFE6g)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

bromosulfophthalein

green tea leaf

  • OATP1B3 (SLCO1B3)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

5 min

37 deg C

1 µM

0.01-10 % v/v

Healthya green tea (Kao Corp., Tokyo, Japan)

Control Experiment Information

15 . Inhibition of P-gp by (-)epigallocatecin gallate (id=NPDI-6qDdNA)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

digoxin 1326303

  • P-gp (ABCB1)

Cell system Caco-2 cells

Results

In the presence of (-)-epigallocatechin gallate, the B-A transport of digoxin significantly decreased from 3.2%/h to 1.6%/h (P < 0.01 to 0.001). In the presence of (-)-epigallocatechin gallate, the B-A transport was greater than the A-B transport.

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

transwell plates

7

Viability and Function

>200 Ω

Assay Conditions

4 h

37 deg C

5 µM

1 µM

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

16 . Inhibition of P-gp by Green Tea (id=NPDI-PRsP6g)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — NA

digoxin 1326303

green tea leaf

  • P-gp (ABCB1)

Cell system Caco-2 cells

Results

In the presence of green tea, the B-A transport of digoxin significantly decreased from 3.2%/h to < 2.4%/h (P < 0.01 to 0.001). In the presence of green tea, the B-A transport was greater than the A-B transport.

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

transwell plates

7

Viability and Function

> 200 Ω

Assay Conditions

4 h

37 deg C

5 µM

1-10% v/v

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

17 . Inhibition of OATP1B3 by (-)-epigallocatechin gallate (id=NPDI-SSGtgQ)

In Vitro Transporter Inhibition Experiment

Non-inhibition was detected.  Cutoff used — NA

atorvastatin

  • OATP1B3 (SLCO1B3)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

10 min

37 deg C

2.5 µM

100 µM

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information

18 . Inhibition of OATP1B1 by (-)-epigallocatechin gallate (id=NPDI-Iy3POQ)

In Vitro Transporter Inhibition Experiment

Non-inhibition was detected.  Cutoff used — NA

  • OATP1B1 (SLCO1B1)

Transfected / siRNA knock-out / injected cells HEK293 transfected cells

Results

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

7.0 x 10^5 cells/well

12-well poly-D-lysine coated plates

48 h

Assay Conditions

5 min

37 deg C

0.05 µM

100 µM

Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)

Control Experiment Information