Green tea (Camellia sinensis)
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 OATP1B1 by (-)-epigallocatechin gallate (id=NPDI-8SDYnQ)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- OATP1B1 (SLCO1B1)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
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
2 . Inhibition of OATP1B3 by Green Tea (id=NPDI-OBEe_w)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- OATP1B3 (SLCO1B3)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
10 min
37 deg C
2.5 µM
0.01-10 % v/v
Healthya green tea (Kao Corp., Tokyo, Japan)
Control Experiment Information
3 . Inhibition of OATP1B1 by Green Tea (id=NPDI-KsBTjg)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- OATP1B1 (SLCO1B1)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
10 min
37 deg C
2.5 µM
0.01-10 % v/v
Healthya green tea (Kao Corp., Tokyo, Japan)
Control Experiment Information
4 . 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
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
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
5 . Inhibition of OATP1B1 by Green Tea Leaf (id=NPDI-VWdSJg)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- OATP1B1 (SLCO1B1)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
5 min
37 deg C
0.05 µM
0.01-10 % v/v
Healthya green tea (Kao Corp., Tokyo, Japan)
Control Experiment Information
6 . Inhibition of OATP1B3 by Green Tea (id=NPDI-wmFE6g)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- OATP1B3 (SLCO1B3)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
5 min
37 deg C
1 µM
0.01-10 % v/v
Healthya green tea (Kao Corp., Tokyo, Japan)
Control Experiment Information
7 . 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.
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
transwell plates
7
>200 Ω
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
8 . Inhibition of P-gp by Green Tea (id=NPDI-PRsP6g)
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 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.
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
transwell plates
7
> 200 Ω
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
9 . Inhibition of OCT1 by (-)-epigallocatechin gallate (id=NPDI-27Q7-A)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- OCT1 (SLC22A1)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
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
10 . Inhibition of MATE1 by (-)-epigallocatechingallate (id=NPDI-GaLUxw)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- MATE1 (SLC47A1)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
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
11 . Inhibition of OCT2 by (-)-epigallocatechin gallate (id=NPDI-bVuJYQ)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- OCT2 (SLC22A2)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
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
12 . Inhibition of MATE2-K by (-)-epigallocatechingallate (id=NPDI-DGEDfw)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
- MATE2-K (SLC47A2)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
5.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
72 h
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
13 . Inhibition of OCT1 by Green Tea (id=NPDI-8cPLyw)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
metformin 1503297
- OCT1 (SLC22A1)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
10 min
37 deg C
10 µM
0.1-30 v/v
Healthya green tea (Kao Corp., Tokyo, Japan)
Control Experiment Information
14 . 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 1503297
- 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.
| Sample | Compound measured | Value | Measurement | 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.
5.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
72 h
10 min
37 deg C
8
10 µM
30% v/v
Healthya green tea (Kao Corp., Tokyo, Japan)
Control Experiment Information
15 . Inhibition of OCT2 by Green Tea (id=NPDI-nm2fYg)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — NA
metformin 1503297
- OCT2 (SLC22A2)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
10 min
37 deg C
10 µM
0.1-30 v/v
Healthya green tea (Kao Corp., Tokyo, Japan)
Control Experiment Information
16 . Inhibition of MATE1 by Green Tea (id=NPDI-cpRc8Q)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used — N/A
metformin 1503297
- MATE1 (SLC47A1)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
10 min
37 deg C
8
10 µM
0.1-30% v/v
Healthya green tea (Kao Corp., Tokyo, 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
- OATP1B3 (SLCO1B3)
Transfected / siRNA knock-out / injected cells HEK293 transfected cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
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
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
7.0 x 10^5 cells/well
12-well poly-D-lysine coated plates
48 h
5 min
37 deg C
0.05 µM
100 µM
Prof. Toshiyuki Kan (Department of Synthetic Organic and Medicinal Chemistry, University of Shizuoka, Japan)