Cannabis L. (Cannabis sativa)
Potent inhibition of human cytochrome P450 3A isoforms by cannabidiol: role of phenolic hydroxyl groups in the resorcinol moiety. CSV JSON

AIMS:

In this study, we examined the inhibitory effects of Δ(9)-tetrahydrocannabinol (Δ(9)-THC), cannabidiol (CBD), and cannabinol (CBN), the three major cannabinoids, on the activity of human cytochrome P450 (CYP) 3A enzymes. Furthermore, we investigated the kinetics and structural requirement for the inhibitory effect of CBD on the CYP3A activity.

MAIN METHODS:

Diltiazem N-demethylase activity of recombinant CYP3A4, CYP3A5, CYP3A7, and human liver microsomes (HLMs) in the presence of cannabinoids was determined.

KEY FINDINGS:

Among the three major cannabinoids, CBD most potently inhibited CYP3A4 and CYP3A5 (IC(50)=11.7 and 1.65 μM, respectively). The IC(50) values of Δ(9)-THC and CBN for CYP3A4 and CYP3A5 were higher than 35 μM. For CYP3A7, Δ(9)-THC, CBD, and CBN inhibited the activity to a similar extent (IC(50)=23-31 μM). CBD competitively inhibited the activity of CYP3A4, CYP3A5, and HLMs (K(i)=1.00, 0.195, and 6.14 μM, respectively). On the other hand, CBD inhibited the CYP3A7 activity in a mixed manner (K(i)=12.3 μM). Olivetol partially inhibited all the CYP3A isoforms tested, whereas d-limonene showed lack of inhibition. The lesser inhibitory effects of monomethyl and dimethyl ethers of CBD indicated that the ability of CYP3A inhibition by the cannabinoid attenuated with the number of methylation on the phenolic hydroxyl groups in the resorcinol moiety.

SIGNIFICANCE:

This study indicated that CBD most potently inhibited catalytic activity of human CYP3A enzymes, especially CYP3A4 and CYP3A5. These results suggest that two phenolic hydroxyl groups in the resorcinol moiety of CBD may play an important role in the CYP3A inhibition.

1 . HLM-CBD (id=NPDI-zkYApA)

In Vitro Enzyme Inhibition Experiment

Inhibition was detected.  Cutoff used — Not specified

diltiazem

cannabidiol

N-Demethyldiltiazem

  • CYP3A

Cell fraction Individual human liver microsomes

Results

Table 2, Table 3

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

50 μg

Commercially available

200 μL

15 min

MgCl2
NADPH regenerating system

50 μM

NADPH with precipitant

0-50 μM

2 . CYP3A4-CBD (id=NPDI-FmAizA)

In Vitro Enzyme Inhibition Experiment

Inhibition was detected.  Cutoff used — Not specified

diltiazem

cannabidiol

N-Demethyldiltiazem

  • CYP3A4 4306811

Recombinant expression system Baculovirus-insect cells
Cytochrome B5 Co-expressed

Results

Table 2, Table 3

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

2 pmol

Commercially available

200 μL

15 min

MgCl2
NADPH regenerating system

50 μM

NADPH with precipitant

0-50 μM

3 . HLM-CBN (id=NPDI-yszOMw)

In Vitro Enzyme Inhibition Experiment

Negligible Inhibition was detected.  Cutoff used — Not specified

diltiazem

cannabinol

N-Demethyldiltiazem

  • CYP3A

Cell fraction Individual human liver microsomes

Results

Table 2

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

50 μg

Commercially available

200 μL

15 min

MgCl2
NADPH regenerating system

50 μM

NADPH with precipitant

0-50 μM

4 . CYP3A4-CBN (id=NPDI-ens_Gw)

In Vitro Enzyme Inhibition Experiment

Negligible Inhibition was detected.  Cutoff used — Not specified

diltiazem

cannabinol

N-Demethyldiltiazem

  • CYP3A4 4306811

Recombinant expression system Baculovirus-insect cells
Cytochrome B5 Co-expressed

Results

Table 2

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

2 pmol

Commercially available

200 μL

15 min

MgCl2
NADPH regenerating system

50 μM

NADPH with precipitant

0-50 μM

5 . CYP3A4-THC (id=NPDI-8vislA)

In Vitro Enzyme Inhibition Experiment

Negligible Inhibition was detected.  Cutoff used — Not specified

diltiazem

N-Demethyldiltiazem

  • CYP3A4 4306811

Recombinant expression system Baculovirus-insect cells
Cytochrome B5 Co-expressed

Results

Table 2

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

2 pmol

Commercially available

200 μL

15 min

MgCl2
NADPH regenerating system

50 μM

NADPH with precipitant

0-50 μM

6 . HLM-THC (id=NPDI-PS8_LQ)

In Vitro Enzyme Inhibition Experiment

Negligible Inhibition was detected.  Cutoff used — Not specified

diltiazem

N-Demethyldiltiazem

  • CYP3A

Cell fraction Individual human liver microsomes

Results

Table 2

Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

50 μg

Commercially available

200 μL

15 min

MgCl2
NADPH regenerating system

50 μM

NADPH with precipitant

0-50 μM