Pogostone inhibits the activity of CYP3A4, 2C9, and 2E1 in vitro

Figures & data

Table 1. Isoforms tested, marker reactions, incubation conditions, and K_m used in the inhibition study.

CYPs	Marker reactions	Substrate concentration (μM)	Protein concentration (mg/mL)	Incubation time (min)	Estimated Km (μM)
1A2	phenacetin O-deethylation	40	0.2	30	48
2A6	coumarin 7-hydroxylation	1.0	0.1	10	1.5
3A4	testosterone 6β-hydroxylation	50	0.5	10	53
2C8	paclitaxel 6α-hydroxylation	10	0.5	30	16
2C9	diclofenac 4′-hydroxylation	10	0.3	10	13
2C19	S-Mephenytoin 4-hydroxylation	100	0.2	40	105
2D6	dextromethorphan O-demethylation	25	0.25	20	4.8
2E1	chlorzoxazone 6-hydroxylation	120	0.4	30	126

Figure 1. Effect of pogostone on the activity of CYP450s. The activity of CYP3A4, 2C9, and 2E1 was inhibited by pogostone. Negative control: without pogostone or positive inhibitors; Pogostone: 100 μM pogostone; Positive control: corresponding positive inhibitors. *p < 0.05.

Figure 2. The inhibition of CYP3A4 by pogostone. A. The Lineweaver-Burk plots of CYP3A4 in the presence of 0, 2, 5, 10, and 30 μM pogostone and 20–100 μM testosterone. The inhibition of CYP3A4 was found to be non-competitive and the K_i value was obtained as 5.69 μM. B. The inhibition of CYP3A4 was affected by the incubation time and the KI and K_inact value were obtained as 5.86/μM and 0.056/min, respectively.

Figure 3. The inhibition of CYP2C9 and 2E1. (A) The Lineweaver-Burk plots of CYP2C9 in the presence of 0, 2, 5, 10, and 30 μM pogostone and 2–20 μM diclofenac. The inhibition of CYP2C9 was found to be competitive and the Ki value was obtained as 6.46 μM. (B) The Lineweaver-Burk plots of CYP2E1 in the presence of 0, 2, 5, 10, and 30 μM pogostone and 25–250 μM chlorzoxazone. The K_i value was obtained as 7.67 μM.