*Article* **Strong and Selective Inhibitory E** ff**ects of the Biflavonoid Selamariscina A against CYP2C8 and CYP2C9 Enzyme Activities in Human Liver Microsomes**

**So-Young Park 1,2, Phi-Hung Nguyen 3, Gahyun Kim 2, Su-Nyeong Jang 1,2, Ga-Hyun Lee 1,2, Nguyen Minh Phuc 2,4, Zhexue Wu 2 and Kwang-Hyeon Liu 1,2,\***


Received: 25 March 2020; Accepted: 9 April 2020; Published: 10 April 2020

**Abstract:** Like flavonoids, biflavonoids, dimeric flavonoids, and polyphenolic plant secondary metabolites have antioxidant, antibacterial, antiviral, anti-inflammatory, and anti-cancer properties. However, there is limited data on their effects on cytochrome P450 (P450) and uridine 5-diphosphoglucuronosyl transferase (UGT) enzyme activities. In this study we evaluate the inhibitory potential of five biflavonoids against nine P450 activities (P450s1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A) in human liver microsomes (HLMs) using cocktail incubation and liquid chromatography-tandem mass spectrometry (LC–MS/MS). The most strongly inhibited P450 activity was CYP2C8-mediated amodiaquine *N*-dealkylation with IC50 ranges of 0.019~0.123 μM. In addition, the biflavonoids—selamariscina A, amentoflavone, robustaflavone, cupressuflavone, and taiwaniaflavone—noncompetitively inhibited CYP2C8 activity with respective *K*i values of 0.018, 0.083, 0.084, 0.103, and 0.142 μM. As selamariscina A showed the strongest e ffects, we then evaluated it against six UGT isoforms, where it showed weaker inhibition (UGTs1A1, 1A3, 1A4, 1A6, 1A9, and 2B7, IC50 > 1.7 μM). Returning to the P450 activities, selamariscina A inhibited CYP2C9-mediated diclofenac hydroxylation and tolbutamide hydroxylation with respective *K*i values of 0.032 and 0.065 μM in a competitive and noncompetitive manner. However, it only weakly inhibited CYP1A2, CYP2B6, and CYP3A with respective *K*i values of 3.1, 7.9, and 4.5 μM. We conclude that selamariscina A has selective and strong inhibitory e ffects on the CYP2C8 and CYP2C9 isoforms. This information might be useful in predicting herb-drug interaction potential between biflavonoids and co-administered drugs mainly metabolized by CYP2C8 and CYP2C9. In addition, selamariscina A might be used as a strong CYP2C8 and CYP2C9 inhibitor in P450 reaction-phenotyping studies to identify drug-metabolizing enzymes responsible for the metabolism of new chemicals.

**Keywords:** biflavonoid; cytochrome P450; drug interactions; selamariscina A; uridine 5-diphosphoglucuronosyl transferase
