In Vitro Glucuronidation of Wushanicaritin by Liver Microsomes, Intestine Microsomes and Expressed Human UDP-Glucuronosyltransferase Enzymes

Abstract

Wushanicaritin, a natural polyphenol compound, exerts many biological activities. This study aimed to characterize wushanicaritin glucuronidation by pooled human liver microsomes (HLM), human intestine microsomes and individual uridine diphosphate-glucuronosyltransferase (UGT) enzyme. Glucuronidation rates were determined by incubating wushanicaritin with uridine diphosphoglucuronic acid-supplemented microsomes. Kinetic parameters were derived by appropriate model fitting. Reaction phenotyping, the relative activity factor (RAF) and activity correlation analysis were performed to identify the main UGT isoforms. Wushanicaritin glucuronidation in HLM was efficient with a high CLint (intrinsic clearance) value of 1.25 and 0.69 mL/min/mg for G1 and G2, respectively. UGT1A1 and 1A7 showed the highest activities with the intrinsic clearance (CLint) values of 1.16 and 0.38 mL/min/mg for G1 and G2, respectively. In addition, G1 was significantly correlated with β-estradiol glucuronidation (r = 0.847; p = 0.0005), while G2 was also correlated with chenodeoxycholic acid glucuronidation (r = 0.638, p = 0.026) in a bank of individual HLMs (n = 12). Based on the RAF approach, UGT1A1 contributed 51.2% for G1, and UGT1A3 contributed 26.0% for G2 in HLM. Moreover, glucuronidation of wushanicaritin by liver microsomes showed marked species difference. Taken together, UGT1A1, 1A3, 1A7, 1A8, 1A9 and 2B7 were identified as the main UGT contributors responsible for wushanicaritin glucuronidation.

Efflux excretion of bisdemethoxycurcumin‐O‐glucuronide in UGT1A1‐overexpressing HeLa cells: Identification of breast cancer resistance protein (BCRP) and multidrug resistance‐associated proteins 1 (MRP1) as the glucuronide transporters

“…D) both followed classical Michaelis–Menten equation. The K m values were simimlar with previous study , while the V max and CL int values showed significant differences ( P < 0.05) by UGT1A1 and HeLa1A1 cell lysate (Table ). Taken together, these results indicated that the engineered HeLa1A1 cells significantly expressed active UGT1A1 protein and could functionally catalyze the glucuronidation reaction.…”

“…Glucuronidation activities of BDMC by expressed UGT1A1 and the HeLa1A1 cell lysate were measured following our published procedures . In brief, the incubation system (200 μL) contained recombinant UGT1A1 (1.0 mg/mL) or HeLa1A1 cell lysate (9.2 mg/mL), MgCl 2 (4 mM), alamethicin (20 μg/mL), and BDMC in 50 mM Tris–HCl buffer (pH = 7.4).…”

Metabolism and disposition of corylifol A from Psoralea corylifolia: metabolite mapping, isozyme contribution, species differences and identification of efflux transporters for corylifol A-O-glucuronide in HeLa1A1 cells

“…As mentioned, paclitaxel was regarded as the most widely used probe substrate for CYP2C8 (Xu et al, 2018), whereas β-estradiol and propofol were recognized as the A c c e p t e d M a n u s c r i p t probe substrate for UGT1A1 and 1A3, respectively (Hong et al, 2017). As shown in Figure S4A, a significant correlation was obtained between CA-mono-oxidation and paclitaxel-6-hydroxylation (r = 0.921, p = 0.001) in a panel of HLMs (n = 9).…”

An investigation of the metabolic activity, isozyme contribution, species differences and potential drug–drug interactions of PI-103, and the identification of efflux transporters for PI-103-O-glucuronide in HeLa1A9 cells

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