In vitro metabolism and inhibitory effects of atractylenolide II on various hepatic CYPs in HLMs

doi:10.5246/jcps.2021.08.052

Abstract

Abstract:

In the present study, we aimed to investigate the interaction between atractylenolide II (AT-II) and CYP450 enzyme in human liver microsomes, and to lay a theoretical foundation for predicting the possible interaction of AT-II in combination with drugs. The chemical inhibition experiment was carried out with specific inhibitors to clarify the CYP450 subtypes affecting the metabolism of AT-II, and the mechanism, kinetics, and type of inhibition of CYP450 enzyme by AT-II were studied by using the probe-based determination method of human liver microsome system with the related data of IC50 and K_i as evaluation indexes. The metabolism of AT-II was affected by CYP1A2, CYP2C9 and CYP3A4 inhibitors, and the highest inhibition rates were 41.35%, 41.97% and 82.45%, respectively. The IC₅₀ values of AT-II to five subtypes of P450 CYP2C9, CYP1A2, CYP2C19, CYP3A4 and CYP2D6 were 69.7, 84.3, 92.4, 173.8 and 190.1 μmol/L, respectively. The K_i values of AT-II to five subtypes of P450 CYP2C9, CYP1A2, CYP2C19, CYP3A4 and CYP2D6 were 190.6, 179.1, > 200, 72.2 and 66.8, respectively. Among these enzymes, AT-II exhibited non-competitive inhibition on CYP1A2, showed competitive inhibition on CYP2C9 and CYP3A4, and displayed mixed AT-II inhibition on CYP2C19 and CYP2D6. CYP1A2, CYP2C9 and CYP3A4 were involved in the AT-II metabolism, and AT-II exhibited different inhibitory mechanisms and strengths for the five subtypes of CYP450.

Key words: Atractylenolide II, Human liver microsomes, Metabolic phenotype, Enzymatic activity, Drug-drug interaction

Supporting:

Xulong Chen, Zhengge Liao, Cheng Li, Guoyong Huang, Yunyan Song, Wei Dong, Abid Naeem, Xinli Liang. In vitro metabolism and inhibitory effects of atractylenolide II on various hepatic CYPs in HLMs[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(8): 645-656.

Figures/Tables 8

Table 1. Inhibition rate of speci?c inhibitors on CYP450-mediated metabolism of AT-II in HLMs (x ± SD, n = 3).

Table 2. The Ki values and inhibition type of AT-II on CYP450 isoforms.

References 26

[1]	Chen, Y.Z.; Yang, W.L.; Guo, L.Y.; Wu, X.L.; Zhang, T.T.; Liu, J.L.; Zhang, J.F. Atractylodes lactone compounds inhibit platelet activation. Platelets. 2017, 28, 194–202.
[2]	Fu, X.Q.; Chou, G.X.; Kwan, H.Y.; Tse, A.K.W.; Zhao, L.H.; Yuen, T.K.; Cao, H.H.; Yu, H.; Chao, X.J.; Su, T.; Cheng, B.C.Y.; Sun, X.G.; Yu, Z.L. Inhibition of STAT3 signalling contributes to the antimelanoma action of atractylenolide II. Exp. Dermatol. 2014, 23, 855–857.
[3]	Singhuber, J.; Baburin, I.; Kählig, H.; Urban, E.; Kopp, B.; Hering, S. GABAA receptor modulators from Chinese herbal medicines traditionally applied against insomnia and anxiety. Phytomedicine. 2012, 19, 334–340.
[4]	Zhang, N.; Liu, C.; Sun, T.M.; Ran, X.K.; Kang, T.G.; Dou, D.Q. Two new compounds from Atractylodes macrocephala with neuroprotective activity. J. Asian Nat. Prod. Res. 2017, 19, 35–41.
[5]	Tian, S.; Yu, H.D. Atractylenolide II inhibits proliferation, motility and induces apoptosis in human gastric carcinoma cell lines HGC-27 and AGS. Molecules. 2017, 22, E1886.
[6]	Wang, T.; Long, F.; Zhang, X.; Yang, Y.; Jiang, X.; Wang, L. Chemopreventive effects of atractylenolide II on mammary tumorigenesis via activating Nrf₂-ARE pathway. Oncotarget. 2017, 8, 77500–77514.
[7]	Xu, M.J.; Jiang, L.F.; Wu, T.; Chu, J.H.; Wei, Y.D.; Aa, J.Y.; Wang, G.J.; Hao, H.P.; Ju, W.Z.; Li, P. Inhibitory effects of Danshen components on CYP2C8 and CYP2J2. Chem. Biol. Interactions. 2018, 289, 5–22.
[8]	Thomford, N.E.; Dzobo, K.; Chopera, D.; Wonkam, A.; Maroyi, A.; Blackhurst, D.; Dandara, C. In vitro reversible and time-dependent CYP450 inhibition profiles of medicinal herbal plant extracts newbouldia laevis and cassia abbreviata: implications for herb-drug interactions. Molecules. 2016, 21, E891.
[9]	MacKenzie, P.I.; Walter Bock, K.; Burchell, B.; Guillemette, C.; Ikushiro, S.I.; Iyanagi, T.; Miners, J.O.; Owens, I.S.; Nebert, D.W. Nomenclature update for the mammalian UDP glycosyltransferase (UGT) gene superfamily. Pharmacogenetics Genom. 2005, 15, 677–685.
[10]	Yang, N.; Sun, R.B.; Liao, X.Y.; Aa, J.Y.; Wang, G.J. UDP-glucuronosyltransferases (UGTs) and their related metabolic cross-talk with internal homeostasis: a systematic review of UGT isoforms for precision medicine. Pharmacol. Res. 2017, 121, 169–183.
[11]	Niu, L.F.; Ding, L.N.; Lu, C.Y.; Zuo, F.F.; Yao, K.; Xu, S.B.; Li, W.; Yang, D.H.; Xu, X. Flavokawain A inhibits Cytochrome P450 in in vitro metabolic and inhibitory investigations. J. Ethnopharmacol. 2016, 191, 350–359.
[12]	Hu, T.; Zhou, X.L.; Wang, L.; Or, P.M.Y.; Yeung, J.H.K.; Kwan, Y.W.; Cho, C.H. Effects of tanshinones from Salvia miltiorrhiza on CYP2C19 activity in human liver microsomes: Enzyme kinetic and molecular docking studies. Chem. Biol. Interactions. 2015, 230, 1–8.
[13]	Habenschus, M.; Moreira, F.; Lopes, N.; de Oliveira, A. In vitro inhibition of human CYP450s 1A2, 2C9, 3A4/5, 2D6 and 2E1 by grandisin. Planta Med. 2017, 83, 727–736.
[14]	Ouyang, D.S.; Huang, W.H.; Chen, D.; Zhang, W.; Tan, Z.R.; Peng, J.B.; Wang, Y.C.; Guo, Y.; Hu, D.L.; Xiao, J.; Chen, Y. Kinetics of cytochrome P450 enzymes for metabolism of sodium tanshinone IIA sulfonate in vitro. Chin. Med. 2016, 11, 11.
[15]	Li, Y.; Liu, J.; Yang, X.W. Four new eudesmane-type sesquiterpenoid lactones from atractylenolide II by biotransformation of rat hepatic microsomes. J. Asian Nat. Prod. Res. 2013, 15, 344–356.
[16]	Ge, J.; Wang, Y.W.; Lu, X.C.; Sun, X.H.; Gong, F.J. Determination of atractylenolide II in rat plasma by reversed-phase high-performance liquid chromatography. Biomed. Chromatogr. 2007, 21, 299–303.
[17]	Mortelé, O.; Vervliet, P.; Gys, C.; Degreef, M.; Cuykx, M.; Maudens, K.; Covaci, A.; van Nuijs, A.L.N.; Lai, F.Y. In vitro Phase I and Phase II metabolism of the new designer benzodiazepine cloniprazepam using liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. J. Pharm. Biomed. Anal. 2018, 153, 158–167.
[18]	Peng, Y.; Wu, H.; Zhang, X.Y.; Zhang, F.Y.; Qi, H.H.; Zhong, Y.X.; Wang, Y.; Sang, H.; Wang, G.J.; Sun, J.G. A comprehensive assay for nine major cytochrome P450 enzymes activities with 16 probe reactions on human liver microsomes by a single LC/MS/MS Run to support reliable in vitro inhibitory drug-drug interaction evaluation. Xenobiotica. 2015, 45, 961–977.
[19]	Bojić, M.; Kondža, M.; Rimac, H.; Benković, G.; Maleš, Ž. The effect of flavonoid aglycones on the CYP1A2, CYP2A6, CYP2C8 and CYP2D6 enzymes activity. Molecules. 2019, 24, 3174.
[20]	Masubuchi, Y.; Horie, T. Dihydralazine-induced inactivation of cytochrome P450 enzymes in rat liver microsomes. Drug Metab. Dispos. 1998, 26, 338–342.
[21]	Liu, Y.; Feng, N.P. Nanocarriers for the delivery of active ingredients and fractions extracted from natural products used in traditional Chinese medicine (TCM). Adv. Colloid Interface Sci. 2015, 221, 60–76.
[22]	Yu, Y.; Liu, Z.Z.; Ju, P.; Zhang, Y.Y.; Zhang, L.H.; Bi, K.S.; Chen, X.H. In vitro metabolism of alisol A and its metabolites’ identification using high-performance liquid chromatography-mass spectrometry. J. Chromatogr. B. 2013, 941, 31–37.
[23]	Ge, J.; Wang, Y.W.; Lu, X.C.; Sun, X.H.; Gong, F.J. Determination of atractylenolide II in rat plasma by reversed-phase high-performance liquid chromatography. Biomed. Chromatogr. 2007, 21, 299–303.
[24]	Jeong, S.; Nguyen, P.D.; Desta, Z. Comprehensive in vitro analysis of voriconazole inhibition of eight cytochrome P450 (CYP) enzymes: major effect on CYPs ₂B6, ₂C9, ₂C19, and ₃A. Antimicrob Agents Chemother. 2009, 53, 541–551.
[25]	Kong, W.M.; Chik, Z.; Ramachandra, M.; Subramaniam, U.; Aziddin, R.E.; Mohamed, Z. Evaluation of the effects of Mitragyna speciosa alkaloid extract on cytochrome P450 enzymes using a high throughput assay. Molecules. 2011, 16, 7344–7356.
[26]	Lu, C.; Suri, A.; Shyu, W.C.; Prakash, S. Assessment of cytochrome P450-mediated drug-drug interaction potential of orteronel and exposure changes in patients with renal impairment using physiologically based pharmacokinetic modeling and simulation. Biopharm. Drug Dispos. 2014, 35, 543–552.