Major metabolites of alpha-asarone in liver microsomes are epoxide-derived side-chain diols. The intermediately formed epoxides are mutagenic and form DNA adducts and thus are likely responsible for the (hepato) carcinogenic effect of alpha-asarone observed in male mice. We here investigated the role of eight human cytochrome P450 enzymes (CYP1A1, 1A2, 2A6, 2B6, 2C19, 2D6, 2E1, and 3A4) in the metabolism of alpha-asarone using Supersomes™. The epoxidation of the side-chain of alpha-asarone was mainly catalyzed by CYP3A4 and to a lesser extent by 2B6 and 1A1 whereas the hydroxylation of the side-chain leading to (E)-3′-hydroxyasarone was catalyzed by all investigated CYPs excluding CYP2A6. O-demethylation was catalyzed by CYP1A1, 2A6, 2B6, and 2C19. Applying relative activity factors (RAF) to the observed formation rates revealed that CYP3A4, at least at lower substrate concentrations, is nearly solely responsible for the formation of the mutagenic side-chain epoxides of alpha-asarone. Comparison of the RAF-corrected formation rates of all metabolites with those found in incubation with human liver microsomes revealed that the side-chain hydroxylation and epoxidation can be explained in good approximation by the tested hepatic CYPs, whereas other CYPs or enzymes may contribute to the O-demethylation of alpha-asarone. Therefore, the capacity for metabolic activation of alpha-asarone has to be expected to be widely present among the general population.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Amato G, Longo V, Mazzaccaro A, Gervasi PG (1998) Chlorzoxazone 6-hydroxylase and p-nitrophenol hydroxylase as the most suitable activities for assaying cytochrome P450 2E1 in cynomolgus monkey liver. Drug Metab Dispos 26:483–489
Belova LF, Alibekov SD, Baginskaia AI, Sokolov SI, Pokrovskaia GV (1985) Asarone and its biological properties. Farmakol Toksikol 48:17–20
Berg K, Bischoff R, Stegmüller S, Cartus A, Schrenk D (2016) Comparative investigation of the mutagenicity of propenylic and allylic asarone isomers in the Ames fluctuation assay. Mutagenesis 31:443–451. https://doi.org/10.1093/mutage/gew007
Cartus AT, Stegmüller S, Simson N, Wahl A, Neef S, Kelm H, Schrenk D (2015) Hepatic metabolism of carcinogenic β-asarone. Chem Res Toxicol 28:1760–1773. https://doi.org/10.1021/acs.chemrestox.5b00223
Cartus AT, Schrenk D (2016) Metabolism of the carcinogen alpha-asarone in liver microsomes. Food Chem Toxicol 87:103–112. https://doi.org/10.1016/j.fct.2015.11.021
Chamorro G, Salazar M, Salazar S (1996) Dominant lethal effects in male and female Wistar rats after short-term alpha-asarone treatment. Toxicol Lett Suppl 1:67. https://doi.org/10.1016/S0378-4274(96)80243-8
Chamorro G, Garduño L, Martínez E, Madrigal E, Tamariz J, Salazar M (1998) Dominant lethal study of alpha-asarone in male mice. Toxicol Lett 99:71–77
Della Greca MD, Monaco P, Previtera L, Aliotta G, Pinto G, Pollio A (1989) Allelochemical activity of phenylpropanes from Acorus gramineus. Phytochemistry 28:2319–2321. https://doi.org/10.1016/S0031-9422(00)97975-5
Evans WE, Relling MV (1999) Pharmacogenomics: translating functional genomics into rational therapeutics. Science 286(5439):487–491
Kim BW, Koppula S, Kumar H, Park JY, Kim IW, More SV, Kim IS, Han SD, Kim SK, Yoon SH, Choi DK (2015) α-Asarone attenuates microglia-mediated neuroinflammation by inhibiting NF kappa B activation and mitigates MPTP-induced behavioral deficits in a mouse model of Parkinson’s disease. Neuropharm 97:46–57. https://doi.org/10.1016/j.neuropharm.2015.04.037
Klein K, Zanger UM (2013) Pharmacogenomics of cytochrome P450 3A4: recent progress toward the “missing heritability” problem. Front Genet 4:12
Kobets T, Cartus AT, Fuhlbrueck JA, Brengel A, Stegmüller S, Duan JD, Brunnemann KD, Williams GM (2019) Assessment and characterization of DNA adducts produced by alkenylbenzenes in fetal turkey and chicken livers. Food Chem Toxicol 129:424–433. https://doi.org/10.1016/j.fct.2019.05.010
Krahulcova A (2003) Chromosome numbers in selected monocotyledons. Preslia Praha 75:97–113
Lee HJ, Ahn SM, Pak ME, Jung DH, Lee SY, Shin HK, Choi BT (2018) Positive effects of α-asarone on transplanted neural progenitor cells in a murine model of ischemic stroke. Phytomedicine 51:151–161. https://doi.org/10.1016/j.phymed.2018.09.230
Meng X, Zhao X, Wang S, Jia P, Bai Y, Liao S, Zheng X (2013) Simultaneous determination of volatile constituents from Acorus tatarinowii Schott in rat plasma by gas chromatography-mass spectrometry with selective ion monitoring and application in pharmacokinetic study. J Anal Methods Chem 2013:949830. https://doi.org/10.1155/2013/949830
Minet EF, Daniela G, Meredith C, Massey ED (2012) A comparative in vitro kinetic study of [14C]-eugenol and [14C]-methyleugenol activation and detoxification in human, mouse, and rat liver and lung fractions. Xenobiotica 42(5):429–441. https://doi.org/10.3109/00498254.2011.637582
Morales-Ramírez P, Madrigal-Bujaidar E, Mercader-Martínez J, Cassini M, González G, Chamorro-Cevallos G, Salazar-Jacobo M (1992) Sister-chromatid exchange induction produced by in vivo and in vitro exposure to alpha-asarone. Mutat Res 279:269–273
Morse MA, Lu J (1998) High-performance liquid chromatographic method for measurement of cytochrome P450-mediated metabolism of 7-ethoxy-4-trifluoromethylcoumarin. J Chromatogr B Biomed Sci Appl 708(1–2):290–293
Mukherjee PK, Kumar V, Mal M, Houghton PJ (2007a) In vitro acetylcholinesterase inhibitory activity of the essential oil from Acorus calamus and its main constituents. Planta Med 73:283–285. https://doi.org/10.1055/s-2007-967114
Mukherjee PK, Kumar V, Mal M, Houghton PJ (2007b) Acorus calamus: scientific validation of Ayurvedic tradition from natural resources. Pharm Biol 45:651–666. https://doi.org/10.1080/13880200701538724
Qin W, Huang S, Li C, Chen S, Peng Z (2010) Biological activity of the essential oil from the leaves of Piper sarmentosum Roxb. (Piperaceae) and its chemical constituents on Brontispa longissima (Gestro) (Coleoptera: Hispidae). Pestic Biochem Physiol 96(3):132–139. https://doi.org/10.1016/j.pestbp.2009.10.006
Rajput SB, Tonge MB, Karuppayil SM (2014) An overview on traditional uses and pharmacological profile of Acorus calamus Linn. (Sweet flag) and other Acorus species. Phytomedicine 21:268–276. https://doi.org/10.1016/j.phymed.2013.09.020
Rana TS, Mahar KS, Pandey MM, Srivastava SK, Rawat AK (2013) Molecular and chemical profiling of ‘sweet flag’ (Acorus calamus L.) germplasm from India. Physiol Mol Biol Plants 19:231–237. https://doi.org/10.1007/s12298-013-0164-8
Rendic S, Guengerich FP (2012) Contributions of human enzymes in carcinogen metabolism. Chem Res Toxicol 25(7):1316–1383. https://doi.org/10.1021/tx300132k
Salazar M, Salazar S, Ulloa V, Mendoza T, Pages N, Chamoro G (1992) Teratogenic action of alpha-asarone in the mouse. J Toxicol Clin Exp 12:149–154
Satyal P, Paudel P, Poudel A, Dosoky NS, Moriarity DM, Vogler B, Setzer WN (2013) Chemical compositions, phytotoxicity, and biological activities of Acorus calamus essential oils from Nepal. Nat Prod Commun 8:1179–1181
Shin JW, Cheong YJ, Koo YM, Kim S, Noh CK, Son YH, Kang C, Sohn NW (2014) α-Asarone ameliorates memory deficit in lipopolysaccharide-treated mice via suppression of pro-inflammatory cytokines and microglial activation. Biomol Ther (Seoul) 22:17–26. https://doi.org/10.4062/biomolther.2013.102
Soars MG, Gelboin HV, Krausz KW, Riley RJ (2003) A comparison of relative abundance, activity factor and inhibitory monoclonal antibody approaches in the characterization of human CYP enzymology. Br J Clin Pharmacol 55(2):175–181. https://doi.org/10.1046/j.1365-2125.2003.01721.x
Stegmüller S, Schrenk D, Cartus AT (2018) Formation and fate of DNA adducts of alpha- and beta-asarone in rat hepatocytes. Food Chem Toxicol 116(Pt B):138–146. https://doi.org/10.1016/j.fct.2018.04.025
Wiseman RW, Miller EC, Miller JA, Liem A (1987) Structure-activity studies of the hepatocarcinogenicities of alkenylbenzene derivatives related to estragole and safrole on administration to preweanling male C57BL/6J x C3H/HeJ F1 mice. Cancer Res 47:2275–2283
Yao H, Peng Y, Zheng J (2016) Identification of glutathione and related cysteine conjugates derived from reactive metabolites of methyleugenol in rats. Chem Biol Interact 253:143–152. https://doi.org/10.1016/j.cbi.2016.05.006
Zuba D, Byrska B (2012) Alpha- and beta-asarone in herbal medicinal products. A case study. Forensic Sci Int 223:e5–e9. https://doi.org/10.1016/j.forsciint.2012.08.015
This work was funded by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG) under grant #SCHR 327/14-1.
Conflict of interest
The authors declare that there are no conflicts of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Cartus, A.T., Schrenk, D. Metabolism of carcinogenic alpha-asarone by human cytochrome P450 enzymes. Naunyn-Schmiedeberg's Arch Pharmacol 393, 213–223 (2020). https://doi.org/10.1007/s00210-019-01724-0
- Cytochrome P450