Design, synthesis and evaluation of the inhibitory selectivity of novel trans-resveratrol analogues on human recombinant CYP1A1, CYP1A2 and CYP1B1 Renata Mikstacka a,⇑ , Agnes M. Rimando b , Zbigniew Dutkiewicz a , Tomasz Stefan ´ ski a , Stanisław Sobiak a a Department of Chemical Technology of Drugs, Poznan University of Medical Sciences, Grunwaldzka 6, 60-780 Poznan, Poland b United States Department of Agriculture, Agricultural Research Service, Natural Products Utilization Research Unit, PO Box 8048, University, MS 38677, USA article info Article history: Received 30 March 2012 Revised 2 July 2012 Accepted 6 July 2012 Available online 17 July 2012 Keywords: P450 CYP1A1 CYP1A2 CYP1B1 trans-Stilbene derivatives Molecular docking abstract A series of trans-stilbene derivatives containing 4 0 -methylthio substituent were synthesized and evalu- ated for inhibitory activities on human recombinant cytochrome P450(s): CYP1A1, CYP1A2, and CYP1B1. CYP1A2-related metabolism of stilbene derivatives was estimated by using NADPH oxidation assay. Addi- tionally, for CYP1A2 and CYP1B1 molecular docking analysis was carried out to provide information on enzyme–ligand interactions and putative site of metabolism. 3,4,5-Trimethoxy-4 0 -methylthio-trans-stilbene, an analogue of DMU-212 (3,4,5,4 0 -tetramethoxy-trans- stilbene) was an effective inhibitor of all CYP1 enzymes. On the other hand, 2,3,4-trimethoxy-4 0 -methyl- thio-trans-stilbene, appeared to be the most selective inhibitor of the isozymes CYP1A1 and CYP1B1, dis- playing extremely low affinity towards CYP1A2. Molecular modeling suggested that the most probable binding poses of the methylthiostilbene derivatives in CYP1A2 active sites are those with the methylthio substituent directed towards the heme iron. Products of CYP1A2-catalyzed oxidation of 2,4,5-trimeth- oxy-4 0 -methylthiostilbene and 3,4,5-trimethoxy-4 0 -methylthiostilbene were identified as monohydroxy- lated compounds. Other studied derivatives appeared to be poor substrates of CYP1A2. Structure–activity relationship analysis rendered better understanding of the mechanism of action of xenobiotic-metaboliz- ing enzymes crucial at the early stage of carcinogenesis. Ó 2012 Elsevier Ltd. All rights reserved. 1. Introduction Chemopreventive, cardioprotective and neuroprotective activi- ties of trans-resveratrol (3,4 0 ,5-trihydroxystilbene), the best known natural stilbene derivative, has been documented in numerous studies on animal models. 1 Its therapeutic action in cancer diseases is also under intensive preclinical and clinical studies. 2–4 In the last decade, other natural compounds with stilbene backbone were shown to possess promising activity concerning cancer preven- tion. 2,5 New stilbene derivatives are designed and synthesized in order to find resveratrol analogues with cancer chemopreventive and/or therapeutic activities superior to that of the parent com- pound. The compounds are examined with respect to different tar- gets such as estrogen receptors, 6 COX 1/2, 7 signal transduction pathways, 8,9 cell proliferation, 10 and inhibitory activity towards cytochromes P450 (CYPs). The studies are focused on structure– activity relationship in order to identify structural determinants responsible for beneficial activities of stilbene derivatives. Other investigations are aimed to find resveratrol analogues that demon- strate better bioavailability than the parent compound that under- goes fast biotransformation to glucuronides and sulfates. 11 Methoxylation of hydroxyl groups is supposed to prevent polyphe- nol metabolism and enhance stilbene bioactivity. 12,13 Cytochrome P450 family 1 includes xenobiotic-metabolizing en- zymes: CYP1A1, CYP1A2 and CYP1B1 that catalyze the activation of environmental procarcinogens such as polycyclic aromatic hydro- carbons, aromatic and heterocyclic amines to carcinogenic forms. Additionally, CYP1B1 metabolizes 17b-estradiol to 4-hydroxyestra- diol that is oxidized by peroxidase to estradiol-3,4-quinone, and forms a quinone-DNA adducts responsible for estrogen-related car- cinogenesis. 14 For these reasons, there is a demand for inhibitors that demonstrate high selectivity toward specific forms of cyto- chrome 450; namely, CYP1A1 and CYP1B1 for blocking CYP-related carcinogenesis. CYP1A2 metabolizes numerous xenobiotics includ- ing drugs such as cafeine, theophylline, methadone, verapamil, pro- pranolol, warfarin, and tamoxifen. Not much is known about CYP1A2; hence its properties, the structure of its active site and substrate specificity need thorough investigation. All members of CYP1A family are expressed in extrahepatic tissues; however, CYP1A2 is the only constitutive form of liver enzyme. In humans CYP1B1 is overexpressed in tumor cells, and this has important 0968-0896/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.bmc.2012.07.012 ⇑ Corresponding author. Tel./fax: +48 618 546 625. E-mail address: rmikstac@ump.edu.pl (R. Mikstacka). Bioorganic & Medicinal Chemistry 20 (2012) 5117–5126 Contents lists available at SciVerse ScienceDirect Bioorganic & Medicinal Chemistry journal homepage: www.elsevier.com/locate/bmc