An evaluation of possible interactions with enzymes of drug metabolism (cytochromes P450, CYP) is an important part of studies on safety and, in general, on the properties of any drug or biologically active compound. The article is focused on the preliminary metabolic study of selected 2,6,9-trisubstituted purine kinase inhibitors with significant anticancer activities which we have developed. The compounds BP-21 and BP-117 represent strong CDK inhibitors and the compound BPA-302 was developed as selective FLT3-ITD kinase inhibitor. Here, emphasis is placed on interactions of these compounds with the nine most important forms of CYP to evaluate the possibility of inhibition of these enzymes. The possibility of their inhibitory effect was studied in vitro on selected human liver microsomal CYP enzymes. The most affected enzyme was CYP2C19. Its activity dropped to 22 % of its original value by BPA 302, to 13 % by BP-21 and to 6 % by BP-117 at the highest concentration tested (250 µmol·l(-1)). The results suggest that the metabolism of concomitantly administered drugs should not be significantly affected at lower doses. Molecular docking of BPA-302 indicated that it can bind to active site of both CYP2C19 and CYP2D6 enzymes above the heme cofactor corroborating the experimental data.

Department of Pharmacology Faculty of Medicine Palacký University Olomouc Czech Republic

Zobrazit více v PubMed

ANZENBACHER P, ANZENBACHEROVÁ E. Cytochromes P450 and metabolism of xenobiotics. Cell Mol Life Sci. 2001;58:737–747. doi: 10.1007/PL00000897. PubMed DOI PMC

ANZENBACHER P, ZANGER UM. Metabolism of Drugs and Other Xenobiotics. Wiley-VCH Verlag & Co KGaA; Germany: 2012. DOI

BRODNEY MA, BECK EM, BUTLER CR, BARREIRO G, JOHNSON EF, RIDDELL D, PARRIS K, NOLAN CE, FAN Y, ATCHISON K, GONZALES C, ROBSHAW AE, DORAN SD, BUNDESMANN MW, BUZON L, DUTRA J, HENEGAR K, LACHAPELLE E, HOU X, ROGERS BN, PANDIT J, LIRA R, MARTINEZ-ALSINA L, MIKOCHIK P, MURRAY JC, OGILVIE K, PRICE L, SAKYA SM, YU A, ZHANG Y, O’NEILL BT. Utilizing structures of CYP2D6 and BACE1 complexes to reduce risk of drug-drug interactions with a novel series of centrally efficacious BACE1 inhibitors. J Med Chem. 2015;58:223–3252. doi: 10.1021/acs.jmedchem.5b00191. PubMed DOI PMC

CICENAS J, KALYAN K, SOROKINAS A, STANKUNAS E, LEVY J, MESKINYTE I, STANKEVICIUS V, KAUPINIS A, VALIUS M. Roscovitine in cancer and other diseases. Ann Transl Med. 2015;3:135. PubMed PMC

CRESPI CL, CHANG TKH, WAXMAN DJ. Determination of CYP2C9-catalyzed diclofenac 4′ hydroxylation by high-performance liquid chromatography. In: PHILLIPS IR, SHEPHARD EA, editors. Cytochrome P450 Protocols. Humana; Totowa, USA: 1998a. pp. 109–113. PubMed DOI

CRESPI CL, CHANG TKH, WAXMAN DJ. CYP 2D6-dependent bufuralol 1′ hydroxylation assayed by reversed-phase ion-pair high-performance liquid chromatography with fluorescence detection. In: PHILLIPS IR, SHEPHARD EA, editors. Cytochrome P450 Protocols. Humana; Totowa, USA: 1998b. pp. 141–146. PubMed DOI

DONATO MT, JIMENEZ N, CASTELL JV, GOMEZ-LECHON MJ. Fluorescencebased assays for screening nine cytochrome P450 (P450) activities in intact cells expressing individual human P450 enzymes. Drug Metab Dispos. 2004;32:699–706. doi: 10.1124/dmd.32.7.699. PubMed DOI

GUCKÝ T, JORDA R, ZATLOUKAL M, BAZGIER V, BERKA K, ŘEZNÍČKOVÁ E, BÉRES T, STRNAD M, KRYŠTOF V. A novel series of highly potent 2,6,9-trisubstituted purine cyclin-dependent kinase inhibitors. J Med Chem. 2013;56:6234–6247. doi: 10.1021/jm4006884. PubMed DOI

GUCKÝ T, ŘEZNÍČKOVÁ E, RADOŠOVÁ MUCHOVÁ T, JORDA R, KLEJOVÁ Z, MALÍNKOVÁ V, BERKA K, BAZGIER V, AJANI H, LEPŠÍK M, DIVOKÝ V, KRYŠTOF V. Discovery of N2-(4-amino-cyclohexyl)-9-cyclopentyl-N6-(4-morpholin-4-ylmethyl-phenyl)-9H-purine-2,6-diamine as a potent FLT3 kinase inhibitor for acute myeloid leukemia with FLT3 mutations. J Med Chem. 2018;61:855–3869. doi: 10.1021/acs.jmedchem.7b01529. PubMed DOI

GUENGERICH FP. Cytochromes P450, drugs, and diseases. Mol Interv. 2003;3:194–204. doi: 10.1124/mi.3.4.194. PubMed DOI

GUENGERICH FP, MARTIN MV, BEAUNNE PH, KTEMERS P, WOLFF T, WAXMAN DJ. Characterization of Rat and human-liver microsomal cytochrome-P-450 forms involved in nifedipine oxidation, a prototype for genetic-polymorphism in oxidative drug-metabolism. J Biol Chem. 1986;261:5051–5060. PubMed

HAIDER C, GRUBINGER M, ŘEZNÍČKOVÁ E, WEISS TS, ROTHENEDER H, MIKLOS W, BERGER W, JORDA R, ZATLOUKAL M, GUCKÝ T, STRNAD M, KRYŠTOF V, MIKULITS W. Novel inhibitors of cyclin-dependent kinases combat hepatocellular carcinoma without inducing chemoresistance. Mol Cancer Ther. 2013;12:1947–1957. doi: 10.1158/1535-7163.MCT-13-0263. PubMed DOI

HANAHAN D, WEINBERG RA. Hallmarks of cancer: the next generation. Cell. 2011;144:646–674. doi: 10.1016/j.cell.2011.02.013. PubMed DOI

CHANG TKH, WAXMAN DJ. Enzymatic analysis of cDNA-expressed human CYP1A1, CYP1A2 and CYP1B1 with ethoxyresorufin as a substrate. In: PHILLIPS IR, SHEPHARD EA, editors. Cytochrome P450 Protocols. Humana; Totowa, USA: 1998. pp. 103–122. PubMed DOI

LUCAS D, MENEZ JF, BERTHOU F. Chlorzoxazone: an in vitro and in vivo substrate probe for liver CYP2E1. Methods Enzymol. 1996;272:115–123. doi: 10.1016/S0076-6879(96)72014-1. PubMed DOI

McCLUE SJ, STUART I. Metabolism of the trisubstituted purine cyclin-dependent kinase inhibitor seliciclib (r-roscovitine) in vitro and in vivo. Drug Metab Dispos. 2008;36:561–570. doi: 10.1124/dmd.107.019232. PubMed DOI

McTAVISH D, BUCKLEY MMT, HEEL RC. Omeprazole. An updated review of its pharmacology and therapeutic use in acid-related disorders. Drugs. 1991;42:138–170. doi: 10.2165/00003495-199142010-00008. PubMed DOI

MORRIS GM, HUEY R, LINDSTROM W, SANNER MF, BELEW RK, GOODSELL DS, OLSON AJ. Autodock4 and AutoDockTools4: automated docking with selective receptor flexibility. J Comp Chem. 2009;16:2785–2791. doi: 10.1002/jcc.21256. PubMed DOI PMC

NUTLEY BP, RAYNAUD FI, WILSON SC, FISCHER PM, HAYES A, GODDARD PM, McCLUE SJ, JARMAN M, LANE DP, WORKMAN P. Metabolism and pharmacokinetics of the cyclin-dependent kinase inhibitor R-roscovitine in the mouse. Mol Cancer Ther. 2005;4:125–139. PubMed

OTTO T, SICINSKI P. Cell cycle proteins as promising targets in cancer therapy. Nat Rev Cancer. 2017;17:93–115. doi: 10.1038/nrc.2016.138. PubMed DOI PMC

ORTIZ De MONTELLANO PR, editor. Cytochrome P450: Structure, Mechanism, and Biochemistry. 3rd edition. New York: Kluwer Academic/Plenum; 2005. DOI

REYNALD RL, SANSEN S, STOUT CD, JOHNSON EF. Structural characterization of human cytochrome P450 2C19: active site differences between P450s 2C8, 2C9, and 2C19. J Bio Chem. 2012;287:44581–44591. doi: 10.1074/jbc.M112.424895. PubMed DOI PMC

SOUČEK P. Novel sensitive high-performance liquid chromatographic method for assay of coumarin 7-hydroxylation. J Chromatogr B. 1999;734:23–29. doi: 10.1016/S0378-4347(99)00325-4. PubMed DOI

ŠILLER M, ANZENBACHER P, ANZENBACHEROVÁ E, DOLEŽAL K, POPA I, STRNAD M. Interactions of olomoucine II with human liver microsomal cytochromes P450. Drug Metab Dispos. 2009;37:1198–1202. doi: 10.1124/dmd.108.025502. PubMed DOI

ŠPIČÁKOVÁ A, BAZGIER V, SKÁLOVÁ L, OTYEPKA M, ANZENBACHER P. β caryophyllene oxide and transnerolidol affect enzyme activity of CYP3A4 - in vitro and in silico studies. Physiol Res. 2019;68(Suppl 1):S51–S58. doi: 10.33549/physiolres.934323. PubMed DOI

ŠPIČÁKOVÁ A, ANZENBACHER P, LIŠKOVÁ B, KUČA K, FUSEK J, ANZENBACHEROVÁ E. Evaluation of possible inhibition of human liver drug metabolizing cytochromes p450 by two new acetylcholinesterase oxime-type reactivators. Food Chem Toxicol. 2016;88:100–104. doi: 10.1016/j.fct.2015.11.024. PubMed DOI

ŠREJBER M, NAVRÁTILOVÁ V, PALONCÝOVÁ M, BAZGIER V, BERKA K, ANZENBACHER P, OTYEPKA M. Membrane-attached mammalian cytochromes P450: An overview of the membrane’s effects on structure, drug binding, and interactions with redox partners. J Inorg Biochem. 2018;183:117–136. doi: 10.1016/j.jinorgbio.2018.03.002. PubMed DOI

TOMÁNKOVÁ V, VLČKOVÁ A, ANZENBACHER P, BACHLEDA P, ANZENBACHEROVÁ E. The use of goldenrod general (Solidago virgaurea) preparations does not influence the metabolism of concomitantly administered drugs. Klin Farmakol Farm. 2016;30:5–10. doi: 10.36290/far.2016.002. DOI

TROTT O, OLSON AJ. Software news and update autodock vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem. 2010;31:455–461. doi: 10.1002/jcc.21334. PubMed DOI PMC

Van der BIESSEN DA, BURGER H, de BRUIJN P, LAMERS CH, NAUS N, LOFERER H, WIEMER EA, MATHIJSSEN RH, De JONGE MJ. Phase I study of RGB-286638, a novel, multitargeted cyclin-dependent kinase inhibitor in patients with solid tumors. Clin Cancer Res. 2014;20:4776–83. doi: 10.1158/1078-0432.CCR-14-0325. PubMed DOI

WANG X, DENG K, WANG C, LI Y, WANG T, HUANG Z, MA Y, SUN P, SHI Y, YANG S, FAN Y, XIANG R. Novel CDKs inhibitors for the treatment of solid tumour by simultaneously regulating the cell cycle and transcription control. J Enzyme Inhib Med Chem. 2020;35:414–423. doi: 10.1080/14756366.2019.1705290. PubMed DOI PMC

YEO W, GOH B, LE TOURNEAU C, GREEN SR, CHIAO JH, SIU LL. A phase II randomized study of oral seliciclib in patients with previously treated nasopharyngeal carcinoma. J Clin Oncol. 2009;27 doi: 10.1200/jco.2009.27.15_suppl.6026. DOI

ZANGER UM, SCHWAB M. Cytochrome P450 enzymes in drug metabolism: regulation of gene expression, enzyme activities, and impact of genetic variation. Pharmacol Ther. 2013;138:103–141. doi: 10.1016/j.pharmthera.2012.12.007. PubMed DOI

ZATLOUKAL M, JORDA R, GUCKÝ T, REZNICKOVA E, VOLLER J, POSPISIL T, MALINKOVA V, ADAMCOVA H, KRYSTOF V, STRNAD M. Synthesis and in vitro biological evaluation of 2,6,9-trisubstituted purines targeting multiple cyclin-dependent kinases. Eur J Med Chem. 2013;61:61–72. doi: 10.1016/j.ejmech.2012.06.036. PubMed DOI