Purine cyclin-dependent kinase inhibitors have been recognized as promising candidates for the treatment of various cancers; nevertheless, data regarding interaction of these substances with drug efflux transporters is still lacking. Recently, we have demonstrated inhibition of breast cancer resistance protein (ABCG2) by olomoucine II and purvalanol A and shown that these compounds are able to synergistically potentiate the antiproliferative effect of mitoxantrone, an ABCG2 substrate. In this follow up study, we investigated whether olomoucine II and purvalanol A are transported by ABCG2 and ABCB1 (P-glycoprotein). Using monolayers of MDCKII cells stably expressing human ABCB1 or ABCG2, we demonstrated that olomoucine II, but not purvalanol A, is a dual substrate of both ABCG2 and ABCB1. We, therefore, assume that pharmacokinetics of olomoucine II will be affected by both ABCB1 and ABCG2 transport proteins, which might potentially result in limited accumulation of the compound in tumor tissues or lead to drug-drug interactions. Pharmacokinetic behavior of purvalanol A, on the other hand, does not seem to be affected by either ABCG2 or ABCB1, theoretically favoring this drug in the potential treatment of efflux transporter-based multidrug resistant tumors. In addition, we observed intensive sulfatation of olomoucine II in MDCKII cell lines with subsequent active efflux of the metabolite out of the cells. Therefore, care should be taken when performing pharmacokinetic studies in MDCKII cells, especially if radiolabeled substrates are used; the generated sulfated conjugate may largely contaminate pharmacokinetic analysis and result in misleading interpretation. With regard to chemical structures of olomoucine II and purvalanol A, our data emphasize that even drugs with remarkable structure similarity may show different pharmacokinetic behavior such as interactions with ABC transporters or biotransformation enzymes.

Department of Pharmacology and Toxicology Charles University Prague Faculty of Pharmacy Hradec Kralove Czech Republic

Zobrazit více v PubMed

Krystof V, Lenobel R, Havlicek L, Kuzma M, Strnad M (2002) Synthesis and biological activity of olomoucine II. Bioorg Med Chem Lett 12: 3283–3286. PubMed

Gray NS, Wodicka L, Thunnissen AM, Norman TC, Kwon S, et al. (1998) Exploiting chemical libraries, structure, and genomics in the search for kinase inhibitors. Science 281: 533–538. PubMed

Krystof V, McNae IW, Walkinshaw MD, Fischer PM, Muller P, et al. (2005) Antiproliferative activity of olomoucine II, a novel 2,6,9-trisubstituted purine cyclin-dependent kinase inhibitor. Cell Mol Life Sci 62: 1763–1771. PubMed PMC

Villerbu N, Gaben AM, Redeuilh G, Mester J (2002) Cellular effects of purvalanol A: a specific inhibitor of cyclin-dependent kinase activities. Int J Cancer 97: 761–769. PubMed

Monaco EA, Beaman-Hall CM, Mathur A, Vallano ML (2004) Roscovitine, olomoucine, purvalanol: inducers of apoptosis in maturing cerebellar granule neurons. Biochem Pharmacol 67: 1947–1964. PubMed

Krystof V, Uldrijan S (2010) Cyclin-dependent kinase inhibitors as anticancer drugs. Curr Drug Targets 11: 291–302. PubMed

Diaz-Padilla I, Siu LL, Duran I (2009) Cyclin-dependent kinase inhibitors as potential targeted anticancer agents. Invest New Drugs. PubMed

Cicenas J, Valius M (2011) The CDK inhibitors in cancer research and therapy. J Cancer Res Clin Oncol 137: 1409–1418. PubMed

Aldoss IT, Tashi T, Ganti AK (2009) Seliciclib in malignancies. Expert Opin Investig Drugs 18: 1957–1965. PubMed

Knockaert M, Gray N, Damiens E, Chang YT, Grellier P, et al. (2000) Intracellular targets of cyclin-dependent kinase inhibitors: identification by affinity chromatography using immobilised inhibitors. Chem Biol 7: 411–422. PubMed

Knockaert M, Meijer L (2002) Identifying in vivo targets of cyclin-dependent kinase inhibitors by affinity chromatography. Biochem Pharmacol 64: 819–825. PubMed

Bain J, McLauchlan H, Elliott M, Cohen P (2003) The specificities of protein kinase inhibitors: an update. Biochem J 371: 199–204. PubMed PMC

Hikita T, Oneyama C, Okada M (2010) Purvalanol A, a CDK inhibitor, effectively suppresses Src-mediated transformation by inhibiting both CDKs and c-Src. Genes Cells 15: 1051–1062. PubMed

Staud F, Ceckova M, Micuda S, Pavek P (2010) Expression and function of p-glycoprotein in normal tissues: effect on pharmacokinetics. Methods Mol Biol 596: 199–222. PubMed

Staud F, Pavek P (2005) Breast cancer resistance protein (BCRP/ABCG2). Int J Biochem Cell Biol 37: 720–725. PubMed

Allen JD, Schinkel AH (2002) Multidrug resistance and pharmacological protection mediated by the breast cancer resistance protein (BCRP/ABCG2). Mol Cancer Ther 1: 427–434. PubMed

Szakacs G, Paterson JK, Ludwig JA, Booth-Genthe C, Gottesman MM (2006) Targeting multidrug resistance in cancer. Nat Rev Drug Discov 5: 219–234. PubMed

Han B, Zhang JT (2004) Multidrug resistance in cancer chemotherapy and xenobiotic protection mediated by the half ATP-binding cassette transporter ABCG2. Curr Med Chem Anticancer Agents 4: 31–42. PubMed

Yang K, Wu J, Li X (2008) Recent advances in the research of P-glycoprotein inhibitors. Biosci Trends 2: 137–146. PubMed

Shukla S, Ohnuma S, Ambudkar SV (2011) Improving cancer chemotherapy with modulators of ABC drug transporters. Curr Drug Targets 12: 621–630. PubMed PMC

Giacomini KM, Huang SM, Tweedie DJ, Benet LZ, Brouwer KL, et al. (2010) Membrane transporters in drug development. Nat Rev Drug Discov 9: 215–236. PubMed PMC

Hofman J, Ahmadimoghaddam D, Hahnova L, Pavek P, Ceckova M, et al. (2012) Olomoucine II and purvalanol A inhibit ABCG2 transporter in vitro and in situ and synergistically potentiate cytostatic effect of mitoxantrone. Pharmacol Res 65: 312–319. PubMed

Rabindran SK, Ross DD, Doyle LA, Yang W, Greenberger LM (2000) Fumitremorgin C reverses multidrug resistance in cells transfected with the breast cancer resistance protein. Cancer Res 60: 47–50. PubMed

Dantzig AH, Shepard RL, Cao J, Law KL, Ehlhardt WJ, et al. (1996) Reversal of P-glycoprotein-mediated multidrug resistance by a potent cyclopropyldibenzosuberane modulator, LY335979. Cancer Res 56: 4171–4179. PubMed

Shepard RL, Cao J, Starling JJ, Dantzig AH (2003) Modulation of P-glycoprotein but not MRP1- or BCRP-mediated drug resistance by LY335979. Int J Cancer 103: 121–125. PubMed

Goh LB, Spears KJ, Yao D, Ayrton A, Morgan P, et al. (2002) Endogenous drug transporters in in vitro and in vivo models for the prediction of drug disposition in man. Biochem Pharmacol 64: 1569–1578. PubMed

Kuteykin-Teplyakov K, Luna-Tortos C, Ambroziak K, Loscher W (2010) Differences in the expression of endogenous efflux transporters in MDR1-transfected versus wildtype cell lines affect P-glycoprotein mediated drug transport. Br J Pharmacol 160: 1453–1463. PubMed PMC

Holcapek M, Kolarova L, Nobilis M (2008) High-performance liquid chromatography-tandem mass spectrometry in the identification and determination of phase I and phase II drug metabolites. Anal Bioanal Chem 391: 59–78. PubMed PMC

Bachmeier CJ, Miller DW (2005) A fluorometric screening assay for drug efflux transporter activity in the blood-brain barrier. Pharm Res 22: 113–121. PubMed

An R, Hagiya Y, Tamura A, Li S, Saito H, et al. (2009) Cellular phototoxicity evoked through the inhibition of human ABC transporter ABCG2 by cyclin-dependent kinase inhibitors in vitro. Pharm Res 26: 449–458. PubMed

Rajnai Z, Mehn D, Beery E, Okyar A, Jani M, et al. (2010) ATP-binding cassette B1 transports seliciclib (R-roscovitine), a cyclin-dependent kinase inhibitor. Drug Metab Dispos 38: 2000–2006. PubMed

Seamon JA, Rugg CA, Emanuel S, Calcagno AM, Ambudkar SV, et al. (2006) Role of the ABCG2 drug transporter in the resistance and oral bioavailability of a potent cyclin-dependent kinase/Aurora kinase inhibitor. Mol Cancer Ther 5: 2459–2467. PubMed

Ng KH, Lim BG, Wong KP (2003) Sulfate conjugating and transport functions of MDCK distal tubular cells. Kidney Int 63: 976–986. PubMed

Ishikawa T, Ikegami Y, Sano K, Nakagawa H, Sawada S (2006) Transport mechanism-based drug molecular design: novel camptothecin analogues to circumvent ABCG2-associated drug resistance of human tumor cells. Curr Pharm Des 12: 313–325. PubMed

Nakagawa H, Saito H, Ikegami Y, Aida-Hyugaji S, Sawada S, et al. (2006) Molecular modeling of new camptothecin analogues to circumvent ABCG2-mediated drug resistance in cancer. Cancer Lett 234: 81–89. PubMed

Purvalanol A, olomoucine II and roscovitine inhibit ABCB1 transporter and synergistically potentiate cytotoxic effects of daunorubicin in vitro