A novel fluorescence-based functional assay for human OATP1A2 and OATP1C1 identifies interaction between third-generation P-gp inhibitors and OATP1A2

被引:28
作者
Bakos, Eva [1 ]
Nemet, Orsolya [1 ]
Patik, Izabel [1 ]
Kucsma, Nora [1 ]
Varady, Gyorgy [2 ]
Szakacs, Gergely [1 ,3 ]
Ozvegy-Laczka, Csilla [1 ]
机构
[1] Hungarian Acad Sci, Inst Enzymol, Membrane Prot Res Grp, Res Ctr Nat Sci, Magyar Tudosok Krt 2, H-1117 Budapest, Hungary
[2] Hungarian Acad Sci, Inst Enzymol, Res Ctr Nat Sci, Mol Cell Biol Lab, Budapest, Hungary
[3] Med Univ Vienna, Inst Canc Res, Vienna, Austria
关键词
central nervous system; drug interaction screen; fluorescent dye; OATP; P-gp inhibitor; ANION-TRANSPORTING POLYPEPTIDES; BLOOD-BRAIN-BARRIER; THYROID-HORMONE TRANSPORTERS; PHASE-I TRIAL; TRIHYDROCHLORIDE LY335979; GLYCOPROTEIN INHIBITOR; SODIUM FLUORESCEIN; CLINICAL-RELEVANCE; CHOROID-PLEXUS; EXPRESSION;
D O I
10.1111/febs.15156
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Organic anion-transporting polypeptide 1A2 (OATP1A2), expressed in the human blood-brain barrier, promotes drug uptake from the blood and hence can be exploited for central nervous system-targeted drug delivery. The thyroid transporter OATP1C1, expressed in the choroid plexus and in astrocytes, is also a potential pharmacological target. Based on their established pharmacological relevance, screening the drug interaction profile of OATP1A2 and OATP1C1 is highly desirable. However, drug interaction screens require suitable model systems and functional assays. In the current study, uptake of a set of cell-impermeable fluorescent dyes was screened in HEK-293 and A431 cell lines overexpressing OATP1A2 and OATP1C1. Based on the uptake of fluorescent dye substrates, a functional assay was developed, which was used to characterize OATP inhibitors/substrates. We identify Live/Dead Green (LDG), Live-or-Dye 488, and sulforhodamines 101, G, and B as novel fluorescent substrates of OATP1A2 and OATP1C1. We show that LDG uptake is proportional to OATP1A2/1C1 expression, allowing the isolation of cells expressing high transporter levels. Additionally, dye uptake can be used to characterize the drug interaction pattern of OATP1A2 and OATP1C1. We demonstrate that third-generation P-glycoprotein inhibitors elacridar, tariquidar, and zosuquidar inhibit OATP1A2 function. Increased toxicity of elacridar in OATP1A2-expressing cells suggests that OATP1A2 may modulate the distribution of this compound. The fluorescence-based assays developed in the current study are a good alternative of radioligand-based tests and pave the way toward high-throughput screens for OATP1A2/1C1 drug interaction studies.
引用
收藏
页码:2468 / 2485
页数:18
相关论文
共 63 条
[1]   Functional Expression of P-glycoprotein and Organic Anion Transporting Polypeptides at the Blood-Brain Barrier: Understanding Transport Mechanisms for Improved CNS Drug Delivery? [J].
Abdullahi, Wazir ;
Davis, Thomas P. ;
Ronaldson, Patrick T. .
AAPS JOURNAL, 2017, 19 (04) :931-939
[2]   Expression of Thyroid Hormone Transporters in the Human Hypothalamus [J].
Alkemade, Anneke ;
Friesema, Edith C. H. ;
Kalsbeek, Andries ;
Swaab, Dick F. ;
Visser, Theo J. ;
Fliers, Eric .
JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM, 2011, 96 (06) :E967-E971
[3]   Naringin is a major and selective clinical inhibitor of organic anion-transporting polypeptide 1A2 (OATP1A2) in grapefruit juice [J].
Bailey, D. G. ;
Dresser, G. K. ;
Leake, B. F. ;
Kim, R. B. .
CLINICAL PHARMACOLOGY & THERAPEUTICS, 2007, 81 (04) :495-502
[4]   Fruit juice inhibition of uptake transport: a new type of food-drug interaction [J].
Bailey, David G. .
BRITISH JOURNAL OF CLINICAL PHARMACOLOGY, 2010, 70 (05) :645-655
[5]   A Call for Systematic Research on Solute Carriers [J].
Cesar-Razquin, Adrian ;
Snijder, Berend ;
Frappier-Brinton, Tristan ;
Isserlin, Ruth ;
Gyimesi, Gergely ;
Bai, Xiaoyun ;
Reithmeier, Reinhart A. ;
Hepworth, David ;
Hediger, Matthias A. ;
Edwards, Aled M. ;
Superti-Furga, Giulio .
CELL, 2015, 162 (03) :478-487
[6]   Therapeutic Potential and Utility of Elacridar with Respect to P-glycoprotein Inhibition: An Insight from the Published In Vitro, Preclinical and Clinical Studies [J].
Dash, Ranjeet Prasad ;
Babu, R. Jayachandra ;
Srinivas, Nuggehally R. .
EUROPEAN JOURNAL OF DRUG METABOLISM AND PHARMACOKINETICS, 2017, 42 (06) :915-933
[7]   Sodium Fluorescein is a Probe Substrate for Hepatic Drug Transport Mediated by OATP1B1 and OATP1B3 [J].
De Bruyn, Tom ;
Fattah, Sarinj ;
Stieger, Bruno ;
Augustijns, Patrick ;
Annaert, Pieter .
JOURNAL OF PHARMACEUTICAL SCIENCES, 2011, 100 (11) :5018-5030
[8]   The organic anion transporting polypeptide 1a5 is a pivotal transporter for the uptake of microcystin-LR by gonadotropin-releasing hormone neurons [J].
Ding, Jie ;
Wang, Jing ;
Xiang, Zou ;
Diao, Weiyi ;
Su, Moxi ;
Shi, Weiwei ;
Wan, Ting ;
Han, Xiaodong .
AQUATIC TOXICOLOGY, 2017, 182 :1-10
[9]   Protein Abundance of Clinically Relevant Multidrug Transporters along the Entire Length of the Human Intestine [J].
Drozdzik, Marek ;
Groeer, Christian ;
Penski, Jette ;
Lapczuk, Joanna ;
Ostrowski, Marek ;
Lai, Yurong ;
Prasad, Bhagwat ;
Unadkat, Jashvant D. ;
Siegmund, Werner ;
Oswald, Stefan .
MOLECULAR PHARMACEUTICS, 2014, 11 (10) :3547-3555
[10]   The impact of Organic Anion-Transporting Polypeptides (OATPs) on disposition and toxicity of antitumor drugs: Insights from knockout and humanized mice [J].
Durmus, Selvi ;
van Hoppe, Stephanie ;
Schinkel, Alfred H. .
DRUG RESISTANCE UPDATES, 2016, 27 :72-88