Fluorescence-Based Assay for the Interaction of Small Molecules with the Human Renal Organic Anion Transporter 1

Cihlář, Tomáš; Ho, Edmund

doi:10.1006/abio.2000.4633

Cited by 74 publications

(37 citation statements)

References 21 publications

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“…Uptake of nonpermeable fluorescein derivates has been ascribed to the activation not only of SLCO/OATP transporters but also of the family of organic anion transporters, SLC22A/OAT (20,42). Accordingly, fluorescein derivatives uptake by SLC22A/OAT transporters has been reported to be inhibited by p-aminohippuric acid and coupled to the Na ϩ gradient across the plasma membrane (42,43). As shown in Fig.…”

Section: Gsh Transportersmentioning

confidence: 99%

SLCO/OATP-like Transport of Glutathione in FasL-induced Apoptosis

Franco

Cidlowski

2006

Journal of Biological Chemistry

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Apoptosis is a ubiquitous genetically encoded pathway that enables cells to undergo highly regulated death in response to specific signals. It has also been observed to occur as a consequence of distinct pathologies, and its deregulation can lead to autoimmune diseases, cancer, and neurodegenerative diseases (1). It is characterized by the activation of precise pathways leading to specific biochemical and morphological alterations, including changes in the intracellular ionic homeostasis, cell shrinkage, phosphatidylserine externalization, chromatin condensation, DNA degradation, membrane blebbing, and apoptotic body formation (1, 2). Fas ligand (FasL) 2 -induced apoptosis by binding to its receptor Fas (CD95/Apo-1) has been reported to play a critical role in immune homeostasis. Fas receptormediated apoptosis has also been implicated in the pathogenesis of fulminant hepatitis, postischemic neuronal degeneration, traumatic brain injury, and inflammatory lung diseases. Its deregulation is also associated with progression and metastasis of tumors and autoimmune lymphoproliferative syndrome (3).GSH is the predominant low molecular weight thiol in animal cells. It participates in many cellular reactions, including antioxidant defense of the cell, drug detoxification, and cell signaling, and has been reported to be necessary for the proliferation of several cells, including lymphocytes (4 -6). A reduction in the intracellular GSH concentration ([GSH] i ) has been reported during cell death in response to different apoptotic stimuli, including death receptor-induced (7-10), stress-induced, (11), and drug-induced cell death (12). Several studies have shown a correlation between GSH efflux and the progression of apoptosis (13,14), and a recent report suggests that inhibition of GSH release in apoptosis is able to rescue cells from apoptosis (8); however, the precise mechanisms involved in intracellular GSH (GSH i ) loss have not been identified. Glutathione synthesis occurs intracellularly, and its catabolism occurs by a series of both enzymatic and plasma membrane export mechanisms. Glutathione degradation occurs extracellularly; therefore, the export of GSH and GSH adducts is also an important step in its turnover (15). To date, two GSH transporter families have been implicated in GSH efflux across the plasma membrane. These include the multidrug-resistant pro-

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Section: Gsh Transportersmentioning

confidence: 99%

SLCO/OATP-like Transport of Glutathione in FasL-induced Apoptosis

Franco

Cidlowski

2006

Journal of Biological Chemistry

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“…In study 1, subjects were given benzylpenicillin alone [400,000 U, by mouth (PO)] or concomitantly with probenecid (750 mg, PO); in study 2, they were given adefovir alone (10 mg, PO) or concomitantly with probenecid (750 mg, PO); and in study 3, they were given metformin alone (100 mg, PO) or concomitantly with pyrimethamine (50 mg, PO). Probenecid is a well known, potent inhibitor of OAT1 and OAT3 (Cihlar and Ho, 2000;Tahara et al, 2005;Nozaki et al, 2007b;Takahara et al, 2013) and significantly reduces the renal clearance of various drugs that undergo tubular secretion by OATs (Masereeuw and Russel, 2010;Kusuhara et al, 2013). Pyrimethamine is a potent MATE inhibitor (Ito et al, 2010), and at its therapeutic dose, it significantly reduces the renal clearance of metformin .…”

Section: Introductionmentioning

confidence: 99%

6β-Hydroxycortisol Is an Endogenous Probe for Evaluation of Drug–Drug Interactions Involving a Multispecific Renal Organic Anion Transporter, OAT3/SLC22A8, in Healthy Subjects

et al. 2014

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6b-Hydroxycortisol (6b-OHF) is a substrate of the organic anion transporter 3 (OAT3) and the multidrug and toxin extrusion proteins MATE1 and MATE-2K in the corresponding cDNA-transfected cells. This study aimed to examine the contribution of OAT3 and MATEs to the urinary excretion of 6b-OHF in humans using the appropriate in vivo inhibitors, probenecid and pyrimethamine, for OAT3 and MATEs, respectively. Oat3(-/-) mice showed significantly reduced renal clearance of 6b-OHF (CL renal, 6b-OHF ) compared with wild-type mice (18.1 6 1.5 versus 7.60 6 1.8 ml/min/kg). 6b-OHF uptake by human kidney slices was inhibited significantly by probenecid to 20-45% of the control values and partly by 1-methyl-4-phenylpyridinium. 6b-OHF plasma concentration and the amount of 6b-OHF excreted into the urine (X 6b-OHF ) were measured in healthy subjects enrolled in drug-drug interaction studies of benzylpenicillin alone or with probenecid (study 1), adefovir alone or with probenecid (study 2), and metformin alone or with pyrimethamine (study 3). Probenecid treatment caused a 57 and 76% increase in the area under the plasma concentration-time curve for 6b-OHF (AUC 6b-OHF ) in studies 1 and 2, respectively, but did not affect X 6b-OHF . Consequently, CL renal, 6b-OHF (milliliters per minute) decreased significantly from 231 6 11 to 135 6 9 and from 225 6 26 to 141 6 12 after probenecid administration in studies 1 and 2, respectively. By contrast, neither AUC 6b-OHF nor CL renal, 6b-OHF was significantly altered by pyrimethamine administration. Taken together, these data suggest that OAT3 plays a significant role in the urinary excretion of 6b-OHF, and that 6b-OHF can be used to investigate the perpetrators of the pharmacokinetic drug interactions involving OAT3 in humans.

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“…A cell line stably expressing functional hOAT1 is currently being explored as an in vitro model to evaluate the interaction of various drugs with hOAT1 Cihlar and Ho, 2000;Ho et al, 2000;Mulato et al, 2000). In this model, hOAT1 mediates transport of p-aminohippuric acid (PAH), a prototypic substrate for the renal organic anion transport system, with a K m of 12.3 M .…”

Section: Introductionmentioning

confidence: 99%

“…Although animal pharmacokinetic studies may provide some insight into the specific drug-drug interactions, their results should be interpreted cautiously because of interspecies difference in organ transport pathways (Dresser et al, 2000) and may require clinical studies for confirmation. It is therefore desirable to develop an in vitro screening system that could provide a quick assessment of potential pharmacokinetic drug interactions subsequent to interference with specific secretion pathways.A cell line stably expressing functional hOAT1 is currently being explored as an in vitro model to evaluate the interaction of various drugs with hOAT1 Cihlar and Ho, 2000;Ho et al, 2000;Mulato et al, 2000). In this model, hOAT1 mediates transport of p-aminohippuric acid (PAH), a prototypic substrate for the renal organic anion transport system, with a K m of 12.3 M .…”

mentioning

confidence: 99%

The Anti-Influenza Drug Oseltamivir Exhibits Low Potential to Induce Pharmacokinetic Drug Interactions via Renal Secretion—Correlation of in Vivo and in Vitro Studies

Hill¹,

Cihlář²,

Oo³

et al. 2002

Drug Metab Dispos

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124

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This paper is available online at http://dmd.aspetjournals.org ABSTRACT:Oseltamivir is an ester prodrug of the active metabolite [3R,4R,5S]-4-acetamido-5-amino-3-(1-ethylpropoxy)-1-cyclohexene-1-carboxylate phosphate (Ro 64-0802), a potent and selective inhibitor of neuraminidase enzyme of influenza virus. Oseltamivir is rapidly hydrolyzed by hepatic carboxylesterases to Ro 64-0802, which is then exclusively excreted by glomerular filtration and active tubular secretion without further metabolism. In vivo and in vitro studies were conducted to evaluate the renal drug-drug interaction potential of oseltamivir. Crossover studies were conducted in healthy subjects in which oral oseltamivir was administered alone and coadministered with probenecid, cimetidine, or amoxicillin. Probenecid completely blocked the renal secretion of Ro 64-0802, increasing systemic exposure (area under the curve) by 2.5-fold, but no interaction was observed with cimetidine or amoxicillin. These in vivo data show that Ro 64-0802 is secreted via an organic anion pathway, but Ro 64-0802 does not inhibit amoxicillin renal secretion. In vitro effects of Ro 64-0802 on the human renal organic anionic transporter 1 (hOAT1) were investigated using novel Chinese hamster ovary cells stably transfected with hOAT1. Ro 64-0802 was found to be a low-efficiency substrate for hOAT1 and a very weak inhibitor of hOAT1-mediated transport of p-aminohippuric acid (PAH). Ro 64-0802 did not inhibit the hOAT1-mediated transport of amoxicillin. In contrast, probenecid effectively inhibited the transport of PAH, Ro 64-0802, and amoxicillin via hOAT1. These in vitro observations are consistent with the in vivo data, validating the usefulness of the in vitro system for evaluating such drugdrug interaction. The study results demonstrate that oseltamivir has a low drug-drug interaction potential at the renal tubular level due to inhibition of hOAT1.Oseltamivir is an orally bioavailable prodrug of Ro 64-0802 1 (GS4071), a potent and selective inhibitor of influenza A and B neuraminidase (Bardsley-Elliot and Noble, 1999). After oral administration, the prodrug is extensively hydrolyzed to its active metabolite Ro 64-0802, which is then extensively excreted by glomerular filtration and renal tubular secretion without further metabolism (He et al., 1999a).Active renal secretion occurs via specific transport proteins located in the basolateral and apical membrane of the proximal tubule (Pritchard and Miller, 1993). Two general drug secretion pathways exist in proximal tubules, one for basic compounds (organic cation transport system) and another for acidic compounds (organic anion transport system). Thus far, two active pathways capable of transporting organic cations have been identified in kidney, OCT1 and OCT2 (Grundemann et al., 1994;Okuda et al., 1996). Similarly, several renal organic anion transporters have been recently cloned and characterized. Of these, organic anion transporter 1 (OAT1) is the main component mediating tubular secretion of organic acids (Sekine, 1...

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Fluorescence-Based Assay for the Interaction of Small Molecules with the Human Renal Organic Anion Transporter 1

Cited by 74 publications

References 21 publications

SLCO/OATP-like Transport of Glutathione in FasL-induced Apoptosis

SLCO/OATP-like Transport of Glutathione in FasL-induced Apoptosis

6β-Hydroxycortisol Is an Endogenous Probe for Evaluation of Drug–Drug Interactions Involving a Multispecific Renal Organic Anion Transporter, OAT3/SLC22A8, in Healthy Subjects

The Anti-Influenza Drug Oseltamivir Exhibits Low Potential to Induce Pharmacokinetic Drug Interactions via Renal Secretion—Correlation of in Vivo and in Vitro Studies

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