Species Difference in the Inhibitory Effect of Nonsteroidal Anti-Inflammatory Drugs on the Uptake of Methotrexate by Human Kidney Slices

Abstract

Simultaneous use of nonsteroidal anti-inflammatory drugs (NSAIDs), probenecid, and other drugs has been reported to delay the plasma elimination of methotrexate in patients. Previously, we have reported that inhibition of the uptake process cannot explain such drug-drug interactions using rats. The present study quantitatively evaluated the possible role of the transporters in such drug-drug interactions using human kidney slices and membrane vesicles expressing human ATP-binding cassette (ABC) transporters. The uptake of methotrexate by human kidney slices was saturable with a K m of 45 to 49 M. Saturable uptake of methotrexate by human kidney slices was markedly inhibited by p-aminohippurate and benzylpenicillin, but only weakly by 5-methyltetrahydrofolate. These transport characteristics are similar to those of a basolateral organic anion transporter (OAT) 3/SLC22A8. NSAIDs and probenecid inhibited the uptake of methotrexate by human kidney slices, and, in particular, salicylate, indomethacin, phenylbutazone, and probenecid were predicted to exhibit significant inhibition at clinically observed plasma concentrations. Among ABC transporters, such as BCRP/ABCG2, multidrug resistance-associated protein (MRP) 2/ABCC2, and MRP4/ABCC4, which are candidates for the luminal efflux of methotrexate, ATPdependent uptake of methotrexate by MRP4-expressing membrane vesicles was most potently inhibited by NSAIDs. Salicylate and indomethacin were predicted to inhibit MRP4 at clinical plasma concentrations. Diclofenac-glucuronide significantly inhibited MRP2-mediated transport of methotrexate in a concentration-dependent manner, whereas naproxen-glucuronide had no effect. Inhibition of renal uptake (via OAT3) and efflux processes (via MRP2 and MRP4) explains the possible sites of drug-drug interaction for methotrexate with probenecid and some NSAIDs, including their glucuronides.Drug-drug interactions involving metabolism and/or excretion processes prolong the plasma elimination half-lives leading to the accumulation of drugs in the body and potentiate pharmacological/adverse effects. Recent progress in molecular biological research has shown that many types of transporters play important roles in the tissue uptake and/or subsequent secretion of drugs in the liver and kidney, and such transporters exhibit a broad substrate specificity with a degree of overlap, suggesting the possibility of transportermediated drug-drug interactions with other substrates (Shitara et al., 2005;Li et al., 2006). Methotrexate (MTX) is an analog of natural folate and has been widely and successfully used for the treatment of neoplastic diseases and autoimmune diseases, including rheu-

MCF-7 Breast Cancer Cell Line, a Model for the Study of the Association Between Inflammation and ABCG2-Mediated Multi Drug Resistance

“…As a result, expression of a MDR phenotype in human malignant cells may not always be sensitive to potentiation of drug cytotoxicity by NSAIDs (Roller et al, 1999). The present results also confirmed other studies that show NSAIDs' effects are cell and efflux transporter specific (Nozaki et al, 2007).…”

Lansoprazole Exacerbates Pemetrexed-Mediated Hematologic Toxicity by Competitive Inhibition of Renal Basolateral Human Organic Anion Transporter 3

“…A previous retrospective study demonstrated that the coadministration of NSAIDs was not a risk factor for the development of hematologic toxicity by pemetrexed (Sakata et al, 2013); however, NSAIDs are known as substrates and/or inhibitors of hOAT3 (Uwai et al, 2004;Nozaki et al, 2007). A clinical study has reported that a 20% increase in the area under the plasma concentration curve of pemetrexed was observed when 400 mg of ibuprofen was administered orally every 6 hours .…”

Transport of Diclofenac by Breast Cancer Resistance Protein (ABCG2) and Stimulation of Multidrug Resistance Protein 2 (ABCC2)-Mediated Drug Transport by Diclofenac and Benzbromarone

“…When MTX is used at high-dose regimens for cancer treatment, interactions with NSAIDs can result in severe toxicity, with sometimes fatal outcome (Thyss et al, 1986). In a recent study, using human kidney slides, it was demonstrated that diclofenac and its acyl-glucuronide can inhibit the luminal urinary efflux of MTX via the ATP-binding cassette (ABC) multidrug transporters Mrp4 and Mrp2, respectively (Nozaki et al, 2007).…”

“…When MTX is used at high-dose regimens for cancer treatment, interactions with NSAIDs can result in severe toxicity, with sometimes fatal outcome (Thyss et al, 1986). In a recent study, using human kidney slides, it was demonstrated that diclofenac and its acyl-glucuronide can inhibit the luminal urinary efflux of MTX via the ATP-binding cassette (ABC) multidrug transporters Mrp4 and Mrp2, respectively (Nozaki et al, 2007).Because it is recognized more and more that many interactions of drugs occur at the level of drug-transporting proteins, we investigated whether diclofenac is a substrate of one of the apical ABC transporters, using MDCK-II cells transfected with human P-gp, MRP2, BCRP, or murine Bcrp1 cDNA. We further used the MDCK-II cells transduced with human and mouse MRP2/Mrp2 to examine whether diclofenac could modulate MRP2-mediated transport of taxane anticancer drugs.…”