Structural Requirements for Drug Inhibition of the Liver Specific Human Organic Cation Transport Protein 1

Ahlin, Gustav; Karlsson, Johan; Pedersen, Jenny M.; Gustavsson, Lena; Larsson, Rolf; Matsson, Pär; Norinder, Ulf; Bergström, Christel A S; Artursson, Per

doi:10.1021/jm8003152

Cited by 181 publications

(237 citation statements)

References 45 publications

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“…Not all but many organic cations (of endogenous and exogenous origin) are substrates of OCTs. Endogenous organic cations include choline, dopamine, histamine, and creatinine, and exogenous organic cations include many drugs (up to 40% of the prescribed drugs are cations at physiological pH), such as vecuronium, procainamide, quinine, and cimetidine, to name a few (5). Platin-like compounds are transported by OCT2, and inhibition of OCT2-mediated transport by cimetidine reduces cisplatin-induced nephrotoxicity and apoptosis (6).…”

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confidence: 99%

Inhibitory Interaction Potential of 22 Antituberculosis Drugs on Organic Anion and Cation Transporters of the SLC22A Family

Parvez

Kaisar

Shin

et al. 2016

Antimicrob Agents Chemother

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b Twenty-two currently marketed antituberculosis drugs were comprehensively evaluated for their inhibitory effect on organic anionic transporter (OAT)-and organic cation transporter (OCT)-mediated uptake using stably transfected HEK293 cells in vitro. We observed moderate to strong inhibitory effects on OAT1-and OAT3-mediated para-aminohippurate (PAH) uptake and OCT1-and OCT2-mediated N-methyl-4-phenylpylidinium acetate (MPP ؉ ) uptake. Ciprofloxacin, linezolid, para-aminosalicylic acid (PAS), and rifampin were observed to have strong inhibitory effects, with the concentrations for a 50% inhibitory effect (IC 50 s) being 35.1, 31.1, 37.6, and 48.1 M, respectively, for OAT1 and >100, 21.9, 24.6, and 30.2 M, respectively, for OAT3. Similarly, pyrazinamide, rifabutin, and levofloxacin were observed to have inhibitory effects, with IC 50 values being 36.5, 42.7, and 30.3 M, respectively, for OCT1 and with the IC 50 value for PAS being 94.2 M for OCT2. In addition, we used zidovudine and metformin as clinically prescribed substrates of OATs and OCTs, respectively, and zidovudine and metformin uptake was also strongly inhibited by the antituberculosis drugs. Among the tested drugs, the highest drug-drug interaction (DDI) indexes were found for PAS, which were 9.3 to 13.9 for OAT1 and 12.0 to 17.7 for OAT3, and linezolid, which were 1.18 to 2.15 for OAT1 and 1.7 to 3.01 for OAT3. Similarly, the DDI indexes of pyrazinamide and levofloxacin were 0.57 and 0.30, respectively, for OCT1, and the DDI index of PAS was 3.8 for OCT2, suggesting a stronger possibility (DDI index value cutoff, >0.1) of in vivo DDIs. This is the first comprehensive report of the inhibitory potential of anti-TB drugs on OAT-and OCT-mediated uptake of prototype and clinically prescribed substrate drugs in vitro, providing an ability to predict DDIs between anti-TB drugs and other coprescribed drugs in clinical studies in vivo. So far, 400 human solute carrier family (SLC) transporter proteins have been identified as a superfamily and have been classified into 52 different subfamilies (1). The organic cation transporter (OCT; SLC22 subfamily) and organic anion transporter (OAT; SLC22 subfamily) families play a major role in the disposition and pharmacology of drug substances (2). Among these, organic cation transporter OCT1 (SLC22A1) appears to be highly expressed in the liver of all species and in many tissues, although intriguing differences between species have been noted. OCT1 expression is restricted to the basolateral membrane of proximal tubular cells, with the greatest expression being apparent in an early segment (segment S1) of superficial, midcortical, and juxtamedullary nephrons (3), whereas OCT2 (SLC22A2) is expressed in the kidney and appears to be restricted to the basolateral membrane of proximal tubular cells (4).The function of OCTs includes mediation of the transport of organic cations into or out of cells. Not all but many organic cations (of endogenous and exogenous origin) are substrates of OCTs. Endogenous organic cations includ...

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confidence: 99%

Inhibitory Interaction Potential of 22 Antituberculosis Drugs on Organic Anion and Cation Transporters of the SLC22A Family

Parvez

Kaisar

Shin

et al. 2016

Antimicrob Agents Chemother

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“…2,15 Contrary to a multitude of endogenous and xenobiotic substances that inhibit OCT-mediated transport without being transported themselves, 2,15,16 the number of proven OCT transport substrates is limited. In addition to prototypic cations (such as tetraethylammonium and 1-methyl-4-phenylpyridinium) 2,15 and endogenous substances (such as acetylcholine [an OCT1 substrate] 17 and catecholamines [OCT3 substrates]), 10,17 several drugs have been identified as OCT substrates, and their number is increasing.…”

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confidence: 99%

Expression of Organic Cation Transporters OCT1 (SLC22A1) and OCT3 (SLC22A3) Is Affected by Genetic Factors and Cholestasis in Human Liver†

et al. 2009

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An important function of hepatocytes is the biotransformation and elimination of various drugs, many of which are organic cations and are taken up by organic cation transporters (OCTs) of the solute carrier family 22 (SLC22). Because interindividual variability of OCT expression may affect response to cationic drugs such as metformin, we systematically investigated genetic and nongenetic factors of OCT1/SLC22A1 and OCT3/SLC22A3 expression in human liver. OCT1 and OCT3 expression (messenger RNA [mRNA], protein) was analyzed in liver tissue samples from 150 Caucasian subjects. Hepatic OCTs were localized by way of immunofluorescence microscopy. Matrixassisted laser desorption/ionization time-of-flight mass spectrometry and genome-wide singlenucleotide polymorphism microarray technology served to genotype 92 variants in the SLC22A1-A3/ OCT1-3 gene cluster. Transport of metformin by recombinant human OCT1 and OCT3 was compared using transfected cells. OCT1 mRNA and protein expression varied 113-and 83-fold, respectively; OCT3 mRNA expression varied 27-fold. OCT1 transcript levels were on average 15-fold higher compared with OCT3. We localized the OCT3 protein to the basolateral hepatocyte membrane and identified metformin as an OCT3 substrate. OCT1 and OCT3 expression are independent of age and sex but were significantly reduced in liver donors diagnosed as cholestatic (P < 0.01). Several haplotypes for OCT1 and OCT3 were identified. Multivariate analysis adjusted for multiple testing showed that only the OCT1-Arg61Cys variant (rs12208357) strongly correlated with decreased OCT1 protein expression (P < 0.0001), and four variants in OCT3 (rs2292334, rs2048327, rs1810126, rs3088442) were associated with reduced OCT3 mRNA levels (P ‫؍‬ 0.03). Conclusion: We identified cholestasis and genetic variants as critical determinants for considerable interindividual variability of hepatic OCT1 and OCT3 expression. This indicates consequences for hepatic elimination of and response to OCT substrates such as metformin.

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“…Human embryonic kidney 293 (HEK293) cells constitutively expressing the Epstein-Barr virus nuclear antigen (HEK293-EBNA) were used to overexpress the human drug efflux transporters P-gp, MRP2, and BCRP. The HEK293-EBNA cell line was chosen because it exhibits a low background of both uptake and efflux transporters (Ahlin et al, 2008(Ahlin et al, , 2009) and has been used widely for expression of various recombinant proteins (Wurm, 2004). Moreover, this cell line is amenable to large-scale polyethylenimine (PEI)-mediated transient transfection for large-scale expression of recombinant protein (Tuvesson et al, 2008).…”

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High-Activity P-Glycoprotein, Multidrug Resistance Protein 2, and Breast Cancer Resistance Protein Membrane Vesicles Prepared from Transiently Transfected Human Embryonic Kidney 293-Epstein-Barr Virus Nuclear Antigen Cells

Karlsson¹,

Heddle²,

Rozkov³

et al. 2010

Drug Metab Dispos

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ABSTRACT:Membrane-bound transporter proteins play an important role in the efflux of drugs from cells and can significantly influence the pharmacokinetics of drug molecules. This study describes the production of large amounts of high-activity transporter membrane vesicles from human embryonic kidney 293-Epstein-Barr virus nuclear antigen cells transiently transfected using a Gateway-adapted pCEP4 plasmid. Transfections were scaled up to 10-liter cell cultures, and vesicle preparations were optimized using ultracentrifugation with a sucrose cushion, which enabled us to produce hundreds of milligrams of membrane vesicles expressing human efflux transporter proteins Pglycoprotein (P-gp)/multidrug resistance 1 (ABCB1), multidrug resistance protein 2 (MRP2) (ABCC2), and breast cancer resistance protein (BCRP) (ABCG2). Assays were developed and optimized for analyzing the ATP-dependent functionality of the transporters using probe substrates and specific inhibitors. Excellent signal/noise ratios of ATP-stimulated uptake for P-gp, MRP2, and BCRP vesicles were obtained, indicating high expression of functioning transporters. The uptake kinetics of the transporters was investigated by determining K m and V max using the model substrates N-methylquinidine (P-gp), estradiol-17␤-glucuronide (MRP2), and estrone-3-sulfate (BCRP). The ATP-dependent transport was inhibited by the model inhibitors verapamil (P-gp), benzbromarone (MRP2), and sulfasalazine (BCRP). The vesicles are thus well suited to screen for possible substrates and inhibitors in high throughput systems or are used for detailed mechanistic investigations of transporter kinetics of specific substances.

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Structural Requirements for Drug Inhibition of the Liver Specific Human Organic Cation Transport Protein 1

Cited by 181 publications

References 45 publications

Inhibitory Interaction Potential of 22 Antituberculosis Drugs on Organic Anion and Cation Transporters of the SLC22A Family

Inhibitory Interaction Potential of 22 Antituberculosis Drugs on Organic Anion and Cation Transporters of the SLC22A Family

Expression of Organic Cation Transporters OCT1 (SLC22A1) and OCT3 (SLC22A3) Is Affected by Genetic Factors and Cholestasis in Human Liver†

High-Activity P-Glycoprotein, Multidrug Resistance Protein 2, and Breast Cancer Resistance Protein Membrane Vesicles Prepared from Transiently Transfected Human Embryonic Kidney 293-Epstein-Barr Virus Nuclear Antigen Cells

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