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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Tocris tamsulosin
Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or <t>tamsulosin</t> (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.
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Image Search Results


Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or tamsulosin (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.

Journal: Journal of medicinal chemistry

Article Title: Stereoselectivity in Cell Uptake by SLC22 Organic Cation Transporters 1, 2, and 3.

doi: 10.1021/acs.jmedchem.3c01436

Figure Lengend Snippet: Figure 6. Combined effects of OCT1 and CYP2D6 on the cellular disposition of racemic drugs. HEK293 cells overexpressing OCT1, CYP2D6, and both, and empty-vector (EV)-transfected control cells were incubated with 1 μM racemic formoterol (A), oxyphenonium (B), or tamsulosin (C). After 90 min, the amount of the substance left over in the cellular supernatant as well as the intracellular concentrations were quantified by chiral LC−MS/MS analysis. Results are shown as mean ± SEM of three independent experiments. Chiral centers of OCT1/CYP2D6 substrates are highlighted with asterisks. Asterisks indicate statistical significance of contributions of OCT1, CYP2D6, or the combination of OCT1 and CYP2D6 in the double-transfected cells to the depletion of the individual enantiomer, *p < 0.05, **p < 0.01, ***p < 0.001. The number signs indicate statistically significant differences between two enantiomers with Student’s t-test and #p < 0.05, ##p < 0.01, ###p < 0.001.

Article Snippet: Compounds were purchased from Santa-Cruz Biotechnology (Darmstadt, Germany; sc-294579, Etilefrine hydrochloride; sc-295159, Homatropine hydrochloride; sc-204086, Milnacipran hydrochloride; and sc-203699, Sotalol hydrochloride), SigmaAldrich (Darmstadt, Germany; SML2868, aclidinium bromide; A2729, amisulpride; B8684, bambuterol hydrochloride; B5274, bupivacaine hydrochloride; BP567; carteolol hydrochloride; 492051, choline chloride−trimethyl-d9; C0414, clidinium bromide; E4630, (S,S)-ethambutol dihydrochloride; PHR2703, formoterol fumarate; 1286402, frovatriptan; 1286413, (R)-frovatriptan succinate; M2398, metaproterenol hemisulfate; M5651, mepenzolate bromide; L5783, N-ethyl lidocaine bromide; O5501, oxyphenonium bromide; 32142, pirbuterol acetate; G7048; proguanil hydrochloride; Y0000653, tamsulosin; T1330, (R)-tamsulosin hydrochloride; T2528, terbutaline hemisulfate; Y0001986, (R)-zolmitriptan; and SML0248, (S)zolmitriptan) and Toronto Research Chemicals (Toronto, Canada; A190150, (S)-aclidinium bromide; A633255, (R)-amisulpride; E889805, (R,R)-ethambutol dihydrochloride; E67805, rel-(R,S)ethambutol dihydrochloride; and X499808; Xamoterol hemifumarate).

Techniques: Plasmid Preparation, Transfection, Control, Incubation, Liquid Chromatography with Mass Spectroscopy