Pharmacokinetic Analysis of Transcellular Transport of Levofloxacin across LLC-PK1 and Caco-2 Cell Monolayers

Takaai, Mari; Suzuki, Honoh; Ishida, Kazuya; Tahara, Katsutoshi; Hashimoto, Yoshiaki

doi:10.1248/bpb.30.2167

Cited by 16 publications

(23 citation statements)

References 19 publications

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“…2,7) The cellular uptake of quinidine in Caco-2 cells grown on plastic dishes was temperature-dependent, and markedly increased by alkalization of the apical medium at 37°C.…”

Section: Discussionmentioning

confidence: 99%

“…]procainamide in LLC-PK 1 cell monolayers prepared on a porous membrane was examined as described previously. 1,7) In brief, the monolayer was pre-incubated for 30 min at 37°C with culture medium (pH 6.3) containing 10 mM unlabeled drug to equilibrate the drug concentration. After the equilibration period, the radio-labeled drug (0.33 mCi/ml) was applied to the apical chamber (0.75 ml) to examine apical-to-basolateral transcellular transport.…”

Section: Methodsmentioning

confidence: 99%

“…After the equilibration period, the radio-labeled drug (0.33 mCi/ml) was applied to the apical chamber (0.75 ml) to examine apical-to-basolateral transcellular transport. 1,7,8) A volume (50 ml) of medium in the basolateral chamber (1.5 ml) was then collected 30, 60, and 90 min after the radio-labeled drug was applied. Cells on the porous membrane were collected following the last collection of medium.…”

Section: Methodsmentioning

confidence: 99%

“…1,7,8) When transcellular drug transport is examined under the condition where the unlabeled drug concentration in the monolayer is equilibrated with that of the incubation medium in the apical and basolateral chambers, transport data for a small amount of radiolabeled drug can be analyzed using the linear pharmacokinetic model. That is, the following mass balance equations were prepared for pharmacokinetic analysis: (1) (2) (3) where X A , X B , and X C are the amounts of radio-labeled drugs in the apical chamber, basolateral chamber, and monolayer determined at time t, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…2,7) In the present study, we investigated whether the cation transport system for quinidine is present in another human intestinal epithelial cell line, LS180. Figure 2A shows the time course of the uptake of 100 mM quinidine at 37°C and 4°C in LS180 cells grown on plastic dishes.…”

Section: Cellular Uptake Of Quinidine and Procainamide In Intestinal mentioning

confidence: 99%

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Membrane Transport Mechanisms of Quinidine and Procainamide in Renal LLC-PK1 and Intestinal LS180 Cells

Masago

Takaai

Sakata

et al. 2010

Biological & Pharmaceutical Bulletin

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We previously evaluated the renal excretion mechanism of quinidine, which is a tertiary amine compound, using porcine kidney epithelial LLC-PK 1 cells and P-glycoprotein (P-gp)-expressed LLC-GA5-COL150 cells.1) The transepithelial transport of quinidine in the basolateral-to-apical direction in LLC-PK 1 cells was similar to that in the opposite direction. In contrast, quinidine was transported actively in the basolateral-to-apical direction in LLC-GA5-COL150 cells. The results suggested that P-gp is mainly responsible for the tubular secretion of quinidine in the kidney. 1) We also evaluated the intestinal absorption mechanism of quinidine using human intestinal epithelial Caco-2 cells.2) The temperaturedependent uptake of quinidine in Caco-2 cells grown on a plastic dish was increased by alkalization of the apical medium, and was inhibited by diphenhydramine and imipramine. The results suggested that a cation transport system was involved in the influx of quinidine at the apical membrane in intestinal epithelial cells. 2)Procainamide, another tertiary amine compound with a pK a value of 9.23, is classified as a type IA antiarrhythmic drug that works by decreasing conduction velocity, and prolonging tissue refractoriness.3) More than 80% of orally administered procainamide is absorbed from the intestine in humans.4) The Kp (octanol/buffer at pH 7.4) value of procainamide is about 0.1, and approximately half of the dose is excreted in the urine as unchanged drug.3-6) However, the mechanisms responsible for the membrane transport of procainamide in intestinal and renal epithelial cells are still unclear.In the present study, the transport characteristics of procainamide in LLC-PK 1 cells were compared with those of quinidine. In addition, we evaluated whether the transport system for quinidine is present in another intestinal cell line, LS180, as well as Caco-2. We also investigated whether the transport system of procainamide in LS180 cells is the same as that of quinidine. Cell Culture and Preparation of Monolayers LLC-PK 1 cells at passage 197 and LS180 cells at passage 38 were obtained from the American Type Culture Collection (Manassas, VA, U.S.A.). These cells were maintained by serial passage in plastic dishes with Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (Biowest Inc., Nuaille, France) in an atmosphere of 5% CO 2 -95% air at 37°C. MATERIALS AND METHODS MaterialsLLC-PK 1 cells were seeded at a density of 5ϫ10 5 cells/ cm 2 on a 1.12 cm 2 porous membrane (3 mm pore size) in a polyester membrane Transwell ® -Clear insert (Costar, Cambridge, MA, U.S.A.) to evaluate the transcellular transport of cationic drugs. The seeded cells were maintained for 6 d to prepare differentiated cell monolayers. The maturity of the monolayer was judged by transepithelial electrical resistance (TEER). TEER was measured using a Millicell-ERS resistance system (Millipore, Bedford, MA, U.S.A.). LLC-PK 1 cell monolayers whose TEER was above 60 W · cm 2 were used to The aim of the present study was t...

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“…2,7) The cellular uptake of quinidine in Caco-2 cells grown on plastic dishes was temperature-dependent, and markedly increased by alkalization of the apical medium at 37°C.…”

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Cellular Uptake Of Quinidine and Procainamide In Intestinal mentioning

confidence: 99%

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Membrane Transport Mechanisms of Quinidine and Procainamide in Renal LLC-PK1 and Intestinal LS180 Cells

Masago

Takaai

Sakata

et al. 2010

Biological & Pharmaceutical Bulletin

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The possible clinical impact of risperidone on P‐glycoprotein‐mediated transport of tacrolimus: A case report and in vitro study

Watanabe

Higashi

Nakamura

et al. 2017

Biopharm & Drug Disp

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The authors encountered the case of an 8-fold increase in the concentration/dose (C/D) ratio of tacrolimus (TAC) following the coadministration of voriconazole (VRCZ) in a hematopoietic stem cell transplantation (HSCT) recipient. The interaction observed was much greater than expected and the patient had also been treated with oral risperidone (RSP). It was hypothesized that cytochrome P450 (CYP)3A inhibition of the small intestine by voriconazole and P-glycoprotein (P-gp) inhibition of the small intestine by risperidone exerted a synergistic effect on the bioavailability of tacrolimus. The aim of the present study was to evaluate the effect of risperidone on the P-gp-mediated transport of tacrolimus. The transcellular transport of P-gp substrates was examined in Caco-2 and P-gp-expressing renal epithelial LLC-GA5-COL150 cells. In Caco-2 cells, the apical-basal (A-B) transport of rhodamine123 (Rh123) after a 120 min incubation was increased by 47.1%, whereas that in the B-A direction was decreased by 61.7% in the presence of risperidone (100 μM). These results indicate that risperidone showed an inhibitory effect on the P-gp-mediated transport of Rh123. In LLC-GA5-COL150 cells, the A-B transport of tacrolimus after 120 min incubation was increased by 21.7% in the presence of risperidone (100 μM), whereas that in the B-A direction was decreased by 10.7%. These results suggest that risperidone was at least partly involved in the mechanism of the marked increase in the C/D ratio of tacrolimus. This case report provides new insights into the diversity of drug interactions of tacrolimus triggered by the combination of two concomitant drugs.

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Temperature‐dependent specific transport of levofloxacin in human intestinal epithelial LS180 cells

Fukumori

Masago

Ishida

et al. 2009

Biopharm & Drug Disp

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It was reported previously that specific levofloxacin uptake in Caco-2 cells was inhibited by nicotine, enalapril, L-carnitine and fexofenadine. The aim of the present study was to characterize the cellular uptake of levofloxacin using another human intestinal cell line, LS180. Levofloxacin uptake in LS180 cells was temperature-dependent and optimal at neutral pH, but was Na(+)-independent. The rank order of inhibitory effects of the four compounds on [(14)C] levofloxacin uptake in LS180 cells was nicotine>enalapril>L-carnitine>fexofenadine, which is consistent with that in Caco-2 cells. The mRNA levels of OATP1A2, 1B1, 1B3 and 2B1 in LS180 cells were markedly different from those in Caco-2 cells, and OATP substrates/inhibitors had no systematic effect on the levofloxacin uptake. The mRNA levels of OCTN1 and 2 in LS180 cells were similar to those in Caco-2 cells. However, the inhibitory effect of nicotine on L-[(3)H]carnitine uptake was much less potent than that of unlabeled L-carnitine. These results indicate that the specific uptake system for levofloxacin in LS180 cells is identical/similar to that in Caco-2 cells, but that OATPs and OCTNs contribute little to levofloxacin uptake in the human intestinal epithelial cells.

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Pharmacokinetic Analysis of Transcellular Transport of Levofloxacin across LLC-PK1 and Caco-2 Cell Monolayers

Cited by 16 publications

References 19 publications

Membrane Transport Mechanisms of Quinidine and Procainamide in Renal LLC-PK1 and Intestinal LS180 Cells

Membrane Transport Mechanisms of Quinidine and Procainamide in Renal LLC-PK1 and Intestinal LS180 Cells

The possible clinical impact of risperidone on P‐glycoprotein‐mediated transport of tacrolimus: A case report and in vitro study

Temperature‐dependent specific transport of levofloxacin in human intestinal epithelial LS180 cells

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