Influence of Efflux Transporters on the Accumulation and Efflux of Four Quinolones (Ciprofloxacin, Levofloxacin, Garenoxacin, and Moxifloxacin) in J774 Macrophages

Abstract

Ciprofloxacin is subject to efflux from J774 macrophages through a multidrug resistance-related protein-like transporter (J. M. Michot, F. Van Bambeke, M. P. Mingeot-Leclercq, and P. M. Tulkens, Antimicrob. Agents Chemother. 48: [2673][2674][2675][2676][2677][2678][2679][2680][2681][2682] 2004). Here, we compare ciprofloxacin to levofloxacin, garenoxacin, and moxifloxacin for transport. At 4 mg/liter, an apparent steady state in accumulation was reached after 30 to 60 min for all quinolones but to quite different levels (approximately 3, 5, 10, and 16 fold). Accumulation of ciprofloxacin was increased (to about 16 to 20 fold) by ATP depletion, increase in extracellular concentration, and the addition of probenecid, gemfibrozil, or MK571 (but not verapamil or GF120918). These treatments did not affect the accumulation of moxifloxacin. Levofloxacin and garenoxacin showed an intermediate behavior. Efflux of ciprofloxacin was slowed down by probenecid (half-life, 7.2 versus 1.6 min). Moxifloxacin efflux was faster and unaffected by probenecid (half-lifes, 0.27 versus 0.33 min). Efflux of levofloxacin and garenoxacin was modestly decreased by probenecid (1.5 and 2.1 fold). Accumulation of 14 C-labeled ciprofloxacin was increased by unlabeled ciprofloxacin and moxifloxacin, but moxifloxacin was two times less potent. Accumulation of moxifloxacin at 4°C was almost identical to that at 37°C, whereas that of ciprofloxacin was minimal (levofloxacin and garenoxacin showed intermediate behaviors). Cells subjected to thermal shock (56°C; 10 min) accumulated all quinolones at a similar level (16 to 23 fold). We conclude that moxifloxacin is apparently not subject to efflux from J774 macrophages, even though it can interact with the ciprofloxacin transporter. Levofloxacin and garenoxacin are partially effluxed. Data suggest that efflux plays an important role in the differential accumulation of quinolones by J774 macrophages.Fluoroquinolones have long been known to accumulate in phagocytic cells (8), but quite significant differences among closely related derivatives have been observed (3, 7, 13) which have so far not received satisfactory explanation. One factor that can modulate antibiotic accumulation in eucaryotic cells is their differential recognition by active efflux transporters (see reference 28 and the references cited therein). Fluoroquinolones are recognized by several eucaryotic multidrug transporters, most notably by two main members of the ATP-binding cassette superfamily, namely the multidrug resistance-related proteins (MRP) and the P-glycoprotein (28). In J774 macrophages, norfloxacin has been shown to be subject to efflux by a probenecid-and gemfibrozil-inhibitable transporter (2), which has been tentatively identified as a member of the MRP family (16). In this context, we have now examined the accumulation and efflux of levofloxacin and moxifloxacin in J774 macrophages in comparison with ciprofloxacin. These quinolones were chosen on the basis of their increasingly lipophilic character and potential clinic...

Contrasting Effects of Acidic pH on the Extracellular and Intracellular Activities of the Anti-Gram-Positive Fluoroquinolones Moxifloxacin and Delafloxacin againstStaphylococcus aureus

“…This suggests a capacity to rapidly redistribute inside the cell to reach their target, which is consistent with their capacity to easily diffuse through membranes (29) and to their strong binding to the DNA-topoisomerase complex in bacteria (45). In infected cells incubated in neutral medium, delafloxacin shows a very low static concentration and reaches a nearbactericidal effect, making it one of the most active drugs against intracellular S. aureus in this model.…”

“…Fractionation studies were performed with J774 murine macrophages. Both cell lines were maintained in our laboratory as previously described, using RPMI medium supplemented with 10% fetal calf serum (15,29).…”

Contribution of Moxifloxacin or Levofloxacin in Second-Line Regimens with or without Continuation of Pyrazinamide in Murine Tuberculosis

“…Furthermore, Malik and colleagues found that M may be unique among marketed fluoroquinolones in its ability to kill M. tuberculosis in the presence of the protein synthesis inhibitor chloramphenicol (33,34). Improved drug delivery to the site of infection may be an additional reason for the greater potency of M. M concentrates inside macrophages and neutrophils to a greater extent than L (35,36). This may lead to higher exposures for M relative to L in our murine model in which M. tuberculosis resides almost entirely intracellularly.…”

Pharmacodynamic Evaluation of the Intracellular Activities of Antibiotics against Staphylococcus aureus in a Model of THP-1 Macrophages

“…For each of these antibiotics, the assay method was checked for linearity (telithromycin, 0.03 to 4 mg/liter; azithromycin, 0.05 to 4 mg/liter; ampicillin, 0.21 to 10 mg/liter, nafcillin, 1.1 to 120 mg/liter; oxacillin, 1.6 to 30 mg/liter; penicillin V, 0.04 to 10 mg/liter; rifampin, 0.2 to 15 mg/liter; linezolid, 12 to 190 mg/liter, vancomycin, 2.7 to 150 mg/liter; teicoplanin, 2.9 to 250 mg/liter; gentamicin, 0.6 to 240 mg/liter) and for reproducibility (coefficient of variation, Ͻ10%). Ciprofloxacin, moxifloxacin, and levofloxacin concentrations were measured by fluorimetry (12,56), and garenoxacin and oritavancin concentrations were measured by radiometry by using 14 C-labeled drugs (41,64). In pilot studies we checked that these assays detected genuine, bioactive drug.…”