Intracellular accumulation and activity of ampicillin used as freedrug and as its phthalimidomethyl or pivaloyloxymethyl ester (pivampicillin) against Listeria monocytogenes in J774 macrophages

Chanteux, Hugues

doi:10.1093/jac/dkg431

Cited by 7 publications

(6 citation statements)

References 24 publications

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“…While in some cases prodrugs offered better intestinal permeability and increased exposure after oral (Huryn et al, 2004a,b;Dyatkin et al, 2005;Venkatraman et al, 2009;Reger et al, 2010), some other attempts remained unfruitful or of limited benefit (Chiba et al, 2006a,b;Gong et al, 2006Gong et al, , 2008. As demonstrated with other ester prodrugs (Chanteux et al, 2003), the present data showed that CDP323 prodrug increased mass transfer across biologic membranes compared with the carboxylic acid derivative CT7758. In addition, CDP323 also efficiently released in vitro (Chanteux et al, 2014) and in vivo its active moiety, CT7758, as described earlier.…”

Section: Discussionsupporting

confidence: 45%

Cross-Species Differences in the Preclinical Pharmacokinetics of CT7758, an α4β1/α4β7 Integrin Antagonist

et al. 2015

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CT7758, a carboxylate containing a4b1/a4/b7 integrin antagonist, was characterized for its pharmacokinetic profile in various in vitro and in vivo assays in support of clinical development. The oral bioavailability of CT7758 was 4% in mice, 2% in rats, 7-55% in dogs, and 0.2% in cynomolgus monkeys. The low bioavailability in rodents and monkey results from low intestinal absorption as evidenced by a low fraction absorbed in the rat portal vein model (3%), low-tomedium permeability in Caco-2 cells (£1.3 3 10 26 cm/s) with evidences of polarized efflux, and high polar surface area (104 Å). In rodents and cynomolgus monkeys, the total plasma clearance was moderate to high ( ‡50% hepatic blood flow Q H ) and associated with a short elimination half-life (£1 hour). This contrast with the dog data which showed a much lower clearance (6% Q H ) and a longer t 1/2 (2.4 hours). The volume of distribution (V z ) also varied significantly across species with value of 5.5, 2.8, 0.24, and 0.93 l/kg in mouse, rat, dog, and cynomolgus monkey, respectively. In vitro assays demonstrated that active hepatic uptake accounted for most of the in vivo clearance and was the source of the large species variability. In vitro uptake assays predicted a total plasma clearance in humans in the low range (33% Q H ), a finding subsequently confirmed in the clinic. Assays in OAPT1B1-transfected cells demonstrated active uptake transport through this transporter. The prospect of limited absorption in human prompted the synthesis an ethyl ester prodrug, CDP323, which demonstrated higher in vitro permeability, increased oral bioavailability, as well as efficient in vivo release of its active moiety CT7758.

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Section: Discussionsupporting

confidence: 45%

Cross-Species Differences in the Preclinical Pharmacokinetics of CT7758, an α4β1/α4β7 Integrin Antagonist

et al. 2015

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“…For the cell infection studies, we used the general procedures described earlier for J774 cells with S. aureus ATCC 25923 (24-h model [31]) or L. monocytogenes hemolysin-producing strain EGD serotype 1/2a (5-h model [7]). For S. aureus, gentamicin (0.5 mg/ liter) was used to prevent extracellular growth in the control cultures.…”

Section: Cell Infection Studies (S Aureus and L Monocytogenes)mentioning

confidence: 99%

“…The persistence of oritavancin in the organism is believed to result from this high level of protein binding, analogous to the protein binding of teicoplanin (21), but it may also suggest the existence of deep storage compartments for the drug in the body. This triggered us to study the cellular pharmacokinetics and disposition of oritavancin in cultured cells by the approaches used previously to study the intracellular fates of other cationic antibiotics such as aminoglycosides (40), macrolides (4), and basic derivatives of ␤-lactam antibiotics (7,27). The mechanism of accumulation was studied by using as comparators chloroquine and azithromycin, which accumulate in lysosomes by diffusion and segregation of their protonated forms (4,12,45), and with horseradish peroxidase (HRP) and small latex beads as markers of endocytosis (26,41).…”

mentioning

confidence: 99%

“…The mechanism of accumulation was studied by using as comparators chloroquine and azithromycin, which accumulate in lysosomes by diffusion and segregation of their protonated forms (4,12,45), and with horseradish peroxidase (HRP) and small latex beads as markers of endocytosis (26,41). Concentrating on macrophages and using previously established models (7,31), we have also assessed the intracellular activities of oritavancin and vancomycin against phagolysosomal (Staphylococcus aureus) and cytosolic (Listeria monocytogenes) organisms using previously established models in an attempt to correlate accumulation, disposition, and expression of activity.…”

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confidence: 99%

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Cellular Pharmacokinetics and Pharmacodynamics of the Glycopeptide Antibiotic Oritavancin (LY333328) in a Model of J774 Mouse Macrophages

Bambeke

Carryn

Seral

et al. 2004

Antimicrob Agents Chemother

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The intracellular pharmacokinetics and pharmacodynamics of oritavancin (LY333328) were studied in cultured cells. Oritavancin was avidly accumulated by J774 and THP-1 macrophages and rat fibroblasts and to a lesser extent by LLC-PK1 and Caco-2 cells. In J774 macrophages, the level of accumulation reached a plateau (at 370-fold the extracellular concentration) within 24 h and was partly defeated by a rise in serum protein levels. Efflux was incomplete (with a plateau at two-thirds of the original level at 6 h). In short-term kinetic studies, oritavancin uptake was linear for up to 4 h (as was the case for horseradish peroxidase and small latex beads, used as markers of the fluid phase and adsorptive endocytosis, respectively), which was in contrast to azithromycin and chloroquine uptake (which accumulate in cells by diffusion and segregation). The rates of clearance of oritavancin and latex beads were comparable (150 and 120 l ؋ mg of protein ؊1 ؋ h ؊1 , respectively) and were approximately 200 times higher than that of horseradish peroxidase. Oritavancin accumulation was partially reduced by monensin but was unaffected by acidic pH (these conditions abolished chloroquine accumulation). Cell-associated oritavancin was found in lysosomal fractions after homogenization of J774 macrophages and fractionation by isopycnic centrifugation. Oritavancin was bactericidal against intracellular Staphylococcus aureus (phagolysosomal infection) but was unable to control the intracellular growth of Listeria monocytogenes (cytosolic infection), even though its cellular concentration largely exceeded the MIC (0.02 mg/liter) and minimal bactericidal concentration (2 mg/liter). We conclude that oritavancin enters cells by adsorptive endocytosis (favored by its lipophilic side chain and/or the presence of three protonatable amines), which drives it to lysosomes, where it exerts antibiotic activity.

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“…When this property was specifically looked for in cultured macrophages, however, we observed that PIVA was merely bound to the pericellular membrane, with no evidence of true intracellular penetration and lysosomal accumulation (11). Third, studies with Listeria-infected macrophages showed that macrophages incubated with low concentrations of PIVA accumulated large amounts of free ampicillin (which was active against this intracellular organism), whereas no ampicillin (and no antibacterial activity) could be seen when cells were incubated with similar concentrations of ampicillin (10).…”

mentioning

confidence: 99%

Accumulation and Oriented Transport of Ampicillin in Caco-2 Cells from Its Pivaloyloxymethylester Prodrug, Pivampicillin

Chanteux

Bambeke

Mingeot‐Leclercq

et al. 2005

Antimicrob Agents Chemother

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Pivampicillin (PIVA), an acyloxymethylester of ampicillin, is thought to enhance the oral bioavailability of ampicillin because of its greater lipophilicity compared to that of ampicillin. The fate of PIVA in intestinal cells and the exact location of its conversion into ampicillin have, however, never been unambiguously established. Polarized Caco-2 cells have been used to examine the handling of PIVA and the release of ampicillin from PIVA by the intestinal epithelium. Experiments were limited to 3 h. Cells incubated with PIVA (apical pole) showed a fast accumulation of ampicillin and transport toward the basolateral medium, whereas PIVA itself was only poorly accumulated and transported. Cells incubated with free ampicillin accumulated and transported only minimal amounts of this drug. Release of ampicillin from cells incubated with PIVA was unaffected by PEPT1 and OCTN2 inhibitors but was sharply decreased after ATP depletion or addition of bis(4-nitrophenyl)-phosphate (BNPP; an esterase inhibitor). PIVA incubated with Caco-2 lysates released free ampicillin, and this release was inhibited by BNPP. Efflux studies showed that the ampicillin that accumulated in cells after incubation with PIVA was preferentially transported out of the cells through the basolateral pole. This efflux was decreased by multidrug resistance-associated protein (MRP) inhibitors (probenecid, MK-571) and by ATP depletion. A phthalimidomethylester of ampicillin that resists cellular esterases failed to cause any significant release (cell lysate) or transport (polarized Caco-2 cells) of ampicillin. These results show that when PIVA is given to Caco-2 cells from their apical pole, ampicillin is released intracellularly and that ampicillin is thereafter preferentially effluxed into the basolateral medium through an MRP-like transporter.

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Intracellular accumulation and activity of ampicillin used as freedrug and as its phthalimidomethyl or pivaloyloxymethyl ester (pivampicillin) against Listeria monocytogenes in J774 macrophages

Abstract: This is the first demonstration that PIVA (a prodrug of ampicillin) can be used to promote ampicillin cellular accumulation and, thereby to increase ampicillin intracellular activity. PIVA could be useful for control of the intracellular multiplication of L. monocytogenes.

Cited by 7 publications

References 24 publications

Cross-Species Differences in the Preclinical Pharmacokinetics of CT7758, an α4β1/α4β7 Integrin Antagonist

Cross-Species Differences in the Preclinical Pharmacokinetics of CT7758, an α4β1/α4β7 Integrin Antagonist

Cellular Pharmacokinetics and Pharmacodynamics of the Glycopeptide Antibiotic Oritavancin (LY333328) in a Model of J774 Mouse Macrophages

Accumulation and Oriented Transport of Ampicillin in Caco-2 Cells from Its Pivaloyloxymethylester Prodrug, Pivampicillin

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