Rapid LC-MS Drug Metabolite Profiling Using Microsomal Enzyme Bioreactors in a Parallel Processing Format

Bajrami, Besnik; Zhao, Ling; Schenkman, John B.; Rusling, James F.

doi:10.1021/ac902800t

Cited by 4 publications

(8 citation statements)

References 26 publications

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“…Our initial approach to the metabolism of PS was to determine its transformation by liver microsomes, which represent an enriched source of the major metabolic enzymes that, in general, render xenobiotic drugs more polar and, therefore, easier to eliminate from the body (Bajrami et al ., 2009). Such oxidative drug metabolism is the principal means of drug clearance.…”

Section: Resultsmentioning

confidence: 99%

The metabolism and pharmacokinetics of phospho‐sulindac (OXT‐328) and the effect of difluoromethylornithine

Xie

Tang

Mackenzie

et al. 2012

British J Pharmacology

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BACKGROUND AND PURPOSEPhospho-sulindac (PS; OXT-328) prevents colon cancer in mice, especially when combined with difluoromethylornithine (DFMO). Here, we explored its metabolism and pharmacokinetics. EXPERIMENTAL APPROACHPS metabolism was studied in cultured cells, liver microsomes and cytosol, intestinal microsomes and in mice. Pharmacokinetics and biodistribution of PS were studied in mice. KEY RESULTSPS undergoes reduction and oxidation yielding PS sulphide and PS sulphone; is hydrolysed releasing sulindac, which generates sulindac sulphide (SSide) and sulindac sulphone (SSone), all of which are glucuronidated. Liver and intestinal microsomes metabolized PS extensively but cultured cells converted only 10% of it to PS sulphide and PS sulphone. In mice, oral PS is rapidly absorbed, metabolized and distributed to the blood and other tissues. PS survives only partially intact in blood; of its three major metabolites (sulindac, SSide and SSone), sulindac has the highest Cmax and SSone the highest t1/2; their AUC0-24h are similar. Compared with conventional sulindac, PS generated more SSone but less SSide, which may contribute to the safety of PS. In the gastroduodenal wall of mice, 71% of PS was intact; sulindac, SSide and SSone together accounted for <30% of the total. This finding may explain the lack of gastrointestinal toxicity by PS. DFMO had no effect on PS metabolism but significantly reduced drug level in mouse plasma and other tissues.

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Section: Resultsmentioning

confidence: 99%

The metabolism and pharmacokinetics of phospho‐sulindac (OXT‐328) and the effect of difluoromethylornithine

Xie

Tang

Mackenzie

et al. 2012

British J Pharmacology

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“…LbL enzyme-DNA film fabrication on 1 μm particles was similar to that reported previously. 31,32,35 Briefly, polycation poly(diallyldimethylammoniumchloride) (PDDA), supersomes and salmon testes dsDNA were assembled in alternate successive steps on the negatively charged magnetic particle surface. 36 Steady-state adsorption times were 20 min for PDDA and DNA solutions and 30 min for supersomes while kept on ice.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

“…29,30 Then, biocolloid reactor particles coated with similar enzyme/DNA films were used to produce metabolites and DNA adducts for LC-MS/MS analysis (Figure 1B). 31,32 Both of these approaches indicate the formation of metabolites that can possibly react with DNA, and that may be linked to genotoxicity. 27,28 Both approaches use thin films of enzymes and DNA deposited on either a graphite chip for the ECL array or on 1 μm magnetic beads processed in 96-well plates for LC-MS/MS analyses.…”

Section: ■ Introductionmentioning

confidence: 99%

Genotoxicity-Related Chemistry of Human Metabolites of Benzo[ghi]perylene (B[ghi]P) Investigated using Electro-Optical Arrays and DNA/Microsome Biocolloid Reactors with LC-MS/MS

Pan

Zhao

et al. 2013

Chem. Res. Toxicol.

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There is limited and sometimes contradictory information about the genotoxicity of polycyclic aromatic hydrocarbon benzo[ghi]perylene (B[ghi]P). Using recently developed metabolic toxicity screening arrays and a biocolloid reactor-LC-MS/MS approach, both featuring films of DNA and human metabolic enzymes, we demonstrated relatively low reactivity of metabolically activated B[ghi]P towards DNA. Electro-optical toxicity screening arrays showed that B[ghi]P metabolites damage DNA at a 3-fold lower rate than benzo[a]pyrene (B[a]P), whose metabolites have a strong and well-understood propensity for DNA damage. Metabolic studies using magnetic bead biocolloid reactors coated with microsomal enzymes in 96-well plates showed that cyt P450s 1A1 and 1B1 provide high activity for B[ghi]P and B[a]P conversion. Consistent with published results, the major metabolism of B[ghi]P involved oxidations at 3,4 and 11,12 positions, leading to formation of B[ghi]P 3,4-oxide and B[ghi]P 3,4,11,12-bisoxide. B[ghi]P 3,4-oxide was synthesized and reacted with deoxyadenosine at N6 and N7 positions and with deoxyguanosine at the N2 position. B[ghi]P 3,4-oxide is hydrolytically unstable and transforms into the 3,4-diol or converts to 3- or 4-hydroxy B[ghi]P. LC-MS/MS of reaction products from the magnetic biocolloid reactor particles coated with DNA and human enzymes revealed for the first time that a major DNA adduct results from reaction between B[ghi]P 3,4,11,12-bisoxide and deoxyguanosine. Results also demonstrated 5-fold lower formation rates of the major DNA adduct for B[ghi]P metabolites compared to B[a]P. Overall, results from both ECL array and biocolloid reactor-LC-MS/MS consistently suggest a lower human genotoxicity profile of B[ghi]P than B[a]P.

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“…Given the efficacy of PI, we explored its metabolism by rat liver microsomes (RLM), an enriched source of major metabolic enzymes (Bajrami et al, 2009). These enzymes, including oxidative enzymes, hydrolases, and transferases, function to make xenobiotic compounds more polar and more water-soluble, thus facilitating their elimination from the body.…”

Section: Pi Inhibits the Growth Of Sw480 Colon Cancer Cellsmentioning

confidence: 99%

Phospho-Ibuprofen (MDC-917) Is a Novel Agent against Colon Cancer: Efficacy, Metabolism, and Pharmacokinetics in Mouse Models

Xie¹,

Sun²,

Tang³

et al. 2011

J Pharmacol Exp Ther

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We have developed a novel chemical modification of conventional nonsteroidal anti-inflammatory drugs to reduce their toxicity and enhance their efficacy. Phospho-ibuprofen [(PI) 2-(4-isobutyl-phenyl)-propionic acid-4-(diethoxy-phosphoryloxy)-butyl ester (MDC-917)], a novel derivative of ibuprofen, strongly inhibited the growth of human colon cancer cells in vitro and SW480 human colon cancer xenografts in nude mice. PI was metabolized minimally by cultured cells, but extensively by liver microsomes and mice, undergoing regioselective oxidation to produce 1-OH-PI and carboxyl-PI, which can be hydrolyzed to 1-OH-ibuprofen and carboxyl-ibuprofen, respectively. PI also can be hydrolyzed to release ibuprofen, which can generate 2-OH-ibuprofen, carboxyl-ibuprofen, and ibuprofen glucuronide. After a single oral administration (400 mg/kg) of PI, ibuprofen and ibuprofen glucuronide are the main plasma metabolites of PI; they have, respectively, C max of 530 and 215 M, T max of 1 and 2 h, elimination t 1/2 of 7.7 and 5.3 h, and area under the concentration-time curve (0 -24 h) of 1816 and 832 M ϫ h. Intact PI was detected in several tissues but not in plasma; at a higher PI dose (1200 mg/kg), PI plasma levels were 12.4 M. PI generated the same metabolites in mouse plasma as conventional ibuprofen, but with much lower levels, perhaps accounting for the enhanced safety of PI. The antitumor effect of PI was significantly associated with plasma ibuprofen levels (p ϭ 0.016) but not with xenograft ibuprofen levels (p ϭ 0.08), suggesting a complex anticancer effect. These results provide a pharmacological basis to explain, at least in part, the anticancer efficacy and safety of this promising compound and indicate that PI merits further evaluation as an anticancer agent.

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Rapid LC-MS Drug Metabolite Profiling Using Microsomal Enzyme Bioreactors in a Parallel Processing Format

Cited by 4 publications

References 26 publications

The metabolism and pharmacokinetics of phospho‐sulindac (OXT‐328) and the effect of difluoromethylornithine

The metabolism and pharmacokinetics of phospho‐sulindac (OXT‐328) and the effect of difluoromethylornithine

Genotoxicity-Related Chemistry of Human Metabolites of Benzo[ghi]perylene (B[ghi]P) Investigated using Electro-Optical Arrays and DNA/Microsome Biocolloid Reactors with LC-MS/MS

Phospho-Ibuprofen (MDC-917) Is a Novel Agent against Colon Cancer: Efficacy, Metabolism, and Pharmacokinetics in Mouse Models

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