Rapid and simple quantitative assay method for diastereomeric flurbiprofen glucuronides in the incubation mixture

Mano, Nariyasu; Nikaido, Ayako; Narui, Takashi; Yamasaki, Daisuke; Goto, Junichi

doi:10.1016/s1570-0232(02)00032-6

Cited by 13 publications

(15 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The peak areas of (R)-and (S)-flurbiprofen glucuronide were analyzed using reverse-phase HPLC according to the reported method (Mano et al, 2002). In brief, the HPLC system consisted of LC-10AS pumps, a SCL-10A system controller, a SIL-10A autosampler, a SPD-10AV UV detector, and a C-R4AX integrator (Shimadzu, Kyoto, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…The mobile phase consisted of 20 mM ammonium acetate (pH 5.6)/ethanol/acetonitrile [20:7:2 (v/v)] and was delivered at a flow rate of 1.0 ml/min. The retention times of (R)-and (S)-glucuronide, detected at a UV wavelength of 240 nm, were approximately 15 and 16 min, respectively, which were identified by reference to the previous report (Mano et al, 2002). Because authentic standards for (R)-and (S)-glucuronide were not available, unknown concentrations of the glucuronides were determined by reference to a standard curve for racemic flurbiprofen.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Predominant Contribution of UDP-Glucuronosyltransferase 2B7 in the Glucuronidation of Racemic Flurbiprofen in the Human Liver

Mano¹,

Usui²,

Kamimura³

2007

Drug Metab Dispos

View full text Add to dashboard Cite

ABSTRACT:Flurbiprofen is a nonsteroidal anti-inflammatory drug used as a racemic mixture. Although glucuronidation is one of its elimination pathways, the role of UDP-glucuronosyltransferase (UGT) in this process remains to be investigated. Thus, the kinetics of the stereoselective glucuronidation of racemic (R,S)-flurbiprofen by recombinant UGT isozymes and human liver microsomes (HLMs) were investigated, and the major human UGT isozymes involved were identified. UGT1A1, 1A3, 1A9, 2B4, and 2B7 showed glucuronidation activity for both (R)-and (S)-glucuronide, with UGT2B7 possessing the highest activity. UGT2B7 formed the (R)-glucuronide at a rate 2.8-fold higher than that for (S)-glucuronide, whereas the other UGTs had similar formation rates. The glucuronidation of racemic flurbiprofen by HLMs also resulted in the formation of (R)-glucuronide as the dominant form, which occurred to a degree similar to that by recombinant UGT2B7 (2.1 versus 2.8). The formation of (R)-glucuronide correlated significantly with morphine 3-OH glucuronidation (r ‫؍‬ 0.96, p < 0.0001), morphine 6-OH glucuronidation (r ‫؍‬ 0.91, p < 0.0001), and 3-azido-3-deoxythymidine glucuronidation (r ‫؍‬ 0.85, p < 0.0001), a reaction catalyzed mainly by UGT2B7, in individual HLMs. In addition, the formation of both glucuronides correlated significantly (r ‫؍‬ 0.99, p < 0.0001). Mefenamic acid inhibited the formation of both (R)-and (S)-glucuronide in HLMs with similar IC 50 values (2.0 and 1.7 M, respectively), which are close to those in recombinant UGT2B7. In conclusion, these findings suggest that the formation of (R)-and (S)-glucuronide from racemic flurbiprofen is catalyzed by the same UGT isozyme, namely UGT2B7. Fig. 1) is commonly used as an analgesic nonsteroidal anti-inflammatory drug. Although the anti-inflammatory activity is almost entirely attributable to (S)-flurbiprofen (Kulmacz and Lands, 1985), it has been suggested that both (R)-and (S)-flurbiprofen may possess antinociceptive activity (Brune et al., 1991;Geisslinger and Schaible, 1996). In addition, both enantiomers have been shown to exhibit antiproliferative effects (McCracken et al., 1996). Flurbiprofen exists as a chiral compound with a stereoselective disposition in humans (Geisslinger et al., 1994) and is metabolized via several drug metabolizing pathways, including cytochromes P450 and UDP-glucuronosyltransferases (UGTs). Approximately 60 to 70% of the administered dose is eliminated as acyl glucuronides (Davies, 1995). After oral administration of racemic flurbiprofen (100 mg/kg), 8.4 and 7.3% of the dose was excreted into the urine as the acyl glucuronide of (R)-and (S)-flurbiprofen, respectively (Patel et al., 2003). However, the flurbiprofen glucuronidation fraction is difficult to determine and may be higher because of the instability of acyl glucuronides in vivo. The data from these reports suggest that the formation of glucuronide constitutes some portion of the metabolic pathway of flurbiprofen. The other major oxidative metabolite (4Ј-hydroxyflurbiprofen) and min...

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Predominant Contribution of UDP-Glucuronosyltransferase 2B7 in the Glucuronidation of Racemic Flurbiprofen in the Human Liver

Mano¹,

Usui²,

Kamimura³

2007

Drug Metab Dispos

View full text Add to dashboard Cite

show abstract

“…For the simultaneous analysis of twelve NSAIDs in human urine, Hirari et al [16] developed a solid-phase extraction (SPE) procedure in combination with a reversed-phase gradient separation method employing octadecylated silica as stationary phase and acetonitrile/phosphate buffer (pH 5) as mobile phase. Flurbiprofen glucuronides originating from incubation mixtures, which are known to be highly active due to the formation of covalently bound adducts with proteins, were analysed quantitatively on a TSK gel ODS-80Ts column employing a 20 mM ammonium acetate buffer (pH 5.6)/ethanol/acetonitrile as the mobile phase and monitoring at 246 nm [17]. Selected NSAIDs, namely racemic flurbiprofen, ibuprofen, fenprofen, tiaprofenic acid, ketoprofen, and tolmetin were resolved by Aboul-Enein on Kromasil tartardiamide-DMB chiral stationary phase [18].…”

Section: Liquid Chromatographymentioning

confidence: 99%

Sample Pretreatment and Determination of Non Steroidal Anti-Inflammatory Drugs (NSAIDs) in Pharmaceutical Formulations and Biological Samples (Blood, Plasma, Erythrocytes) by HPLC-UV-MS and μ-HPLC

Sultan¹,

Stecher²,

Stöggl³

et al. 2005

Current Medicinal Chemistry

View full text Add to dashboard Cite

The article discusses the qualitative and quantitative determination of non-steroidal antiinflammatory drugs like salicin, salicylic acid, tenoxicam, ketorolac, piroxicam, tolmetin, naproxen, flurbiprofen, diclofenac and ibuprofen by reversed phase high performance liquid chromatography (RP-HPLC) and micro-HPLC (µ-HPLC) hyphenated with UV-absorbance and mass spectrometric detection. Both detection methods delivered calibration plots with good linearity (r 2 > 0.9800), limits of detection in the low nanogram range and recovery rates between 94 and 104 %. For the analysis of biological samples such as blood, plasma and erythrocytes liquid-liquid extraction (LLE) and solid phase extraction (SPE) on the basis of new synthesized glycidylmethacrylate/divinylbenzene copolymer (GMA/DVB) particles and commercially available material on the basis of poly(divinylbenzene-co-N-vinylpyrrolidone) copolymer were investigated. Finally the use of a µ-HPLC system with separation columns in the range of 8 cm × 200 µm I.D. for the determination of non-steroidal anti-inflammatory drugs (NSAIDs) is presented, emphasizing on the type of column and sample amount needed.

show abstract

“…The measurement of the acyl glucuronides in plasma/serum has been used to establish toxicity‐exposure correlations. Numerous methods using high‐performance liquid chromatography (HPLC) have been reported3–19 for the indirect or direct measurement of acyl glucuronides.…”

mentioning

confidence: 99%

“…Mano et al . developed a rapid analytical method for the separation and determination of diastereoisomeric glucuronides of flurbiptofen in the incubation mixture of the hepatic microsomal preparation 18. More recently, Khoschsorur and Erwa reported the simultaneous determination of mycophenolic acid (MPA) and its metabolic phenolic MPA‐glucuronide and acyl glucuronide in human plasma 19…”

mentioning

confidence: 99%

Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450_BM3 for Selective Hydroxylation of Large Substrates

et al. 2010

View full text Add to dashboard Cite

Combinatorial alanine substitution of active site residues in a thermostable cytochrome P450BM3 (BM3) variant was used to generate BM3 variants with activity on large substrates. Selective hydroxylation of methoxymethylated monosaccharides, alkaloids, and steroids was thus made possible. This approach could be generally useful for improving the activity of enzymes that show only limited activity on larger substrates.

show abstract

Rapid and simple quantitative assay method for diastereomeric flurbiprofen glucuronides in the incubation mixture

Cited by 13 publications

References 15 publications

Predominant Contribution of UDP-Glucuronosyltransferase 2B7 in the Glucuronidation of Racemic Flurbiprofen in the Human Liver

Predominant Contribution of UDP-Glucuronosyltransferase 2B7 in the Glucuronidation of Racemic Flurbiprofen in the Human Liver

Sample Pretreatment and Determination of Non Steroidal Anti-Inflammatory Drugs (NSAIDs) in Pharmaceutical Formulations and Biological Samples (Blood, Plasma, Erythrocytes) by HPLC-UV-MS and μ-HPLC

Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450_BM3 for Selective Hydroxylation of Large Substrates

Contact Info

Product

Resources

About

Rapid and simple quantitative assay method for diastereomeric flurbiprofen glucuronides in the incubation mixture

Cited by 13 publications

References 15 publications

Predominant Contribution of UDP-Glucuronosyltransferase 2B7 in the Glucuronidation of Racemic Flurbiprofen in the Human Liver

Predominant Contribution of UDP-Glucuronosyltransferase 2B7 in the Glucuronidation of Racemic Flurbiprofen in the Human Liver

Sample Pretreatment and Determination of Non Steroidal Anti-Inflammatory Drugs (NSAIDs) in Pharmaceutical Formulations and Biological Samples (Blood, Plasma, Erythrocytes) by HPLC-UV-MS and μ-HPLC

Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450BM3 for Selective Hydroxylation of Large Substrates

Contact Info

Product

Resources

About

Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450_BM3 for Selective Hydroxylation of Large Substrates