6,563 results on '"Glucuronide"'
Search Results
2. Preparation of glucuronides using liver microsomes and their characterization by 1D/2D NMR spectroscopy and mass spectrometry: Application to fentanyl metabolites.
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Kanamori, Tatsuyuki, Okada, Yuki, Segawa, Hiroki, Yamamuro, Tadashi, Kuwayama, Kenji, Tsujikawa, Kenji, and Iwata, Yuko T.
- Abstract
A simple, low‐cost method for preparing glucuronic acid‐conjugated metabolites was developed using fentanyl, a potent synthetic opioid, as a model drug. Five glucuronic acid‐conjugated metabolites of fentanyl were measured in the culture medium of fresh human hepatocytes incubated with fentanyl. These glucuronides were also formed by incubation of their corresponding substrates (e.g., 4′‐hydroxy‐fentanyl and β‐hydroxy‐fentanyl) with uridine 5′‐diphosphoglucuronic acid and human liver microsomes (HLM). Experiments using liver microsomes of several animals revealed that significant species differences exist in the glucuronide formation patterns; fentanyl glucuronide was only formed in HLM, and 4′‐hydroxy‐fentanyl glucuronide was formed much more in rat liver microsomes (RLM) than HLM and dog liver microsomes. Furthermore, surprisingly, HLM and RLM showed opposite substrate selectivity for the enantiomers of β‐hydroxy‐fentanyl. Submilligram amounts of three of these metabolites, namely, 4′‐hydroxy‐fentanyl glucuronide and two glucuronides of β‐hydroxy‐fentanyl, were prepared by using HLM or RLM. The products were readily purified with a reversed‐phase/anion‐exchange mixed‐mode solid‐phase extraction cartridge, and then, their chemical structures were confirmed by 1D/2D nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry data. In addition, the products were quantitated by quantitative NMR, and the yields were 3.6–69%. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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- View/download PDF
3. The Active Glucuronide Metabolite of the Brain Protectant IMM-H004 with Poor Blood–Brain Barrier Permeability Demonstrates a High Partition in the Rat Brain via Multiple Mechanisms.
- Author
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Jiang, Jianwei, Luo, Lijun, Zhang, Ziqian, Liu, Xiao, Chen, Naihong, Li, Yan, and Sheng, Li
- Subjects
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BLOOD-brain barrier , *ACTIVE biological transport , *RATS , *PERMEABILITY , *ISCHEMIC stroke ,BRAIN metabolism - Abstract
Background: Glucuronidation is an essential metabolic pathway for a variety of drugs. IMM-H004 is a novel neuroprotective agent against ischemic stroke, and its glucuronide metabolite IMM-H004G exhibits similar pharmacological activity. Despite possessing a higher molecular weight and polarity, brain exposure of IMM-H004G is much higher than that of IMM-H004. This study aimed to investigate the brain metabolism and transport mechanisms of IMM-H004 and IMM-H004G. Methods: First, the possibility of IMM-H004 glucuronidation in the brain was evaluated in several human brain cell lines and rat homogenate. Subsequently, the blood–brain barrier carrier-mediated transport mechanism of IMM-H004 and IMM-H004G was studied using overexpression cell models. In addition, intracerebroventricular injection, in situ brain perfusion model, and microdialysis/microinjection techniques were performed to study the distribution profiles of IMM-H004 and IMM-H004G. Results: IMM-H004 could be metabolized to IMM-H004G in both rat brain and HEB cells mediated by UGT1A7. However, IMM-H004G could not be hydrolyzed back into IMM-H004. Furthermore, the entry and efflux of IMM-H004 in the brain were mediated by the pyrilamine-sensitive H+/OC antiporter and P-gp, respectively, while the transport of IMM-H004G from the blood to the brain was facilitated by OATP1A2 and OATP2B1. Ultimately, stronger concentration gradients and OATP-mediated uptake played a critical role in promoting greater brain exposure of IMM-H004G. Conclusions: The active glucuronide metabolite of the brain protectant IMM-H004 with poor blood–brain barrier permeability demonstrates a high partition in the rat brain via multiple mechanisms, and our findings deepen the understanding of the mechanisms underlying the blood–brain barrier metabolism and transport of active glucuronide conjugates. [ABSTRACT FROM AUTHOR]
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- 2024
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4. A critical examination of human data for the biological activity of quercetin and its phase-2 conjugates.
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Williamson, Gary and Clifford, Michael N.
- Abstract
AbstractThis critical review examines evidence for beneficial effects of quercetin phase-2 conjugates from clinical intervention studies, volunteer feeding trials, and in vitro work. Plasma concentrations of quercetin-3-
O -glucuronide (Q3G) and 3′-methylquercetin-3-O- glucuronide (3′MQ3G) after supplementation may produce beneficial effects in macrophages and endothelial cells, respectively, especially if endogenous deglucuronidation occurs, and lower blood uric acid concentration via quercetin-3′-O -sulfate (Q3′S). Unsupplemented diets produce much lower concentrations (<50 nmol/l) rarely investigated in vitro. At 10 nmol/l, Q3′S and Q3G stimulate or suppress, respectively, angiogenesis in endothelial cells. Statistically significant effects have been reported at 100 nmol/l in breast cancer cells (Q3G), primary neuron cultures (Q3G), lymphocytes (Q3G and3′MQ3G) and HUVECs (QG/QS mixture), but it is unclear whether these translate to a health benefit in vivo. More sensitive and more precise methods to measure clinically significant endpoints are required before a conclusion can be drawn regarding effects at normal dietary concentrations. Future requirements include better understanding of inter-individual and temporal variation in plasma quercetin phase-2 conjugates, their mechanisms of action including deglucuronidation and desulfation both in vitro and in vivo, tissue accumulation and washout, as well as potential for synergy or antagonism with other quercetin metabolites and metabolites of other dietary phytochemicals. [ABSTRACT FROM AUTHOR]- Published
- 2023
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5. The Active Glucuronide Metabolite of the Brain Protectant IMM-H004 with Poor Blood–Brain Barrier Permeability Demonstrates a High Partition in the Rat Brain via Multiple Mechanisms
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Jianwei Jiang, Lijun Luo, Ziqian Zhang, Xiao Liu, Naihong Chen, Yan Li, and Li Sheng
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blood–brain barrier ,glucuronide ,conjugate metabolite ,uridine diphosphate-glucuronosyltransferase ,organic anion-transporting polypeptide ,Pharmacy and materia medica ,RS1-441 - Abstract
Background: Glucuronidation is an essential metabolic pathway for a variety of drugs. IMM-H004 is a novel neuroprotective agent against ischemic stroke, and its glucuronide metabolite IMM-H004G exhibits similar pharmacological activity. Despite possessing a higher molecular weight and polarity, brain exposure of IMM-H004G is much higher than that of IMM-H004. This study aimed to investigate the brain metabolism and transport mechanisms of IMM-H004 and IMM-H004G. Methods: First, the possibility of IMM-H004 glucuronidation in the brain was evaluated in several human brain cell lines and rat homogenate. Subsequently, the blood–brain barrier carrier-mediated transport mechanism of IMM-H004 and IMM-H004G was studied using overexpression cell models. In addition, intracerebroventricular injection, in situ brain perfusion model, and microdialysis/microinjection techniques were performed to study the distribution profiles of IMM-H004 and IMM-H004G. Results: IMM-H004 could be metabolized to IMM-H004G in both rat brain and HEB cells mediated by UGT1A7. However, IMM-H004G could not be hydrolyzed back into IMM-H004. Furthermore, the entry and efflux of IMM-H004 in the brain were mediated by the pyrilamine-sensitive H+/OC antiporter and P-gp, respectively, while the transport of IMM-H004G from the blood to the brain was facilitated by OATP1A2 and OATP2B1. Ultimately, stronger concentration gradients and OATP-mediated uptake played a critical role in promoting greater brain exposure of IMM-H004G. Conclusions: The active glucuronide metabolite of the brain protectant IMM-H004 with poor blood–brain barrier permeability demonstrates a high partition in the rat brain via multiple mechanisms, and our findings deepen the understanding of the mechanisms underlying the blood–brain barrier metabolism and transport of active glucuronide conjugates.
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- 2024
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6. Enzymatic Synthesis of α-Glucosyl-Baicalin through Transglucosylation via Cyclodextrin Glucanotransferase in Water.
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Lambert, Carole, Lemagnen, Perrine, Don Simoni, Eglantine, Hubert, Jane, Kotland, Alexis, Paulus, Chantal, De Bizemont, Audrey, Bernard, Sylvie, Humeau, Anne, Auriol, Daniel, and Reynaud, Romain
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CYCLODEXTRINS , *PARTITION chromatography , *GLUCOAMYLASE , *MOIETIES (Chemistry) , *GLUCOSIDASES , *BACILLUS (Bacteria) , *GLUCOSIDES - Abstract
Baicalin is a biologically active flavone glucuronide with poor water solubility that can be enhanced via glucosylation. In this study, the transglucosylation of baicalin was successfully achieved with CGTases from Thermoanaerobacter sp. and Bacillus macerans using α-cyclodextrin as a glucosyl donor. The synthesis of baicalin glucosides was optimized with CGTase from Thermoanaerobacter sp. Enzymatically modified baicalin derivatives were α-glucosylated with 1 to 17 glucose moieties. The two main glucosides were identified as Baicalein-7-O-α-D-Glucuronidyl-(1→4′)-O-α-D-Glucopyranoside (BG1) and Baicalein-7-O-α-D-Glucuronidyl-(1→4′)-O-α-D-Maltoside (BG2), thereby confirming recent findings reporting that glucuronyl groups are acceptors of this CGTase. Optimized conditions allowed for the attainment of yields above 85% (with a total glucoside content higher than 30 mM). BG1 and BG2 were purified via centrifugal partition chromatography after an enrichment through deglucosylation with amyloglucosidase. Transglucosylation increased the water solubility of BG1 by a factor of 188 in comparison to that of baicalin (molar concentrations), while the same value for BG2 was increased by a factor of 320. Finally, BG1 and BG2 were evaluated using antioxidant and anti-glycation assays. Both glucosides presented antioxidant and anti-glycation properties in the same order of magnitude as that of baicalin, thereby indicating their potential biological activity. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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7. Investigation of buprenorphine‐related deaths using urinary metabolite concentrations.
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Mariottini, Claudia, Kriikku, Pirkko, and Ojanperä, Ilkka
- Abstract
Quantitative analysis of postmortem urine, instead of blood, for buprenorphine and metabolites may provide additional evidence for the diagnosis of fatal buprenorphine poisoning. In this study, 247 autopsy urine samples, previously testing positive for buprenorphine or norbuprenorphine, were quantitatively reanalysed with a recently developed liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method for unconjugated buprenorphine (BUP), norbuprenorphine (NBUP), naloxone (NAL), and their respective conjugated metabolites, buprenorphine glucuronide (BUPG), norbuprenorphine glucuronide (NBUPG), and naloxone glucuronide (NALG). The cases were divided, according to medical examiners' decision, to buprenorphine poisonings and other causes of death. The groups were compared for urinary concentrations and metabolite concentration ratios of the six analytes. All median concentrations were higher in the buprenorphine poisoning group. The median concentration of BUPG was significantly higher and the median metabolite ratios NBUP/BUP, NBUPG/BUPG, and NBUPtotal/BUPtotal were significantly lower in poisonings than in other causes of death. Naloxone‐related concentrations and ratios were not significantly different between the groups. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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8. Enzymatic Synthesis of α-Glucosyl-Baicalin through Transglucosylation via Cyclodextrin Glucanotransferase in Water
- Author
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Carole Lambert, Perrine Lemagnen, Eglantine Don Simoni, Jane Hubert, Alexis Kotland, Chantal Paulus, Audrey De Bizemont, Sylvie Bernard, Anne Humeau, Daniel Auriol, and Romain Reynaud
- Subjects
baicalin ,polyphenol ,glucuronide ,cyclodextrin glucanotransferase ,α-cyclodextrin ,enzymatic glucosylation ,Organic chemistry ,QD241-441 - Abstract
Baicalin is a biologically active flavone glucuronide with poor water solubility that can be enhanced via glucosylation. In this study, the transglucosylation of baicalin was successfully achieved with CGTases from Thermoanaerobacter sp. and Bacillus macerans using α-cyclodextrin as a glucosyl donor. The synthesis of baicalin glucosides was optimized with CGTase from Thermoanaerobacter sp. Enzymatically modified baicalin derivatives were α-glucosylated with 1 to 17 glucose moieties. The two main glucosides were identified as Baicalein-7-O-α-D-Glucuronidyl-(1→4′)-O-α-D-Glucopyranoside (BG1) and Baicalein-7-O-α-D-Glucuronidyl-(1→4′)-O-α-D-Maltoside (BG2), thereby confirming recent findings reporting that glucuronyl groups are acceptors of this CGTase. Optimized conditions allowed for the attainment of yields above 85% (with a total glucoside content higher than 30 mM). BG1 and BG2 were purified via centrifugal partition chromatography after an enrichment through deglucosylation with amyloglucosidase. Transglucosylation increased the water solubility of BG1 by a factor of 188 in comparison to that of baicalin (molar concentrations), while the same value for BG2 was increased by a factor of 320. Finally, BG1 and BG2 were evaluated using antioxidant and anti-glycation assays. Both glucosides presented antioxidant and anti-glycation properties in the same order of magnitude as that of baicalin, thereby indicating their potential biological activity.
- Published
- 2023
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9. Stereoselective Covalent Adduct Formation of Acyl Glucuronide Metabolite of Nonsteroidal Anti-Inflammatory Drugs with UDP-Glucuronosyltransferase.
- Author
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Kawase, Atsushi, Yamashita, Rio, Yoshizato, Tsubasa, Yoshikawa, Mashiro, Shimada, Hiroaki, and Iwaki, Masahiro
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ANTI-inflammatory agents , *GLUCURONOSYLTRANSFERASE , *DNA adducts , *PROPIONIC acid , *CARRIER proteins , *ACID derivatives - Abstract
A reactive metabolite of nonsteroidal anti-inflammatory drugs (NSAIDs), acyl-β-D-glucuronide (AG), covalently binds to endogenous proteins. The covalent adduct formation of NSAIDs-AG may lead to the dysfunction of target proteins. Therefore, it is important to clarify the detailed characterization of the formation of covalent protein adducts of NSAID-AG. UDP-glucuronosyltransferase (UGT) catalyzes the conversion of NSAIDs to NSAIDs-AG. The aim of this study was to perform a quantitative analysis of the covalent adduct formation of NSAIDs-AG with UGT. Diclofenac-AG and ketoprofen-AG formed covalent adducts with organelle proteins. Next, the number of covalent adducts formed between NSAIDs-AG and UGT isoforms (UGT1A1, UGT1A9, UGT2B4, and UGT2B9) was determined. The capacity of diclofenac-AG to form covalent adducts with UGT1A9 or UGT2B7 was approximately 10 times higher than that of mefenamic acid-AG. The amounts of covalent adducts of AG of propionic acid derivative NSAIDs with UGT2B were higher than those with UGT1A. Stereoselectivity was observed upon covalent binding to UGT. A significant negative correlation between the half-lives of NSAIDs-AG in phosphate buffers and the amount of covalent adduct with UGT2B7 was observed, suggesting the more labile NSAID-AG forms higher irreversible bindings to UGT. This report provides comprehensive information on the covalent adduct formation of NSAIDs-AGs with UGT. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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10. Stability of UV filter 2-ethylhexyl salicylate and its glucuronide in human urine – A challenge for robust human biomonitoring.
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Kuhlmann, Laura, Hiller, Julia, and Göen, Thomas
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SALICYLATES , *BIOLOGICAL monitoring , *URINE , *HYGIENE products , *GLUCURONIC acid , *QUALITY control - Abstract
[Display omitted] • Unconjugated EHS is significantly less stable in urine compared to glucuronidated EHS. • Lower stability of unconjugated EHS occurred at both 8 °C and −20 °C. • The hydroxylated metabolites of EHS showed no relevant stability issues. • Only small ratio of unconjugated EHS found in urine samples after dermal exposure. • Low stability of unconjugated EHS shall be considered for calibration and quality control samples. 2-Ethylhexyl salicylate (EHS) is used as a UV filter in personal care products, especially sunscreens. The elimination of EHS and its metabolite 2-ethyl-5-hydroxyhexyl salicylate (5OH–EHS) after dermal exposure has been previously investigated and showed that both are excreted in high amounts. However, the low stability of the reference substance for EHS highly affected the analysis of this parameter. Since EHS was assumed to be excreted primarily as a conjugate to glucuronic acid in vivo , we compared the stability of unconjugated EHS to that of glucuronidated EHS (EHS-GlcA) in urine at 8 °C and —20 °C. The relative recovery of unconjugated EHS decreases significantly when stored at both 8 °C and –20 °C. Stored as EHS-GlcA, the relative recovery remains steady over the course of the experiment. In addition to the stability experiment, the fraction of unconjugated EHS in human urine samples directly after dermal exposure to EHS was assessed, and only a low effective portion of unconjugated EHS was found. The same samples were reassessed after several months of storage at —20 °C and showed relative recoveries within the acceptance criteria, which also endorses the low initial level of unconjugated EHS. The hydroxylated EHS metabolites, 5OH-EHS, 4OH-EHS and 2OH-EHS, were also included in the experiment and showed slightly decreasing relative recoveries at 8 °C, but stable relative recoveries at –20 °C. Despite unconjugated EHS being excreted only in a low portion in vivo , its low stability in urine may affect the effectiveness of EHS biomonitoring, because only the use of EHS glucuronide guarantees a stable and reliable calibration in urine. However, the hydroxylated EHS metabolites have proven to be suitable standards for calibration as well as parameters for biomonitoring when stored at —20 °C. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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11. Effect of glucuronides on metabolic enzymes and active hepatic uptake : in vitro assessment and prediction of drug-drug interaction risk
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Sullivan, Rebecca and Galetin, Aleksandra
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615.1 ,Drug-drug interaction ,Glucuronide - Abstract
The potential contribution of drug metabolites to drug-drug interactions (DDIs) is increasingly recognised. The latest FDA guidance recommends investigation of the effect of metabolites on CYP450 enzymes if present at greater than or equal to25% of parent systemic exposure. In addition, the clinical relevance of transporter mediated DDIs is highlighted in the latest EMA and FDA guidelines. Glucuronidation is a major conjugation pathway and increasing numbers of glucuronides have been reported to inhibit metabolic enzymes and/or uptake transporters e.g., gemfibrozil glucuronide inhibits both CYP2C8 and OATP1B1. However, their inhibitory effects on a range of metabolic enzymes and transporters has not been characterised in a systematic manner and their potency in relation to parent compounds is unknown. The aim of this Thesis was to comprehensively investigate the inhibitory potential of 18 compounds in total including 10 glucuronides, parent compounds and reference inhibitors of interest against CYP2C8, CYP3A4 and UGT1A1 in human liver microsomes, using repaglinide as a probe substrate. Similar studies were performed on OATP1B1 in HEK293 cells, using estradiol 17β glucuronide (E17βG) as an initial probe substrate. For a subset of glucuronides and parent drugs, additional studies were conducted using pitavastatin as a clinically relevant OATP1B1 probe. A pre-incubation with inhibitor was included in both transporter and metabolism experiments to assess potential for time-dependent inhibition. In addition, the clinical exposure of glucuronides was assessed in relation to FDA guidance exposure limits. The in vitro data generated and inhibitor clinical exposure data collated were used to predict glucuronide DDI risk initially using basic models (1 + I/Ki). Subsequently, the extent of DDI was predicted using static mechanistic models incorporating the fraction of victim drug metabolised (fmCYP2C8) or transported (ftOATP1B1) by the enzyme or transporter of interest using repaglinide and pitavastatin as relevant victim drugs. CYP2C8 IC50 values were characterised for 5/10 glucuronides (IC50 8.6 - 54.1 μM for telmisartan and diclofenac glucuronides, respectively) which demonstrated similar or greater inhibitory potency to their parent compounds except for mefenamic acid which was 3-fold more potent than its glucuronide. The choice of enzyme co-factor conditions influenced the CYP2C8 inhibitory potential of mefenamic acid and clopidogrel glucuronides, no changes were seen for other glucuronides. Time-dependent increase in potency was observed for clopidogrel and gemfibrozil glucuronides resulting in 3 to 10-fold more potent inhibition than parent compounds. Minimal inhibition of CYP3A4 and UGT1A1 by glucuronides was observed. OATP1B1 IC50 values with E17βG as a probe substrate were obtained for 8/10 glucuronides (IC50 1.2 - 55.1 μM for telmisartan and raloxifene glucuronides, respectively) and all 5 parent compounds investigated (IC50 0.73 - 47.8 μM). For repaglinide, diclofenac and gemfibrozil glucuronides, OATP1B1 IC50 values were comparable to parents. Ezetimibe glucuronide was 4-fold more potent than its parent but the opposite was observed for telmisartan. The OATP1B1 inhibitory potency of parents and glucuronides was in the same rank order; however, differential pre-incubation effects were seen between glucuronide-parent pairs. Using pitavastatin as a probe, IC50 values were within 2-fold of those obtained using E17βG except for gemfibrozil, diclofenac and ezetimibe where up to 12-fold more potent OATP1B1 inhibition was observed using E17βG. A mean increase in OATP1B1 inhibitory potency of 1.7 and 2.3-fold was observed following pre-incubation with inhibitor using E17βG and pitavastatin as probe substrates, respectively. In conclusion, glucuronides were found to inhibit both CYP2C8 and OATP1B1 in vitro causing comparable or more potent inhibition than their parent drugs in the majority of cases. Inclusion of a pre-incubation step with inhibitor is recommended for both OATP1B1 and CYP2C8 inhibition studies to obtain the most conservative estimate and assess the mechanism of inhibition. The inhibition of metabolising enzymes or OATP1B1 by glucuronides was not predicted to cause a clinically relevant DDI using the 1 + I/Ki approach, except in the case of gemfibrozil and clopidogrel glucuronides and the reference inhibitors. Using static mechanistic equations, only gemfibrozil glucuronide and reference inhibitors were predicted to cause clinically relevant inhibition of OATP1B1 and CYP2C8. The paucity of glucuronide clinical exposure data limited the prediction of the DDI risk of glucuronides. However, where clinical exposure data were available, glucuronides exceeded 25% of parent drug exposure. Improved understanding of glucuronide DDI risk resulting from inhibition of multiple clearance mechanisms (CYP2C8 and OATP1B1) by glucuronides and their parent drugs in conjunction with glucuronide exposure at the site of inhibition is required to more accurately predict DDI risk associated with glucuronides.
- Published
- 2016
12. In silico deconjugation of glucuronide conjugates enhances tandem mass spectra library annotation of human samples.
- Author
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Huber, Carolin, Krauss, Martin, Reinstadler, Vera, Denicolò, Sara, Mayer, Gert, Schulze, Tobias, Brack, Werner, and Oberacher, Herbert
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MASS spectrometry , *TANDEM mass spectrometry , *MOLECULAR structure , *ANNOTATIONS - Abstract
Mass spectral library annotation of liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS) data is a reliable approach for fast identification of organic contaminants and toxicants in complex environmental and biological matrices. While determining the exposure of humans or mammals, it is indispensable to include phase I and phase II metabolites (conjugates) along with the parent compounds, but often, tandem mass spectra for these are unavailable. In this study, we present and evaluate a strategy for annotating glucuronide conjugates in LC-HRMS/MS scans by applying a neutral loss search for detection, then truncating the spectra which we refer to as in silico deconjugation, and finally searching these against mass spectral libraries of the aglycones. The workflow was tested on a dataset of in vitro–generated glucuronides of reference standard mixtures and a dataset of 51 authentic urine samples collected from patients with known medication status, acquired on different instrumentations. A total number of 75 different glucuronidated molecular structures were identified by in silico deconjugation and spectral library annotation. We also identified specific molecular structures (sulfonamides, ether bonds, di-glucuronides), which resulted in slightly different fragmentation patterns between the glucuronide and the unconjugated compound. This led to a decreased spectral matching score and in some cases to a false-negative identification. Still, by applying this method, we revealed a reliable annotation of most common glucuronides, leading to a new strategy reducing the need for deconjugation steps or for recording many reference glucuronide spectra for screening approaches. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
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13. A rapid ultra high performance liquid chromatography–tandem mass spectrometry method for the quantification of daidzein, its valine carbamate prodrug, and glucuronide in rat plasma samples: Comparison of the pharmacokinetic behavior of daidzine valine carbamate prodrugs
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Li, Yingchao, Lu, Farong, Zhang, Yawei, Liu, Xiaoyu, Lin, Longyi, Jiang, Qikun, and Zhang, Tianhong
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LIQUID chromatography-mass spectrometry , *VALINE , *DAIDZEIN , *PHARMACOKINETICS , *PRODRUGS - Abstract
Two valine carbamate prodrugs of daidzein were designed to improve its bioavailability. To compare the pharmacokinetic behavior of these prodrugs with different protected phenolic hydroxyl groups of daidzein, a rapid and sensitive method for simultaneous quantification of daidzein, its valine carbamate prodrug, and daidzein‐7‐O‐glucuronide in rat plasma was developed and validated in this study. The samples were processed using a fast one‐step protein precipitation method with methanol added to 50 μL of plasma and were analyzed by ultra‐high performance liquid chromatography with tandem mass spectrometry. To improve the selectivity, peak shape, and peak elution, several key factors, especially stationary phase and the composition of the mobile phase, were tested, and the analysis was performed using the Kinetex® C18 column (100 × 2.1 mm, 2.6 μm) within only 2.6 min under optimal conditions. The established method exhibited good linearity over the concentration range of 2.0–1000 ng/mL for daidzein, and 8.0–4000 ng/mL for the prodrug and daidzein‐7‐O‐glucuronide. The accuracy of the quality control samples was between 95.5 and 110.2% with satisfactory intra‐ and interday precision (relative standard deviation values < 10.85%), respectively. This sensitive, rapid, low‐cost, and high‐throughput method was successfully applied to compare the pharmacokinetic behavior of different daidzein carbamate prodrugs. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
14. Stanozolol‐N‐glucuronide metabolites in human urine samples as suitable targets in terms of routine anti‐doping analysis.
- Author
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Göschl, Lorenz, Gmeiner, Günter, Gärtner, Peter, Stadler, Georg, Enev, Valentin, Thevis, Mario, Schänzer, Wilhelm, Guddat, Sven, and Forsdahl, Guro
- Abstract
The exogenous anabolic‐androgenic steroid (AAS) stanozolol stays one of the most detected substances in professional sports. Its detection is a fundamental part of doping analysis, and the analysis of this steroid has been intensively investigated for a long time. This contribution to the detection of stanozolol doping describes for the first time the unambiguous proof for the existence of 17‐epistanozolol‐1′N‐glucuronide and 17‐epistanozolol‐2′N‐glucuronide in stanozolol‐positive human urine samples due to the access to high‐quality reference standards. Examination of excretion study samples shows large detection windows for the phase‐II metabolites stanozolol‐1′N‐glucuronide and 17‐epistanozolol‐1′N‐glucuronide up to 12 days and respectively up to almost 28 days. In addition, we present appropriate validation parameters for the analysis of these metabolites using a fully automatic method online solid‐phase extraction (SPE) method already published before. Limits of identification (LOIs) as low as 100 pg/ml and other validation parameters like accuracy, precision, sensitivity, robustness, and linearity are given. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
15. Synthetic Artificial Apoptosis‐Inducing Receptor for On‐Demand Deactivation of Engineered Cells.
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Monge, Pere, Løvschall, Kaja Borup, Søgaard, Ane Bretschneider, Walther, Raoul, Golbek, Thaddeus W., Schmüser, Lars, Weidner, Tobias, and Zelikin, Alexander N.
- Subjects
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SYNTHETIC receptors , *CELL receptors , *CELLULAR signal transduction , *CELL populations , *MOLECULES , *TOXINS , *CELL culture , *BILAYER lipid membranes - Abstract
The design of a fully synthetic, chemical "apoptosis‐inducing receptor" (AIR) molecule is reported that is anchored into the lipid bilayer of cells, is activated by the incoming biological input, and responds with the release of a secondary messenger—a highly potent toxin for cell killing. The AIR molecule has four elements, namely, an exofacial trigger group, a bilayer anchor, a toxin as a secondary messenger, and a self‐immolative scaffold as a mechanism for signal transduction. Receptor installation into cells is established via a robust protocol with minimal cell handling. The synthetic receptor remains dormant in the engineered cells, but is effectively triggered externally by the addition of an activating biomolecule (enzyme) or in a mixed cell population through interaction with the surrounding cells. In 3D cell culture (spheroids), receptor activation is accessible for at least 5 days, which compares favorably with other state of the art receptor designs. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
16. Synthetic Artificial Apoptosis‐Inducing Receptor for On‐Demand Deactivation of Engineered Cells
- Author
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Pere Monge, Kaja Borup Løvschall, Ane Bretschneider Søgaard, Raoul Walther, Thaddeus W. Golbek, Lars Schmüser, Tobias Weidner, and Alexander N. Zelikin
- Subjects
artificial receptors ,glucuronide ,prodrugs ,self‐immolative linkers ,signal transduction ,Science - Abstract
Abstract The design of a fully synthetic, chemical “apoptosis‐inducing receptor” (AIR) molecule is reported that is anchored into the lipid bilayer of cells, is activated by the incoming biological input, and responds with the release of a secondary messenger—a highly potent toxin for cell killing. The AIR molecule has four elements, namely, an exofacial trigger group, a bilayer anchor, a toxin as a secondary messenger, and a self‐immolative scaffold as a mechanism for signal transduction. Receptor installation into cells is established via a robust protocol with minimal cell handling. The synthetic receptor remains dormant in the engineered cells, but is effectively triggered externally by the addition of an activating biomolecule (enzyme) or in a mixed cell population through interaction with the surrounding cells. In 3D cell culture (spheroids), receptor activation is accessible for at least 5 days, which compares favorably with other state of the art receptor designs.
- Published
- 2021
- Full Text
- View/download PDF
17. Outlook on Next Generation Technologies and Strategy Considerations for ADC Process Development and Manufacturing
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Marcq, Olivier, Teicher, Beverly A., Series Editor, and Damelin, Marc, editor
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- 2018
- Full Text
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18. Drug Metabolism
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Talevi, Alan, Bellera, Carolina Leticia, Talevi, Alan, editor, and Quiroga, Pablo A. M., editor
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- 2018
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19. Association of NQO2 With UDP-Glucuronosyltransferases Reduces Menadione Toxicity in Neuroblastoma Cells
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Monivan Chhour, Pierre Perio, Regis Gayon, Hélène Ternet-Fontebasso, Gilles Ferry, Françoise Nepveu, Jean A. Boutin, Jan Sudor, and Karine Reybier
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NQO2 ,UGT1A6 ,conjugation ,glucuronide ,menadiol ,cellular toxicity ,Therapeutics. Pharmacology ,RM1-950 - Abstract
The balance between detoxification and toxicity is linked to enzymes of the drug metabolism Phase I (cytochrome P450 or oxidoreductases) and phase II conjugating enzymes (such as the UGTs). After the reduction of quinones, the product of the reaction, the quinols—if not conjugated—re-oxidizes spontaneously to form the substrate quinone with the concomitant production of the toxic reactive oxygen species (ROS). Herein, we documented the modulation of the toxicity of the quinone menadione on a genetically modified neuroblastoma model cell line that expresses both the quinone oxidoreductase 2 (NQO2, E.C. 1.10.5.1) alone or together with the conjugation enzyme UDP-glucuronosyltransferase (UGT1A6, E.C. 2.4.1.17), one of the two UGT isoenzymes capable to conjugate menadione. As previously shown, NQO2 enzymatic activity is concomitant to massive ROS production, as previously shown. The quantification of ROS produced by the menadione metabolism was probed by electron-paramagnetic resonance (EPR) on cell homogenates, while the production of superoxide was measured by liquid chromatography coupled to mass spectrometry (LC-MS) on intact cells. In addition, the dysregulation of the redox homeostasis upon the cell exposure to menadione was studied by fluorescence measurements. Both EPR and LCMS studies confirmed a significant increase in the ROS production in the NQO2 overexpressing cells due to the fast reduction of quinone into quinol that can re-oxidize to form superoxide radicals. However, the effect of NQO2 inhibition was drastically different between cells overexpressing only NQO2 vs. both NQO2 and UGT. Whereas NQO2 inhibition decreases the amount of superoxide in the first case by decreasing the amount of quinol formed, it increased the toxicity of menadione in the cells co-expressing both enzymes. Moreover, for the cells co-expressing QR2 and UGT the homeostasis dysregulation was lower in presence of menadione than for the its counterpart expressing only QR2. Those results confirmed that the cooperation of the two enzymes plays a fundamental role during the cells’ detoxification process. The fluorescence measurements of the variation of redox homeostasis of each cell line and the detection of a glucuronide form of menadiol in the cells co-expressing NQO2 and UGT1A6 enzymes further confirmed our findings.
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- 2021
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20. Stability of diazepam's phase II metabolites in dried blood spots on filter paper.
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Wang, Lele, Wang, Xuezhi, Li, Wenyue, Liu, Jiajia, Yao, Xiukun, Wei, Zhiwen, and Yun, Keming
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FILTER paper , *DIAZEPAM , *SODIUM fluoride , *METABOLITES , *STATISTICAL software - Abstract
Phase II metabolites play an important role in diazepam-related cases. The study aimed to assess the stability of diazepam's phase II metabolites in dried blood spots on filter paper. A piece of filter paper was spotted with 100 µL of whole blood (added 1% sodium fluoride as needed) obtained from participant who received 5 mg diazepam orally, air dried for 2 h at room temperature, and then stored at different conditions. Whole spots were cut at 0.1 cm from the outer edge of blood spots at post-consumption time-points of prior (zero), 5, 16, 35, 61, 120 days and 1, 1.5 years. Analytes were extracted with methanol/water mixture (8:2, v/v) and determined using HPLC-MS/MS. Decomposition rules were analyzed by a statistical software "SPSS". Temazepam glucuronide remained stable (0.5–18.6% loss) at 20 ℃ and at 20 ℃ with 1% sodium fluoride for 16 days, while it was unstable after 5 days at 4 ℃ (21.1–26.2% loss) and − 20 ℃ (28.9 - 34.4% loss). After 35 days, temazepam glucuronide concentrations began to fluctuate significantly under all conditions, and an obvious increase (290.4–355.1%) was observed in 1.5 years. Oxazepam glucuronide was always unstable after 5 days, the percentage loss was even 100% when it was stored for 61 days and 1.5 years. Dried blood spots on ordinary filter paper are recommended to be stored at 20 ℃ or 20 ℃ with 1% sodium fluoride within 16 days. Samples should be analyzed immediately or stored in sterile and dry media. • TG was stable on filter paper at 20 ℃ with or without 1 % NaF within 16 days, and it is a more reliable marker in real cases. • OG was always unstable after 5 days, the percentage loss was even 100% when it was stored for 61 days and 1.5 years. • DBS on filter paper in diazepam-related cases are recommended to be stored at 20 ℃ with or without 1 % NaF within 16 days. [ABSTRACT FROM AUTHOR]
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- 2024
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21. Stereoselective Covalent Adduct Formation of Acyl Glucuronide Metabolite of Nonsteroidal Anti-Inflammatory Drugs with UDP-Glucuronosyltransferase
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Atsushi Kawase, Rio Yamashita, Tsubasa Yoshizato, Mashiro Yoshikawa, Hiroaki Shimada, and Masahiro Iwaki
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NSAIDs ,glucuronidation ,glucuronide ,stereoselective ,liver injury ,covalent adduct ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
A reactive metabolite of nonsteroidal anti-inflammatory drugs (NSAIDs), acyl-β-D-glucuronide (AG), covalently binds to endogenous proteins. The covalent adduct formation of NSAIDs-AG may lead to the dysfunction of target proteins. Therefore, it is important to clarify the detailed characterization of the formation of covalent protein adducts of NSAID-AG. UDP-glucuronosyltransferase (UGT) catalyzes the conversion of NSAIDs to NSAIDs-AG. The aim of this study was to perform a quantitative analysis of the covalent adduct formation of NSAIDs-AG with UGT. Diclofenac-AG and ketoprofen-AG formed covalent adducts with organelle proteins. Next, the number of covalent adducts formed between NSAIDs-AG and UGT isoforms (UGT1A1, UGT1A9, UGT2B4, and UGT2B9) was determined. The capacity of diclofenac-AG to form covalent adducts with UGT1A9 or UGT2B7 was approximately 10 times higher than that of mefenamic acid-AG. The amounts of covalent adducts of AG of propionic acid derivative NSAIDs with UGT2B were higher than those with UGT1A. Stereoselectivity was observed upon covalent binding to UGT. A significant negative correlation between the half-lives of NSAIDs-AG in phosphate buffers and the amount of covalent adduct with UGT2B7 was observed, suggesting the more labile NSAID-AG forms higher irreversible bindings to UGT. This report provides comprehensive information on the covalent adduct formation of NSAIDs-AGs with UGT.
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- 2022
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22. Lack of impairment due to confirmed codeine use prior to a motor vehicle accident: role of pharmacogenomics.
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Wu, Alan HB and Kearney, Thomas
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Humans ,Fluoxetine ,Bupropion ,Morphine ,Codeine ,Cytochrome P-450 CYP2D6 ,Glucuronosyltransferase ,Dopamine Uptake Inhibitors ,Serotonin Uptake Inhibitors ,Analgesics ,Opioid ,Pharmacogenetics ,Accidents ,Traffic ,Genotype ,Phenotype ,Female ,Migraine Disorders ,Forensic Toxicology ,Genetic Carrier Screening ,Cytochrome P450 ,Glucuronide ,UDP glucuronosyltransferase ,UDP glucuronosyltransferase pharmacogenomics ,pharmacogenomics ,Patient Safety ,Genetics ,Clinical Sciences ,Legal & Forensic Medicine - Abstract
BackgroundWe examined forensic serum toxicology and pharmacogenomics data from a woman on codeine shortly before she caused a motor vehicle accident.MethodsA woman driving erratically collided with a parked car of a highway seriously injuring 2 men working to repair the parked vehicle. The woman tested positive for codeine, acetaminophen and barbital. She had been taking these medications for 20 years due to migraine headache. Serum toxicology and genotype analysis for cytochrome P450, UDP glucuronosyltransferase, and other metabolizing enzymes were measured.ResultsThe woman was tried and convicted of driving under the influence resulting in bodily harm and was sentenced to 6 years. Toxicology results on peripheral blood showed a total and free codeine of 840 and 348 μg/L, respectively, and total morphine of 20 μg/L (17, 3, and 0 μg/L for morphine-3-glucuronide, morphine-6-glucuronide, and free morphine, respectively). She was heterozygous for CYP 2D6 *2/*4 (extensive/poor metabolism) and heterozygous for UGT 2B7 *1/*2 (extensive/ultra-rapid metabolism). The woman was also taking fluoxetine and bupropion which are strong inhibitors of CYP 2D6.ConclusionsBased on her genotype and phenotype and reports by the arresting officer, we suggest that the subject in question was not intoxicated by opiates at the time of her motor vehicle accident and may have been falsely incarcerated.
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- 2013
23. A critical examination of human data for the biological activity of quercetin and its phase-2 conjugates.
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Williamson G and Clifford MN
- Abstract
This critical review examines evidence for beneficial effects of quercetin phase-2 conjugates from clinical intervention studies, volunteer feeding trials, and in vitro work. Plasma concentrations of quercetin-3- O -glucuronide (Q3G) and 3'-methylquercetin-3- O- glucuronide (3'MQ3G) after supplementation may produce beneficial effects in macrophages and endothelial cells, respectively, especially if endogenous deglucuronidation occurs, and lower blood uric acid concentration via quercetin-3'- O -sulfate (Q3'S). Unsupplemented diets produce much lower concentrations (<50 nmol/l) rarely investigated in vitro. At 10 nmol/l, Q3'S and Q3G stimulate or suppress, respectively, angiogenesis in endothelial cells. Statistically significant effects have been reported at 100 nmol/l in breast cancer cells (Q3G), primary neuron cultures (Q3G), lymphocytes (Q3G and3'MQ3G) and HUVECs (QG/QS mixture), but it is unclear whether these translate to a health benefit in vivo. More sensitive and more precise methods to measure clinically significant endpoints are required before a conclusion can be drawn regarding effects at normal dietary concentrations. Future requirements include better understanding of inter-individual and temporal variation in plasma quercetin phase-2 conjugates, their mechanisms of action including deglucuronidation and desulfation both in vitro and in vivo, tissue accumulation and washout, as well as potential for synergy or antagonism with other quercetin metabolites and metabolites of other dietary phytochemicals.
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- 2024
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24. Equine uridine diphospho-glucuronosyltransferase 1A1, 2A1, 2B4, 2B31: cDNA cloning, expression and initial characterization of morphine metabolism.
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Hamamoto-Hardman, Briana D., Baden, Russell W., McKemie, Dan S., and Knych, Heather K.
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ANTISENSE DNA , *LIQUID chromatography-mass spectrometry , *LIVER microsomes , *MORPHINE , *ENZYME kinetics , *DRUG metabolism - Abstract
Uridine diphospho-glucuronosyltransferases (UGTs) are membrane-bound enzymes that catalyze the conjugation of glucuronic acid onto a diverse set of xenobiotics. Horses efficiently and extensively glucuronidate a number of xenobiotics, including opioids, making UGTs an important group of drug-metabolizing enzymes for the clearance of drugs. Recombinant enzymes have allowed researchers to characterize the metabolism of a variety of drugs. The primary objective was to clone, express and characterize equine UGTs using drugs characterized as UGT substrates in other species. A secondary objective was to characterize the in vitro metabolism of morphine in horses. In vitro drug metabolism study using liver microsomes and recombinant enzyme systems. Liver microsomes and RNA from tissue collected from two Thoroughbred mares euthanized for other reasons. Based on homology to the human UGT2B7, four equine UGT variants were expressed: UGT1A1, UGT2A1, UGT2B31 and UGT2B4. cDNA sequences were cloned and resulting protein expressed in a baculovirus expression system. Functionality of the enzymes was assessed using 4-methylumbelliferone, testosterone, diclofenac and ketoprofen. Recombinant enzyme, control cells, equine liver microsomes and human UGT2B7 supersomes were then incubated with morphine. Concentrations of metabolites were measured using liquid chromatography–tandem mass spectrometry and enzyme kinetics determined. 4-Methylumbelliferone was glucuronidated by all expressed equine UGTs. Testosterone glucuronide was not produced by any of the expressed enzymes, and diclofenac glucuronide and ketoprofen glucuronide were produced by UG2A1 and UGT1A1, respectively. UGT2B31 metabolized morphine to morphine-3-glucuronide and low concentrations of morphine-6-glucuronide. This is the first successful expression of functional recombinant equine UGTs. UGT2B31 contributes to the glucuronidation of morphine; however, it is probably not the main metabolizing enzyme. These results warrant further investigation of equine UGTs, including expression of additional enzymes and further characterization of UGT2B31 as a contributor to morphine metabolism. [ABSTRACT FROM AUTHOR]
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- 2020
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25. Curcumin and piperine supplementation of obese mice under caloric restriction modulates body fat and interleukin-1β
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Taiki Miyazawa, Kiyotaka Nakagawa, Sharon H. Kim, Michael J. Thomas, Ligi Paul, Jean-Marc Zingg, Gregory G. Dolnikowski, Susan B. Roberts, Fumiko Kimura, Teruo Miyazawa, Angelo Azzi, and Mohsen Meydani
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Caloric restriction ,Curcumin ,Glucuronide ,High fat diet ,Inflammation ,Meso scale discovery ,Nutrition. Foods and food supply ,TX341-641 ,Nutritional diseases. Deficiency diseases ,RC620-627 - Abstract
Abstract Background Dietary bioactive compounds capable of improving metabolic profiles would be of great value, especially for overweight individuals undergoing a caloric restriction (CR) regimen. Curcumin (Cur), a possible anti-obesity compound, and piperine (Pip), a plausible enhancer of Cur’s bioavailability and efficacy, may be candidate agents for controlling body fat, metabolism and low grade inflammation. Methods 47 eight-week-old male C57BL/6 mice were fed a high fat diet (HFD) for 23 weeks to induce obesity. Then, mice were divided into 5 groups. Group 1 continued on HFD ad libitum. The other 4 groups underwent CR (reduced 10% HFD intake for 10 weeks, 20% for 20 weeks) with Cur, Pip, Cur + Pip or none of these. Percent body fat, plasma inflammatory markers associated with obesity (interferon (IFN)-γ, interleukin (IL)-10, IL-12 p70, IL-1β, IL-6 and KC/GRO), plasma Cur metabolites and liver telomere length were measured. Results Compared to the other groups, obese mice who underwent CR and received Cur + Pip in their diet lost more fat and had significantly lower IL-1β and KC/GRO. Tandem mass spectrometry analysis of plasma from obese mice under CR showed no difference in Cur metabolite levels between groups supplemented with Cur alone or combined with Pip. However, plasma IL-1β levels were inversely correlated with curcumin glucuronide. Minor modulation of telomere length were observed. Conclusions It is plausible that supplementing the high fat diet of CR mice with Cur + Pip may increase loss of body fat and suppresses HFD induced inflammation. Combination of Cur and Pip has potential to enhance CR effects for the prevention of metabolic syndrome.
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- 2018
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26. Conjugated Metabolites of Hydroxytyrosol and Tyrosol Contribute to the Maintenance of Nitric Oxide Balance in Human Aortic Endothelial Cells at Physiologically Relevant Concentrations
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Gabriele Serreli, Melanie Le Sayec, Camilla Diotallevi, Alice Teissier, Monica Deiana, and Giulia Corona
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olive oil ,conjugate metabolites ,sulfate ,glucuronide ,hydroxytyrosol ,tyrosol ,Organic chemistry ,QD241-441 - Abstract
Nitric oxide (NO) is an important signaling molecule involved in many pathophysiological processes. NO mediates vasodilation and blood flow in the arteries, and its action contributes to maintaining vascular homeostasis by inhibiting vascular smooth muscle contraction and growth, platelet aggregation, and leukocyte adhesion to the endothelium. Dietary antioxidants and their metabolites have been found to be directly and/or indirectly involved in the modulation of the intracellular signals that lead to the production of NO. The purpose of this study was to investigate the contribution of conjugated metabolites of hydroxytyrosol (HT) and tyrosol (TYR) to the release of NO at the vascular level, and the related mechanism of action, in comparison to their parental forms. Experiments were performed in human aortic endothelial cells (HAEC) to evaluate the superoxide production, the release of NO and production of cyclic guanosine monophosphate (cGMP), the activation of serine/threonine-protein kinase 1 (Akt1), and the activation state of endothelial nitric oxide synthase (eNOS). It was observed that the tested phenolic compounds enhanced NO and cGMP concentration, inhibiting its depletion caused by superoxide overproduction. Moreover, some of them enhanced the activation of Akt (TYR, HT metabolites) and eNOS (HT, HVA, TYR-S, HT-3S). Overall, the obtained data showed that these compounds promote NO production and availability, suggesting that HT and TYR conjugated metabolites may contribute to the effects of parental extra virgin olive oil (EVOO) phenolics in the prevention of cardiovascular diseases.
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- 2021
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27. 姜黄素及其代谢修饰产物对PC12细胞 氧化损伤的保护作用.
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李浩铭, 黄永杰, 王永丽, 李大鹏, and 李 锋
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REACTIVE oxygen species ,SUPEROXIDE dismutase ,PROTEIN expression ,CELL survival ,WESTERN immunoblotting ,CURCUMINOIDS - Abstract
Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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- 2020
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28. Analysis of morphine, morphine-3-glucuronide, morphine-6-glucuronide, codeine and codeine-6-glucuronide using HPLC.
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Almoterie, Abdullataif Aaied, Alshugeer, Mohammad Suliman, Binbaz, Abdulmajeed Saad, Alangari, Waleed Abdulaziz, Alsakran, Sultan Hamdan, Almutairi, Turki Ayidh, Alblaihi, Ahmed Abdulaziz, and Alenzi, Mohammed Mosier
- Subjects
MORPHINE ,HIGH performance liquid chromatography ,FORENSIC toxicology ,GAS chromatography/Mass spectrometry (GC-MS) ,ULTRAVIOLET detectors - Abstract
Copyright of Journal of Medical & Pharmaceutical Sciences is the property of Arab Journal of Sciences & Research Publishing (AJSRP) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
- Published
- 2020
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29. Disposition of asciminib, a potent BCR-ABL1 tyrosine kinase inhibitor, in healthy male subjects.
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Tran, Phi, Hanna, Imad, Eggimann, Fabian Kurt, Schoepfer, Joseph, Ray, Tapan, Zhu, Bing, Wang, Lai, Priess, Petra, Tian, Xianbin, Hourcade-Potelleret, Florence, and Einolf, Heidi J.
- Subjects
- *
PROTEIN-tyrosine kinases , *CHRONIC myeloid leukemia , *KINASE inhibitors , *CYTOCHROME P-450 , *LYMPHOBLASTIC leukemia , *CD19 antigen - Abstract
Asciminib is a potent, specific BCR-ABL1 inhibitor being developed for the treatment of patients with chronic myelogenous leukemia (CML) and Philadelphia chromosome positive acute lymphoblastic leukemia (Ph + ALL). Here, we present the results of human oral absorption, distribution, metabolism, excretion (ADME) and in vitro studies that together provide an overall understanding of the metabolism, distribution and clearance of asciminib in humans. Asciminib was rapidly absorbed with a maximum plasma concentration at two hours post-dose. Total radioactivity and asciminib showed similar terminal half-lives in plasma. Oral asciminib absorption ranged between a minimum of 33%, and a maximum of 57% based on the metabolite profiles of late time-point feces collections. Asciminib was eliminated mainly through feces via unchanged asciminib excretion and metabolism. Direct glucuronidation and oxidation were major metabolic pathways in human that were catalyzed predominantly by UDP-glucuronosyltransferase (UGT)2B7 and cytochrome P450 (CYP)3A4, respectively. The relative contribution of the glucuronidation pathway to the total clearance of asciminib via metabolism is estimated to range ∼28–58%, whereas the relative contribution of the oxidative pathway is estimated to range ∼37–64%, based upon the maximum oral absorption in humans. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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30. Predictability of human pharmacokinetics of diisononyl phthalate (DINP) using chimeric mice with humanized liver.
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Iwata, Hiroshi, Goto, Masatomo, Sakai, Norifumi, Suemizu, Hiroshi, and Yamazaki, Hiroshi
- Subjects
- *
MICE , *PHARMACOKINETICS , *LIVER , *PHTHALATE esters , *RADIOACTIVITY , *LIVER cells - Abstract
1. In order to investigate the pharmacokinetics of diisononyl phthalate (DINP) in humans, we administered [phenyl-U-14C]DINP at a dose of 50.0 mg/kg orally to chimeric mice (humanized-liver mice) in which the liver of TK-NOG mice (control mice) was replaced with human hepatocytes. 2. The plasma radioactivity concentrations peaked (18.0 and 59.9 µg equivalent of DINP/mL, respectively) at 2 h after administration in control and humanized-liver mice. Concentrations rose again at 8 h in controls, but not in humanized-liver mice. 3. The cumulative excretion rates in urine and feces, respectively, were 58.1% and 37.3% of the doses in controls up to 48 h, but were 86.0% and 7.7% in humanized-liver mice. 4. The main circulating metabolites in control and humanized-liver mice were monoisononyl phthalate (MINP) and the glucuronide of oxidized MINP, respectively. The urinary excretion ratio of the glucuronide of oxidized MINP in control mice was one-third of that in humanized-liver mice. 5. The present results suggested that the oxidation rates of the primary metabolite of DINP and their excretion routes to urine/feces were different for control and humanized-liver mice. Species differences in liver activities could be a determinant factor in the in vivo metabolism and disposition of diallyl phthalates such as DINP. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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31. 4-(Hydroxymethylnitrosamino)-1-(3-pyridyl)-1-butanone glucuronide has the potential to form 2'-deoxyguanosine and N-acetylcysteine adducts.
- Author
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Takahito Nishiyama, Nahoko Hayashi, Hiromi Yanagita, Tomokazu Ohnuma, Kenichiro Ogura, and Akira Hiratsuka
- Subjects
- *
CHEMICAL adducts , *DNA adducts , *NITROSOAMINES , *CYTOCHROME P-450 , *BLADDER cancer , *SMOKING , *GLUCURONIDES , *DEOXYGUANOSINE , *TARGET organs (Anatomy) - Abstract
- Cigarette smoking is a risk factor for the development of various cancers, such as lung, nasal, liver and bladder cancers. 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco-specific nitrosamine, is implicated in human lung cancer. NNK-induced DNA adducts are found in target tissues for NNK carcinogenesis. NNK is activated by cytochrome P450 dependent a-hydroxylation at either the methylene carbon or methyl carbon adjacent to the N-nitroso group. The former leads to the formation of the methylating agent, and the latter produce the pyridyloxobutylating agent. NNK and some of its metabolites are further metabolized by UDP-glucuronosyltransferases (UGTs). Glucuronides generally are much less active than the parent aglycon therefore the glucuronides of NNK-related metabolites are thought to be inactive. However, 4-(hydroxymethylnitrosamino)-1-(3-pyridyl)-1-butanone glucuronide (HO-methyl NNK glucuronide) can be transported to the target organs of NNK carcinogenesis where subsequent hydrolysis causes the release of the reactive intermediate. Regeneration of HO-methyl NNK could play an important role in the tissue-specific carcinogenicity of NNK. In the present study, we investigated the reactivity of HO-methyl NNK glucuronide toward 2'-deoxyguanosine (dGuo) and N-acetylcysteine (NAC; used as a models for thiol groups on proteins). The reaction mixtures of HOmethyl NNK glucuronide and dGuo or NAC were analyzed by LCMS-IT-TOF-MS. We also employed 4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone, a pyridyloxobutylating agent, to confirm the formation of pyridyloxobutylated adducts. Thus, we determined the production of pyridyloxobutylated dGuo and NAC adducts. Our results suggest HO-methyl NNK glucuronide could generate a reactive intermediate in the tissues and then form adducts with proteins and DNA. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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32. In Vitro and In Vivo Analysis of Ochratoxin A-Derived Glucuronides and Mercapturic Acids as Biomarkers of Exposure
- Author
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Raphael Dekant, Michael Langer, Maria Lupp, Cynthia Adaku Chilaka, and Angela Mally
- Subjects
ochratoxin A ,biomarker of exposure ,glucuronide ,mercapturic acid ,mycotoxin ,Medicine - Abstract
Ochratoxin A (OTA) is a widespread food contaminant, with exposure estimated to range from 0.64 to 17.79 ng/kg body weight (bw) for average consumers and from 2.40 to 51.69 ng/kg bw per day for high consumers. Current exposure estimates are, however, associated with considerable uncertainty. While biomarker-based approaches may contribute to improved exposure assessment, there is yet insufficient data on urinary metabolites of OTA and their relation to external dose to allow reliable estimates of daily intake. This study was designed to assess potential species differences in phase II biotransformation in vitro and to establish a correlation between urinary OTA-derived glucuronides and mercapturic acids and external exposure in rats in vivo. In vitro analyses of OTA metabolism using the liver S9 of rats, humans, rabbits and minipigs confirmed formation of an OTA glucuronide but provided no evidence for the formation of OTA-derived mercapturic acids to support their use as biomarkers. Similarly, OTA-derived mercapturic acids were not detected in urine of rats repeatedly dosed with OTA, while indirect analysis using enzymatic hydrolysis of the urine samples prior to LC–MS/MS established a linear relationship between urinary glucuronide excretion and OTA exposure. These results support OTA-derived glucuronides but not mercapturic acids as metabolites suitable for biomonitoring.
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- 2021
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33. Automated Annotation of Microbial and Human Flavonoid-Derived Metabolites
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Mihaleva, Velitchka V., Ünlü, Fatma Yelda, Vervoort, Jacques, Ridder, Lars, Dietert, Rodney R., Series editor, Kochhar, Sunil, editor, and Martin, François-Pierre, editor
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- 2015
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34. An ultrafast ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry method for cannabidiol monitoring in pediatric refractory epilepsy
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Cintia V Cruz, Paula Schaiquevich, Carlos A Pérez Montilla, Gabriela Reyes Valenzuela, Paulo Cáceres Guido, Facundo García Bournissen, and Roberto Caraballo
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Drug Resistant Epilepsy ,Metabolite ,Tandem mass spectrometry ,Mass spectrometry ,digestive system ,030226 pharmacology & pharmacy ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Pharmacokinetics ,Limit of Detection ,Tandem Mass Spectrometry ,medicine ,Cannabidiol ,Humans ,Pharmacology (medical) ,Dronabinol ,Child ,Chromatography, High Pressure Liquid ,Pharmacology ,Detection limit ,Chromatography ,Selected reaction monitoring ,digestive system diseases ,surgical procedures, operative ,chemistry ,Glucuronide ,medicine.drug - Abstract
Background: Cannabidiol (CBD) is a nonpsychoactive natural product that has been increasingly used as a promising new drug for the management of neurological conditions such as refractory epilepsy. Development of rapid and sensitive methods to quantitate CBD is essential to evaluate its pharmacokinetics in humans, particularly in children. The objective of this work was to develop and validate an ultrafast ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) method for CBD quantitation that is capable of detecting major CBD and tetrahydrocannabinol (THC) metabolites in the plasma of pediatric refractory epilepsy patients. Methods: Eight-point CBD calibration curves were prepared using 60 µL of plasma from healthy volunteers. Samples were analyzed in a Shimadzu Nexera X2 UHPLC system, which was coupled to a Sciex QTRAP 6500 mass spectrometer. Chromatography was optimized in acetonitrile (ACN)/water with a 70%–90% gradient of ACN in 2 minutes. Multiple reaction monitoring transitions of major CBD and THC metabolites were optimized in patient plasma. Results: The optimized UHPLC-MS/MS method was validated for the linear range (1–300 ng/mL) of CBD (r2 = 0.996). The limit of quantification and limit of detection were 0.26 and 0.86 ng/mL, respectively. Accuracy and precision met the acceptable validation limits. CBD recovery and matrix effects were 83.9 ± 13.9% and 117.4 ± 4.5%, respectively. The method was successfully applied to quantify CBD and detect the major CBD and THC metabolites in clinical samples. 7-COOH-CBD was the most intensely detected metabolite followed by glucuronide conjugates. Conclusions: A simple and sensitive method for rapidly monitoring CBD and identifying relevant metabolites was developed. Its applicability in samples from children treated for epilepsy was demonstrated, making it an excellent alternative for performing pharmacokinetic studies.
- Published
- 2023
35. Whole Cell-Dependent Biosynthesis of Drug Metabolites Using Genetically Engineered Budding Yeast
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Ikushiro, Shinichi, Nishikawa, Miyu, Sakaki, Toshiyuki, and Yamazaki, Hiroshi, editor
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- 2014
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36. Ethylene glycol: Evidence of glucuronidation in vivo shown by analysis of clinical toxicology samples.
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Pedersen, Daniel Sejer, Bélanger, Patrick, Frykman, Mikael, Andreasen, Kirsten, Goudreault, Danielle, Pedersen, Henrik, Hindersson, Peter, and Breindahl, Torben
- Abstract
In the search for improved laboratory methods for the diagnosis of ethylene glycol poisoning, the in vivo formation of a glucuronide metabolite of ethylene glycol was hypothesized. Chemically pure standards of the β‐O‐glucuronide of ethylene glycol (EG‐GLUC) and a deuterated analog (d4‐EG‐GLUC) were synthesized. A high‐performance liquid chromatography and tandem mass spectrometry method for determination of EG‐GLUC in serum after ultrafiltration was validated. Inter‐assay precision (%RSD) was 3.9% to 15.1% and inter‐assay %bias was −2.8% to 12.2%. The measuring range was 2–100 μmol/L (0.48–24 mg/L). Specificity testing showed no endogenous amounts in routine clinical samples (n = 40). The method was used to analyze authentic, clinical serum samples (n = 31) from patients intoxicated with ethylene glycol. EG‐GLUC was quantified in 15 of these samples, with a mean concentration of 6.5 μmol/L (1.6 mg/L), ranging from 2.3 to 15.6 μmol/L (0.55 to 3.7 mg/L). In five samples, EG‐GLUC was detected below the limit of quantification (2 μmol/L) and it was below the limit of detection in 11 samples (1 μmol/L). Compared to the millimolar concentrations of ethylene glycol present in blood after intoxications and potentially available for conjugation, the concentrations of EG‐GLUC found in clinical serum samples are very low, but comparable to concentrations of ethyl glucuronide after medium dose ethanol intake. In theory, EG‐GLUC has a potential value as a biomarker for ethylene glycol intake, but the pharmacokinetic properties, in vivo/vitro stability and the biosynthetic pathways of EG‐GLUC must be further studied in a larger number of patients and other biological matrices. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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37. Identification of equol-7- glucuronide- 4'- sulfate, monoglucuronides and monosulfates in human plasma of 2 equol producers after administration of kinako by LC-ESI-MS.
- Author
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Aki Obara, Mizuki Kinoshita, Kaori Hosoda, Akitomo Yokokawa, Hiromi Shibasaki, and Kazuo Ishii
- Subjects
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GLUCURONIDES , *SULFATES , *PROSTATE diseases , *IDENTIFICATION , *DAIDZEIN , *PROSTATE cancer - Abstract
Equol is a product formed during the biotransformation of the naturally occurring isoflavone daidzein by intestinal bacteria. The role of equol in the prevention of several hormone-dependent diseases such as prostate cancer and osteoporosis as well as vasomotor symptoms has been extensively investigated. Equol primarily occurs in the form of major metabolites such as glucuronides and sulfates, while intact equol has been detected at only ca. 1% in human plasma. However, to date, conjugated metabolites have been evaluated by measuring the free equol obtained after selective enzymatic hydrolysis. Thus, the precise types of conjugates circulating in vivo and the position(s) of the conjugation sites on the equol skeleton have yet to be clarified. Our study describes the identification of polar equol metabolites in the plasma of 2 equol-producers obtained at 8 hours after consuming 50 g of kinako (approximately 37 mg of daidzein). The structural identification of these conjugated metabolites in plasma was performed by comparison to the LC-ESI- MSn and 1H-NMR spectral data of the corresponding chemically synthesized compounds. The results of the LC-ESI- MS/MS analysis indicated that the main conjugated metabolite in plasma was (S)-equol- 7- glucuronide- 4'- sulfate along with lower amounts of 7-and 4'-monoglucuronides as well as 7-and 4'-monosulfates. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
38. Identification and Directed Development of Non‐Organic Catalysts with Apparent Pan‐Enzymatic Mimicry into Nanozymes for Efficient Prodrug Conversion.
- Author
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Walther, Raoul, Winther, Anna K., Fruergaard, Anne Sofie, van den Akker, Wouter, Sørensen, Lise, Nielsen, Signe Maria, Jarlstad Olesen, Morten T., Dai, Yitao, Jeppesen, Henrik S., Lamagni, Paolo, Savateev, Aleksandr, Pedersen, Søren Lykke, Frich, Camilla Kaas, Vigier‐Carrière, Cécile, Lock, Nina, Singh, Mandeep, Bansal, Vipul, Meyer, Rikke L., and Zelikin, Alexander N.
- Subjects
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PRODRUGS , *NANOSTRUCTURED materials , *BIOCATALYSIS , *ENZYMES , *GLUCURONIDES - Abstract
Nanozymes, nanoparticles that mimic the natural activity of enzymes, are intriguing academically and are important in the context of the Origin of Life. However, current nanozymes offer mimicry of a narrow range of mammalian enzymes, near‐exclusively performing redox reactions. We present an unexpected discovery of non‐proteinaceous enzymes based on metals, metal oxides, 1D/2D‐materials, and non‐metallic nanomaterials. The specific novelty of these findings lies in the identification of nanozymes with apparent mimicry of diverse mammalian enzymes, including unique pan‐glycosidases. Further novelty lies in the identification of the substrate scope for the lead candidates, specifically in the context of bioconversion of glucuronides, that is, human metabolites and privileged prodrugs in the field of enzyme‐prodrug therapies. Lastly, nanozymes are employed for conversion of glucuronide prodrugs into marketed anti‐inflammatory and antibacterial agents, as well as "nanozyme prodrug therapy" to mediate antibacterial measures. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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39. An LC-MS/MS method for quantification of abiraterone, its active metabolites D(4)-abiraterone (D4A) and 5α-abiraterone, and their inactive glucuronide derivatives.
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Caron, Patrick, Turcotte, Véronique, Lévesque, Eric, and Guillemette, Chantal
- Subjects
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ABIRATERONE acetate , *DRUG metabolism , *PROSTATE cancer treatment , *LIQUID chromatography-mass spectrometry , *ANDROGEN receptors , *CHEMICAL derivatives - Abstract
Abstract Abiraterone acetate (AA) is a prodrug of abiraterone, a selective and potent steroidal cytochrome P450 17alpha- hydroxylase-17,20-lyase (CYP17A1) blocking androgen synthesis in the treatment of advanced prostate cancer. Abiraterone (Abi) is metabolized to D(4)-abiraterone (D4A) directly blocking CYP17A1 and other steroidogenic enzymes and antagonizing the androgen receptor (AR). D4A is converted by 5α-reductase to 3-keto-5α-abiraterone (5α-Abi), an AR agonist. Our recent work suggests phase II biotransformation of Abi, D4A and 5α-Abi conjugated to glucuronic acid in vitro leading to four glucuronides (G). We developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using a 6500 Qtrap mass analyzer coupled with a Shimadzu Nexera system for quantification of Abi, its active metabolites and their G derivatives in human plasma samples with deuterated internal standards. Validation was carried out according to FDA guidelines for bioanalytical method and results were within the acceptance limits. Analytes were extracted from 50 μL of plasma using a solid phase extraction procedure. Multiple reaction monitoring was used with electrospray ionization in a positive mode. Linearity, precision, and accuracy were validated over a large range of concentrations for each compound (range of 0.5–100 ng/mL for Abi and for metabolites and 0.05–10.00 ng/mL for glucuronides). The method could measure all seven analytes with sensitivity, accuracy (87–106%), and reproducibility (CV < 10.7%). Its clinical application was further examined with plasma samples obtained from prostate cancer patients under AA treatment. This reliable and validated LC-MS/MS method could be a useful tool for human biomonitoring studies. Highlights • Validated LC-MS/MS assay for abiraterone acetate metabolites in human plasma • Simultaneous quantification of active metabolites abiraterone, D4A, 5α-abiraterone • First analytical method for quantification of downstream glucuronide metabolites • Clinical application in plasma samples from patients treated with abiraterone acetate • Interindividual variability in steady state levels of all metabolites was observed [ABSTRACT FROM AUTHOR]
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- 2019
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40. Curcumin, but not curcumin-glucuronide, inhibits Smad signaling in TGFβ-dependent bone metastatic breast cancer cells and is enriched in bone compared to other tissues.
- Author
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Kunihiro, Andrew G, Brickey, Julia A, Frye, Jennifer B, Luis, Paula B, Schneider, Claus, and Funk, Janet L
- Subjects
- *
CURCUMIN , *GLUCURONIDES , *BONE metastasis , *BREAST cancer , *CANCER cells - Abstract
Breast cancer (BCa) bone metastases (BMETs) drive osteolysis via a feed-forward loop involving tumoral secretion of osteolytic factors (e.g., PTHrP) induced by bone-matrix-derived growth factors (e.g., TGFβ). In prior experiments, turmeric-derived curcumin inhibited in vivo BMET progression and in vitro TGFβ/Smad-signaling in a TGFβ-stimulated PTHrP-dependent human xenograft BCa BMET model (MDA-SA cells). However, it is unclear whether curcumin or curcumin-glucuronide mediates in vivo protection since curcumin-glucuronide is the primary circulating metabolite in rodents and in humans. Thus, effects of curcumin vs. curcumin-glucuronide on Smad-dependent TGFβ signaling were compared in a series of BCa cell lines forming TGFβ-dependent BMET in murine models, and tissue-specific metabolism of curcumin in mice was examined by LC-MS. While curcumin inhibited TGFβ-receptor-mediated Smad2/3 phosphorylation in all BCa cells studied (human MDA-SA, MDA-1833, MDA-2287 and murine 4T1 cells), curcumin-glucuronide did not. Similarly, curcumin, but not curcumin-glucuronide, blocked TGFβ-stimulated secretion of PTHrP from MDA-SA and 4T1 cells. Because the predominant serum metabolite, curcumin-glucuronide, lacked bioactivity, we examined tissue-specific metabolism of curcumin in mice. Compared to serum and other organs, free curcumin (both absolute and percentage of total) was significantly increased in bone, which was also a rich source of enzymatic deglucuronidation activity. Thus, curcumin, and not curcumin-glucuronide, appears to inhibit bone-tropic BCa cell TGFβ-signaling and to undergo site-specific activation (deconjugation) within the bone microenvironment. These findings suggest that circulating curcumin-glucuronide may act as a prodrug that preferentially targets bone, a process that may contribute to the bone-protective effects of curcumin and other highly glucuronidated dietary polyphenols. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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41. Simultaneous collection of the portal and superior vena cava blood in conscious rats defined that intestinal epithelium is the major site of glucuronidation, but not sulfation and methylation, of quercetin.
- Author
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Tanaka, Seiya, Oyama, Manami, Nishikawa, Miyu, Ikushiro, Shinichi, and Hara, Hiroshi
- Subjects
- *
FLAVONOIDS , *QUERCETIN - Abstract
Quercetin is a flavonoid with many physiological effects. Absorbed quercetin is rapidly conjugated in the intestinal epithelium and liver. Different positional isomers of quercetin conjugates have different physiological properties. However, the mechanisms of quercetin conjugation in the intestine are not fully clarified. We examined the regioselective quercetin conjugate formation in the intestine after oral administration of quercetin glycosides, by simultaneous sampling of blood from the portal vein and superior vena cava, and quantifying various positional isomers of quercetin glucuronides and sulfates in conscious rats. Concentrations of quercetin glucuronides were higher in blood from the portal vein than the superior vena cava, showing that glucuronidation mainly occurred in the intestine. Such differences were not observed for quercetin sulfates. Regioselectivity of the intestinal glucuronidation in quercetin hydroxyl groups were 7- >3′- >3- >4′-OH. Quercetin was mainly sulfated on 3′-OH at 30 min, but on 4′-OH at 240 min. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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42. Combatting implant-associated biofilms through localized drug synthesis.
- Author
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Walther, Raoul, Nielsen, Signe Maria, Christiansen, Rikke, Meyer, Rikke L., and Zelikin, Alexander N.
- Subjects
- *
BIOFILMS , *BACTERIAL contamination , *DRUG synthesis , *BIOMATERIALS , *FLUOROQUINOLONES , *ANTIBIOTICS - Abstract
Abstract Bacterial contamination of implantable biomaterials is a significant socioeconomic and healthcare burden. Indeed, bacterial colonization of implants after surgery has a high rate of incidence whereas concurrent prophylaxis using systemic antibiotics has limited clinical success. In this work, we develop enzyme-prodrug therapy (EPT) to prevent and to treat bacteria at interfaces. Towards the overall goal, novel prodrugs for fluoroquinolone antibiotics were developed on a privileged glucuronide scaffold. Whereas carbamoyl prodrugs were not stable and not suitable for EPT, glucuronides containing self-immolative linker between glucuronic acid masking group and the antibiotic were stable in solution and readily underwent bioconversion in the presence of β-glucuronidase. Surface coatings for model biomaterials were engineered using sequential polymer deposition technique. Resulting coatings afforded fast prodrug conversion and mediated antibacterial measures against planktonic species as evidenced by pronounced zone of bacterial growth inhibition around the biomaterial surface. These biomaterials coupled with the glucuronide prodrugs also effectively combatted bacteria within established biofilms and also successfully prevented bacterial colonization of the surface. To our knowledge, this is the first report of EPT engineered to the surface of biomaterials to mediate antibacterial measures. Graphical abstract Unlabelled Image [ABSTRACT FROM AUTHOR]
- Published
- 2018
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43. An optimized solvent extraction and characterization of unidentified flavonoid glucuronide derivatives from spinach by UHPLC-HR-QTOF-MS.
- Author
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Singh, Jashbir, Jayaprakasha, G.K., and Patil, Bhimanagouda S.
- Subjects
- *
SOLVENT extraction , *FLAVONOIDS , *GLUCURONIDES , *COMPOSITION of spinach , *LIQUID chromatography-mass spectrometry - Abstract
A rapid, sensitive analytical method using ultra-high-pressure liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-HR-QTOF-MS) was developed for the identification and quantification of flavonoids from spinach. The extraction efficiency of flavonoids was evaluated by different solvents such as acetone, ethanol, methanol, acetone: water (70:30), ethanol: water (70:30) and methanol: water (70:30). Flavonoid identification was achieved by UV spectra, high resolution accurate mass and their fragmentation pattern. The precursor and product ions were recorded by both broadband collision ion dissociation (bbCID) and multiple reaction monitoring (MRM) techniques. Different collision energies (5, 10, 15, 20, 40, and 70 eV) were optimized to obtain the mass spectra of flavonoids in positive and negative ionization modes. For the first time, five minor flavonoid glucuronide derivatives were identified in spinach. MRM and bbCID provided glucuronide fingerprint ions at m/z 175.0278 and m/z 113.0257 respectively in negative ionization mode. The quantification of identified flavonoids was achieved by 5,3',4'-trihydroxy-3-methoxy-6:7-methylen-dioxyflavone-4'- β -D-glucuronide which was purified by semi-preparatory HPLC. The purity of the isolated compound was confirmed by NMR analysis. The identified 5,3',4'-trihydroxy-3-methoxy-6:7-methylen-dioxyflavone-4'-β-D-(2′-O-feurloyl-glucuronide) was the prominent flavonoid and the level was significantly higher in the acetone fraction (2.95 ± 0.16 µg/g FW). This study demonstrates the systematic identification of potential bioactive compounds especially glucuronide derivatives from spinach. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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- View/download PDF
44. Quercetin-3-O-β-D-glucuronide inhibits mitochondria pathway-mediated platelet apoptosis via the phosphatidylinositol-3-kinase/AKT pathway in immunological bone marrow failure
- Author
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Aiping Zhang, Lemin Xia, Qin Zheng, Zhe Jin, Wan-Hui Wong, Hai Yu, Jie Ding, and He-Ping Yu
- Subjects
Medicine (General) ,Chemistry ,Bone marrow failure ,Mitochondrion ,medicine.disease ,chemistry.chemical_compound ,R5-920 ,Complementary and alternative medicine ,Apoptosis ,medicine ,Cancer research ,Phosphatidylinositol 3 kinase akt ,Platelet ,Glucuronide ,Quercetin ,apoptosis ,mitochondrial pathway ,phosphatidylinositol-3-kinase/akt ,platelets ,quercetin-3-o-β-d-glucuronide - Abstract
Objective: Quercetin-3-O-β-D-glucuronide (QG) can alleviate immunological bone marrow failure (BMF) by increasing platelet counts. However, the principal mechanism is less known. This study aimed at deciphering the possible underlying mechanism of QG that is indicated in thrombocytopenic purpura. Methods: In vitro and in vivo experiments were carried out for investigating the mechanism behind QG-facilitated inhibition of mitochondrial pathway-mediated excessive apoptosis of platelets through the phosphatidylinositol-3-kinase (PI3K)/AKT pathway. Results: Our results revealed that QG, the main effective ingredient of Herba Sarcandrae, increases the number of platelets and decreases the expression of Bax, Bad, Bid, and caspase-9 in immunological BMF, indicating the inhibition of mitochondrial pathway-mediated apoptosis. Moreover, we found that the protein and mRNA expressions, as well as the phosphorylated levels of PI3K and AKT, were increased significantly by QG, suggesting the activation of the PI3K/AKT pathway. Furthermore, the inhibition of the PI3K/AKT pathway by LY294002 antagonizes the effects of QG on platelet counts and mitochondrial pathway-mediated apoptosis. Conclusion: We demonstrate that QG inhibits the mitochondria pathway-mediated platelet apoptosis via the PI3K/AKT pathway in immunological BMF. This study thus sheds light on exploring the possible regulatory mechanism of traditional Chinese medicine in the treatment of thrombocytopenia induced by BMF.
- Published
- 2022
45. Development of a quick preparation method for the analysis of 11-nor-9-carboxy-∆9-tetrahydrocannabinol in human urine by phenylboronic-acid solid-phase extraction.
- Author
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Ishii, Ayumu, Sato, Kazuki, Kusakabe, Kosuke, Kato, Noriyuki, and Wada, Takeshi
- Subjects
- *
SOLID phase extraction , *TANDEM mass spectrometry , *LIQUID-liquid extraction , *ALKALINE solutions , *URINE , *HUMAN error - Abstract
A rapid preparation method for the analysis of the urine from a cannabis user was established. Generally, 11-nor-9-carboxy-∆9-tetrahydrocannabinol (THC-COOH), which is one of the main metabolites of ∆9-tetrahydorocannabinol (THC), must be detected from a user's urine to verify cannabis use. However, existing preparation methods are usually multistep and time-consuming processes. Before the analysis by liquid-chromatography tandem mass spectrometry (LC-MS/MS), deconjugation by treatment with β-glucuronidase or alkaline solution, liquid-liquid extraction or solid-phase extraction (SPE), and evaporation are generally performed. In addition, subsequent derivatization (silylation or methylation) are certainly necessary for gas-chromatography mass spectrometry (GC/MS) analysis. Here, we focused on the phenylboronic-acid (PBA) SPE, which selectively binds compounds with a cis -diol moiety. THC-COOH is metabolized as a glucuronide conjugate (THC-COOGlu) which has cis -diol moieties, therefore, we investigated the conditions of its retention and elution to reduce the operating time. We developed four elution conditions, which afford the following derivatives: acidic elution for THC-COOGlu, alkaline elution for THC-COOH, methanolysis elution for the THC-COOH methyl ester (THC-COOMe), and methanolysis elution and following methyl etherification for O -methyl-THC-COOMe (O -Me-THC-COOMe). All repeatability and recovery rates were evaluated by LC-MS/MS in this study. As a result, these four pathways required short times (within 10–25 min) and exhibited good repeatability and recovery rates. Detection limits of pathway I-IV were 10.8, 1.7, 18.9, and 13.8 ng mL-1, respectively. Lower limits of quantification were 62.5, 31.25, 57.3, and 62.5 ng mL-1, respectively. When proof of cannabis use is required, any elution condition can be selected to match the possessing reference standards and analytical instruments. To our knowledge, this is the first report of using PBA SPE for the preparation of the urine samples containing cannabis and achieving partial derivatization when eluting from a PBA carrier. Our method can provide a new and practical solution for the preparation of the urine samples from cannabis users. Although the PBA SPE method cannot recover THC-COOH in urine because of its lack of a 1,2-diol moiety, this method has technological advantages for simplifying the process and reducing the operating time, thereby avoiding human errors. • We developed a quick preparation method for the analysis of urinary cannabis by phenylboronic acid solid-phase extraction. • Four pathways can provide four different kinds of 11-nor-9-carboxy-∆9-tetrahydrocannabinol derivatives, respectively. • All pathways showed good recovery rates and repeatability. • This method can simplify the process and shorten the operating time, resulting in contributing to reduce human errors. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
46. Drug-drug interactions that alter the exposure of glucuronidated drugs: Scope, UDP-glucuronosyltransferase (UGT) enzyme selectivity, mechanisms (inhibition and induction), and clinical significance.
- Author
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Miners, John O., Polasek, Thomas M., Hulin, Julie-Ann, Rowland, Andrew, and Meech, Robyn
- Subjects
- *
DRUG interactions , *GLUCURONOSYLTRANSFERASE , *DRUG accessibility , *ENZYMES , *DRUG development - Abstract
Drug-drug interactions (DDIs) arising from the perturbation of drug metabolising enzyme activities represent both a clinical problem and a potential economic loss for the pharmaceutical industry. DDIs involving glucuronidated drugs have historically attracted little attention and there is a perception that interactions are of minor clinical relevance. This review critically examines the scope and aetiology of DDIs that result in altered exposure of glucuronidated drugs. Interaction mechanisms, namely inhibition and induction of UDP-glucuronosyltransferase (UGT) enzymes and the potential interplay with drug transporters, are reviewed in detail, as is the clinical significance of known DDIs. Altered victim drug exposure arising from modulation of UGT enzyme activities is relatively common and, notably, the incidence and importance of UGT induction as a DDI mechanism is greater than generally believed. Numerous DDIs are clinically relevant, resulting in either loss of efficacy or an increased risk of adverse effects, necessitating dose individualisation. Several generalisations relating to the likelihood of DDIs can be drawn from the known substrate and inhibitor selectivities of UGT enzymes, highlighting the importance of comprehensive reaction phenotyping studies at an early stage of drug development. Further, rigorous assessment of the DDI liability of new chemical entities that undergo glucuronidation to a significant extent has been recommended recently by regulatory guidance. Although evidence-based approaches exist for the in vitro characterisation of UGT enzyme inhibition and induction, the availability of drugs considered appropriate for use as 'probe' substrates in clinical DDI studies is limited and this should be a research priority. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
47. Mechanistic Study on the Species Differences in Excretion Pathway of HR011303 in Humans and Rats
- Author
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Xiaoyan Chen, Guangze Li, Xingxing Diao, Zitao Guo, Mengling Liu, Jinghua Yu, Qi Huang, Jian Meng, Yaru Xue, Dafang Zhong, Yuandong Zheng, Zhendong Chen, and Yali Wu
- Subjects
Male ,medicine.medical_specialty ,Organic anion transporter 1 ,Metabolite ,Organic Anion Transporters ,Pharmaceutical Science ,Excretion ,chemistry.chemical_compound ,Glucuronides ,Species Specificity ,Internal medicine ,medicine ,Animals ,Humans ,Pharmacology ,Kidney ,biology ,Chemistry ,Multidrug resistance-associated protein 2 ,Metabolism ,Multidrug Resistance-Associated Protein 2 ,Rats ,medicine.anatomical_structure ,Endocrinology ,Liver ,Hepatocytes ,biology.protein ,Microsome ,Multidrug Resistance-Associated Proteins ,Glucuronide - Abstract
Excretion of [14C]HR011303-derived radioactivity showed significant species difference. Urine (81.50% of dose) was the main excretion route in healthy male subjects, whereas feces (87.16% of dose) was the main excretion route in rats. To further elucidate the underlying cause for excretion species differences of HR011303, studies were conducted to uncover its metabolism and excretion mechanism. M5, a glucuronide metabolite of HR011303, is the main metabolite in humans and rats. Results of rat microsomes incubation study suggested that HR011303 was metabolized to M5 in the rat liver. According to previous studies, M5 is produced in both human liver and kidney microsomes. We found M5 in human liver can be transported to the blood by multidrug resistance-associated protein (MRP) 3 and then the majority of M5 can be hydrolyzed to HR011303. HR011303 enters the human kidney or liver through passive diffusion, whereas M5 is taken up through organic anion transporter (OAT) 3, organic anion-transporting polypeptide (OATP) 1B1, and OATP1B3. When HR011303 alone was present, it can be metabolized to M5 in both sandwich-cultured rat hepatocytes (SCRH) and sandwich-cultured human hepatocytes (SCHH) and excreted into bile as M5 in SCRH. Using transporter inhibitors in sandwich-cultured model and membrane vesicles that expressing MRP2 or Mrp2, we found M5 was substance of MRP2/Mrp2 and the bile efflux of M5 mainly mediated by MRP2/Mrp2. Considering the significant role of MRP3/Mrp3 and MRP2/Mrp2 in the excretion of glucuronides, the competition between them for M5 was possibly the determinant for the different excretion routes in humans and rats. Significance Statement Animal experiments are necessary to predict dosage and safety of candidate drugs prior to clinical trials. However, extrapolation results often differ from actual situation. For HR011303, excretory pathways exhibited a complete reversal, through urine in humans and feces in rats. Such phenomena have been observed in several drugs, but no in-depth studies have been conducted to date. In the present study, the excretion species differences of HR011303 can be explained by the competition for M5 between MRP2/Mrp2 and MRP3/Mrp3.
- Published
- 2021
48. Identification and Quantification of MIDD0301 Metabolites
- Author
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Leggy A. Arnold, Nicolas M Zahn, James M. Cook, Daniel A. Webb, Brandon N Mikulsky, Yeunus Mian, Margaret L. Guthrie, M S Rashid Roni, Douglas C. Stafford, and Daniel E. Knutson
- Subjects
Metabolite ,Clinical Biochemistry ,Glucuronidation ,Administration, Oral ,Urine ,Pharmacology ,Kidney ,Article ,Mice ,chemistry.chemical_compound ,Dogs ,Pharmacokinetics ,Tandem Mass Spectrometry ,In vivo ,Oral administration ,Microsomes ,Animals ,Humans ,Tissue Distribution ,Anti-Asthmatic Agents ,Lung ,Chemistry ,Imidazoles ,Azepines ,Rats ,Microsomes, Liver ,Microsome ,Administration, Intravenous ,Female ,Glucuronide ,Injections, Intraperitoneal ,Chromatography, Liquid - Abstract
Background: MIDD0301 is an oral asthma drug candidate that binds GABAA receptors on airway smooth muscle and immune cells. Objective: The objective of this study is to identify and quantify MIDD0301 metabolites in vitro and in vivo and determine the pharmacokinetics of oral, IP, and IV administered MIDD0301. Methods: In vitro conversion of MIDD0301 was performed using liver and kidney microsomes/S9 fractions followed by quantification using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A LC-MS/MS method was developed using synthesized standards to quantify MIDD0301 and its metabolites in urine and feces. Blood, lung, and brain were harvested from animals that received MIDD0301 by oral, IP, and IV administration, followed by LCMS/ MS quantification. Imaging mass spectrometry was used to demonstrate the presence of MIDD0301 in the lung after oral administration. Results: MIDD0301 is stable in the presence of liver and kidney microsomes and S9 fractions for at least two hours. MIDD0301 undergoes conversion to the corresponding glucuronide and glucoside in the presence of conjugating cofactors. For IP and IV administration, unconjugated MIDD0301 together with significant amounts of MIDD0301 glucoside and MIDD0301 taurine were found in urine and feces. Less conjugation was observed following oral administration, with MIDD0301 glucuronide being the main metabolite. Pharmacokinetic quantification of MIDD0301 in blood, lung, and brain showed very low levels of MIDD0301 in the brain after oral, IV, or IP administration. The drug half-life in these tissues ranged between 4-6 hours for IP and oral and 1-2 hours for IV administration. Imaging mass spectrometry demonstrated that orally administered MIDD0301 distributes uniformly in the lung parenchyma. Conclusion: MIDD0301 undergoes no phase I and moderate phase II metabolism.
- Published
- 2021
49. Inhibitory Activity of 4-O-Benzoyl-3′-O-(OMethylsinapoyl) Sucrose from Polygala tenuifolia on Escherichia coli β-Glucuronidase
- Author
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Sung-Cheol Koo, Mok Hur, Yoon Jeong Lee, Le Ba Vinh, Youn-Ho Moon, Yun-Chan Huh, Young Ho Kim, Seo Young Yang, Jang Hoon Kim, and Woo Tae Park
- Subjects
chemistry.chemical_classification ,Sucrose ,biology ,General Medicine ,medicine.disease_cause ,biology.organism_classification ,Applied Microbiology and Biotechnology ,In vitro ,Glucuronidase ,chemistry.chemical_compound ,Enzyme ,chemistry ,Biochemistry ,Polygala tenuifolia ,medicine ,Uncompetitive inhibitor ,Glucuronide ,Escherichia coli ,Biotechnology - Abstract
Bacterial β-glucuronidase in the intestine is involved in the conversion of 7-ethyl-10- hydroxycamptochecin glucuronide (derived from irinotecan) to 7-ethyl-10-hydroxycamptothecin, which causes intestinal bleeding and diarrhea (side effects of anti-cancer drugs). Twelve compounds (1-12) from Polygala tenuifolia were evaluated in terms of β-glucuronidase inhibition in vitro. 4-O-Benzoyl-3'-O-(O-methylsinapoyl) sucrose (C3) was highly inhibitory at low concentrations. C3 (an uncompetitive inhibitor) exhibited a ki value of 13.4 μM; inhibitory activity increased as the substrate concentration rose. Molecular simulation revealed that C3 bound principally to the Gln158-Tyr160 enzyme loop. Thus, C3 will serve as a lead compound for development of new β- glucuronidase inhibitors.
- Published
- 2021
50. Functional characterization of Clonorchis sinensis sodium-bile acid co-transporter (CsSBAT) as a steroid sulfate transporter
- Author
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Jeong Yeon Won, Jin-Hee Han, Yun-Kyu Park, Haneul Jung, and Seok Ho Cha
- Subjects
medicine.drug_class ,Metabolite ,Biology ,Bile Acids and Salts ,chemistry.chemical_compound ,Dehydroepiandrosterone sulfate ,Estrone sulfate ,medicine ,Animals ,Steroid sulfate ,Sulfate ,Clonorchis sinensis ,Symporters ,General Veterinary ,Bile acid ,Sodium ,Transporter ,General Medicine ,Infectious Diseases ,chemistry ,Biochemistry ,Sulfate Transporters ,Insect Science ,Oocytes ,Parasitology ,Glucuronide - Abstract
Clonorchis sinensis (Cs) is a common trematode in Asian countries. Infection by Cs can result in many clinical symptoms. Here, a cDNA encoding a Cs apical sodium-dependent bile acid transporter (CsSBAT) was isolated from a Cs cDNA library, and functional characterization was performed using Xenopus laevis oocyte expression system. When expressed in Xenopus laevis oocytes, CsSBAT mediated the transport of radiolabeled estrone sulfate and dehydroepiandrosterone sulfate. No trans-uptake of carnitine, estradiol 17 β-D glucuronide, prostaglandin E2, p-aminohippuric acid, α-ketoglutaric acid, and tetraethylammonium was observed. CsSBAT-mediated estrone sulfate uptake was in a time- and sodium-dependent manner. CsSBAT showed no exchange properties in efflux experiments. Concentration-dependent results showed saturable kinetics consistent with the Michaelis–Menten equation. Nonlinear regression analyses yielded a Km value of 0.3 ± 0.04 μM for [3H]estrone sulfate. CsSBAT-mediated estrone sulfate uptake was strongly inhibited by sulfate conjugates but not glucuronide conjugates. These findings contribute to our understanding of CsSBAT transport properties and the cascade of estrogen metabolite movement in Cs.
- Published
- 2021
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