[{"id":124,"uid":"NPDI-9C7QHg","name":"Effects of mitragynine and 7-hydroxymitragynine (the alkaloids of Mitragyna speciosa Korth) on 4-methylumbelliferone glucuronidation in rat and human liver microsomes and recombinant human uridine 5’- diphospho-glucuronosyltransferase isoforms","napdiIdentifier":null,"overallSummary":"

Background:  Glucuronidation catalyzed by uridine 5’- diphospho-glucuronosyltransferase (UGT) is a major phase II drug metabolism reaction which facilitates drug elimination. Inhibition of UGT activity can cause drug-drug interaction. Therefore, it is important to determine the inhibitory potentials of drugs on glucuronidation. 

\r\n

Objective: The objective was to evaluate the inhibitory potentials of mitragynine, 7-hydroxymitragynine, ketamine and buprenorphine, respectively on 4-methylumbelliferone (4-MU) glucuronidation in rat liver microsomes, human liver microsomes and recombinant human UGT1A1 and UGT2B7 isoforms.

\r\n

Materials and Methods: The effects of the above four compounds on the formation of 4-MU glucuronide from 4-MU by rat liver microsomes, human liver microsomes, recombinant human UGT1A1 and UGT2B7 isoforms were determined using high-performance liquid chromatography with ultraviolet detection. 

\r\n

Results: For rat liver microsomes, ketamine strongly inhibited 4-MU glucuronidation with an IC50 value of 6.21 ± 1.51 μM followed by buprenorphine with an IC50 value of 73.22 ± 1.63 μM. For human liver microsomes, buprenorphine strongly inhibited 4-MU glucuronidation with an IC50 value of 6.32 ± 1.39 μM. For human UGT1A1 isoform, 7-hydroxymitragynine strongly inhibited 4-MU glucuronidation with an IC50 value of 7.13 ± 1.16 μM. For human UGT2B7 isoform, buprenorphine strongly inhibited 4-MU glucuronidation followed by 7-hydroxymitragynine and ketamine with respective IC50 values of 5.14 ± 1.30, 26.44 ± 1.31, and 27.28 ± 1.18 μM.

\r\n

Conclusions: These data indicate the possibility of drug-drug interaction if 7-hydroxymitragynine, ketamine, and buprenorphine are co-administered with drugs that are UGT2B7 substrates since these three compounds showed significant inhibition on UGT2B7 activity. In addition, if 7-hydroxymitragynine is to be taken with other drugs that are highly metabolized by UGT1A1, there is a possibility of drug-drug interaction to occur.

","pubmedId":26692748,"embaseId":2015485354,"croIdentifier":"Centre for Drug Research","croInformation":"Universiti Sains Malaysia","dateStart":null,"dateEnd":null,"internalComment":"Positive data entered","status":"published","compoundId":null,"naturalProductUid":"NP-00ed1235-cbd8-4117-85df-298b8b3cdcad","naturalProductSampleId":null,"studySourceTypeId":1,"naturalProduct":{"uid":"NP-00ed1235-cbd8-4117-85df-298b8b3cdcad","binomial":"Mitragyna speciosa","name":"Kratom","itis":null,"srs":"d469b67d-e9a6-459f-b209-c59451936336","source_id":"","conceptId":null},"compound":null,"studySourceType":{"id":1,"name":"Published report"},"experiments":[{"id":578,"uid":"NPDI-Nl2Cww","name":"Negligible inhibition of UGT in human liver microsomes by mitragynine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"Pooled human liver microsomes were purchased from Sigma-Aldrich; The enzyme activity assay mixture (250 μL) contained Tris-HCl (100 mM) (pH 7.4), MgCl (5 mM), microsomal protein (0.1 mg/mL), Triton X-100 (0.005%), UDPGA (3 mM), and 4-MU (100 μM).

Tested concentrations of precipitant only up to 100 μM due to limited solubility

Control=no inhibitor","resultsComment":"The IC50 values were all greater than the highest concentrations used (IC50 > 100 μM) since inhibition at more than 50% did not occur at the highest concentration.

Concentration of precipitant and % inhibition of pooled human liver microsomes (intermediate values estimated from figure 3) (p < 0.05 for all values)
0.01 μM: 16 ± 3%
0.1 μM: 26 ± 3%
1 μM: 29 ± 3%
10 μM:30 ± 2%
100 μM: See above data","internalComment":null,"objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":94,"cytochromeB5Id":null,"studyId":124,"experimentTypeId":1,"testSystemId":6,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":94,"name":"mitragynine","unii":null,"inChIKey":"LELBFTMXCIIKKX-QVRQZEMUSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":27,"name":"UGT","conceptId":null,"experiment_enzyme_xref":{"enzymeId":27,"experimentId":578}}],"transporters":[],"quantifiedMetabolites":[]},{"id":579,"uid":"NPDI-Q5Hs5A","name":"No inhibition of UGT in human liver microsome activity by 7-hydroxymitragynine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"Pooled human liver microsomes were purchased from Sigma-Aldrich; The enzyme activity assay mixture (250 μL) contained Tris-HCl (100 mM) (pH 7.4), MgCl (5 mM), microsomal protein (0.1 mg/mL), Triton X-100 (0.005%), UDPGA (3 mM), and 4-MU (100 μM).

Tested concentrations of precipitant only up to 100 μM due to limited solubility

Control=no inhibitor","resultsComment":"The IC50 values were all greater than the highest concentrations used (IC50 > 100 μM) since inhibition at more than 50% did not occur at the highest concentration.

Concentration of precipitant and % inhibition of pooled human liver microsomes (intermediate values estimated from figure 3)
0.01 μM: 5 ± 1%
0.1 μM: 9 ± 5%
1 μM: 10 ± 2% (P < 0.05)
10 μM:10 ± 3% (P < 0.05)
100 μM: See above data","internalComment":null,"objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":104,"cytochromeB5Id":null,"studyId":124,"experimentTypeId":1,"testSystemId":6,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":104,"name":"7-hydroxymitragynine","unii":null,"inChIKey":"RYENLSMHLCNXJT-CYXFISRXSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":27,"name":"UGT","conceptId":null,"experiment_enzyme_xref":{"enzymeId":27,"experimentId":579}}],"transporters":[],"quantifiedMetabolites":[]},{"id":580,"uid":"NPDI-ipamgg","name":"No inhibition of UGT in human liver microsomes by ketamine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"Pooled human liver microsomes were purchased from Sigma-Aldrich; The enzyme activity assay mixture (250 μL) contained Tris-HCl (100 mM) (pH 7.4), MgCl (5 mM), microsomal protein (0.1 mg/mL), Triton X-100 (0.005%), UDPGA (3 mM), and 4-MU (100 μM).

Control=no inhibitor","resultsComment":"The IC50 values were all greater than the highest concentrations used (IC50 > 1000 μM) since inhibition at more than 50% did not occur at the highest concentration.

Concentration of precipitant and % inhibition of pooled human liver microsomes (intermediate values estimated from figure 3)
0.01 μM: 8 ± 3%
0.1 μM: 16 ± 1% (p<0.05)
1 μM: 18 ± 2% (p<0.05)
10 μM: 21 ± 4% (p<0.05)
100 μM: 21 ± 2% (p<0.05)
1000 μM: See above data","internalComment":null,"objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":191,"cytochromeB5Id":null,"studyId":124,"experimentTypeId":1,"testSystemId":6,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":191,"name":"ketamine","unii":null,"inChIKey":"YQEZLKZALYSWHR-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":27,"name":"UGT","conceptId":null,"experiment_enzyme_xref":{"enzymeId":27,"experimentId":580}}],"transporters":[],"quantifiedMetabolites":[]},{"id":581,"uid":"NPDI-pMOhyQ","name":"Inhibition of UGT in human liver microsomes by buprenorphine","overallEffect":1,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"Pooled human liver microsomes were purchased from Sigma-Aldrich; The enzyme activity assay mixture (250 μL) contained Tris-HCl (100 mM) (pH 7.4), MgCl (5 mM), microsomal protein (0.1 mg/mL), Triton X-100 (0.005%), UDPGA (3 mM), and 4-MU (100 μM).

Control=no inhibitor","resultsComment":"Concentration of precipitant and % inhibition of pooled human liver microsomes (values estimated from figure 3) (p<0.05 for all values)
0.01 μM: 27 ± 2%
0.1 μM: 39 ± 1%
1 μM: 43 ± 3%
10 μM: 55 ± 1%
100 μM: 61 ± 2%
1000 μM: See above data","internalComment":null,"objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":192,"cytochromeB5Id":null,"studyId":124,"experimentTypeId":1,"testSystemId":6,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":192,"name":"buprenorphine","unii":null,"inChIKey":"RMRJXGBAOAMLHD-IHFGGWKQSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":27,"name":"UGT","conceptId":null,"experiment_enzyme_xref":{"enzymeId":27,"experimentId":581}}],"transporters":[],"quantifiedMetabolites":[]},{"id":582,"uid":"NPDI-u96XKg","name":"Negligible inhibition of UGT in human liver microsomes by diclofenac","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"Pooled human liver microsomes were purchased from Sigma-Aldrich; The enzyme activity assay mixture (250 μL) contained Tris-HCl (100 mM) (pH 7.4), MgCl (5 mM), microsomal protein (0.1 mg/mL), Triton X-100 (0.005%), UDPGA (3 mM), and 4-MU (100 μM).

Control=no inhibitor","resultsComment":"Concentration of precipitant and % inhibition of pooled human liver microsomes (values estimated from figure 3) (p<0.05 for all values)
0.01 μM: 8 ± 3%
0.1 μM: 9 ± 2%
1 μM: 20 ± 1%
10 μM: 21 ± 1%
100 μM: 41 ± 1%
1000 μM: See above data","internalComment":null,"objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":14,"cytochromeB5Id":null,"studyId":124,"experimentTypeId":1,"testSystemId":6,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":14,"name":"diclofenac","unii":"144O8QL0L1","inChIKey":"DCOPUUMXTXDBNB-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":1124300,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":27,"name":"UGT","conceptId":null,"experiment_enzyme_xref":{"enzymeId":27,"experimentId":582}}],"transporters":[],"quantifiedMetabolites":[]},{"id":583,"uid":"NPDI-5VhYlA","name":"Negligible inhibition of UGT1A1 by mitragynine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Tested concentrations of precipitant only up to 100 μM due to limited solubility

Control=no inhibitor","resultsComment":"The IC50 values were all greater than the highest concentrations used (IC50 > 100 μM) since inhibition at more than 50% did not occur at the highest concentration.

Concentration of precipitant and % inhibition of UGT1A1 (some values estimated from figure 4)
0.01 μM: -4 ± 0%
0.1 μM: 8 ± 6%
1 μM: 15 ± 4% (p < 0.05)
10 μM:19 ± 0% (p < 0.05)
100 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":94,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":94,"name":"mitragynine","unii":null,"inChIKey":"LELBFTMXCIIKKX-QVRQZEMUSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":17,"name":"UGT1A1","conceptId":40782950,"experiment_enzyme_xref":{"enzymeId":17,"experimentId":583}}],"transporters":[],"quantifiedMetabolites":[]},{"id":584,"uid":"NPDI-4do0FA","name":"Inhibition of UGT1A1 by 7-hydroxymitragynine","overallEffect":1,"isControlData":false,"isIc50Shift":false,"croCutoff":"IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Tested concentrations of precipitant only up to 100 μM due to limited solubility

Control=no inhibitor","resultsComment":"Concentration of precipitant and % inhibition of UGT1A1 (values estimated from figure 4)
0.01 μM: 6 ± 1%
0.1 μM: 17 ± 1% (p < 0.05)
1 μM: 26 ± 2% (p < 0.05)
10 μM:49 ± 3% (p < 0.05)
100 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":104,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":104,"name":"7-hydroxymitragynine","unii":null,"inChIKey":"RYENLSMHLCNXJT-CYXFISRXSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":17,"name":"UGT1A1","conceptId":40782950,"experiment_enzyme_xref":{"enzymeId":17,"experimentId":584}}],"transporters":[],"quantifiedMetabolites":[]},{"id":585,"uid":"NPDI-Ve-SYA","name":"No inhibition of UGT1A1 by ketamine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Control=no inhibitor","resultsComment":"The IC50 values were all greater than the highest concentrations used (IC50 > 1000 μM) since inhibition at more than 50% did not occur at the highest concentration.

Concentration of precipitant and % inhibition of UGT1A1 (intermediate values estimated from figure 4)
0.01 μM: 9 ± 1%
0.1 μM: 14 ± 2% (p < 0.05)
1 μM: 17 ± 5% (p < 0.05)
10 μM:18 ± 3% (p < 0.05)
100 μM: 20 ± 1% (p < 0.05)
1000 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":191,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":191,"name":"ketamine","unii":null,"inChIKey":"YQEZLKZALYSWHR-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":17,"name":"UGT1A1","conceptId":40782950,"experiment_enzyme_xref":{"enzymeId":17,"experimentId":585}}],"transporters":[],"quantifiedMetabolites":[]},{"id":586,"uid":"NPDI-3eovVg","name":"Negligible inhibition of UGT1A1 by buprenorphine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Control=no inhibitor","resultsComment":"The IC50 values were all greater than the highest concentrations used (IC50 > 1000 μM) since inhibition at more than 50% did not occur at the highest concentration.

Concentration of precipitant and % inhibition of UGT1A1 (intermediate values estimated from figure 4)
0.01 μM: 12 ± 4%
0.1 μM: 24 ± 2% (p < 0.05)
1 μM: 25 ± 3% (p < 0.05)
10 μM:32 ± 2% (p < 0.05)
100 μM: 35 ± 3% (p < 0.05)
1000 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":192,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":192,"name":"buprenorphine","unii":null,"inChIKey":"RMRJXGBAOAMLHD-IHFGGWKQSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":17,"name":"UGT1A1","conceptId":40782950,"experiment_enzyme_xref":{"enzymeId":17,"experimentId":586}}],"transporters":[],"quantifiedMetabolites":[]},{"id":587,"uid":"NPDI-4_m2Gw","name":"Inhibition of UGT1A1 by diclofenac","overallEffect":1,"isControlData":false,"isIc50Shift":false,"croCutoff":"IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Control=no inhibitor","resultsComment":"Concentration of precipitant and % inhibition of UGT1A1 (values estimated from figure 4) (p < 0.05 for all values)
0.01 μM: 17 ± 4%
0.1 μM: 32 ± 2%
1 μM: 47 ± 2%
10 μM:55 ± 3%
100 μM: 69 ± 2%
1000 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":14,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":14,"name":"diclofenac","unii":"144O8QL0L1","inChIKey":"DCOPUUMXTXDBNB-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":1124300,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":17,"name":"UGT1A1","conceptId":40782950,"experiment_enzyme_xref":{"enzymeId":17,"experimentId":587}}],"transporters":[],"quantifiedMetabolites":[]},{"id":588,"uid":"NPDI-9spGvg","name":"Negligible inhibition of UGT2B7 by mitragynine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Tested concentrations of precipitant only up to 100 μM due to limited solubility

Control=no inhibitor","resultsComment":"The IC50 values were all greater than the highest concentrations used (IC50 > 100 μM) since inhibition at more than 50% did not occur at the highest concentration.

Concentration of precipitant and % inhibition of UGT2B7 (intermediate values estimated from figure 5)
0.01 μM: 8 ± 4%
0.1 μM: 11 ± 2%
1 μM: 23 ± 0% (p < 0.05)
10 μM:36 ± 5% (p < 0.05)
100 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":94,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":94,"name":"mitragynine","unii":null,"inChIKey":"LELBFTMXCIIKKX-QVRQZEMUSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":22,"name":"UGT2B7","conceptId":null,"experiment_enzyme_xref":{"enzymeId":22,"experimentId":588}}],"transporters":[],"quantifiedMetabolites":[]},{"id":589,"uid":"NPDI-3C6gmg","name":"Negligible inhibition of UGT2B7 by 7-hydroxymitragynine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Tested concentrations of precipitant only up to 100 μM due to limited solubility

Control=no inhibitor","resultsComment":"Concentration of precipitant and % inhibition of UGT2B7 (values estimated from figure 5)
0.01 μM: 3 ± 5%
0.1 μM: 13 ± 2% (p < 0.05)
1 μM: 26 ± 5% (p < 0.05)
10 μM:33 ± 5% (p < 0.05)
100 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":104,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":104,"name":"7-hydroxymitragynine","unii":null,"inChIKey":"RYENLSMHLCNXJT-CYXFISRXSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":22,"name":"UGT2B7","conceptId":null,"experiment_enzyme_xref":{"enzymeId":22,"experimentId":589}}],"transporters":[],"quantifiedMetabolites":[]},{"id":590,"uid":"NPDI-bX8XPA","name":"No inhibition of UGT2B7 by ketamine","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Control=no inhibitor","resultsComment":"Concentration of precipitant and % inhibition of UGT2B7 (values estimated from figure 5)
0.01 μM: -5 ± 6%
0.1 μM: 4 ± 5%
1 μM: 11 ± 5%
10 μM: 34 ± 3% (p < 0.05)
100 μM: 70 ± 1% (p < 0.05)
1000 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":191,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":191,"name":"ketamine","unii":null,"inChIKey":"YQEZLKZALYSWHR-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":22,"name":"UGT2B7","conceptId":null,"experiment_enzyme_xref":{"enzymeId":22,"experimentId":590}}],"transporters":[],"quantifiedMetabolites":[]},{"id":591,"uid":"NPDI-ZTY8Qw","name":"Inhibition of UGT2B7 by buprenorphine","overallEffect":1,"isControlData":false,"isIc50Shift":false,"croCutoff":"IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Control=no inhibitor","resultsComment":"Concentration of precipitant and % inhibition of UGT2B7 (values estimated from figure 5)
0.01 μM: -8 ± 4%
0.1 μM: 19 ± 3% (p < 0.05)
1 μM: 31 ± 7% (p < 0.05)
10 μM: 61 ± 3% (p < 0.05)
100 μM: 81± 3% (p < 0.05)
1000 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":192,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":192,"name":"buprenorphine","unii":null,"inChIKey":"RMRJXGBAOAMLHD-IHFGGWKQSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":22,"name":"UGT2B7","conceptId":null,"experiment_enzyme_xref":{"enzymeId":22,"experimentId":591}}],"transporters":[],"quantifiedMetabolites":[]},{"id":592,"uid":"NPDI-NZQ3aw","name":"Negligible inhibition of UGT2B7 by diclofenac","overallEffect":0,"isControlData":false,"isIc50Shift":false,"croCutoff":"

IC50 > 10 μM are low potential inhibitors.
1 μM < IC50 < 10 μM are moderate potential inhibitors. 
IC50 < 1 μM are high potential inhibitors.

","croIdentifier":null,"comment":null,"experimentalConditionsComment":"The amount of protein, the incubation time and the concentration of 4-MU used in the measurement of 4-MU glucuronidation were described in the method by Uchaipichat et al.

Microsomes expressing recombinant human UGT1A1 and UGT2B7 were obtained from BD Biosciences (Woburn, MA, United States);incubations (total volume 250 μL) contained phosphate buffer (100 mM) (pH 7.4), MgCl 2 (5 mM), cell lysate (83.3 μg/mL for UGT1A1 or 62.5 μg/mL for UGT2B7), UDPGA (5 mM), and 4-MU (substrate) (100 μM for UGT1A1 assay or 350 μM for UGT2B7 assay).

Control=no inhibitor","resultsComment":"Concentration of precipitant and % inhibition of UGT2B7 (values estimated from figure 5)
0.01 μM: -1 ± 3%
0.1 μM: 3 ± 5%
1 μM: 8 ± 4%
10 μM: 38 ± 2% (p < 0.05)
100 μM: 84± 1% (p < 0.05)
1000 μM: See above data","internalComment":"Experimental conditions tab: Uchaipichat et al  can be found here http://dmd.aspetjournals.org/content/dmd/32/4/413.full.pdf","objectCompoundId":70,"objectMetaboliteCompoundId":73,"precipitantCompoundId":14,"cytochromeB5Id":4,"studyId":124,"experimentTypeId":1,"testSystemId":14,"ic50ShiftExperimentId":null,"controlDataExperimentId":null,"controlDataForExperimentId":null,"naturalProductSampleId":null,"experimentType":{"id":1,"name":"In Vitro Enzyme Inhibition","isInVitro":true,"isTransporter":false,"isEnzyme":true,"purl":"http://purl.obolibrary.org/obo/DIDEO_00000058"},"objectCompound":{"id":70,"name":"4-methylumbelliferone","unii":null,"inChIKey":"HSHNITRMYYLLCV-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"precipitantCompound":{"id":14,"name":"diclofenac","unii":"144O8QL0L1","inChIKey":"DCOPUUMXTXDBNB-UHFFFAOYSA-N","publicDescription":null,"internalComment":null,"conceptId":1124300,"enantiomerOfId":null},"objectMetaboliteCompound":{"id":73,"name":"4-methylumbelliferone glucuronide","unii":null,"inChIKey":"ARQXEQLMMNGFDU-JHZZJYKESA-N","publicDescription":null,"internalComment":null,"conceptId":null,"enantiomerOfId":null},"enzymes":[{"id":22,"name":"UGT2B7","conceptId":null,"experiment_enzyme_xref":{"enzymeId":22,"experimentId":592}}],"transporters":[],"quantifiedMetabolites":[]}]}]