FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2015215157> ?p ?o ?g. }

• W2015215157 endingPage "226" @default.
• W2015215157 startingPage "215" @default.
• W2015215157 abstract "The standard method to evaluate CYP3A inhibition is to study the conversion of the specific CYP3A probe testosterone to its 6 beta-hydroxy metabolite in human liver microsomes, in the absence and presence of potential inhibitors. Quantification of the 6 beta-hydroxy metabolite is achieved by HPLC resulting in a tedious and time-consuming assay. In order to increase the P450 inhibition throughput, efforts were made to find a CYP3A probe that would produce a fluorescent metabolite. This paper reports the discovery of DFB as a potential CYP3A fluorescent probe. DFB was significantly metabolized in human microsomes (approximately 1-2 nmol/(min. mg protein)) to give the fluorescent compound DFH. The involvement of CYP3A in the metabolism of DFB was determined using multiple approaches. First, incubations conducted with microsomes made from cell lines expressing single CYPs (Gentest Supersomes) indicated that CYP3A played a major role in the metabolism of DFB. Secondly, immunoinhibition studies conducted with CYP3A antibody resulted in >95% inhibition of DFB metabolism in HLM. Thirdly, inhibition studies with specific CYP1A1, 1A2, 2C8/9, 2C19, 2D6, and 2E1 chemical inhibitors did not suppress DFB activity in HLM. However, ketoconazole, miconazole, nicardipine, and nifedipine, all known CYP3A inhibitors, completely abolished the formation of DFH in HLM. The potency of several inhibitors determined using DFB and testosterone as CYP3A probes was consistent (R = 0.98). Finally, a good agreement was obtained for the formation of DFH and production of 6 beta-hydroxytestosterone when DFB and testosterone were incubated separately with various human liver microsome preparations (R = 0.94, N = 11). In order to use DFH as a fluorescent CYP3A marker in a 96-well plate format, it was important to remove the excess of NADPH at the end of the incubation because the fluorescence of NADPH interferes with DFH detection. This was achieved by adding oxidized glutathione and glutathione reductase to convert NADPH to NADP(+) which is not fluorescent. The liquid-handling steps were fully automated in a 96-well plate format and a template was designed to generate IC(50) curves and to address potential fluorescent interferences from the test compounds. The assay was found to be reproducible (intraday variability <10% and interday variability indicated less than a 2-fold variation in the IC(50) values) and is now routinely used in our laboratory to evaluate CYP3A inhibition of NCEs." @default.
• W2015215157 created "2016-06-24" @default.
• W2015215157 creator A5005665758 @default.
• W2015215157 creator A5010374726 @default.
• W2015215157 creator A5012243268 @default.
• W2015215157 creator A5028459087 @default.
• W2015215157 creator A5032579475 @default.
• W2015215157 creator A5051128694 @default.
• W2015215157 creator A5071176619 @default.
• W2015215157 creator A5075874590 @default.
• W2015215157 date "1999-12-01" @default.
• W2015215157 modified "2023-10-16" @default.
• W2015215157 title "Description of a 96-Well Plate Assay to Measure Cytochrome P4503A Inhibition in Human Liver Microsomes Using a Selective Fluorescent Probe" @default.
• W2015215157 cites W173148493 @default.
• W2015215157 cites W1775749144 @default.
• W2015215157 cites W2009649574 @default.
• W2015215157 cites W2021022873 @default.
• W2015215157 cites W2030089759 @default.
• W2015215157 cites W2032794920 @default.
• W2015215157 cites W2043209699 @default.
• W2015215157 cites W2044695596 @default.
• W2015215157 cites W2051175552 @default.
• W2015215157 cites W2060365395 @default.
• W2015215157 cites W2106245429 @default.
• W2015215157 cites W2151724311 @default.
• W2015215157 doi "https://doi.org/10.1006/abio.1999.4348" @default.
• W2015215157 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/10603245" @default.
• W2015215157 hasPublicationYear "1999" @default.
• W2015215157 type Work @default.
• W2015215157 sameAs 2015215157 @default.
• W2015215157 citedByCount "92" @default.
• W2015215157 countsByYear W20152151572012 @default.
• W2015215157 countsByYear W20152151572013 @default.
• W2015215157 countsByYear W20152151572014 @default.
• W2015215157 countsByYear W20152151572015 @default.
• W2015215157 countsByYear W20152151572016 @default.
• W2015215157 countsByYear W20152151572017 @default.
• W2015215157 countsByYear W20152151572018 @default.
• W2015215157 countsByYear W20152151572019 @default.
• W2015215157 countsByYear W20152151572020 @default.
• W2015215157 countsByYear W20152151572021 @default.
• W2015215157 countsByYear W20152151572022 @default.
• W2015215157 countsByYear W20152151572023 @default.
• W2015215157 crossrefType "journal-article" @default.
• W2015215157 hasAuthorship W2015215157A5005665758 @default.
• W2015215157 hasAuthorship W2015215157A5010374726 @default.
• W2015215157 hasAuthorship W2015215157A5012243268 @default.
• W2015215157 hasAuthorship W2015215157A5028459087 @default.
• W2015215157 hasAuthorship W2015215157A5032579475 @default.
• W2015215157 hasAuthorship W2015215157A5051128694 @default.
• W2015215157 hasAuthorship W2015215157A5071176619 @default.
• W2015215157 hasAuthorship W2015215157A5075874590 @default.
• W2015215157 hasConcept C11824378 @default.
• W2015215157 hasConcept C185592680 @default.
• W2015215157 hasConcept C202751555 @default.
• W2015215157 hasConcept C2777477808 @default.
• W2015215157 hasConcept C2779548794 @default.
• W2015215157 hasConcept C2781064554 @default.
• W2015215157 hasConcept C526171541 @default.
• W2015215157 hasConcept C55493867 @default.
• W2015215157 hasConcept C62231903 @default.
• W2015215157 hasConcept C86803240 @default.
• W2015215157 hasConcept C87644729 @default.
• W2015215157 hasConcept C89423630 @default.
• W2015215157 hasConcept C98274493 @default.
• W2015215157 hasConceptScore W2015215157C11824378 @default.
• W2015215157 hasConceptScore W2015215157C185592680 @default.
• W2015215157 hasConceptScore W2015215157C202751555 @default.
• W2015215157 hasConceptScore W2015215157C2777477808 @default.
• W2015215157 hasConceptScore W2015215157C2779548794 @default.
• W2015215157 hasConceptScore W2015215157C2781064554 @default.
• W2015215157 hasConceptScore W2015215157C526171541 @default.
• W2015215157 hasConceptScore W2015215157C55493867 @default.
• W2015215157 hasConceptScore W2015215157C62231903 @default.
• W2015215157 hasConceptScore W2015215157C86803240 @default.
• W2015215157 hasConceptScore W2015215157C87644729 @default.
• W2015215157 hasConceptScore W2015215157C89423630 @default.
• W2015215157 hasConceptScore W2015215157C98274493 @default.
• W2015215157 hasIssue "2" @default.
• W2015215157 hasLocation W20152151571 @default.
• W2015215157 hasLocation W20152151572 @default.
• W2015215157 hasOpenAccess W2015215157 @default.
• W2015215157 hasPrimaryLocation W20152151571 @default.
• W2015215157 hasRelatedWork W1988423359 @default.
• W2015215157 hasRelatedWork W2015972641 @default.
• W2015215157 hasRelatedWork W2036206840 @default.
• W2015215157 hasRelatedWork W2042554554 @default.
• W2015215157 hasRelatedWork W2053532436 @default.
• W2015215157 hasRelatedWork W2064563583 @default.
• W2015215157 hasRelatedWork W2069964545 @default.
• W2015215157 hasRelatedWork W2079895414 @default.
• W2015215157 hasRelatedWork W2138650407 @default.
• W2015215157 hasRelatedWork W2380122696 @default.
• W2015215157 hasVolume "276" @default.
• W2015215157 isParatext "false" @default.
• W2015215157 isRetracted "false" @default.
• W2015215157 magId "2015215157" @default.
• W2015215157 workType "article" @default.