{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T01:00:34Z","timestamp":1777424434163,"version":"3.51.4"},"reference-count":25,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2020,9,7]],"date-time":"2020-09-07T00:00:00Z","timestamp":1599436800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Vet Pharm &amp; Therapeutics"],"published-print":{"date-parts":[[2020,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>In humans, the cytochrome P450 3A (CYP3A) subfamily is involved in midazolam (MDZ) biotransformation into 1\u2032\u2010 and 4\u2010hydroxy metabolites, and the former serves as a probe for CYP3A catalytic activity. In veterinary species is still crucial to identify enzyme\u2010 and species\u2010specific CYP substrates; thus, the aim of this study was to characterize MDZ oxidation in cattle liver. A HPLC\u2010UV method was used to measure 1\u2032\u2010 and 4\u2010hydroxy MDZ (1\u2032\u2010 and 4\u2010OHMDZ, respectively) formation in cattle liver microsomes and assess the role of CYP3A by an immunoinhibition study. Moreover, MDZ hydroxylation was evaluated in 300 cattle liver samples and results were correlated with testosterone hydroxylation. Formation of both metabolites conformed to a single\u2010enzyme Michaelis\u2013Menten kinetics. Values of<jats:italic>V<jats:sub>max<\/jats:sub><\/jats:italic>and<jats:italic>K<jats:sub>m<\/jats:sub><\/jats:italic>were 0.67\u00a0nmol\/min\/mg protein and 6.16\u00a0\u03bcM for 4\u2010OHMDZ, and 0.06\u00a0nmol\/min\/mg protein and 10.08\u00a0\u03bcM for 1\u2032\u2010OHMDZ. An anti\u2010rat CYP3A1 polyclonal antibody inhibited up to 50% and 94% 1\u2032\u2010 and 4\u2010OHMDZ formation, respectively. MDZ oxidation in liver microsomes was poorly correlated with testosterone hydroxylation. In conclusion, cattle metabolized MDZ to 1\u2032\u2010OHMDZ and 4\u2010OHMDZ. The immunoinhibition results indicated a major contribution of CYP3As to 4\u2010OHMDZ formation and the involvement of other CYPs in 1\u2032\u2010OHMDZ production, paving the way for further investigations.<\/jats:p>","DOI":"10.1111\/jvp.12906","type":"journal-article","created":{"date-parts":[[2020,11,9]],"date-time":"2020-11-09T19:43:26Z","timestamp":1604951006000},"page":"608-613","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Midazolam oxidation in cattle liver microsomes: The role of cytochrome P450 3A"],"prefix":"10.1111","volume":"43","author":[{"given":"Alberto","family":"Nassi","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical and Pharmacological Sciences University of Padua Padua Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luigi","family":"Quintieri","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical and Pharmacological Sciences University of Padua Padua Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberta","family":"Merlanti","sequence":"additional","affiliation":[{"name":"Department of Comparative Biomedicine and Food Science University of Padua Padua Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Pezzato","sequence":"additional","affiliation":[{"name":"Department of Comparative Biomedicine and Food Science University of Padua Padua Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Capolongo","sequence":"additional","affiliation":[{"name":"Department of Comparative Biomedicine and Food Science University of Padua Padua Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marianna","family":"Pauletto","sequence":"additional","affiliation":[{"name":"Department of Comparative Biomedicine and Food Science University of Padua Padua Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3637-5643","authenticated-orcid":false,"given":"Mauro","family":"Dacasto","sequence":"additional","affiliation":[{"name":"Department of Comparative Biomedicine and Food Science University of Padua Padua Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0015-3800","authenticated-orcid":false,"given":"Mery","family":"Giantin","sequence":"additional","affiliation":[{"name":"Department of Comparative Biomedicine and Food Science University of Padua Padua Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,9,7]]},"reference":[{"key":"e_1_2_5_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejphar.2008.03.013"},{"key":"e_1_2_5_3_1","first-page":"940","article-title":"Inhibition and kinetics of cytochrome P4503A activity in microsomes from rat, human, and cdna\u2010expressed human cytochrome P450","volume":"24","author":"Ghosal A.","year":"1996","journal-title":"Drug Metabolism and Disposition"},{"key":"e_1_2_5_4_1","doi-asserted-by":"publisher","DOI":"10.1124\/dmd.107.019042"},{"key":"e_1_2_5_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-56271-8"},{"key":"e_1_2_5_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-2952(94)90543-6"},{"key":"e_1_2_5_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/jvp.12163"},{"key":"e_1_2_5_8_1","doi-asserted-by":"publisher","DOI":"10.1124\/dmd.30.1.63"},{"key":"e_1_2_5_9_1","doi-asserted-by":"publisher","DOI":"10.3109\/00498254.2014.1002122"},{"key":"e_1_2_5_10_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2885.2001.00362.x"},{"key":"e_1_2_5_11_1","doi-asserted-by":"publisher","DOI":"10.3109\/00498250903540875"},{"key":"e_1_2_5_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tvjl.2008.08.006"},{"key":"e_1_2_5_13_1","first-page":"618","article-title":"Midazolam and triazolam biotransformation in mouse and human liver microsomes: relative contribution of CYP3A and CYP2C isoforms","volume":"292","author":"Perloff M. 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