Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking

Kweh, Jia Rong; Ming, Nicholas Kai; Ngoh, Le Min; Yan, Cynthia Jing; Tan, Bao Jie; Tan, Wee Kiat; Mettu, Vijaya Saradhi; Austin-Muttitt, Karl; Mullins, Jonathan; Lau, Aik Jiang

doi:10.1016/j.molpha.2025.100097

Journal article 36 views 8 downloads

Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking

Jia Rong Kweh, Nicholas Kai Ming Ng, Le Min Ngoh, Cynthia Jing Yan Li, Bao Jie Tan, Wee Kiat Tan

, Vijaya Saradhi Mettu, Karl Austin-Muttitt, Jonathan Mullins

, Aik Jiang Lau

Molecular Pharmacology, Volume: 108, Issue: 1, Start page: 100097

Swansea University Authors: Karl Austin-Muttitt, Jonathan Mullins

PDF | Version of Record

© 2025 The Authors. Published by Elsevier Inc. on behalf of American Society for Pharmacology and Experimental Therapeutics. This is an open access article under the CC BY license.
Download (6.7MB)

Check full text

DOI (Published version): 10.1016/j.molpha.2025.100097

Abstract

Aldehyde oxidase (AOX1) is a cytosolic molybdo-flavoenzyme that metabolizes azaheterocyclic drugs. Erlotinib and gefitinib are azaheterocyclic drugs. We deployed structural analogs to investigate the molecular interaction between these drugs and AOX1. Erlotinib, O-desmethylerlotinib, and O-didesmeth...

Full description

Published in:	Molecular Pharmacology
ISSN:	0026-895X
Published:	Elsevier BV 2026
Online Access:	Check full text
URI:	https://cronfa.swan.ac.uk/Record/cronfa71215

first_indexed	2026-01-08T14:34:47Z
last_indexed	2026-01-09T05:32:19Z
id	cronfa71215
recordtype	SURis
fullrecord	<?xml version="1.0"?><rfc1807><datestamp>2026-01-08T14:36:12.3473731</datestamp><bib-version>v2</bib-version><id>71215</id><entry>2026-01-08</entry><title>Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking</title><swanseaauthors><author><sid>fafc0917b48af4eaec154646854867f8</sid><firstname>Karl</firstname><surname>Austin-Muttitt</surname><name>Karl Austin-Muttitt</name><active>true</active><ethesisStudent>false</ethesisStudent></author><author><sid>4cf2dddedbe1dacb506ec925fdbd5b40</sid><ORCID>0000-0003-0144-2962</ORCID><firstname>Jonathan</firstname><surname>Mullins</surname><name>Jonathan Mullins</name><active>true</active><ethesisStudent>false</ethesisStudent></author></swanseaauthors><date>2026-01-08</date><deptcode>MEDS</deptcode><abstract>Aldehyde oxidase (AOX1) is a cytosolic molybdo-flavoenzyme that metabolizes azaheterocyclic drugs. Erlotinib and gefitinib are azaheterocyclic drugs. We deployed structural analogs to investigate the molecular interaction between these drugs and AOX1. Erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib, but not gefitinib, O-desmethylgefitinib, or O-desmorpholinopropylgefitinib, decreased carbazeran 4-oxidation by liver cytosol (human, rat, and mouse) and human recombinant AOX1. Erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib exhibited time- and concentration-dependent inactivation with unbound inactivation potency (KI,u) of 1.52, 4.41, and 1.67 μM, respectively. The inactivation was not reversed after dialysis, not protected by nucleophilic trapping agents or scavengers of reactive oxygen species, not affected by an oxidizing or reducing agent, but was attenuated by an alternative AOX1 substrate (O6-benzylguanine) and competitive AOX1 inhibitor (gefitinib). The terminal alkyne group of erlotinib was essential for AOX1 inactivation, as suggested by the findings for 3-vinylerlotinib (less potent inactivator) and tetrahydroerlotinib (no inactivation). Molecular docking results predicted covalent binding of erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib to the molybdenum cofactor. Adding a 4′-methyl group to erlotinib increased the inactivation potency but decreased inactivation efficiency, whereas blocking the C2-position of erlotinib with a hydroxy group or a methyl group decreased inactivation potency and efficiency, suggesting that the C2-position of erlotinib plays a role in AOX1 inactivation. In mice, erlotinib increased carbazeran (Aox substrate) and decreased 4-oxo-carbazeran (metabolite) levels in blood, liver, and kidneys. Overall, our study provides molecular insights into the mechanism-based inactivation of AOX1 by erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib and the irreversible AOX1 inactivation by erlotinib on the pharmacokinetics of AOX1-metabolized drugs. Significance Statement: This study shows that erlotinib and select metabolites are mechanism-based inactivators of AOX1, provides insights into the mechanism of the inactivation by deploying structural analogs and molecular docking, and demonstrates the in vivo impact on AOX1-mediated drug metabolism.</abstract><type>Journal Article</type><journal>Molecular Pharmacology</journal><volume>108</volume><journalNumber>1</journalNumber><paginationStart>100097</paginationStart><paginationEnd/><publisher>Elsevier BV</publisher><placeOfPublication/><isbnPrint/><isbnElectronic/><issnPrint/><issnElectronic>0026-895X</issnElectronic><keywords>Aldehyde oxidase; Erlotinib; Structural analogs; Mechanism-based inactivation; Molecular docking; Pharmacokinetics</keywords><publishedDay>1</publishedDay><publishedMonth>1</publishedMonth><publishedYear>2026</publishedYear><publishedDate>2026-01-01</publishedDate><doi>10.1016/j.molpha.2025.100097</doi><url/><notes/><college>COLLEGE NANME</college><department>Medical School</department><CollegeCode>COLLEGE CODE</CollegeCode><DepartmentCode>MEDS</DepartmentCode><institution>Swansea University</institution><apcterm>Another institution paid the OA fee</apcterm><funders>This research was supported in part by the Natural Sciences and Engineering Research Council of Canada (NSERC) [Grant RGPIN-2022-03342] and in part by the Singapore Ministry of Education and the Singapore Ministry of Health’s National Medical Research Council [Grant NMRC/BNIG/2044/2016]. The molecular docking was undertaken at the Supercomputing Wales research facility, Swansea University.</funders><projectreference/><lastEdited>2026-01-08T14:36:12.3473731</lastEdited><Created>2026-01-08T14:26:53.2688902</Created><path><level id="1">Faculty of Medicine, Health and Life Sciences</level><level id="2">Swansea University Medical School - Biomedical Science</level></path><authors><author><firstname>Jia Rong</firstname><surname>Kweh</surname><order>1</order></author><author><firstname>Nicholas Kai Ming</firstname><surname>Ng</surname><order>2</order></author><author><firstname>Le Min</firstname><surname>Ngoh</surname><order>3</order></author><author><firstname>Cynthia Jing Yan</firstname><surname>Li</surname><order>4</order></author><author><firstname>Bao Jie</firstname><surname>Tan</surname><order>5</order></author><author><firstname>Wee Kiat</firstname><surname>Tan</surname><orcid>0009-0002-8146-3498</orcid><order>6</order></author><author><firstname>Vijaya Saradhi</firstname><surname>Mettu</surname><order>7</order></author><author><firstname>Karl</firstname><surname>Austin-Muttitt</surname><order>8</order></author><author><firstname>Jonathan</firstname><surname>Mullins</surname><orcid>0000-0003-0144-2962</orcid><order>9</order></author><author><firstname>Aik Jiang</firstname><surname>Lau</surname><orcid>0000-0003-0742-5223</orcid><order>10</order></author></authors><documents><document><filename>71215__35925__32bdfbabf8c140cebe4cfa8aa482eea5.pdf</filename><originalFilename>71215.VOR.pdf</originalFilename><uploaded>2026-01-08T14:33:47.3778050</uploaded><type>Output</type><contentLength>7025917</contentLength><contentType>application/pdf</contentType><version>Version of Record</version><cronfaStatus>true</cronfaStatus><documentNotes>© 2025 The Authors. Published by Elsevier Inc. on behalf of American Society for Pharmacology and Experimental Therapeutics. This is an open access article under the CC BY license.</documentNotes><copyrightCorrect>true</copyrightCorrect><language>eng</language><licence>http://creativecommons.org/licenses/by/4.0/</licence></document></documents><OutputDurs/></rfc1807>
spelling	2026-01-08T14:36:12.3473731 v2 71215 2026-01-08 Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking fafc0917b48af4eaec154646854867f8 Karl Austin-Muttitt Karl Austin-Muttitt true false 4cf2dddedbe1dacb506ec925fdbd5b40 0000-0003-0144-2962 Jonathan Mullins Jonathan Mullins true false 2026-01-08 MEDS Aldehyde oxidase (AOX1) is a cytosolic molybdo-flavoenzyme that metabolizes azaheterocyclic drugs. Erlotinib and gefitinib are azaheterocyclic drugs. We deployed structural analogs to investigate the molecular interaction between these drugs and AOX1. Erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib, but not gefitinib, O-desmethylgefitinib, or O-desmorpholinopropylgefitinib, decreased carbazeran 4-oxidation by liver cytosol (human, rat, and mouse) and human recombinant AOX1. Erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib exhibited time- and concentration-dependent inactivation with unbound inactivation potency (KI,u) of 1.52, 4.41, and 1.67 μM, respectively. The inactivation was not reversed after dialysis, not protected by nucleophilic trapping agents or scavengers of reactive oxygen species, not affected by an oxidizing or reducing agent, but was attenuated by an alternative AOX1 substrate (O6-benzylguanine) and competitive AOX1 inhibitor (gefitinib). The terminal alkyne group of erlotinib was essential for AOX1 inactivation, as suggested by the findings for 3-vinylerlotinib (less potent inactivator) and tetrahydroerlotinib (no inactivation). Molecular docking results predicted covalent binding of erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib to the molybdenum cofactor. Adding a 4′-methyl group to erlotinib increased the inactivation potency but decreased inactivation efficiency, whereas blocking the C2-position of erlotinib with a hydroxy group or a methyl group decreased inactivation potency and efficiency, suggesting that the C2-position of erlotinib plays a role in AOX1 inactivation. In mice, erlotinib increased carbazeran (Aox substrate) and decreased 4-oxo-carbazeran (metabolite) levels in blood, liver, and kidneys. Overall, our study provides molecular insights into the mechanism-based inactivation of AOX1 by erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib and the irreversible AOX1 inactivation by erlotinib on the pharmacokinetics of AOX1-metabolized drugs. Significance Statement: This study shows that erlotinib and select metabolites are mechanism-based inactivators of AOX1, provides insights into the mechanism of the inactivation by deploying structural analogs and molecular docking, and demonstrates the in vivo impact on AOX1-mediated drug metabolism. Journal Article Molecular Pharmacology 108 1 100097 Elsevier BV 0026-895X Aldehyde oxidase; Erlotinib; Structural analogs; Mechanism-based inactivation; Molecular docking; Pharmacokinetics 1 1 2026 2026-01-01 10.1016/j.molpha.2025.100097 COLLEGE NANME Medical School COLLEGE CODE MEDS Swansea University Another institution paid the OA fee This research was supported in part by the Natural Sciences and Engineering Research Council of Canada (NSERC) [Grant RGPIN-2022-03342] and in part by the Singapore Ministry of Education and the Singapore Ministry of Health’s National Medical Research Council [Grant NMRC/BNIG/2044/2016]. The molecular docking was undertaken at the Supercomputing Wales research facility, Swansea University. 2026-01-08T14:36:12.3473731 2026-01-08T14:26:53.2688902 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Biomedical Science Jia Rong Kweh 1 Nicholas Kai Ming Ng 2 Le Min Ngoh 3 Cynthia Jing Yan Li 4 Bao Jie Tan 5 Wee Kiat Tan 0009-0002-8146-3498 6 Vijaya Saradhi Mettu 7 Karl Austin-Muttitt 8 Jonathan Mullins 0000-0003-0144-2962 9 Aik Jiang Lau 0000-0003-0742-5223 10 71215__35925__32bdfbabf8c140cebe4cfa8aa482eea5.pdf 71215.VOR.pdf 2026-01-08T14:33:47.3778050 Output 7025917 application/pdf Version of Record true © 2025 The Authors. Published by Elsevier Inc. on behalf of American Society for Pharmacology and Experimental Therapeutics. This is an open access article under the CC BY license. true eng http://creativecommons.org/licenses/by/4.0/
title	Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking
spellingShingle	Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking Karl Austin-Muttitt Jonathan Mullins
title_short	Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking
title_full	Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking
title_fullStr	Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking
title_full_unstemmed	Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking
title_sort	Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking
author_id_str_mv	fafc0917b48af4eaec154646854867f8 4cf2dddedbe1dacb506ec925fdbd5b40
author_id_fullname_str_mv	fafc0917b48af4eaec154646854867f8_*_Karl Austin-Muttitt 4cf2dddedbe1dacb506ec925fdbd5b40_*_Jonathan Mullins
author	Karl Austin-Muttitt Jonathan Mullins
author2	Jia Rong Kweh Nicholas Kai Ming Ng Le Min Ngoh Cynthia Jing Yan Li Bao Jie Tan Wee Kiat Tan Vijaya Saradhi Mettu Karl Austin-Muttitt Jonathan Mullins Aik Jiang Lau
format	Journal article
container_title	Molecular Pharmacology
container_volume	108
container_issue	1
container_start_page	100097
publishDate	2026
institution	Swansea University
issn	0026-895X
doi_str_mv	10.1016/j.molpha.2025.100097
publisher	Elsevier BV
college_str	Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id	facultyofmedicinehealthandlifesciences
hierarchy_top_title	Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id	facultyofmedicinehealthandlifesciences
hierarchy_parent_title	Faculty of Medicine, Health and Life Sciences
department_str	Swansea University Medical School - Biomedical Science{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Biomedical Science
document_store_str	1
active_str	0
description	Aldehyde oxidase (AOX1) is a cytosolic molybdo-flavoenzyme that metabolizes azaheterocyclic drugs. Erlotinib and gefitinib are azaheterocyclic drugs. We deployed structural analogs to investigate the molecular interaction between these drugs and AOX1. Erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib, but not gefitinib, O-desmethylgefitinib, or O-desmorpholinopropylgefitinib, decreased carbazeran 4-oxidation by liver cytosol (human, rat, and mouse) and human recombinant AOX1. Erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib exhibited time- and concentration-dependent inactivation with unbound inactivation potency (KI,u) of 1.52, 4.41, and 1.67 μM, respectively. The inactivation was not reversed after dialysis, not protected by nucleophilic trapping agents or scavengers of reactive oxygen species, not affected by an oxidizing or reducing agent, but was attenuated by an alternative AOX1 substrate (O6-benzylguanine) and competitive AOX1 inhibitor (gefitinib). The terminal alkyne group of erlotinib was essential for AOX1 inactivation, as suggested by the findings for 3-vinylerlotinib (less potent inactivator) and tetrahydroerlotinib (no inactivation). Molecular docking results predicted covalent binding of erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib to the molybdenum cofactor. Adding a 4′-methyl group to erlotinib increased the inactivation potency but decreased inactivation efficiency, whereas blocking the C2-position of erlotinib with a hydroxy group or a methyl group decreased inactivation potency and efficiency, suggesting that the C2-position of erlotinib plays a role in AOX1 inactivation. In mice, erlotinib increased carbazeran (Aox substrate) and decreased 4-oxo-carbazeran (metabolite) levels in blood, liver, and kidneys. Overall, our study provides molecular insights into the mechanism-based inactivation of AOX1 by erlotinib, O-desmethylerlotinib, and O-didesmethylerlotinib and the irreversible AOX1 inactivation by erlotinib on the pharmacokinetics of AOX1-metabolized drugs. Significance Statement: This study shows that erlotinib and select metabolites are mechanism-based inactivators of AOX1, provides insights into the mechanism of the inactivation by deploying structural analogs and molecular docking, and demonstrates the in vivo impact on AOX1-mediated drug metabolism.
published_date	2026-01-01T05:33:31Z
_version_	1856805810094473216
score	11.09611

Mechanism-based inactivation of human aldehyde oxidase by erlotinib: Mechanistic insights from structural analogs and molecular docking

Similar Items