Inhibitory effects of drugs on the metabolic activity of mouse and human aldehyde oxidases and influence on drug–drug interactions

Takaoka, Naoki; Sanoh, Seigo; Okuda, Kunio; Kotake, Yaichiro; Sugahara, Go; Yanagi, Ami; Ishida, Yuji; Tateno, Chise; Tayama, Yoshitaka; Sugihara, Kazumi; Kawa, Shigeyuki; Kurosaki, Mami; Terao, Mineko; Garattini, Enrico; Ohta, Shigeru

doi:10.1016/j.bcp.2018.04.017

Cited by 20 publications

(12 citation statements)

References 37 publications

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“…Raloxifene is also an inhibitor of the mouse isoforms mAOX1 and mAOX3 but with 20-fold higher IC50 values when compared with hAOX1. 14,17 Structural analysis and multiple sequence alignments explain this large difference. Although the negatively charged residues that make hydrogen bonds with the raloxifene phenolic groups in hAOX1 (Asp783 and Glu882) are replaced by Glu779 and Asp878 in mAOX3 (and mAOX1), respectively, Gate 1 is shorter in mAOX3, with no aromatic residues to stabilize raloxifene (Figure 8A).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…Out of those 239 compounds, 36 revealed ≥80% enzymatic inhibition at a concentration of 50 μM. In addition to the important role of hAOX1 in drug metabolism, there is also evidence of its involvement in the context of drug–drug interactions (DDI) as reported recently …”

Section: Introductionmentioning

confidence: 82%

“…In addition to the important role of hAOX1 in drug metabolism, there is also evidence of its involvement in the context of drug−drug interactions (DDI) as reported recently. 14 hAOX1 is a complex and large enzyme that belongs to the xanthine oxidase (XO) family, a well-studied and characterized enzyme involved in the last steps of purine catabolism. 15 As found for other enzymes of the XO family, hAOX1 is active as a homodimer (2 × 150 kDa) and harbors, in different domains, a molybdenum active site (Moco catalytic domain), two [2Fe-2S] (FeS domain), and a FAD cofactor (flavin domain) (Figure 1).…”

Section: ■ Introductionmentioning

confidence: 99%

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Interrogating the Inhibition Mechanisms of Human Aldehyde Oxidase by X-ray Crystallography and NMR Spectroscopy: The Raloxifene Case

et al. 2021

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Human aldehyde oxidase (hAOX1) is mainly present in the liver and has an emerging role in drug metabolism, since it accepts a wide range of molecules as substrates and inhibitors. Herein, we employed an integrative approach by combining NMR, X-ray crystallography, and enzyme inhibition kinetics to understand the inhibition modes of three hAOX1 inhibitorsthioridazine, benzamidine, and raloxifene. These integrative data indicate that thioridazine is a noncompetitive inhibitor, while benzamidine presents a mixed type of inhibition. Additionally, we describe the first crystal structure of hAOX1 in complex with raloxifene. Raloxifene binds tightly at the entrance of the substrate tunnel, stabilizing the flexible entrance gates and elucidating an unusual substrate-dependent mechanism of inhibition with potential impact on drug−drug interactions. This study can be considered as a proof-of-concept for an efficient experimental screening of prospective substrates and inhibitors of hAOX1 relevant in drug discovery.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 82%

Section: ■ Introductionmentioning

confidence: 99%

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Interrogating the Inhibition Mechanisms of Human Aldehyde Oxidase by X-ray Crystallography and NMR Spectroscopy: The Raloxifene Case

et al. 2021

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“…Future clinical studies would be needed to determine whether these interactions occur in vivo. To date, studies have been reported on AOX1 protein structure-drug metabolism relationships to predict human AOX1 substrates (Lepri et al, 2017;Cruciani et al, 2018), but limited information on human AOX1 structure-enzyme inhibitor relationships determined based on the crystal structure of human AOX1 (Takaoka et al, 2018;Deris-Abdolahpour et al, 2019). The molecular-docking approach developed in this study, by virtue of its consistent performance and the ability it allows for meaningful comparative analyses of chemical inhibitors, provides a robust in silico framework for future investigation of the binding of chemical inhibitors with AOX1.…”

Section: Discussionmentioning

confidence: 99%

In Vitro and In Silico Analyses of the Inhibition of Human Aldehyde Oxidase by Bazedoxifene, Lasofoxifene, and Structural Analogues

Chen

Austin-Muttitt

Zhang

et al. 2019

J Pharmacol Exp Ther

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Tamoxifen, raloxifene, and nafoxidine are selective estrogen receptor modulators (SERMs) reported to inhibit the catalytic activity of human aldehyde oxidase 1 (AOX1). How these drugs interact with AOX1 and whether other SERMs inhibit this drugmetabolizing enzyme are not known. Therefore, a detailed in vitro and in silico study involving parent drugs and their analogs was conducted to investigate the effect of specific SERMs, particularly acolbifene, bazedoxifene, and lasofoxifene on AOX1 catalytic activity, as assessed by carbazeran 4-oxidation, an AOX1-selective catalytic marker. The rank order in the potency (based on IC 50 values) of AOX1 inhibition by SERMs was raloxifene. bazedoxifene ∼ lasofoxifene. tamoxifen. acolbifene. Inhibition of liver cytosolic AOX1 by bazedoxifene, lasofoxifene, and tamoxifen was competitive, whereas that by raloxifene was noncompetitive. Loss of 1-azepanylethyl group increased the inhibitory potency of bazedoxifene, whereas the N-oxide group decreased it. The 7-hydroxy group and the substituted pyrrolidine ring attached to the tetrahydronaphthalene structure contributed to AOX1 inhibition by lasofoxifene. These results are supported by molecular-docking simulations in terms of predicted binding modes, encompassing binding orientation and efficiency, and analysis of key interactions, particularly hydrogen bonds. The extent of AOX1 inhibition by bazedoxifene was increased by estrone sulfate and estrone. In summary, SERMs differentially inhibited human AOX1 catalytic activity. Structural features of bazedoxifene and lasofoxifene contributed to AOX1 inhibition, whereas those of acolbifene rendered it considerably less susceptible to AOX1 inhibition. Overall, our novel biochemical findings and molecular-docking analyses provide new insights into the interaction between SERMs and AOX1. SIGNIFICANCE STATEMENT Aldehyde oxidase (AOX1) is a molybdo-flavoprotein and has emerged as a drug-metabolizing enzyme of potential therapeutic importance because drugs have been identified as AOX1 substrates. Selective estrogen receptor modulators (SERM), which are drugs used to treat and prevent various conditions, differentially inhibit AOX1 catalytic activity. Structural features of bazedoxifene and lasofoxifene contribute to AOX1 inhibition, whereas those of acolbifene render it considerably less susceptible to AOX1 inhibition. Our novel biochemical findings, together with molecular-docking analyses, provide new insights into the differential inhibitory effect of SERMs on the catalytic activity of human AOX1, how SERMs bind to AOX1, and increase our understanding of the AOX1 pharmacophore in the inhibition of AOX1 by drugs and other chemicals.

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“…Humanized liver chimeric mice have been produced by transplantation of normal HHs into genetically engineered immunodeficient/liver injured host mice [ 14 ]. We reported that chimeric mice generated from albumin enhancer/promoter-driven cDNA urokinase-type plasminogen activator/severe combined immunodeficiency (cDNA-uPA/SCID) mice (PXB-mice) exhibited stable repopulation rates of HHs for several months [ 15 ], and were useful for studies on drug metabolism and pharmacokinetics, drug-drug interactions [ 16 – 18 ], and chronic infection by hepatitis viruses [ 19 ]. PXB-mice and the other type of mice with humanized livers have been utilized for in vivo toxicology studies, with human specific cytotoxicity reported for troglitazone, fialuridine, and bosentan [ 20 – 23 ].…”

Section: Introductionmentioning

confidence: 99%

Detection of acute toxicity of aflatoxin B1 to human hepatocytes in vitro and in vivo using chimeric mice with humanized livers

et al. 2020

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Aflatoxin B1 (AFB1), a mycotoxin, is acutely hepatotoxic to many animals including humans. However, there are marked interspecies differences in sensitivity to AFB1-induced toxicity depending on bioactivation by cytochrome P450s (CYPs). In the present study, we examined the applicability of chimeric mice with humanized livers and derived fresh human hepatocytes for in vivo and vitro studies on AFB1 cytotoxicity to human hepatocytes. Chimeric mice with highly humanized livers and SCID mice received daily injections of vehicle (corn oil), AFB1 (3 mg/kg), and carbon tetrachloride (50 mg/kg) for 2 days. Histological analysis revealed that AFB1 promoted hepatocyte vacuolation and inflammatory cell infiltration in the area containing human hepatocytes. A novel human alanine aminotransferase 1 specific enzyme-linked immunosorbent assay demonstrated the acute toxicity of AFB1 to human hepatocytes in the chimeric mouse livers. The sensitivity of cultured fresh human hepatocytes isolated from the humanized liver mice for AFB1 cytotoxicity was comparable to that of primary human hepatocytes. Long-term exposure to AFB1 (6 or 14 days) produced a more severe cytotoxicity. The half-maximal lethal concentration was 10 times lower in the 2week treatment than after 2 days of exposure. Lastly, the significant reduction of AFB1 cytotoxicity by a pan-CYP inhibitor or transfection with CYP3A4 specific siRNA clearly suggested that bioactivation of AFB1 catalyzed by CYPs was essential for AFB1 cytotoxicity to the human hepatocytes in our mouse model. Collectively, our results implicate the humanized liver mice and derived fresh human hepatocytes are useful models for studies of AFB1 cytotoxicity to human hepatocytes.

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Inhibitory effects of drugs on the metabolic activity of mouse and human aldehyde oxidases and influence on drug–drug interactions

Cited by 20 publications

References 37 publications

Interrogating the Inhibition Mechanisms of Human Aldehyde Oxidase by X-ray Crystallography and NMR Spectroscopy: The Raloxifene Case

Interrogating the Inhibition Mechanisms of Human Aldehyde Oxidase by X-ray Crystallography and NMR Spectroscopy: The Raloxifene Case

In Vitro and In Silico Analyses of the Inhibition of Human Aldehyde Oxidase by Bazedoxifene, Lasofoxifene, and Structural Analogues

Detection of acute toxicity of aflatoxin B1 to human hepatocytes in vitro and in vivo using chimeric mice with humanized livers

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