Human aldehyde oxidase (AOX) has emerged as a key enzyme activity for consideration in modern drug discovery. The enzyme catalyzes the oxidation of a wide variety of compounds, most notably azaheterocyclics that often form the building blocks of small molecule therapeutics. Failure to consider and assess AOX drug exposure early in the drug development cycle can have catastrophic consequences for novel compounds entering the clinic. AOX is a complex molybdopterin-containing iron-sulfur flavoprotein comprised of two identical 150 kDa subunits that has proven difficult to produce in recombinant form, and a commercial source of the purified human enzyme is currently unavailable. Thus, the potential exposure of novel drug development candidates to human AOX metabolism is usually assessed by using extracts of pooled human liver cytosol as a source of the enzyme. This can complicate the assignment of AOX-specific compound exposure due to its low activity and the presence of contaminating enzymes that may have overlapping substrate specificities. Herein is described a two-step process for the isolation of recombinant human AOX dimers to near homogeneity following production in the baculovirus expression vector system (BEVS). The deployment of this BEVS-produced recombinant human AOX as a substitute for human liver extracts in a fraction-of-control AOX compound-exposure screening assay is described. The ability to generate this key enzyme activity readily in a purified recombinant form provides for a more accurate and convenient approach to the assessment of new compound exposure to bona fide AOX drug metabolism.

人醛氧化酶（AOX）已经成为现代药物开发中考虑的关键酶活性。该酶催化多种化合物的氧化，最显着的是杂杂环化合物，它们通常构成小分子治疗剂的组成部分。在药物开发周期的早期不考虑和评估AOX药物暴露可能会对进入临床的新型化合物造成灾难性后果。AOX是一种复杂的含钼蝶呤的铁硫黄素蛋白，由两个相同的150 kDa亚基组成，已证明难以以重组形式生产，目前尚无纯化的人类酶的商业来源。从而，通常通过使用合并的人肝细胞溶质提取物作为酶的来源来评估新药开发候选物对人AOX代谢的潜在暴露。由于其活性低和存在可能具有重叠底物特异性的污染酶，这会使AOX特异性化合物暴露的分配复杂化。本文描述了在杆状病毒表达载体系统（BEVS）中产生后用于将重组人AOX二聚体分离至接近同质的两步过程。描述了这种BEVS生产的重组人AOX在对照AOX化合物暴露筛选试验中作为人肝提取物的替代物的用途。真正的AOX药物代谢。