Characterization of fluorescent probe substrates to develop an efficient high-throughput assay for neonatal hepatic CYP3A7 inhibition screening.
Characterization of fluorescent probe substrates to develop an efficient high-throughput assay for neonatal hepatic CYP3A7 inhibition screening.
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DOI：
10.1038/s41598-021-98219-x
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发表时间：
2021-09-30
影响因子：
4.6
通讯作者：
Lampe JN
中科院分区：
文献类型：
--
作者：
Work HM;Kandel SE;Lampe JN
CYP 3A 7是细胞色素P450（CYP 3A）3A酶亚家族的成员，在胎儿和新生儿中表达。除了代谢维甲酸和内源性类固醇硫酸脱氢表雄酮（DHEA-S）的作用外，它还在生命最初几周的药物代谢和处置中发挥关键作用。尽管如此，在新药候选物的临床前测试中，它通常被忽视。这增加了新生儿发生药物相互作用（DDI）和毒性的风险。因此，筛选抑制CYP 3A 7的候选药物对于识别对新生儿具有潜在毒性风险的化学实体至关重要。目前，没有有效的高通量筛选（HTS）试验来评估CYP 3A 7抑制。在这里，我们报告了我们的各种荧光探针的测试，以高通量的方式评估CYP 3A 7活性。我们确定，荧光化合物二苄基荧光素（DBF）是上级的其他化合物，在满足标准考虑一个有效的HTS测定。此外，HIV/HCV抗病毒药物微型库的初步筛选证明了DBF在HTS检测系统中的实用性。我们预计，这一工具将是非常有益的筛选药物，可能会在未来的新生儿人口。
CYP3A7 is a member of the cytochrome P450 (CYP) 3A enzyme sub-family that is expressed in the fetus and neonate. In addition to its role metabolizing retinoic acid and the endogenous steroid dehydroepiandrosterone sulfate (DHEA-S), it also has a critical function in drug metabolism and disposition during the first few weeks of life. Despite this, it is generally ignored in the preclinical testing of new drug candidates. This increases the risk for drug-drug interactions (DDI) and toxicities occurring in the neonate. Therefore, screening drug candidates for CYP3A7 inhibition is essential to identify chemical entities with potential toxicity risks for neonates. Currently, there is no efficient high-throughput screening (HTS) assay to assess CYP3A7 inhibition. Here, we report our testing of various fluorescent probes to assess CYP3A7 activity in a high-throughput manner. We determined that the fluorescent compound dibenzylfluorescein (DBF) is superior to other compounds in meeting the criteria considered for an efficient HTS assay. Furthermore, a preliminary screen of an HIV/HCV antiviral drug mini-library demonstrated the utility of DBF in a HTS assay system. We anticipate that this tool will be of great benefit in screening drugs that may be used in the neonatal population in the future.
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