Schmidt Monika, Vaskova Michaela, Rotterova Aneta, Fiandova Pavlina, Miskerikova Marketa, Zemanova Lucie, Benek Ondrej, Musilek Kamil
Department of Chemistry, Faculty of Science, University of Hradec Kralove, Hradec Kralove, Czech Republic.
J Neurochem. 2023 Oct;167(2):154-167. doi: 10.1111/jnc.15917. Epub 2023 Jul 17.
Mitochondrial enzyme 17β-hydroxysteroid dehydrogenase type 10 (HSD10) is a potential molecular target for treatment of mitochondrial-related disorders such as Alzheimer's disease (AD). Its over-expression in AD brains is one of the critical factors disturbing the homeostasis of neuroprotective steroids and exacerbating amyloid beta (Aβ)-mediated mitochondrial toxicity and neuronal stress. This study was focused on revalidation of the most potent HSD10 inhibitors derived from benzothiazolyl urea scaffold using fluorescent-based enzymatic assay with physiologically relevant substrates of 17β-oestradiol and allopregnanolone. The oestradiol-based assay led to the identification of two nanomolar inhibitors (IC 70 and 346 nM) differing from HSD10 hits revealed from the formerly used assay. Both identified inhibitors were found to be effective also in allopregnanolone-based assay with non-competitive or uncompetitive mode of action. In addition, both inhibitors were confirmed to penetrate the HEK293 cells and they were able to inhibit the HSD10 enzyme in the cellular environment. Both molecules seem to be potential lead structures for further research and development of HDS10 inhibitors.
线粒体酶17β-羟类固醇脱氢酶10型（HSD10）是治疗线粒体相关疾病如阿尔茨海默病（AD）的潜在分子靶点。它在AD大脑中的过度表达是扰乱神经保护类固醇稳态并加剧淀粉样β蛋白（Aβ）介导的线粒体毒性和神经元应激的关键因素之一。本研究聚焦于使用基于荧光的酶促测定法，以生理相关底物17β-雌二醇和别孕烯醇酮对源自苯并噻唑基脲支架的最有效的HSD10抑制剂进行重新验证。基于雌二醇的测定法鉴定出两种纳摩尔级抑制剂（IC分别为70和346 nM），与先前使用的测定法所揭示的HSD10活性位点不同。发现这两种鉴定出的抑制剂在基于别孕烯醇酮的测定法中也有效，作用模式为非竞争性或反竞争性。此外，证实这两种抑制剂均可穿透HEK293细胞，并能够在细胞环境中抑制HSD10酶。这两种分子似乎都是进一步研发HDS10抑制剂的潜在先导结构。
