In Vitro Assay Development and HTS of Small-Molecule Human ABAD/17β-HSD10 Inhibitors as Therapeutics in Alzheimer’s Disease

Aitken, Laura; Baillie, Gemma; Pannifer, Andrew; Morrison, Angus J.; Jones, Philip S.; Smith, Terry; McElroy, Stuart P.; Gunn-Moore, Frank J.

doi:10.1177/2472555217697964

Cited by 17 publications

(19 citation statements)

References 27 publications

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“…To establish the reaction properties correctly, the enzyme kinetic parameters were determined. The Michaelis constant (K m ) obtained for acetoacetyl-CoA as a substrate, 79.2 ± 15.0 µM at pH 7.4, corresponded to the previously reported values of 117 ± 28 µM [38] and 89 ± 5.4 µM [3] for the same substrate. The maximum reaction rate, V max , for the enzyme, was estimated to be 27.7 ± 1.8 µM•min −1 .…”

Section: β-Hsd10 Reductase Assay and Enzyme Kineticssupporting

confidence: 87%

“…The enzyme activity of the human recombinant 17β-HSD10 was determined from previously published methods using acetoacetyl-CoA (AAC) as a substrate [4,19,38,39]. The method was based on the decrease in nicotinamide adenine dinucleotide (NADH) cofactor absorbance at 340 nm during its utilization by the enzyme.…”

Section: β-Hsd10 Reductase Assay and Enzyme Kineticsmentioning

confidence: 99%

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Benzothiazolyl Ureas are Low Micromolar and Uncompetitive Inhibitors of 17β-HSD10 with Implications to Alzheimer’s Disease Treatment

Schmidt

Benek

Vinklářová

et al. 2020

IJMS

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Human 17β-hydroxysteroid dehydrogenase type 10 is a multifunctional protein involved in many enzymatic and structural processes within mitochondria. This enzyme was suggested to be involved in several neurological diseases, e.g., mental retardation, Parkinson’s disease, or Alzheimer’s disease, in which it was shown to interact with the amyloid-beta peptide. We prepared approximately 60 new compounds based on a benzothiazolyl scaffold and evaluated their inhibitory ability and mechanism of action. The most potent inhibitors contained 3-chloro and 4-hydroxy substitution on the phenyl ring moiety, a small substituent at position 6 on the benzothiazole moiety, and the two moieties were connected via a urea linker (4at, 4bb, and 4bg). These compounds exhibited IC50 values of 1–2 μM and showed an uncompetitive mechanism of action with respect to the substrate, acetoacetyl-CoA. These uncompetitive benzothiazolyl inhibitors of 17β-hydroxysteroid dehydrogenase type 10 are promising compounds for potential drugs for neurodegenerative diseases that warrant further research and development.

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Section: β-Hsd10 Reductase Assay and Enzyme Kineticssupporting

confidence: 87%

Section: β-Hsd10 Reductase Assay and Enzyme Kineticsmentioning

confidence: 99%

Benzothiazolyl Ureas are Low Micromolar and Uncompetitive Inhibitors of 17β-HSD10 with Implications to Alzheimer’s Disease Treatment

Schmidt

Benek

Vinklářová

et al. 2020

IJMS

Self Cite

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“…In order to reduce attrition rates and improve assay reproducibility we have developed a high throughput screening (HTS) pipeline (Figure 3 [19]). In brief, compounds are screened in the recombinant 17β-HSD10 enzyme activity assay (Table 5, Figure 4).…”

Section: Resultsmentioning

confidence: 99%

“…During our enzymatic assay development, it was noted that the assay was susceptible to false positives through redox cycling and aggregation mechanism [19], therefore, two orthogonal counter screens have been implemented to validate the primary screen results. The addition of the detergent Triton X-100 to the assay buffer prevents the hydrophobic interactions required for aggregation, by which a reduction in inhibition in the presence of Triton X-100 indicates the undesirable inhibitory mode of action whereby, the compound could be potentially inhibiting the enzyme through the indirect sequestration of the protein.…”

Section: Resultsmentioning

confidence: 99%

Novel Benzothiazole-Based Ureas as 17β-HSD10 Inhibitors, A Potential Alzheimer’s Disease Treatment

et al. 2019

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: It has long been established that mitochondrial dysfunction in Alzheimer’s disease (AD) patients can trigger pathological changes in cell metabolism by altering metabolic enzymes such as the mitochondrial 17β-hydroxysteroid dehydrogenase type 10 (17β-HSD10), also known as amyloid-binding alcohol dehydrogenase (ABAD). We and others have shown that frentizole and riluzole derivatives can inhibit 17β-HSD10 and that this inhibition is beneficial and holds therapeutic merit for the treatment of AD. Here we evaluate several novel series based on benzothiazolylurea scaffold evaluating key structural and activity relationships required for the inhibition of 17β-HSD10. Results show that the most promising of these compounds have markedly increased potency on our previously published inhibitors, with the most promising exhibiting advantageous features like low cytotoxicity and target engagement in living cells.

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“…A combination of frentizole and rasagiline scaffolds for inhibiting both 17β‐HSD10 and MAO (monoamine oxidase) revealed one compound exhibiting good potency for MAO (IC 50 6.34 μM), but no activity towards 17β‐HSD10 (Hroch et al., 2017). Searching for novel lead structures for 17β‐HSD10 inhibitors using high‐throughput screening of more than 6,500 compounds from different collections and libraries (Aitken et al., 2017) revealed 16 hits. These inhibitors were identified to be in the micromolar range and might be capable to cross the blood–brain barrier.…”

Section: Inhibition Of 17β‐hsd10‐aβ Interaction or Enzymatic Activitymentioning

confidence: 99%

Friend or enemy? Review of 17β‐HSD10 and its role in human health or disease

Vinklářová

Schmidt

Benek

et al. 2020

Journal of Neurochemistry

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17β-HSD10 is a mitochondrial enzyme involved in the metabolism of a wide range of substrates, including neurosteroids (He, Dobkin, & Yang, 2019) and sex steroids, maintaining their physiological level (Shafqat et al., 2003). However, it also plays an important role in tRNA processing as a structural component of RNase P (Holzmann et al., 2008). Its abnormal function including inherited mutations leads to disruption in mitochondrial physiology and is thought to be one of the underlying pathological causes for diseases such as Alzheimer's disease (He, Isaacs, & Yang, 2018) and some forms of cancer (Yang, He, & Schulz, 2005). This paper discusses the role of 17β-HSD10 in physiology, as well as its connections to various diseases. The first section gives an overview of 17β-HSD10 structure, localization and physiological functions. In the second section, the role of 17β-HSD10 in disease, specifically in neurodegenerative disorders and cancer are discussed, which together are attracting more and broader interest in this enzyme.

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In Vitro Assay Development and HTS of Small-Molecule Human ABAD/17β-HSD10 Inhibitors as Therapeutics in Alzheimer’s Disease

Cited by 17 publications

References 27 publications

Benzothiazolyl Ureas are Low Micromolar and Uncompetitive Inhibitors of 17β-HSD10 with Implications to Alzheimer’s Disease Treatment

Benzothiazolyl Ureas are Low Micromolar and Uncompetitive Inhibitors of 17β-HSD10 with Implications to Alzheimer’s Disease Treatment

Novel Benzothiazole-Based Ureas as 17β-HSD10 Inhibitors, A Potential Alzheimer’s Disease Treatment

Friend or enemy? Review of 17β‐HSD10 and its role in human health or disease

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