Developing selective histone deacetylases (HDACs) inhibitors through ebselen and analogs
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Updated: Mar 2, 2026

Assays for Validating Histone Acetyltransferase Inhibitors
Published on: August 6, 2020
Yuren Wang1, Jason Wallach2, Stephanie Duane1
1Reaction Biology Corp., Malvern.
Abstract:
Histone deacetylases (HDACs) are key regulators of gene expression in cells and have been investigated as important therapeutic targets for cancer and other diseases. Different subtypes of HDACs appear to play disparate roles in the cells and are associated with specific diseases. Therefore, substantial effort has been made to develop subtype-selective HDAC inhibitors. In an effort to discover existing scaffolds with HDAC inhibitory activity, we screened a drug library approved by the US Food and Drug Administration and a National Institutes of Health Clinical Collection compound library in HDAC enzymatic assays. Ebselen, a clinical safe compound, was identified as a weak inhibitor of several HDACs, including HDAC1, HDAC3, HDAC4, HDAC5, HDAC6, HDAC7, HDAC8, and HDAC9 with half maximal inhibitory concentrations approximately single digit of µM. Two ebselen analogs, ebselen oxide and ebsulfur (a diselenide analog of ebselen), also inhibited these HDACs, however with improved potencies on HDAC8. Benzisothiazol, the core structure of ebsulfur, specifically inhibited HDAC6 at a single digit of µM but had no inhibition on other HDACs. Further efforts on structure-activity relationship based on the core structure of ebsulfur led to the discovery of a novel class of potent and selective HDAC6 inhibitors with RBC-2008 as the lead compound with single-digit nM potency. This class of histone deacetylase inhibitor features a novel pharmacophore with an ebsulfur scaffold selectively targeting HDAC6. Consistent with its inhibition on HDAC6, RBC-2008 significantly increased the acetylation levels of α-tubulin in PC-3 cells. Furthermore, treatment with these compounds led to cell death of multiple tumor cell lines in a dose-dependent manner. These results demonstrated that ebselen and ebsulfur analogs are inhibitors of HDACs, supporting further preclinical development of this class of compounds for potential therapeutic applications.
Insights
Ebselen and its analogs were found to inhibit histone deacetylases (HDACs). Further development led to potent and selective HDAC6 inhibitors, like RBC-2008, showing promise for cancer therapy.
Area of Science:
- Biochemistry and Molecular Biology
- Medicinal Chemistry
- Pharmacology
Background:
- Histone deacetylases (HDACs) are crucial regulators of gene expression and are implicated in various diseases, including cancer.
- Developing subtype-selective HDAC inhibitors is essential due to the diverse roles of different HDACs.
- Existing drug libraries offer potential scaffolds for discovering novel HDAC inhibitors.
Purpose of the Study:
- To identify existing compounds with histone deacetylase (HDAC) inhibitory activity.
- To discover novel, potent, and selective HDAC inhibitors for potential therapeutic applications.
Main Methods:
- Screening of FDA-approved drug libraries and NIH Clinical Collection compounds in HDAC enzymatic assays.
- Structure-activity relationship (SAR) studies based on identified scaffolds.
- In vitro assays to determine inhibitory concentrations (IC50) and target selectivity.
- Cell-based assays measuring α-tubulin acetylation and tumor cell line viability.
Main Results:
- Ebselen was identified as a weak inhibitor of multiple HDACs (HDAC1-9) with IC50 in the µM range.
- Ebselen analogs, ebselen oxide and ebsulfur, also inhibited HDACs, with improved potency against HDAC8.
- Benzisothiazol, a core structure of ebsulfur, selectively inhibited HDAC6.
- SAR studies led to the discovery of RBC-2008, a novel class of potent HDAC6 inhibitors with nM potency.
- RBC-2008 increased α-tubulin acetylation in PC-3 cells and induced dose-dependent cell death in tumor cell lines.
Conclusions:
- Ebselen and its analogs are effective inhibitors of various histone deacetylases (HDACs).
- The ebsulfur scaffold can be optimized to yield potent and selective HDAC6 inhibitors, exemplified by RBC-2008.
- These novel HDAC6 inhibitors demonstrate potential for preclinical development in cancer therapy.

