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Updated: Jul 31, 2025
Assays for Validating Histone Acetyltransferase Inhibitors
Published on: August 6, 2020
First-in-Class Selective Inhibitors of the Lysine Acetyltransferase KAT8
Francesco Fiorentino1, Sara Sementilli2, Martina Menna1
1Department of Drug Chemistry and Technologies, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy.
Abstract:
KAT8 is a lysine acetyltransferase primarily catalyzing the acetylation of Lys16 of histone H4 (H4K16). KAT8 dysregulation is linked to the development and metastatization of many cancer types, including non-small cell lung cancer (NSCLC) and acute myeloid leukemia (AML). Few KAT8 inhibitors have been reported so far, none of which displaying selective activity. Based on the KAT3B/KDAC inhibitor C646, we developed a series of N-phenyl-5-pyrazolone derivatives and identified compounds 19 and 34 as low-micromolar KAT8 inhibitors selective over a panel of KATs and KDACs. Western blot, immunofluorescence, and CETSA experiments demonstrated that both inhibitors selectively target KAT8 in cells. Moreover, 19 and 34 exhibited mid-micromolar antiproliferative activity in different cancer cell lines, including NSCLC and AML, without impacting the viability of nontransformed cells. Overall, these compounds are valuable tools for elucidating KAT8 biology, and their simple structures make them promising candidates for future optimization studies.
Insights
Researchers developed novel, selective KAT8 inhibitors (compounds 19 and 34) targeting cancer cells, including non-small cell lung cancer (NSCLC) and acute myeloid leukemia (AML), with potential for further optimization.
Area of Science:
- Epigenetics and Cancer Biology
- Medicinal Chemistry and Drug Discovery
Background:
- Lysine acetyltransferase 8 (KAT8) dysregulation is implicated in cancer development and metastasis, notably in non-small cell lung cancer (NSCLC) and acute myeloid leukemia (AML).
- Existing KAT8 inhibitors lack selectivity, hindering their utility in research and therapeutic development.
Purpose of the Study:
- To design and synthesize novel KAT8 inhibitors with enhanced selectivity.
- To evaluate the efficacy of these inhibitors in targeting cancer cells and elucidating KAT8 biology.
Main Methods:
- Development of N-phenyl-5-pyrazolone derivatives based on the KAT3B/KDAC inhibitor C646.
- In vitro screening for KAT8 inhibitory activity and selectivity against a panel of KATs and KDACs.
- Cellular target engagement validation using Western blot, immunofluorescence, and CETSA.
- Assessment of antiproliferative activity in various cancer cell lines (NSCLC, AML) and non-transformed cells.
Main Results:
- Compounds 19 and 34 were identified as low-micromolar selective KAT8 inhibitors.
- Both compounds demonstrated selective KAT8 targeting within cells.
- Compounds 19 and 34 exhibited mid-micromolar antiproliferative effects against NSCLC and AML cell lines without toxicity to normal cells.
Conclusions:
- Compounds 19 and 34 represent valuable, selective chemical tools for studying KAT8 function in biological systems.
- The simple structures of these inhibitors offer a promising foundation for future optimization and drug development efforts targeting KAT8-driven cancers.
