Abstract
Dysregulation of histone H3 lysine 4 (H3K4) methylation has been implicated in the pathogenesis of several neurodevelopmental disorders. Targeting lysine-specific demethylase 1 (LSD1), an H3K4 demethylase, is therefore a promising approach to treat these disorders. However, LSD1 forms complexes with cofactors including growth factor independent 1B (GFI1B), a critical regulator of hematopoietic differentiation. Known tranylcypromine-based irreversible LSD1 inhibitors bind to coenzyme flavin adenine dinucleotide (FAD) and disrupt the LSD1-GFI1B complex, which is associated with hematotoxicity such as thrombocytopenia, representing a major hurdle in the development of LSD1 inhibitors as therapeutic agents. To discover LSD1 inhibitors with potent epigenetic modulation and lower risk of hematotoxicity, we screened small molecules that enhance H3K4 methylation by the inhibition of LSD1 enzyme activity in primary cultured rat neurons but have little impact on LSD1-GFI1B complex in human TF-1a erythroblasts. Here we report the discovery of a specific inhibitor of LSD1 enzyme activity, T-448 (3-((1S,2R)-2-(cyclobutylamino)cyclopropyl)-N-(5-methyl-1,3,4-thiadiazol-2-yl)benzamide fumarate). T-448 has minimal impact on the LSD1-GFI1B complex and a superior hematological safety profile in mice via the generation of a compact formyl-FAD adduct. T-448 increased brain H3K4 methylation and partially restored learning function in mice with NMDA receptor hypofunction. T-448-type LSD1 inhibitors with improved safety profiles may provide unique therapeutic approaches for central nervous system disorders associated with epigenetic dysregulation.
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Acknowledgements
The authors thank Dr. Takayuki Niimura and Mr. Ryota Maeda for performing the pharmacological analysis, Mr. Yasushi Hattori and Dr. Shigemitsu Matsumoto for compound synthesis, Mr. Takashi Ito and Ms. Yumi Zama for protein purification, as well as Dr. Gyorgy Snell and Dr. Lane Scott for data collection and processing for crystal structure analysis. This work was funded by the Takeda Pharmaceutical Company Limited.
Author contributions
S.Matsuda and H.Kimura conceived and designed the experiments. S.Matsuda, R.B., H.O., S.Igaki, Y.K., S.Iwasaki, K.M., R.Hibino, H.Kamada, T.H., M.Iwatani, K.T., and R.Hara performed the experiments and analyzed the data. S.Morimoto, M.T., and M.Ito designed and synthesized T-448. S.Matsuda and H.Kimura wrote the manuscript.
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The authors are present employees of the Takeda Pharmaceutical Company Limited or were employees of Takeda Pharmaceutical Company Limited while engaged in this research.
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Matsuda, S., Baba, R., Oki, H. et al. T-448, a specific inhibitor of LSD1 enzyme activity, improves learning function without causing thrombocytopenia in mice. Neuropsychopharmacol. 44, 1505–1512 (2019). https://doi.org/10.1038/s41386-018-0300-9
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DOI: https://doi.org/10.1038/s41386-018-0300-9
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