T-448, a specific inhibitor of LSD1 enzyme activity, improves learning function without causing thrombocytopenia in mice

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.

LSD1 enzyme inhibitor TAK-418 unlocks aberrant epigenetic machinery and improves autism symptoms in neurodevelopmental disorder models

“…S1, A to D, and table S1). We have previously found that LSD1 inhibition could increase histone methylation at Ucp2 and Bdnf genes ( 13 ). Thus, histone methylation levels of these genes were used as markers for LSD1 inhibition by TAK-418.…”

“…We focused on LSD1, which demethylates mono- and dimethylated H3K4 (H3K4me1/2) and histone H3 lysine 9 (H3K9me1/2) ( 10 , 11 ). LSD1 has been considered to regulate gene expression through two functions: histone demethylation and scaffolding activity for cofactors, including growth factor independent 1B (GFI1B), a critical regulator of hematopoietic differentiation ( 12 , 13 ). In the previous study, characterization of two LSD1 inhibitors [ N -(4-( trans -2-((cyclopropylmethyl)amino)cyclopropyl)pheny)bipheny-4-carbox amide hydrochloride (T-711) and 3-((1 S ,2 R )-2-(cyclobutylamino)cyclopropyl)- N -(5-methyl-1,3,4-thiadiazol-2-yl)benzamide fumarate (T-448)] revealed that T-711 modulates both of these functions and causes thrombocytopenia, while T-448 selectively inhibits histone demethylation with low hematological toxicity ( 13 ).…”

Pharmacologic inhibition of lysine specific demethylase-1 (LSD1) as a therapeutic and immune-sensitization strategy in diffuse intrinsic pontine glioma (DIPG)

“…Another consideration is the pharmacokinetic (PK) properties of currently available LSD1 inhibitors, which have recently been shown to be ineffective against intracranial medulloblastoma despite very potent in vitro activity 35 . Two separate groups in Japan recently described new catalytic LSD1 inhibitors with favorable PK properties for potential use in neuro-oncology 36,37 . The transcription factor GFI1B is not disrupted by their compounds, reducing peripheral toxicity and sparing immune cells that would be critical for our combination therapy.…”

First-in-Human Randomized Trial to Assess Safety, Tolerability, Pharmacokinetics and Pharmacodynamics of the KDM1A Inhibitor Vafidemstat

“…KDM1A is a histone demethylase that has been proposed as a target for the treatment of neurodegenerative or neurodevelopmental diseases [6][7][8].…”