Design and Synthesis of Highly Potent and Specific ABHD6 Inhibitors

Abstract

Fine‐tuning than complete disruption of 2‐arachidonoylglycerol (2‐AG) metabolism in the brain represents a promising pharmacological approach to limit potential untoward effects associated with complete blockade of monoacylglycerol lipase (MGL), the primary hydrolase of 2‐AG. This could be achieved through a/b‐hydrolase domain containing 6 (ABHD6) inhibition, which will provide a smaller and safer contribution to 2‐AG regulation in the brain. Pharmacological studies with ABHD6 inhibitors have recently been reported, where modulation of ABHD6 activity either through CB1R‐dependent or CB1R‐independent processes showed promise in preclinical models of epilepsy, neuropathic pain and inflammation. Furthermore in the periphery, ABHD6 modulates 2‐AG and other fatty acid monoacylglycerols (MAGs) and is implicated in Type‐2 diabetes, metabolic syndrome and potentially other diseases. Herein, we report the discovery of single‐digit nanomolar potent and highly specific ABHD6 inhibitors with >1000‐fold selectivity against MGL and FAAH. The new ABHD6 inhibitors provide early leads to develop therapeutics for neuroprotection and the treatment of inflammation and diabetes.

P2X7 receptor‐dependent increase in endocannabinoid 2‐arachidonoyl glycerol production by neuronal cells in culture: Dynamics and mechanism

“…ABHD6 is a multifunctional enzyme that produces 2‐AG from DAG under basal conditions in N2a cells (van Esbroeck et al, 2019) but hydrolyses 2‐AG in stimulated neurons (Marrs et al, 2010). To test if ABHD6 controls the P2X 7 R‐dependent increase in 2‐AG production in N2a cells, we used its recently developed inhibitor, AM12100 (Figure 6a) (Kokona et al, 2021; Malamas et al, 2021). ABPP analysis showed that AM12100 inhibited ABHD6 activity with an IC 50 = 19 nM and reduced ABHD6 activity by 74% at 30 nM without affecting DAGLβ activity (Figure 6j and Figure S6f,h).…”

Investigating the Effects of Exogenous and Endogenous 2-Arachidonoylglycerol on Retinal CB1 Cannabinoid Receptors and Reactive Microglia in Naive and Diseased Retina

“…However, there are very few studies in the literature pertaining to their role in the retina. Acute treatment with the novel inhibitors of the 2-AG hydrolytic enzymes, AM12100 (ABHD6) and AM11920 (dual MAGL/ABHD6) [63] attenuated the AMPA-induced microglial/macroglial activation (Iba1-IR/GFAP-IR) and produced dosedependent partial neuroprotection, with the dual inhibitor being more efficacious [31]. These results suggest that the pharmacological blockade of the 2-AG hydrolytic enzymes and the subsequent increase in the endogenous 2-AG levels may provide a beneficial therapeutic strategy for neurodegenerative retinopathies.…”

Research Advances Through Activity‐Based Lipid Hydrolase Profiling

“…Many inhibitors of the ABHD protein family are summarized by Bononi et al ., [62] several of which have been identified through ABPP. Recently, ABPP enabled the discovery of a new ABHD6 inhibitor that improves selectivity over MGL and FAAH [63] …”

“…Recently, ABPP enabled the discovery of a new ABHD6 inhibitor that improves selectivity over MGL and FAAH. [63] In a recent publication on the immunomodulatory lipid networks in innate immune cells, the Cravatt lab used DAGLB inhibitors DO34 and DH376 as well as the MGL inhibitor MJN110 to investigate the activity and contribution of these enzymes in human monocyte-derived macrophages. [6a] They employed competitive ABPP to confirm that these inhibitors work in primary cell lines as well as to rule out off-targets.…”