Donated Chemical Probes

Target	MGLL
Targeted domain	Azetidine-amide carbonyl forms H-bonds with NH of Ala51 and Met123 in oxyanion hole
Mode of action	Non-covalent, competitive, reversible inhibitor
Control	JNJ-8034
Recommended cellular usage concentration	≤ 10 µM
In vivo use	Yes
Donated by	Johnson & Johnson

Inhibitor/agonist potency: goal is < 100 nM (IC₅₀, K_d)	Surpasses criterion: Fluorometric Assay (Inhibition of 4MU-B cleavage): IC₅₀ < 5 nM (n=10)
Selectivity within target family: > 30-fold	Serine protease panel (15 proteases) at 10 µM: clean Panel of endocannabinoid related targets: Closest hit is hFAAH with IC₅₀ > 4 µM
Selectivity outside target family	Kinase Panel: 50 targets, 10 µM ATP at 10 µM, all < 20% inhibition CEREP panel of 50 binding assays at 10 µM (ion channels, receptors, including CB1 and CB2): binding > 50%: rat OPRK1 (66%), rat HTR1B HTR1B (56%), rat chloride ion channel (55%), human SLC6A3 (54%). Follow up in cellular assays (10 µM): OPRK1 (no activity), HTR1B (inhibition 50%) and inhibited dopamine uptake by 35%. Clean PDSP scan (43 targets at 10 µM).
On target cell activity for cell-based targets: goal is < 1 µM IC₅₀/EC₅₀	Surpasses criterion: [3H] 2-OG cleavage activity assay: IC₅₀ = 1.13 ± 0.05 nM (n> 10, human HeLa cells); IC₅₀ = 1.88 ± 0.41 nM (n = 6, human PBMC); IC₅₀ = 0.67 ± 0.11 nM (n = 9, mouse brain); IC₅₀ = 0.97 ± 0.12 nM (n = 10, rat brain)
Control compound (100 times less potent than the probe)	JNJ-8034: Fluorometric Assay (Inhibition of 4MU-B cleavage): IC₅₀ > 10 µM (n=6) Clean PDSP scan (43 targets at 10 µM).