MLi-2
Probe criteria
| Inhibitor/agonist potency: goal is < 100 nM (IC50, Kd) | Surpasses criterion: LRRK2 potency is < 5 nM in MSD and Invitrogen assays: LRRK2 (IC50 = 1.8 nM), LRRK2 with 5mM ATP (IC50 = 6.3 nM) |
| Selectivity within target family: > 30-fold | Surpasses criterion: > 100-fold kinome selective with IC50 < 1 µM for the following kinases: CLK4 (225 nM), MAP3K14 (244 nM), MAP3K5 (428 nM), CLK2 (605 nM), and TTK (935 nM). NanoBRET results (SGC Frankfurt): CLK4 (EC50 = 24 ± 8.8 µM), MAP3K14 (EC50 > 50 µM), MAP3K5 (EC50 = 12 ± 3.1 µM), CLK2 (EC50 = 2 ± 0.63 µM), TTK (EC50 > 50 µM) |
| Selectivity outside target family | PanLabs Profile shows 5 hits > 50% inhibition (@ 10 μM): HTR2B (1.2 µM), SLC6A2 (3.8 µM), CHRM2 (6.4 µM), PPARG (6.5 µM), adenosine transporter (9.7 µM).Clean PDSP scan except for weak activity on TMEM97 (Ki = 4600.45 nM) |
| On target cell activity for cell-based targets: goal is < 1 µM IC50/EC50 | Surpasses criterion: IC50 = 3.5 nM in LRRK2 PS935; NanoBRET (SGC Frankfurt): LRRK2 EC50 = 2.5 ± 0.13 nM |
| Control compound (100 times less potent than the probe) | Surpasses criterion: LRRK2 potency 1000x less: MLi-2-NC (IC50 = 6480 nM)NanoBRET assay (SGC Frankfurt) (EC50 [µM]; n = 3): LRRK2 (> 50), CLK4 (6.2 ± 1.2), MAP3K14 (> 50), MAP3K5 (4.1 ± 1.7), CLK2 (11 ± 1.7), TTK (> 50) Clean PDSP scan except for activity on TMEM97 (Ki = 1074.98 nM) and SIGMAR1 (Ki = 1593.68 nM) |