TBPB, an allosteric M1 mAChR agonist, is an [iso]propyl (2-{4-methoxy-3-[3-(4-methylpiperidin-1-yl)propoxy]phenyl}ethyl)carbamate exhibiting a molar mass of 430.6 g/mol.

M1 mAChR (allosteric):289 nM(ec50)

in vitro kinase assay: MELK recombinant protein (0.4 μg) is mixed with 5 μg of each substrate in 20 μL of kinase buffer containing 30 mM Tris-HCl (pH), 10 mM DTT, 40 mM NaF, 10 mM MgCl2, 0.1 mM EGTA with 50 μM cold-ATP and 10 Ci of [γ-32P]ATP for 30 min at 30 °C. The reaction is terminated by addition of SDS sample buffer and boiled for 5 min prior to SDS-PAGE. The gel is dried and autoradiographed with intensifying screens at room temperature. OTSSP167 (final concentration of 10 nM) is dissolved in DMSO and added to kinase buffer before the incubation.