ACY-738 demonstrates inhibitory activity against recombinant HDAC6 (IC50: 1.7 nM), with respective average selectivity over class I HDACs being 100-fold.

HDAC6:1.7 nM

ACY-738在浓度为2.5μM时，能够增加RN46A-B14细胞中的α-tubulin的乙酰化（赖氨酸40）分子部分。ACY-738（10 μM）引发细胞死亡的效果与LBH589和FK228相似。

ACY-738 (5mg/kg) 显著增加整个大脑裂解物中α-tubulin的乙酰化。ACY-738 (50mg/kg) 在家庭饲养环境中对WT小鼠的运动活动没有增强作用。ACY-738 (5 mg/kg BW) 改变BM B细胞分化，但在NZB/W小鼠中对IgG和C3沉积没有显著影响。ACY-738 (5 mg/kg) 在治疗后的0.0830小时达到最大血浆浓度1310 ng/mL。ACY-738 (20 mg/kg) 显著减轻NZB/W F1小鼠的蛋白尿严重程度。ACY-738 (5 mg/kg) 随着NZB/W小鼠年龄的增长，对抗dsDNA产生显示出显著的减少。它还显著减少了glomerular IL-6和IL-10 mRNA水平超过50%，而使用20 mg/kg的ACY-738治疗将IL-6和IL-10 mRNA降至不可检测水平。ACY-738 (5, 20 mg/kg) 随着NZB/W小鼠年龄的增长，减轻了血清IL-1β的产生。

Recombinant kinase domain of EGFR L858R and T790M-L858R mutants are incubated with EGF816 to confirm covalent modification of EGFR and site of adduction. Recombinant enzyme is incubated at room temperature with a 20-fold molar excess of compound in 40 mM Tris, pH 8, 500 mM NaCl, 1% glycerol, 5 mM TCEP for 1 h. The reaction is quenched by addition of dithiothreitol (DTT, 80-fold excess to compound) and transfer to ice. A third of the reaction (10 μL) is processed for intact MS by adding an equal volume of 6 M Guan HCl, 100 mM Tris, pH 8, 20 mM DTT, 10 mM TCEP and incubating at room temperature for 15 min. Intact MS analysis is performed on an Agilent 6520 QToF mass spectrometer equipped with a dual spray ion source (IS of 4500 V, fragmentor of 250 V, fas temp of 350°C, and skimmer of 75 V). The samples are injected onto a PLRP-S column (2.1 mm × 50 mm), heated to 60°C, and desalted for 2 min at 500 μL/min and 3% B prior to elution with a fast gradient of 3-50% B in 3 min (B, 0.1% formic acid). The data are analyzed in MassHunter for automatic peak selection, integration, and spectral deconvolution with a mass range of 15?000-75?000 Da.