CUDC-101 is a potent inhibitor of HDAC, EGFR and HER2 with IC50s of 4.4, 2.4 and 15.7 nM, respectively.

HER2:15.7 nM, HDAC:4.4 nM, EGFR:2.4 nM

CUDC-101以剂量依赖的方式抑制埃罗替尼敏感的H358 NSCLC移植瘤的生长.CUDC-101也显示在埃罗替尼抗性A549NSCLC移植瘤模型中对肿瘤生长的有效抑制.CUDC-101在拉帕替尼耐药,HER2阴性,EGFR过表达MDA-MB-468乳腺癌模型和EGFR过表达CAL-27头颈部鳞状细胞癌（HNSCC）模型中显著促进肿瘤消退.每天给药120 mg/kg CUDC-101处理Hep-G2肝肿瘤模型能够诱导肿瘤的退化,比埃罗替尼所用的最大耐药剂量（25 mg/kg /天）以及vorinostat等摩尔剂量(72 mg/kg/天)时的抑制效果都更有效.

CUDC-101在很多人癌细胞中表现出广谱的抗增殖活性,IC50在0.04 μM到0.80 μM之间。CUDC-101与厄洛替尼,拉帕替尼以及伏立诺他与埃罗替尼或拉帕替尼的联用相比,在大多数情况下显示出更高的效力。CUDC-101能够抑制具有拉帕替尼和埃罗替尼抗性的癌细胞系的增殖。在各种癌细胞系中,CUDC-101处理以剂量依赖性方式增加组蛋白H3和H4的乙酰化以及HDAC的非组蛋白底物如p53和α-微管蛋白的乙酰化。CUDC-101同时还能够抑制肿瘤细胞中HER3的表达,Met的扩增以及AKT再激活。

HDAC, EGFR and HER2 inhibition assays: The activities of Class I and II HDACs are assessed using the Biomol Color de Lys system. Briefly, HeLa cell nuclear extracts are used as a source of HDACs. Different concentrations of CUDC-101 are added to HeLa cell nuclear extracts in the presence of a colorimetric artificial substrate. Developer is added at the end of the assay and enzyme activity is measured in the Wallac Victor II 1420 microplate reader at 405 nM. EGFR and HER2 kinase activity are measured using HTScan EGF receptor and HER2 kinase assay kits. Briefly, the GST-EGFR fusion protein is incubated with synthetic biotinylated peptide substrate and varying concentrations of CUDC-101 in the presence of 400 mM ATP. Phosphorylated substrate is captured with strapavidin-coated 96-well plates. The level of phosphorylation is monitored by antiphospho-tyrosine- and europium-labeled secondary antibodies. The enhancement solution is added at the end of the assay and enzyme activity is measured in the Wallac Victor II 1420 microplate reader at 615 nM.

Cancer cell lines are plated at 5000 to 10000 cells per well in 96-well flatbottomed plates with varying concentrations of CUDC-101. The cells are incubated with CUDC-101 for 72 hours in the presence of 0.5% of fetal bovine serum. Growth inhibition is assessed by an adenosine triphosphate (ATP) content assay using the Perkin-Elmer ATPlite kit. Apoptosis is routinely assessed by measuring the activities of Caspase-3 and -7 using Apo-ONE Homogeneous Assay Kit.(Only for Reference)