Y06036, a potent and selective BET inhibitor, can bind to the BRD4(1) bromodomain (Kd: 82 nM).

BRD4(1):82 nM (Kd, cell free)

Y06036与BRD4(1)溴结构域结合，其Kd值为82 nM。Y06036在前列腺癌细胞系中强效抑制细胞增长、集落形成以及AR、AR调控基因和MYC的表达。

小鼠在肿瘤体积达到约100 mm3时，随机分组并通过腹膜内(i.p.)给药，接受安慰剂或BET抑制剂Y06036（50 mg/kg，每周5次）治疗。在25天的治疗期间，Y06036表现出强大的抗肿瘤活性，肿瘤生长抑制(TGI)达到70%。

LNCaP, C4-2B, 22Rv1, and VCaP prostate cancer cells were cultured in RPMI 1640 with 10% FBS at 37 °C and an atmosphere of 5% CO2. For cell viability, cells were seeded in 384-well plates at 500?1000 cells per well (optimum density for growth) in a total volume of 20 μL of media. After 12 h, 10 μL of chemical compounds with 2-fold or 3-fold serial dilution was added to each well with final concentration from 5 nM to 100 μM. The measurement was conducted 96 h after seeded for LNCaP, C4-2B, and 22Rv1 and 144 h after seeded for VCaP. Then, 25 μL of CellTiter-GLO reagents was added, and luminescence was measured on GLOMAX microplate luminometer, according to the manufacturer's instructions. The estimated in vitro half-maximal inhibitory concentration (IC50) values were calculated using Prism 6 software.

Four-week-old male mice (strain: C.B-17/IcrHsd-Prkdcscid for C4-2B) were used for tumor inoculation. Each mouse was inoculated subcutaneously at the dorsal flank on both sides of the mice with C4-2B tumor cells (2 × 10^6 cells) in a mixture of 100 μL PBS and Matrigel (1:1). When the tumor volume reached approximately 100 mm3, the mice were randomized into groups (n = 5?7 per group) and then treated intraperitoneally (ip) with 100 μL of either vehicle or Y06036 and 7m (in a formulation of 15% Cremophor EL, Calbiochem, 82.5% PBS, and 2.5% DMSO) five times per week. The length (L) and width (W) of the tumor mass were monitored by calipers, and volume was expressed in mm3 calculated with the equation V = (π/6)(L × W2). Tumor growth inhibition (TGI) was calculated using the equation TGI = [1 ? (T ? T0)/(C ? C0)] × 100, wherein T and T0 are the mean tumor volumes on a specific experimental day and on the first day of treatment, respectively, for the test groups; and likewise C and C0 are the mean tumor volumes for the vehicle group.