Apalutamide (ARN-509) is a small molecule and androgen receptor (AR) antagonist with potential antineoplastic activity.

Androgen receptor:16 nM

每天口服10 mg/kg ARN-509,会抑制成年雄性狗前列腺组织的细胞增殖.在含有LNCaP/AR-luc异种移植肿瘤的阉割雄性免疫缺陷小鼠中,ARN-509（10 mg/kg/d,口服）抑制肿瘤生长,增殖指数降低并且凋亡率增加.在携带LNCaP/AR-luc异种移植肿瘤的阉割雄性免疫缺陷小鼠中,ARN-509剂量依赖性地抑制肿瘤生长.以10 mg/kg/天给药28天的ARN-509导致成年雄性狗缺乏腺分泌活性和附睾重量减少1.7倍,与此相关,前列腺重量减少3倍.30到300 mg范围内的ARN-509在带有转移性CRPC 病人中,会导致PSA的持续下降.ARN-509在阉割抗前列腺癌小鼠模型中,具有很强的抗癌症活性并能诱导治疗完成后长时间的症状缓解.ARN-509在24例已经接受了前期治疗的带有转移性CRPC病人中是安全的,耐受性良好,血药峰值出现在用药后2到3小时.

10 μM ARN-509在表达AR-EYFP的LNCaP 细胞中,会破坏AR的核定位并因此降低可用于结合表达AR-EYFP的LNCaP细胞中雄激素应答元件（ARE）的AR浓度。ARN-509在表达VP16-AR融合蛋白和ARE驱动的萤光素酶报道分子的Hep-G2细胞中抑制R1881诱导的VP16-AR介导的转录,IC50为0.2 μM。< 10 μM ARN-509在LNCaP/AR 前列腺癌细胞系中,抑制雄激素介导的13种内源基因（包括PSA和TMPRSS2）mRNA表达水平的诱导或抑制。在LNCaP/AR前列腺癌细胞系中,ARN-509（<10 μM）抑制R1881（30 pM）的增殖作用。10 μM ARN-509可以有效地与1 nM R1881竞争从而阻止AR与启动子区域结合。

Competitor assay kits (green) are used to determine relative in vitro binding affinities of ARN-509 for the rat AR ligand binding domain (LBD), human progesterone receptor (PR) LBD, and full-length human estrogen receptor-alpha (ERα) and human glucocorticoid receptor (GR). Each hormone dose is performed in triplicate, relative error is calculated from the standard error of the mean (SEM), and binding curves are fit using a single binding site competition model (Prism statistical analysis software package) with R2>0.8. Experiments are conducted multiple times with SEM<0.3 log units from the average logIC50 value. Ki values are calculated as averages across experiments with SEM, and binding affinities are reported as a percentage relative to the tight-binding ligand control for that receptor[1].

Cells are incubated for 48 hours, after which ARN-509 is added in a 16 μL volume to the RPMI culture medium. For the antagonist mode assay, the ARN-509 is diluted in culture medium also containing 30 pM R1881. After 7 days' incubation, 16 μL of CellTiter-Glo Luminescent Cell Viability Assay is added and Relative Luminescence Units (RLUs) measured.(Only for Reference)