Enzalutamide (MDV3100) is an androgen receptor (AR) antagonist (IC50=36 nM in LNCaP). Enzalutamide activates autophagy, has antitumor activity, and is commonly used in the treatment of desmoplasia-resistant prostate cancer.

Androgen receptor:36 nM (LNCaP cells)

方法：人前列腺癌细胞 LNCaP、PC3 和人骨肉瘤细胞 SJSA-1 用 Enzalutamide (0.01-100 μM) 处理 1 h，使用 MTT 检测细胞活力。
结果：Enzalutamide 显示降低了所有细胞系的生存能力，对 LNCaP、PC3 和 SJSA-1 细胞系的 LD50 分别为 12 μM、23.4 μM 和 34.7 μM。[1]
方法：人前列腺癌细胞 VCaP 用 Enzalutamide (10 μM) 处理 1-3 天，使用 Western Blot 方法检测靶点蛋白表达水平。
结果：Enzalutamide 上调 cleaved-PARP 表达，诱导细胞凋亡。[2]

方法：为检测体内抗肿瘤活性，将 Enzalutamide (1-50 mg/kg) 灌胃给药给携带人去势抵抗性肿瘤 (CRPC) LNCaP-AR-Lux 的 CB17SCID 小鼠，每天一次，持续二十八天。
结果：Enzalutamide 在 CRPC 异种移植物模型中诱导肿瘤体积的消退，并在 AR 过度扩张的前列腺癌症细胞中诱导细胞凋亡。[3]
方法：为评估 AR 在性成熟后骨代谢和骨骼生长中的作用，将 Enzalutamide (10-100 mg/kg，1% carboxymethyl cellulose+0.5% Tween 80+5% dimethylsulfoxide) 口服给药给 C57BL/6N 小鼠，每天一次，持续二十一天。
结果：Enzalutamide 降低了轴骨中的骨量，Enzalutamide 显著降低了轴心骨骼的机械强度。性成熟后，Enzalutamide 可减少雄性小鼠中轴骨骼的骨量，但不能减少阑尾骨骼的骨量。[4]

For in vitro experiments, LNCaP or LNCaP/AR cells (10^4 cells/well) were androgen-starved by growth in media containing 5-10% charcoal-stripped serum for 3-5 days. Then the cells were challenged with various concentrations of R1881, bicalutamide, RD162 or MDV3100 in media containing 5-10% charcoal-stripped serum [1].

In vivo tumorigenicity experiments were done by subcutaneous injection of 10^6 cells (100 uL in 50% Matrigel and 50% growth media) into the flanks of castrated male SCID mice. Daily gavage treatment (using a formulation of 1% carboxymethyl cellulose, 0.1% Tween-80, 5% DMSO) was initiated when tumor size reached ~100 mm3. Tumor size was measured weekly in three dimensions (l x w x d) with calipers. For in vivo luciferase imaging, d-luciferin substrate (100 μL, 15 mg/mL) was injected intraperitoneally. After 5 minutes, mice were anesthetized using isofluorane and imaged using a cooled charged-coupled device IVIS camera. Data were analyzed using Living Image 2.30 software [1].