Prostate cancer is the leading cause of cancer and second leading cause of cancer-related death in men in the United States. Twenty percent of patients receiving the standard of care androgen deprivation therapy (ADT) eventually progress to metastatic and incurable castration-resistant prostate cancer (CRPC). Current FDA-approved drugs for CRPC target androgen receptor (AR) binding or androgen production, but only provide a 2- to 5-month survival benefit due to the emergence of resistance. Overexpression of AR coactivators and the emergence of AR splice variants, both promote continued transcriptional activation under androgen-depleted conditions and represent drug resistance mechanisms that contribute to CRPC progression. The AR contains two transactivation domains, activation function 2 (AF-2) and activation function 1 (AF-1), which serve as binding surfaces for coactivators involved in the transcriptional activation of AR target genes. Full-length AR contains both AF-2 and AF-1 surfaces, whereas AR splice variants only have an AF-1 surface. We have recently prosecuted a high-content screening campaign to identify hit compounds that can inhibit or disrupt the protein-protein interactions (PPIs) between AR and transcriptional intermediary factor 2 (TIF2), one of the coactivators implicated in CRPC disease progression. Since an ideal inhibitor/disruptor of AR-coactivator PPIs would target both the AF-2 and AF-1 surfaces, we describe here the development and validation of five AF-2- and three AF-1-focused assays to interrogate and prioritize hits that disrupt both transactivation surfaces. The assays were validated using a test set of seven known AR modulator compounds, including three AR antagonists and one androgen synthesis inhibitor that are FDA-approved ADTs, two investigational molecules that target the N-terminal domain of AR, and an inhibitor of the Hsp90 (heat shock protein) molecular chaperone.

中文翻译：

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询问化合物抑制雄激素受体的AF-2或AF-1反式激活域的能力的测定法。

在美国，前列腺癌是癌症的主要原因，也是与癌症相关的死亡的第二主要原因。接受标准的雄激素剥夺疗法（ADT）的患者中有20％最终发展成转移性和不可治愈的去势抵抗性前列腺癌（CRPC）。目前FDA批准的用于CRPC靶向雄激素受体（AR）结合或雄激素生成的药物，但由于耐药性的出现，只能提供2到5个月的生存获益。AR共激活因子的过度表达和AR剪接变体的出现，都促进了雄激素耗尽条件下的持续转录激活，并代表了导致CRPC进展的耐药机制。AR包含两个反激活域，激活功能2（AF-2）和激活功能1（AF-1），它们作为参与AR靶基因转录激活的共激活因子的结合表面。全长AR包含AF-2和AF-1曲面，而AR拼接变体仅具有AF-1曲面。我们最近发起了一场高含量的筛选活动，以鉴定可以抑制或破坏AR与转录中间因子2（TIF2）之间的蛋白-蛋白质相互作用（PPI）的命中化合物，后者是CRPC疾病进展中涉及的共激活因子之一。由于理想的AR-coactivator PPI抑制剂/干扰物将同时靶向AF-2和AF-1表面，因此我们在这里描述了五个AF-2和三个AF-1聚焦检测法的开发和验证，以检验命中率并确定其优先级破坏两个反式激活表面。使用七个已知的AR调节剂化合物的测试集验证了这些测定，