Chemical, structural and molecular rules for fully antagonizing the estrogen receptor

完全拮抗雌激素受体的化学、结构和分子规则

• 批准号: 10595881
• 负责人: BENITA S KATZENELLENBOGEN
• 金额: --
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10595881
• 关键词: Chemical; structural; molecular; rules; fully

Specific Aims. Our goal is use integrated structural, chemical and molecular systems biology approaches to develop estrogen receptor (ER) antagonists with improved efficacy in targeting distinct mechanisms of hormone-resistant breast cancer, enabling more personalized medicine. As 70-80% of women present with ERα positive disease and up to 50% fail on endocrine therapies with disease recurrence, most deaths are of ERα-positive patients, highlighting a significant unmet clinical need for improved therapies. Importantly, patients who fail on one hormone therapy typically respond to a different one having an alternate mechanism of action. Aim 1. Use structural and chemical systems biology approaches to identify rules for antagonizing ERα in the context of different ERα -based mechanisms of resistance, and as co-treatments with other non-ERα resistance targeting agents. These approaches aim to overcome resistance and synergize with ERα antagonists in ER(+) breast cancer. We have developed a systems biology approach enabling crystallization of the ERα ligand-binding domain in parallel with many ligands. Analysis of dozens of crystal structures in parallel allows us to implement an unbiased approach to identify subtle structural perturbations in the sub-Å range (within the noise of the individual structures) that contribute significantly to ERα-regulated proliferation, which we call super-resolution x-ray crystallography. We will use this approach to identify structural features that drive transcriptional repression, receptor degradation, and therapeutic efficacy of structurally diverse ligands in different resistance models, and in synergy studies with co-treatment regimens, including PI3K, mTOR, and CDK4/6 inhibitors. We have developed a quantitative high-throughput screening assay for tracking receptor degradation. Key ERα-driven target genes, coregulator interaction and quantitative degradation assays will be used to mechanistically tie receptor structure to anti- proliferative effects in treatment-sensitive and resistant breast cancer models, including constitutively active mutant ERα, hyperactive growth factor signaling, and overexpression of SRC3/AIB1, in a chemical systems biology approach that we call ligand class analysis. Aim 2. Design and synthesize ERα full antagonists with novel pharmacophores that produce distinct structural perturbations of the receptor, and characterize their in vivo pharmacology in animal models of treatment-resistant ERα(+) breast cancer. The adamantyl scaffold provides a stable core with which we will explore how different, novel side chains perturb ERα structure to mediate transcriptional repression and overcome resistance. Because we found that the alternative OBHS-N core compounds produced full ER antagonism and degradation without side chains, by acting as indirect antagonists through perturbations within the pocket, this OBHS-N core now provides a novel, isosteric platform for adding side chains to access distinct and combined direct and indirect mechanisms of antagonism. We will optically barcode a number of resistance models, allowing for a multiplexed in vivo assessment of ligand efficacy. With both classes, we will evaluate them against wild type and tamoxifen-resistant ERα (+) models in vivo. On-target activity will be verified with ERα knockout and ER-negative breast cancer cells as control, with validation of effects on key ERα transcripts in vivo.

具体目标。我们的目标是使用综合结构、化学和分子系统生物学方法来开发 雌激素受体（ER）拮抗剂在针对激素抵抗性乳腺的不同机制方面具有更高的功效 癌症，实现更加个性化的医疗。由于 70-80% 的女性患有 ERα 阳性疾病，高达 50% 的女性内分泌治疗失败且疾病复发，因此大多数死亡是 ERα 阳性患者，这凸显了对改进治疗的重大未满足的临床需求。重要的是，一种激素疗法失败的患者通常会对具有替代作用机制的另一种激素疗法产生反应。目标 1. 使用结构和化学系统生物学 在不同的基于 ERα 的耐药机制的背景下确定拮抗 ERα 的规则的方法， 以及与其他非 ERα 耐药靶向药物的联合治疗。这些方法旨在克服阻力 并与 ERα 拮抗剂在 ER(+) 乳腺癌中发挥协同作用。我们开发了一种系统生物学方法，能够 ERα 配体结合结构域与许多配体平行结晶。数十种晶体结构分析 并行使我们能够实施一种公正的方法来识别亚 Å 范围内（在单个结构的噪声内）的微妙结构扰动，这些扰动对 ERα 调节的增殖有显着贡献，我们称之为超分辨率 X 射线晶体学。我们将使用这种方法来识别驱动转录的结构特征 不同耐药模型中结构多样的配体的抑制、受体降解和治疗功效， 以及与联合治疗方案（包括 PI3K、mTOR 和 CDK4/6 抑制剂）的协同研究。我们开发了 用于追踪受体降解的定量高通量筛选测定。关键 ERα 驱动的靶基因， 核心调节剂相互作用和定量降解测定将用于机械地将受体结构与抗- 治疗敏感性和耐药性乳腺癌模型中的增殖效应，包括组成型活性突变体 ERα、过度活跃的生长因子信号传导和 SRC3/AIB1 的过度表达，采用我们称为配体类别分析的化学系统生物学方法。目标 2. 设计并合成具有新颖药效团的 ERα 完全拮抗剂， 产生受体的独特结构扰动，并表征其在动物体内的药理学 难治性 ERα(+) 乳腺癌模型。金刚烷基支架提供了一个稳定的核心，我们可以用它来 探索不同的新颖侧链如何扰乱 ERα 结构以介导转录抑制并克服 反抗。因为我们发现替代的 OBHS-N 核心化合物在没有侧链的情况下产生完全的 ER 拮抗作用和降解，通过口袋内的扰动充当间接拮抗剂，该 OBHS-N 核心现在提供了一个新颖的等排平台，用于添加侧链以获得不同的和组合的直接和间接拮抗机制。我们将对许多耐药模型进行光学条形码标记，从而对配体功效进行多重体内评估。对于这两个类别，我们将在体内针对野生型和他莫昔芬耐药 ERα (+) 模型对它们进行评估。将以 ERα 敲除和 ER 阴性乳腺癌细胞作为对照来验证靶向活性，并验证体内对关键 ERα 转录物的影响。