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: 10199959
• 负责人: BENITA S KATZENELLENBOGEN
• 金额: $ 55.94万
• 依托单位: SCRIPPS 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 2022-04-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10199959
• 关键词: AF2; Agonist; Animal Model; Aromatase Inhibitors; Bar Codes; Binding; Biological Assay; Breast; Breast Cancer Cell; Breast Cancer Model; Breast Cancer therapy; CDK4 gene; Cessation of life; Chemical Structure; Chemicals; Clinical; Color; Crystallization; Cyclic AMP-Dependent Protein Kinases; DNA Binding; Development; Disease; Drug Kinetics; Endocrine; Estradiol; Estrogen Antagonists; Estrogen Receptor alpha; Estrogen Receptors; Estrogen receptor negative; FRAP1 gene; Failure; Fulvestrant; Genes; Genetic Transcription; Goals; Growth Factor; Growth and Development function; Hormones; Hydrogen Bonding; Hyperactivity; Individual; Injections; Knock-out; Ligand Binding Domain; Ligands; Mediating; Modeling; Molecular; NCOA2 gene; NCOA3 gene; Noise; Optics; Pain; Patients; Peptides; Pharmacology; Phosphorylation; Postmenopause; Production; Raloxifene; Recurrence; Repression; Research Personnel; Resistance; Resolution; Selective Estrogen Receptor Modulators; Side; Signal Transduction; Speed; Structure; Surface; Systems Biology; Tamoxifen; Testing; Tissues; Transcript; Treatment Efficacy; Treatment Protocols; Uterus; Validation; Visualization software; Woman; Work; X-Ray Crystallography; base; cell growth; clinical candidate; design; gene repression; genetic corepressor; high throughput screening; hormone therapy; improved; in vivo; in vivo Model; inhibitor/antagonist; innovation; insight; malignant breast neoplasm; mutant; novel; overexpression; personalized approach; personalized medicine; pharmacophore; prevent; receptor; recruit; resistance mechanism; scaffold; synergism; targeted agent; therapy resistant

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.

明确的目标。我们的目标是使用集成的结构、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乳房不同机制方面的有效性得到改善 癌症，使更个性化的药物成为可能。由于70%-80%的ERα阳性患者和高达50%的内分泌治疗因疾病复发而失败，大多数死亡病例是ERα阳性患者，这突显了对改进治疗的重大临床需求未得到满足。重要的是，一种激素治疗失败的患者通常会对另一种具有替代作用机制的激素产生反应。目标1.使用结构和化学系统生物学 在不同的基于ERα的抗性机制的背景下确定拮抗ERα的规则的方法， 作为与其他非ERα耐药靶向剂的联合治疗。这些方法旨在克服阻力。 并与ERα拮抗剂协同治疗ER(+)乳腺癌。我们已经开发了一种系统生物学方法，使 与许多配体平行的ERα配体结合域的结晶。几十种晶体结构的分析 并行使我们能够实施一种公正的方法来识别亚范围内(在单个结构的噪声范围内)的微妙结构扰动，这些扰动对ERα调节的增殖有很大贡献，我们称之为超分辨率X射线结晶学。我们将使用这种方法来确定驱动转录的结构特征 不同耐药模型中结构不同的配体的抑制、受体降解和治疗效果 以及联合治疗方案的协同研究，包括PI3K、mTOR和CDK4/6抑制剂。我们已经开发出 一种跟踪受体降解的定量高通量筛选试验。ERα驱动的关键靶基因， 辅调节因子相互作用和定量降解分析将被用来机械地将受体结构与抗逆转录因子联系起来。 在对治疗敏感和耐药的乳腺癌模型中，通过一种我们称为配基类别分析的化学系统生物学方法，研究了乳腺癌模型中的增殖效应，包括结构性活性突变ERα、高活性生长因子信号转导和SRC3/AIB1的过表达。目的2.设计和合成具有新型药效团的ER-α全拮抗剂 产生明显的受体结构扰动，并在动物体内表征其药理作用 耐药ERα(+)乳腺癌模型。牙釉质支架提供了一个稳定的核心，我们将 探索不同的、新的侧链如何干扰ERα结构来调节转录抑制和克服 抵抗。因为我们发现替代的OBHS-N核心化合物产生了完全的内质网拮抗和降解，没有侧链，通过口袋内的扰动作为间接拮抗剂，这个OBHS-N核心现在提供了一个新的，等量纲的平台，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编码，从而允许对配体有效性进行多路体内评估。通过这两个类别，我们将在体内评估它们与野生型和三苯氧胺耐药ERα(+)模型的对比。将以ERα基因敲除和ER阴性乳腺癌细胞作为对照来验证靶向活性，并验证对体内关键ERα转录本的影响。