Identification of TR4 Modulators for Treatment of Cushing Disease.

用于治疗库欣病的 TR4 调节剂的鉴定。

• 批准号: 10199436
• 负责人: ANTHONY P HEANEY
• 金额: $ 40.11万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10199436
• 关键词: Adrenal Glands; Adrenalectomy; Affect; Bilateral; Biochemical; Biological Assay; Cells; Chemicals; Clinical Research; Clinical Trials; Corticotropin; Databases; Disease; Dose; Elements; Evaluation; Excision; Future; Genetic Transcription; Glucocorticoids; Goals; Health Care Costs; Homeostasis; Hormone secretion; Human; Hybrids; Hydrocortisone; Immunoassay; In Vitro; Lead; Length; Libraries; Life; Ligand Binding Domain; Luciferases; Medical; Morbidity - disease rate; Mus; Nuclear Orphan Receptor; Orphan; POMC gene; Patients; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Pituitary Corticotropin Secreting Adenoma; Pituitary Gland; Pituitary Neoplasms; Pituitary-dependent Cushing' s disease; Plasmids; Primary Neoplasm; Pro-Opiomelanocortin; Radiation therapy; Rare Diseases; Renilla Luciferases; Repeat Surgery; Reporter; Research Personnel; Residual Tumors; Series; Speed; Structure; System; Testing; Transactivation; Translating; Validation; Variant; Yeasts; base; cheminformatics; clinical investigation; cloud based; cost effectiveness; cross reactivity; cytotoxic; drug discovery; efficacious treatment; experience; high throughput screening; hypothalamic-pituitary-adrenal axis; in vivo; inhibitor/antagonist; internal control; multidisciplinary; novel; polypeptide; promoter; receptor; response; scaffold; small molecule; small molecule inhibitor; small molecule libraries; statistics; tool; transcription factor; tumor

ABSTRACT Cushing Disease (CD) is caused by an adrenocorticotropic hormone (ACTH)-secreting pituitary tumor that causes excess adrenal-derived cortisol. It is a life-threatening “orphan disease” with a staggering annual health care cost that is >7-fold higher than average patients. Surgical removal is the current first-line therapy but the disease frequently recurs. Repeat surgery, radiation therapy and bilateral adrenalectomy are not always successful and associated with major morbidity. Currently available drugs or those in clinical trials for CD do not target the pituitary corticotroph tumor itself and escape from control is common with long-term use. A clear unmet need for efficacious and safe therapies that offer biochemical and tumor control exists. We hypothesize that direct targeting of corticotroph tumors to modulate ACTH and in turn, glucocorticoid secretion is the optimal way to treat CD. We recently demonstrated that the orphan testicular receptor 4 (TR4, also known as NR2C2) is a potent regulator of hypothalamic-pituitary-adrenal (HPA) axis function and directly regulates pro- opiomelanocortin (POMC) gene transcription and ACTH secretion. We hypothesize that small molecule inhibitors of TR4 action would be potent inhibitors of corticotroph tumor hormone (ACTH) secretion. Such a discovery would be a transformative therapy for Cushing disease as no similar therapies exist. To identify and characterize TR4 small molecule modulators, we will perform a large scale small molecule library screen using our unique series of transactivation assays that include a TR4-directed GAL4-LBD system and POMC-promoter reporters as well as a secondary screen in human corticotroph tumor primary cultures. Our first aim will use a mammalian one-hybrid GAL4-LBD system to identify TR4 modulators from a library of 200,000 distinct compounds. Potential “hit” compounds will be confirmed by dose-response evaluation assays to calculate their EC50. Compounds identified in aim 1 will be further validated in aim 2 using a POMC transactivation assay using full-length and truncated TR4 variant plasmids together with a series of POMC promoter-driven luciferase reporters in murine corticotroph tumor AtT20 cells. A secondary screen will also be performed in human corticotroph tumor primary cultures to confirm their in vitro effects on ACTH secretion. We expect our proposal will identify and rigorously validate compounds that efficiently and specifically abrogate TR4 actions to inhibit ACTH secretion and lead to the discovery of safe and efficacious lead TR4 inhibitory compounds that can be further advanced as potential drug therapies for CD.

摘要 库欣病（CD）是由分泌促肾上腺皮质激素（ACTH）的垂体肿瘤引起的 导致肾上腺皮质醇分泌过多这是一种威胁生命的“孤儿病”， 医疗费用比普通患者高出7倍以上。手术切除是目前的一线治疗方法， 这种病经常复发。重复手术，放射治疗和双侧肾上腺切除术并不总是 成功并伴有严重的发病率。目前可用的药物或那些在临床试验的CD不 靶向垂体促肾上腺皮质激素细胞瘤本身而失控是长期使用的常见情况。一个明显的未满足 需要提供生物化学和肿瘤控制的有效和安全的治疗。我们假设 直接靶向促肾上腺皮质激素细胞肿瘤以调节ACTH，进而调节糖皮质激素分泌是最佳途径 治疗CD。我们最近证明，孤儿睾丸受体4（TR 4，也称为NR 2C 2）是一种免疫调节因子。 下丘脑-垂体-肾上腺（HPA）轴功能的有效调节剂，并直接调节促肾上腺皮质激素的分泌。 阿黑皮素（POMC）基因转录和ACTH分泌。我们假设小分子抑制剂 TR 4的作用将是促肾上腺皮质激素肿瘤激素（ACTH）分泌的有效抑制剂。这样的发现 将是库欣病的变革性疗法，因为没有类似的疗法存在。以鉴定和表征 TR 4小分子调节剂，我们将使用我们独特的小分子筛选技术进行大规模的小分子文库筛选。 一系列反式激活试验，包括TR 4-指导的GAL 4-LBD系统和POMC-启动子报告基因 以及在人促肾上腺皮质激素细胞肿瘤原代培养物中的二次筛选。我们的第一个目标是利用哺乳动物 单杂交GAL 4-LBD系统，以从200，000种不同化合物的文库中鉴定TR 4调节剂。潜在 “命中”化合物将通过剂量-反应评价测定来确认，以计算其EC 50。化合物 将在目标2中使用使用全长的POMC反式激活试验进一步验证目标1中鉴定的目的， 小鼠中截短的TR 4变体质粒与一系列POMC启动子驱动的荧光素酶报告基因 促肾上腺皮质激素瘤AtT 20细胞。还将在人促肾上腺皮质激素细胞肿瘤原发性 培养物以确认其对ACTH分泌的体外作用。我们希望我们的提案将确定并严格 验证有效和特异性地消除TR 4作用以抑制ACTH分泌并导致 安全有效的TR 4抑制先导化合物的发现， CD的药物治疗。