Universal positive allosteric modulators of CCK1R without intrinsic agonist activity for the treatment of obesity

用于治疗肥胖症的无内在激动剂活性的 CCK1R 通用正变构调节剂

• 批准号: 9918929
• 负责人: LAURENCE J MILLER
• 金额: $ 41.5万
• 依托单位: MAYO CLINIC ARIZONA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-13 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9918929
• 关键词: Accounting; Acute; Affect; Afferent Neurons; Agonist; Animals; Behavior; Bile fluid; Binding; Biological; Biological Assay; Body Weight decreased; Calcium; Cell Line; Cells; Chemicals; Chinese Hamster Ovary Cell; Cholecystokinin; Cholecystokinin Receptor; Cholesterol; Clinic; Clinical Trials; Coupled; Coupling; Data; Defect; Desire for food; Diet; Diet Modification; Eating; Ensure; Environment; Epidemic; Feedback; G-Protein-Coupled Receptors; Gastrointestinal Hormones; General Population; Genomics; Goals; Hormonal; Hormones; Human; Individual; Laboratories; Lead; Libraries; Life Style; Mediating; Membrane; Metabolic syndrome; Molecular; Molecular Conformation; Morbid Obesity; Neurons; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Patients; Peripheral; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Physiological; Population; Postprandial Period; Process; Property; Reagent; Satiation; Signal Transduction; Stimulus; Surveys; Testing; Therapeutic; Time; Toxic effect; Validation; Weight Gain; analog; bariatric surgery; base; cheminformatics; clinical development; comorbidity; cytotoxicity; density; design; drug action; drug candidate; environmental enrichment for laboratory animals; experience; high throughput screening; hormone sensitivity; interest; metabolic phenotype; mortality; mutant; negative affect; novel; obesity treatment; patient subsets; positive allosteric modulator; primary endpoint; receptor; receptor function; reduced food intake; response; scaffold; screening; side effect; small molecule; small molecule libraries; success

PROJECT SUMMARY/ABSTRACT Obesity has reached epidemic proportions, contributing to similar increases in type 2 diabetes mellitus and its multiple co-morbidities, accounting for immense expense, suffering, and early mortality. Modification of diet and lifestyle, while effective for acute weight loss, is not durable, and bariatric surgery, while effective for morbid obesity, is not scalable for this need. Therefore, new safe and effective pharmacotherapies for obesity are needed. This proposal is responsive to PAR-16-374 soliciting assays for discovery of such therapeutics. Our OBJECTIVE is to identify molecules with activity as positive allosteric modulators (PAMs) of the type 1 cholecystokinin receptor (CCK1R) that do not exhibit intrinsic agonist activity and that can be safe and effective across the full spectrum of potential patients, from those tending to gain weight to the morbidly obese with metabolic syndrome. This type of drug does not now exist. It would increase signaling responses to endogenous hormone released after a meal, when a greater biological response can enhance satiety, while overcoming side effects and potential toxicity caused by full agonists. It would also have the unique property of correcting a defect in stimulus-activity coupling now recognized to be present in a subset of these patients. Sequential dual screening strategies will be directed toward the natural wild type (WT) CCK1R, as well as this receptor in an abnormal conformation induced by elevated cholesterol in the membrane, with the latter mimicked by a CCK1R mutant. Enhancing and/or recalibrating CCK1R to be normally responsive to CCK in these settings could accentuate the hormonal effect on satiety in a gentle and safe manner. This represents a partnership between Dr. Miller at Mayo Clinic, with expertise in molecular pharmacology of CCK1R, and Dr. Sergienko at Sanford Burnham Prebys, with expertise in high throughput screening (HTS) and chemical genomics. There are 3 specific AIMS: (i) Primary HTS of a small molecule library using intracellular calcium response assays in CHO cells expressing WT CCK1R, performed in two modes to examine (a) intrinsic agonist activity and (b) PAM activity that enhances the responsiveness to CCK. Hits will be confirmed in orthogonal IP- 1 assays and counter-screened using parental CHO cells; (ii) Hit validation and functional profiling will be performed using analogous assays with CHO cells expressing a mutant CCK1R that mimics the abnormal conformation of this receptor in a high cholesterol environment. Also, assays will be performed to ensure that compounds do not stimulate CCK1R internalization, interfere with natural hormone binding and biological activity, or cause cytotoxicity; and (iii) Lead identification and functional characterization will be performed using a neuronal cell line that naturally expresses a physiologic density of CCK1R, CHP212 cells, to ensure absence of intrinsic agonist and trophic activity, and to reconfirm ability to enhance the responses to CCK. Other characterization will include studying effects on CCK-58, and those at CCK1R from various species and on CCK2R, as well as a panel of GPCRs, and a broad survey of signaling responses and ADME/Tox assays.

项目总结/摘要 肥胖已经达到流行病的比例，导致2型糖尿病及其并发症的类似增加。 多种合并症，造成巨大的费用、痛苦和早期死亡。饮食改良 和生活方式，而有效的急性减肥，是不持久的，减肥手术，而有效的 病态肥胖症，对于这种需要是不可扩展的。因此，新的安全和有效的药物治疗肥胖 是必要的。该提议响应PAR-16-374请求用于发现此类治疗剂的测定。 我们的目的是鉴定具有1型正变构调节剂（PAM）活性的分子， 胆囊收缩素受体（CCK 1 R）不表现出内在激动剂活性， 在所有潜在患者中，从那些倾向于增加体重的人到病态肥胖的人， 代谢综合征这种药物目前还不存在。它会增加信号反应， 餐后释放的内源性激素，当生物反应较大时可增强饱腹感，同时 克服了完全激动剂引起的副作用和潜在毒性。它还具有独特的性质， 纠正了现在被认为存在于这些患者的子集中的刺激-活动耦合中的缺陷。 连续的双重筛选策略将针对天然野生型（WT）CCK 1 R，以及这种 受体在异常构象诱导胆固醇在膜，后者 由CCK 1 R突变体模仿。增强和/或重新校准CCK 1 R，使其正常响应CCK， 这些设置可以以温和和安全的方式强调激素对饱腹感的影响。这表示 马约诊所的米勒博士在CCK 1 R的分子药理学方面具有专长， Sergienko在Sanford Burnham Prebys，拥有高通量筛选（HTS）和化学 基因组学有3个特定的AIMS：（i）使用细胞内钙的小分子库的初级HTS 在表达WT CCK 1 R的CHO细胞中进行的反应测定，以两种模式进行，以检查（a）内在激动剂 活性和（B）增强对CCK的反应性的PAM活性。命中将在正交IP中确认- 1测定和使用亲本CHO细胞的反筛选;（ii）命中验证和功能谱分析将在 使用表达突变型CCK 1 R的CHO细胞进行类似测定， 在高胆固醇环境中这种受体的构象。此外，还将进行试验，以确保 化合物不刺激CCK 1 R内化，干扰天然激素结合和生物学活性， 活性，或引起细胞毒性;和（iii）将进行电极导线识别和功能表征 使用天然表达生理密度CCK 1 R的神经元细胞系，CHP 212细胞，以确保 缺乏内在激动剂和营养活性，并重新确认增强对CCK反应的能力。 其他表征将包括研究对CCK-58的影响，以及来自各种物种和 以及一组GPCR，以及对信号应答和ADME/Tox测定的广泛调查。

项目成果

Screening for positive allosteric modulators of cholecystokinin type 1 receptor potentially useful for management of obesity.

• DOI: 10.1016/j.slasd.2022.07.001
• 发表时间: 2022-10
• 期刊: SLAS DISCOVERY
• 影响因子: 3.1
• 作者: Dengler, Daniela G.;Sun, Qing;Harikumar, Kaleeckal G.;Miller, Laurence J.;Sergienko, Eduard A.
• 通讯作者: Sergienko, Eduard A.

Discovery of a Positive Allosteric Modulator of Cholecystokinin Action at CCK1R in Normal and Elevated Cholesterol.

• DOI: 10.3389/fendo.2021.789957
• 发表时间: 2021
• 期刊: Frontiers in endocrinology
• 影响因子: 5.2
• 作者: Harikumar KG;Coudrat T;Desai AJ;Dong M;Dengler DG;Furness SGB;Christopoulos A;Wootten D;Sergienko EA;Sexton PM;Miller LJ
• 通讯作者: Miller LJ