The Development of TGR5 Antagonists for the Treatment of Cholangiopathies

用于治疗胆管病的 TGR5 拮抗剂的开发

• 批准号: 10431962
• 负责人: Nicholas F. LaRusso
• 金额: $ 45.39万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10431962
• 关键词: 3-Dimensional; Acids; Agonist; Animal Model; Autosomal Dominant Polycystic Kidney; Autosomal Recessive Polycystic Kidney; Bile Acids; Bile duct carcinoma; Biological Assay; Biology; Breast; Cell Proliferation; Cells; Chemicals; Chemistry; Cholangiocarcinoma; Coupled; Coupling; Cyclic AMP; Cyst; Development; Dialysis procedure; Disease; Disease Progression; Diuretics; Dose; Drug Kinetics; Drug Targeting; Effectiveness; Epithelial Cells; Excretory function; Family; G-Protein-Coupled Receptors; GPBAR1 gene; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Genetic; Genetic Diseases; Growth; Hepatic; Hepatic Cyst; Human; Impairment; In Vitro; Intrahepatic bile duct; Kidney; Kidney Failure; Kidney Transplantation; Knowledge; Lead; Ligands; Link; Liver; Liver Failure; Liver diseases; MEKs; Malignant - descriptor; Malignant neoplasm of pancreas; Metabolism; Mus; Mutant Strains Mice; Oral; Pancreas; Pathogenesis; Pathologic; Pathway interactions; Pharmacology; Play; Production; Proteins; Rattus; Reporting; Rodent; Role; Signal Pathway; Signal Transduction; Stomach; Structure; Survival Rate; Therapeutic; Time; Transplantation; Work; absorption; analog; antagonist; base; cholangiocyte; chronic pain; design; druggable target; efficacy evaluation; efficacy study; experimental study; high throughput screening; improved; in vitro Model; in vivo; in vivo evaluation; innovation; lead optimization; liver function; new therapeutic target; novel; overexpression; polycystic liver disease; receptor; small hairpin RNA; small molecule; three dimensional cell culture

PROJECT SUMMARY The objective of this proposal is to develop antagonists of the bile acid receptor TGR5 as a potential treatment for polycystic liver disease (PLD), a genetic cholangiopathy characterized by hepatic cystogenesis. Cholangiopathies are a group of liver diseases in which cholangiocytes, the epithelial cells lining intrahepatic bile ducts, are the primary target. PLD, is incurable and exists as isolated Autosomal Dominant PLD or co-exists with Autosomal Dominant Polycystic Kidney Disease (ADPKD) and Autosomal Recessive PKD. Despite recent advances in the pathogenesis of PLD, there are no therapies for these devastating conditions. Our previous work implicated cAMP (increased in cystic cholangiocytes) as an important central component in the network of dysregulated signaling pathways in PLD and led us to cAMP-targeted strategies for disease treatment. We discovered TGR5, a G protein-coupled bile acid receptor (GPCR) linked to cAMP signaling, plays an important role in cAMP-driven hepatic cystogenesis in PLD. We demonstrated that TGR5 is overexpressed in cystic cholangiocytes in vitro and in vivo, and found TGR5 agonists increase intracellular cAMP, triggering cholangiocyte hyper-proliferation and enhancing cyst growth both in vitro and in PCK rats (worsening disease progression). Further findings support TGR5's role in the pathogenesis of hepatic cystogenesis. Thus, our objective is to develop antagonists to inhibit TGR5 activity, thus reducing hepatic cystogenesis in PLD. As no selective small molecule TGR5 antagonists have been reported, we completed an HTS identifying several promising leads including SBI-319. We propose this lead optimization campaign to develop orally available compounds for in vivo testing to validate TGR5 as a potential target for PLD. Central Hypothesis. Selective TGR5 antagonists will inhibit cAMP levels in cystic cholangiocytes, thus reducing cAMP-driven cell proliferation and hepatic cystogenesis, yielding a novel therapeutic target for PLD. To explore our central hypothesis, we will perform the following specific aims: Aim 1. To design and synthesize TGR5 antagonists orally active in vivo. Through iterative cycles of chemistry, we will identify compounds suitable for in vivo efficacy studies. Aim 2. To clarify the mechanisms of action of SBI-319 analogs in PLD. We will 1) assess SBI-319 analog effects on: (i) cAMP production in cystic cholangiocytes; (ii) cholangiocyte proliferation; and (iii) growth of hepatic cystic structures in 3D cultures; 2) examine the expression of TGR5 and Gαs proteins, and their coupling upon treatment with SBI-319. Aim 3. To evaluate the efficacy of TGR5 antagonists as a treatment for PLD. We will assess effects of SBI-319 analogs on hepatic and renal cystogenesis and clarify the role of TGR5 inhibition in disease progression in vivo. In addition to PLD as a potential therapeutic application, findings show TGR5 is over-expressed in cholangiocarcinoma and is up-regulated in non-biliary cancers (breast, gastric, pancreas). Development of effective TGR5 antagonists would likely have therapeutic application beyond PLD.

项目摘要 该提案的目的是开发胆汁酸受体TGR5的拮抗剂作为潜在的治疗 对于多囊性肝病（PLD），这是一种以肝囊这症为特征的遗传胆管疾病。 胆管病是一组肝脏疾病，其中胆管细胞，上皮细胞衬里ep骨内胆汁 管道是主要目标。 PLD是无法治愈的，并且以孤立的常染色体显性pld或与 常染色体显性多囊肾脏疾病（ADPKD）和常染色体隐性PKD。尽管最近 PLD发病机理的进展，对于这些毁灭性疾病没有疗法。我们以前的工作 实施营地（增加囊性胆管细胞）是网络网络中的重要核心组成部分 PLD中的信号通路失调，导致我们采取了针对营地的疾病治疗策略。我们 发现的TGR5是与CAMP信号相关的G蛋白偶联的胆汁酸受体（GPCR） 在PLD中，在cAMP驱动的肝囊肿发生中的作用。我们证明了TGR5在囊性中过表达 体外和体内胆管细胞，发现TGR5激动剂会增加细胞内营地，触发 体外和PCK大鼠（恶化疾病）的胆管细胞高增殖和增强囊肿的生长 进展）。进一步的发现支持TGR5在肝囊肿发生的发病机理中的作用。那，我们的 目的是发展拮抗剂以抑制TGR5活性，从而减少PLD中的肝囊生。否 据报道，选择性小分子TGR5拮抗剂，我们完成了一个HTS，识别了几个 有希望的线索，包括SBI-319。我们提出了这项铅优化运动以开发口头 体内测试的化合物验证TGR5作为PLD的潜在靶标。 中央假设。选择性TGR5拮抗剂将抑制囊性胆管细胞中的cAMP水平，从而减少 营地驱动的细胞增殖和肝囊生，为PLD提供了一种新型的治疗靶标。探索 我们的中心假设，我们将执行以下特定目的：目标1。设计和合成TGR5 拮抗剂在体内口服活性。通过化学的迭代循环，我们将确定适合的化合物 体内效率研究。目的2。阐明PLD中SBI-319类似物的作用机理。我们将1） 评估SBI-319模拟效应：（i）囊性胆管细胞中的cAMP产生； （ii）胆管细胞增殖； （iii）3D培养物中肝囊性结构的生长； 2）检查TGR5和GαS蛋白的表达， 以及他们在SBI-319治疗时的耦合。目标3。评估TGR5拮抗剂的效率 PLD的处理。我们将评估SBI-319类似物对肝脏和肾脏囊生的影响，并阐明 TGR5抑制在体内疾病进展中的作用。 除了PLD作为潜在的治疗应用外，调查结果表明TGR5在 胆管癌，并在非细胞癌（乳腺癌，胃，胰腺）中上调。发展 有效的TGR5拮抗剂可能会超出PLD的治疗应用。