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: 10201582
• 负责人: Nicholas F. LaRusso
• 金额: $ 49.19万
• 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/10201582
• 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; 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 本提案的目的是开发胆汁酸受体TGR 5的拮抗剂作为潜在的治疗方法 多囊肝病（PLD），一种以肝囊肿形成为特征的遗传性胆管病。 胆管病是一组肝脏疾病，其中胆管细胞，肝内胆汁衬里的上皮细胞， 管道是主要目标PLD是不可治愈的，作为孤立的常染色体显性PLD存在或与 常染色体显性多囊肾病（ADPKD）和常染色体隐性PKD。尽管最近 尽管PLD发病机制的研究取得了进展，但对于这些破坏性的病症还没有治疗方法。我们以前的工作 提示cAMP（在囊性胆管细胞中增加）是肝纤维化网络中的重要中心成分， PLD中的信号通路失调，并引导我们采用cAMP靶向策略进行疾病治疗。我们 TGR 5是一种G蛋白偶联胆汁酸受体（GPCR），与cAMP信号转导有关， 在PLD中cAMP驱动的肝囊肿形成中的作用。我们证明了TGR 5在囊性癌中过表达， 胆管细胞在体外和体内，并发现TGR 5激动剂增加细胞内cAMP，触发 在体外和PCK大鼠中，胆管细胞过度增殖并促进囊肿生长（疾病恶化 进展）。进一步的研究结果支持TGR 5在肝囊肿形成的发病机制中的作用。所以我们 目的是开发抑制TGR 5活性的拮抗剂，从而减少PLD中的肝囊肿形成。因为没有 选择性小分子TGR 5拮抗剂已经报道，我们完成了一个HTS鉴定几个 包括SBI-319在内的重要线索我们提出这个铅优化运动，以开发口服 用于体内测试以验证TGR 5作为PLD的潜在靶标的化合物。 中心假设选择性TGR 5拮抗剂将抑制囊性胆管细胞中的cAMP水平，从而减少 cAMP驱动的细胞增殖和肝囊肿形成，产生PLD的新的治疗靶点。探讨 我们的中心假设，我们将执行以下具体目标：目标1。设计合成TGR 5 拮抗剂口服活性体内。通过反复的化学循环，我们将确定适合于 体内功效研究。目标2.阐明SBI-319类似物在PLD中的作用机制。我们将（1） 评估SBI-319类似物对：（i）囊性胆管细胞中cAMP产生;（ii）胆管细胞增殖; 和（iii）在3D培养物中肝囊性结构的生长; 2）检测TGR 5和Gαs蛋白的表达， 以及它们在用SBI-319处理时的偶联。目标3.为了评价TGR 5拮抗剂作为抗肿瘤药物的功效， PLD的治疗我们将评估SBI-319类似物对肝和肾囊肿形成的影响，并阐明其作用机制。 TGR 5抑制在体内疾病进展中的作用。 除了PLD作为一种潜在的治疗应用外，研究结果显示TGR 5在人乳腺癌中过表达。 在胆管癌中表达上调，在非胆管癌（乳腺癌、胃癌、胰腺癌）中表达上调。发展 有效的TGR 5拮抗剂可能具有超出PLD的治疗应用。