Darifenacin Hydrobromide

EGFR-TKIs目前已成为EGFR敏感突变非小细胞肺癌（NSCLC）患者的标准治疗方案[1]。 EGFR-TKIs目前已成为EGFR敏感突变非小细胞肺癌（NSCLC）患者的标准治疗方案[1]。 多项III期临床试验已经证实：与铂类化疗相比，第一代TKI和第二代中位缓解率为70-75%，无进展生存期（PFS）从10-14个月不等。第三代TKI奥希替尼在NSCLC的治疗中显示出更加优异的疗效，PFS和OS均显著改善[2]。 然而，尽管取得了这些重大的治疗进展，但仍然有部分患者不可避免地会出现对EGFR-TKI的耐药，并导致疾病进展，最终影响患者生存。因此迫切需要开发新的策略来改善NSCLC患者EGFR-TKI治疗耐药性。 目前越来越多的证据表明，治疗后残留的耐药持久性（DTP）肿瘤细胞，在诱导耐药过程中发挥重要作用，可通过多种机制（如转录、表观遗传和代谢重编程等）在初始治疗后存活，并在长期药物治疗后驱动不可逆的获得性耐药性[3]。因此，靶向DTP细胞可能是一种改善患者耐药性，并延长治疗反应的潜在策略。 近日，清华大学药学院胡泽平教授在The Journal of Clinical Investigation杂志发表重要研究成果，阐述了DTP通过异常代谢调控参与TKI治疗耐药的机制。清华大学药学院聂萌、陈娜和庞欢欢，以及同济大学附属上海市肺科医院蒋涛为论文的共同第一作者。 他们发现EGFR-TKI治疗可通过YAP介导的乙酰胆碱生物合成酶胆碱乙酰转移酶（ChAT）的上调表达，导致乙酰胆碱（ACh）的累积，乙酰胆碱以乙酰胆碱毒蕈碱受体-3（M3R）依赖性方式通过WNT信号通路介导药物耐受。 值得注意的是，EGFR-TKI与ACh/M3R信号抑制剂Darifenacin联合应用，可改善小鼠对EGFR-TKI治疗的耐药，为控制药物耐受性，改善NSCLC治疗现状提供了一种潜在的联合治疗策略[4]。 论文首页截图 首先，研究人员通过体外实验，将EGFR突变的人NSCLC细胞系PC9和HCC827暴露于致死剂量的EGFR-TKIs中，以诱导DTP细胞的产生。 随后通过靶向代谢组学比较了亲代细胞与DTP细胞的代谢差异，发现PC9和HCC827衍生的DTP细胞中胆碱代谢产物的丰度显著改变。与亲代细胞相比，乙酰胆碱（ACh）在DTP细胞中显著累积。 乙酰胆碱（ACh）在DTP细胞中显著累积 同时，通过对亲代细胞和DTP细胞的转录组测序发现，DTP细胞与参与调节神经递质生物合成过程的基因表达升高。这表明乙酰胆碱代谢的重编程在诱导NSCLC EGFR-TKI治疗耐药中发挥着重要的作用。 为进一步验证以上发现，作者结合临床样本建立了EGFR突变NSCLC患者奥希替尼耐药的异种移植瘤（PDX）模型，检测其中乙酰胆碱代谢水平的变化并得到了相似的结论。 接下来，作者用奥希替尼处理PC9细胞11天后停药，继续培养14天后重新给药。结果发现停用奥希替尼后，细胞恢复了对奥希替尼的敏感性，重新给药后又新产生了一小群DTP细胞。初始DTP细胞和再给药后再生的DTP细胞中的乙酰胆碱水平较高，而亲代细胞和奥希替尼停药阶段的细胞乙酰胆碱水平相对较低。 那么EGFR-TKI治疗是如何导致细胞内乙酰胆碱水平变化的呢？ 乙酰胆碱由胆碱和乙酰辅酶A通过限速酶ChAT合成，然后通过乙酰胆碱转运体转运到突触囊泡，分泌到细胞外环境与靶细胞上的烟碱乙酰胆碱受体（nAChR）和毒蕈碱乙酰胆碱酯受体（mAChR）结合发挥其生物学作用。 为了阐明DTP细胞中乙酰胆碱累积的可能原因，作者评估了与乙酰胆碱代谢和信号传导相关的蛋白表达，并发现一种短ChAT亚型（41 kDa）表达显著上调。 有趣的是，与EGFR-TKI治疗导致细胞内乙酰胆碱水平的动态改变一致，这种短ChAT异构体表达也表现出可逆动态变化。 ChAT随TKI处理表现出可逆的动态变化 接下来，作者试图确定EGFR-TKI上调短ChAT亚型表达的机制。 通过分析已有文献，作者发现YAP通路的激活，在肿瘤细胞逃避EGFR抑制剂治疗过程中发挥重要作用[5]。同时分析公共染色质免疫沉淀测序（ChIP-seq）数据发现，在ChAT的远端调节区有三个潜在的YAP结合位点，依据这些可推测YAP可能通过结合远端增强子来调节ChAT转录。 实验结果也表明，奥希替尼治疗显著增加了ChAT增强子（#1和#3）上的YAP结合。同时YAP敲除导致增强子（#1和#3）的荧光素酶报告活性显著降低。进一步，YAP敲除在mRNA和蛋白质水平显著降低了奥希替尼诱导的短CHAT亚型的上调。此外，奥希替尼诱导的乙酰胆碱累积也被YAP敲除减弱。 这些发现表明，EGFR-TKI可通过YAP介导的转录激活上调ChAT表达，进一步导致Ach的累积。 YAP介导ChAT的表达上调 基于上述实验结果，作者推断或许可以通过干预乙酰胆碱产生、分泌或乙酰胆碱信号传导，来调节肿瘤对于药物的耐受性。 首先，作者发现外源性乙酰胆碱的处理可促进DTP的形成，并降低PC9和HCC827细胞对奥希替尼的敏感性。 进一步研究发现，短ChAT亚型的表达水平与奥希替尼敏感性之间也存在正相关关系。在奥希替尼治疗后，与ChAT水平低的单细胞克隆相比，ChAT水平高的单细胞克隆DTP形成更多。此外，短ChAT亚型的过表达也导致乙酰胆碱水平显著增加，并显著增加DTP的形成和药物耐受性，进一步导致更快速的肿瘤复发。 相反，奥希替尼诱导的乙酰胆碱增加可被ChAT敲除阻断，同时，ChAT敲除显著抑制了DTP细胞的形成，增强了对奥希替尼的药物敏感性，抑制肿瘤复发，改善生存结果。这种效应可通过补充外源性乙酰胆碱而得以挽救。 敲除ChAT导致肿瘤耐药性的降低 Darifenacin一种M3R抑制剂，使用其处理可显著降低DTP细胞形成。奥希替尼和Darifenacin的联合治疗可改善肿瘤细胞对奥希替尼的耐受性，这些证据均证明了乙酰胆碱代谢和信号传导在介导药物耐受形成中的重要作用。 在以上基础上，作者进一步通过体内实验探索了Darifenacin在肿瘤治疗中的潜在应用价值。在注射PC9细胞后14天，对小鼠单独使用Darifenacin、单独使用奥希替尼或使用奥希替尼和Darifenacin联合治疗。 单独使用奥希替尼，以及奥希替尼和Darifenacin的联合治疗，在9天内使肿瘤显著消退。为了评估Darifenacin对肿瘤复发的特异性影响，第9天后停止奥希替尼治疗。与单独接受奥希替尼的小鼠相比，联合治疗的小鼠在停止奥希替尼治疗后显示出延迟的肿瘤复发。 进一步，在PC9异种移植小鼠接受单独奥希替尼的初始治疗9天，将小鼠随机分为两组：一组单独用奥希替尼治疗，另一组给予奥希替尼与Darifenacin联合治疗。与单独使用奥希替尼治疗相比，联合使用奥希替尼和Darifenacin治疗时，奥希消退的残留肿瘤复发率显著降低。此外，与单独给予奥希替尼的小鼠相比，给予联合治疗的小鼠具有显著改善的生存结果。 总之，这些体内结果表明，EGFR-TKI和ACh/M3R抑制药物联合治疗可以减少残留肿瘤的形成并延缓肿瘤复发。 EGFR-TKI和ACh/M3R抑制药物联合治疗可以减少残留肿瘤的形成并延缓肿瘤复发 那么乙酰胆碱是如何驱动肿瘤对于TKI药物耐受的呢？ 作者回顾了来自亲代PC9细胞和PC9衍生的DTP细胞的RNA测序数据，发现一些WNT信号分子在DTP细胞中上调，而WNT信号通路的几种负调控因子表达下调。因此，作者认为乙酰胆碱可能通过激活WNT信号传导来调节药物耐受。 进一步研究发现，添加WNT信号激动剂CHIR99021或WNT3A后，显著降低了Darifenacin减少DTP形成的能力。相反，与单独的Darifenacin治疗相比，与WNT信号抑制剂LGK974和ICG-001联合使用可进一步抑制DTP的形成。 总之，这些发现证实EGFR-TKI诱导的乙酰胆碱增加，可通过激活WNT信号传导介导药物耐受的出现。 EGFR-TKI诱导的乙酰胆碱增加可通过激活WNT信号传导介导药物耐受的出现 最后，作者探索了患者血浆ACh水平是否与对EGFR-TKIs的临床反应相关。由此设计了一项队列研究，分析EGFR突变的NSCLC患者（n=78）治疗前血浆ACh水平。 根据治疗前的基线ACh水平，将患者分为低表达组、中间组和高表达组。低ACh水平患者的EGFR TKI应答率相对较高（84%），而高和中ACh水平的患者的应答率相对较低（分别为58%和63%），表明患者治疗前血浆ACh水平可能与临床应答率呈负相关。 此外，与治疗前ACh水平较低和/或中等的患者相比，治疗前高ACh水平的患者显示出较短的无进展生存期。 EGFR-TKI前血浆ACh水平高与不良反应高度相关 总之，通过这项研究，确定了NSCLC患者中EGFR-TKI诱导的ACh代谢重编程途径，为评估NSCLC对于EGFR-TKI的治疗效果，改善EGFR-TKI治疗耐药，提升治疗效果提供了潜在的策略。但仍需要通过前瞻性随机临床试验证明其在临床治疗中的有效性和安全性。 参考文献： [1]Hirsch FR, Scagliotti GV, Mulshine JL, et al. Lung cancer: current therapies and new targeted treatments[J]. Lancet, 2017, 389 (10066):299-311. DOI:10.1016/s0140-6736(16)30958-8. [2]Passaro A, Jänne PA, Mok T, et al. Overcoming therapy resistance in EGFR-mutant lung cancer[J]. Nat Cancer, 2021, 2 (4):377-391. DOI:10.1038/s43018-021-00195-8. [3]Oren Y, Tsabar M, Cuoco MS, et al. Cycling cancer persister cells arise from lineages with distinct programs[J]. Nature, 2021, 596 (7873):576-582. DOI:10.1038/s41586-021-03796-6. [4]Nie M, Chen N, Pang H, et al. Targeting acetylcholine signaling modulates persistent drug tolerance in EGFR-mutant lung cancer and impedes tumor relapse[J]. J Clin Invest, 2022. DOI:10.1172/jci160152. [5]Kurppa KJ, Liu Y, To C, et al. Treatment-Induced Tumor Dormancy through YAP-Mediated Transcriptional Reprogramming of the Apoptotic Pathway[J]. Cancer Cell, 2020, 37 (1):104-122.e112. DOI:10.1016/j.ccell.2019.12.006.

Los Angeles, USA, June 07, 2021 (GLOBE NEWSWIRE) -- Overactive Bladder Pipeline Shows Rapid Advancements in the Clinical Trials: A Review by DelveInsight Overactive Bladder therapies with different mechanisms of action are currently being studied with excellent market potential. Ongoing scientific research has provided with greater sophistication and a clearer understanding of the mechanisms of therapeutic interventions, combined with improved outcome measures to assess the efficacy and patient satisfaction, as well as high-quality Overactive Bladder clinical trials with proper design, which would translate into more refined and directed therapies in future years. DelveInsight’s “Overactive Bladder Pipeline Insight” report provides comprehensive insights about 15+ companies and 15+ pipeline drugs in the Overactive Bladder pipeline landscapes. It comprises Overactive Bladder pipeline drug profiles, including clinical and non-clinical stage products. It also includes the Overactive Bladder therapeutics assessment by product type, stage, route of administration, and molecule type and further highlights the inactive Overactive Bladder pipeline products. Some of the key takeaways from the Overactive Bladder Pipeline Report Get an overview of pipeline landscape @ Overactive Bladder Clinical Trials Analysis Overactive Bladder (OAB) causes a frequent and sudden urge to urinate that may be difficult to control. One may need to pass urine many times during the day and night and may also experience unintentional loss of urine. There are six antimuscarinic agents available for the OAB treatment worldwide- oxybutynin, tolterodine, fesoterodine, trospium, darifenacin, and solifenacin. Overactive Bladder Emerging Drugs Neuronox, an acetylcholine inhibitor is being developed by Medy-Tox for the treatment of the Overactive Bladder. The active ingredients involve Clostridium botulinum toxin type A. It is currently in the phase III stage of development. In September 2020, US drugmaker Allergan entered into a license agreement with South Korea’s Medytox under the terms of which, on closing, Allergan would pay Medytox an upfront of USD 65 million and Medytox would grant Allergan exclusive rights, worldwide outside of Korea, to develop and, if approved, commercialise certain neurotoxin product candidates currently in development, including a potential liquid-injectable product. In May 2020, AbbVie announced that it had finished its acquisition of Allergan plc. following receipt of regulatory approval from all government authorities needed by the transaction agreement and approval by the Irish High Court. Research and DevelopmentPhase III NCT04113941: In August 2019, Medy-Tox initiated a phase III trial, a randomised, double-blind, placebo-controlled, multi-centre study to assess the efficacy and safety of Meditoxin in subject with Idiopathic Overactive Bladder. This study is for patients who have idiopathic overactive bladder symptoms. This study will assess the efficacy and safety of Neuronox against placebo. The trial is currently in the active not recruiting stage with an estimated enrollment of 216 participants and anticipated to be completed in August 2021. Urovant Sciences is developing URO-902 for the treatment of the Overactive Bladder. URO-902 has the potential to be the first gene therapy for OAB patients. This innovative treatment has the capability to tackle an unmet need for patients who have failed oral pharmacologic therapies and are concerned with potential urinary retention or surgical interventions pertinent to existing third-line OAB treatments. It is currently in the phase II stage of development. Research and DevelopmentPhase II NCT04211831: In December 2019, Urovant Sciences GmbH initiated a phase II trial titled “An exploratory phase IIa study evaluating the efficacy and safety of URO-902 in subjects with Overactive Bladder and Urge Urinary Incontinence”. This study will investigate the efficacy, safety, and tolerability of a single dose of URO-902 24 milligrams (mg) and 48 mg (administered via intradetrusor injection), compared with placebo, in participants with overactive bladder (OAB) and urge urinary incontinence (UUI) up to 48 weeks post-dose. The trial is currently in recruiting stage with an estimated enrollment of 78 participants and expected to be completed in September 2022. Solabegron is a highly potent and selective beta-3 adrenoceptor agonist that is being developed for Overactive Bladder treatment. Solabegron has been tested in over 800 study subjects in a twice-a-day formulation and demonstrated efficacy in treating OAB and Irritable bowel syndrome (IBS). A once-daily formulation designed to optimise patient convenience and effectiveness has been developed and is currently being evaluated in phase 2b dose-ranging clinical trials. Research and DevelopmentPhase II NCT03594058: In July 2018, Velicept Therapeutics initiated a phase IIb, multicenter, randomised, double-blind, placebo-controlled, parallel-group study to assess the efficacy and safety of oral solabegron modified-release tablets in the treatment of Overactive Bladder in adult female subjects. The trial enrolled 1413 participants and was completed in May 2019. Results Velicept Therapeutics announced that its next-generation beta-3 adrenoceptor agonist solabegron met the primary endpoint in VEL-2002, a phase IIb study in Overactive Bladder patients. In the study, twice-daily administration of solabegron showed a statistically major improvement compared to placebo at week 12, as measured by the mean change in the number of micturitions per day, the study’s primary endpoint. Solabegron also showed statistical significance across various secondary endpoints, comprising a percent reduction of urge urinary incontinence episodes, dry rate, and urgency episodes. For further information, refer to the detailed report @ Overactive Bladder Pipeline Therapeutics Scope of Overactive Bladder Pipeline Drug Insight Key Questions regarding Current Overactive Bladder Treatment Landscape and Emerging Therapies Answered in the Pipeline Report Table of Contents Get a customised pipeline report @ Overactive Bladder Drugs Pipeline Report Related Reports DelveInsight’s Overactive Bladder Market Insights, Epidemiology and Market Forecast report provides a detailed overview of the disease and a depth understanding of historical data. DelveInsight's Overactive Bladder - Epidemiology Forecast 2030 report delivers an in-depth understanding of the disease, historical, and forecasted epidemiology of Overactive Bladder in the United States, EU5 (Germany, France, Italy, Spain, and the United Kingdom), and Japan. DelveInsight's "Interstitial Lung Disease - Market Insights, Epidemiology, and Market Forecast-2030" report delivers an in-depth understanding of the Interstitial Lung Disease, historical and forecasted epidemiology. DelveInsight's Interstitial Cystitis - Market Insights, Epidemiology and Market Forecast - 2030 report provides a detailed overview of the disease. DelveInsight's Bronchiolitis Obliterans Syndrome - Market Insights, Epidemiology and Market Forecast - 2030 report provides a detailed overview of the disease and a depth understanding of historical and forecasted epidemiology. Checkpoint Kinase Inhibitor Pipeline Insights, 2021 report by DelveInsight offers comprehensive insights of the pipeline (under development) therapeutics scenario and growth prospects across Checkpoint Kinase Inhibitor development. Browse ASCO 2021 Abstracts here:- About DelveInsightDelveInsight is a leading Business Consultant and Market Research firm focused exclusively on life sciences. It supports Pharma companies by providing comprehensive end-to-end solutions to improve their performance. It also provides Healthcare Consulting services comprising credible market analysis that will help accelerate the business growth and overcome challenges with a practical approach.