Nitazoxanide as a Novel Therapy for Rare Disease Lymphangioleiomyomatosis and Tuberous Sclerosis

硝唑尼特作为罕见疾病淋巴管平滑肌瘤病和结节性硬化症的新疗法

• 批准号: 10258194
• 负责人: VERA P KRYMSKAYA
• 金额: $ 25.64万
• 依托单位: BIOSPUTNIK LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-10 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10258194
• 关键词: 1 year old; Address; Adverse effects; Affect; Age; Animals; Antibodies; Antiparasitic Agents; Antiprotozoal Agents; Apoptosis; Authorization documentation; Benign; Binding; Brain; Businesses; Categories; Cell Culture Techniques; Cell Line; Cell Proliferation; Cell Survival; Cell model; Cells; Cessation of life; Clinical; Complex; Cryptosporidium parvum; Cyst; Development; Diarrhea; Disadvantaged; Disease; Dose; Enhancers; Evaluation; Eye; FDA approved; FRAP1 gene; Female of child bearing age; Fibroblasts; Future; Genetic; Genetic Diseases; Giardia lamblia; Goals; Grant; Growth; Heart; Hematoxylin and Eosin Staining Method; Human; Human Cell Line; Immunodeficient Mouse; Immunosuppressive Agents; In Vitro; Incentives; Individual; Inherited; Injections; Investigation; Kidney; Laboratories; Legal patent; Lesion; Life; Lung; Lung Neoplasms; Lung Transplantation; Lung diseases; Lymphangiogenesis; Lymphangioleiomyomatosis; Lymphatic Obstruction; Mass Spectrum Analysis; Medical; Modeling; Molecular Target; Morphology; Mus; Mutation; NQO1 gene; Neoplasms; Nodule; Nude Mice; Null Lymphocytes; Organ; Orphan Drugs; Oxidoreductase; Pathologic; Pathway interactions; Patients; Pennsylvania; Pregnancy; Prevention; Property; Quinones; Rare Diseases; Recurrence; Research; Resort; SLC12A3 gene; Safety; Serious Adverse Event; Shortness of Breath; Signal Pathway; Signal Transduction; Sirolimus; Skin; Smooth Muscle Myocytes; Stains; Structure; Structure of parenchyma of lung; TSC1 gene; TSC2 gene; Technology; Therapeutic; Tissues; Titrations; Toxic effect; Translating; Treatment Efficacy; Tuberous Sclerosis; Tuberous sclerosis protein complex; Tumor Suppressor Genes; United States; Universities; Vascular Endothelial Growth Factor D; Western Blotting; Woman; base; cell immortalization; child bearing; clinical development; commercial application; commercialization; direct application; experimental study; gene function; improved; in vivo; inhibitor/antagonist; mTOR Inhibitor; mouse model; neoplastic; nitazoxanide; novel; novel therapeutics; prevent; standard of care; therapy development; treatment comparison; treatment effect; tuberous sclerosis patients; tumor

ABSTRACT Tuberous sclerosis (TS), also called tuberous sclerosis complex, is a rare, multi-systemic genetic and often life- threatening disease that causes benign tumors to grow in the brain and on other vital organs such as the kidneys, heart, eyes, lungs, and skin. Lymphangioleiomyomatosis (LAM) is a TS-related tumor-like disorder. Both occur as a consequence of an inherited or sporadic mutation in either the TSC1 or TSC2 gene, which function as negative regulators of the mTOR pathway. Uncontrolled mTORC1 activity leads to the neoplastic proliferation of abnormal smooth muscle cells (LAM cells) in the lungs, progressive shortness of breath, recurrent pneumothoraxes, and loss of pulmonary tissue structure and function. TS affects as many as 25,000 to 40,000 individuals in the United States and about 1 to 2 million individuals worldwide. LAM is a rare disease affecting women in childbearing age. The first and only FDA-approved treatment for LAM is the immunosuppressant sirolimus (Rapamune©), marketed since 2015 by Pfizer. It is the current standard-of-care and acts by inhibiting mTORC1. Sirolimus has several clinical disadvantages, including a considerable number of non-responders, severe adverse events due to its immunosuppressive properties and pregnancy category C, limiting its use in women of childbearing age. Thus, there is a high unmet medical need to develop alternative and safer treatment options for LAM and TS. We have identified Nitazoxanide (hereafter NTZ; Alinia©), an antiprotozoal FDA-approved and safe therapy for the treatment of diarrhea caused by Giardia lamblia or Cryptosporidium parvum in patients 1 year of age or older, as a potential novel therapy for LAM/TS. Using Capture Compound Mass Spectrometry technology, we discovered NAD(P)H quinone oxidoreductase (NQO1) as the human target of Nitazoxanide. Furthermore, we have shown that NTZ is effective in inhibiting mTORC1 related cellular signaling in three different LAM related murine and human cell lines addressing the underlying pathological mechanism of LAM/TS. Our goal for this grant is the investigation of NTZ’s efficacy for the treatment of LAM/TS. Aim 1 will be the in vivo proof of concept comparing three dose levels of NTZ in a TSC2-null murine model developed at the laboratory of Prof. Krymskaya at the University of Pennsylvania. It will evaluate prevention of TSC2-null lesion growth, immunohistochemical and lung morphology changes and animal survival in a murine LAM model. Aim 2 will cover an in vitro study, investigating underlying cellular effects on mTOR and related signaling pathways. The in vitro study will investigate NTZ's effects in LAM-derived AML (LAMD) cells and immortalized TSC2-null cell cultures of human origin followed by more detailed mTORC1 interactions studies. In addition to the monotherapeutic NTZ approach, dose reduction potential for dose-toxicity prone rapamycine with NTZ add-on will be explored. This project aims to develop a novel therapy for patients with the devastating diseases of LAM/TS. Based on NTZ's excellent safety profile as an approved antiprotozoal drug, positive efficacy results in our studies would enable an accelerated clinical development under FDA section 505 (2)(b) approval pathway and orphan disease designation.

摘要 结节性硬化症（TS），也称为结节性硬化症，是一种罕见的，多系统的遗传性，往往是终身的， 威胁性疾病，导致良性肿瘤生长在大脑和其他重要器官，如肾脏， 心脏眼睛肺和皮肤淋巴管平滑肌瘤病（LAM）是一种与TS相关的肿瘤样疾病。两者都是作为 TSC 1或TSC 2基因中的遗传或散发突变的结果，其功能为阴性 mTOR通路的调节剂。不受控制的mTORC 1活性导致异常细胞的肿瘤性增殖 肺中的平滑肌细胞（LAM细胞），进行性呼吸短促，复发性气胸，以及肺内平滑肌细胞（LAM细胞）的丧失。 肺组织结构和功能。TS影响美国多达25，000至40，000人， 全世界大约有1到2百万人。LAM是一种影响育龄女性的罕见疾病。第一和 唯一FDA批准的LAM治疗是免疫抑制剂西罗莫司（Rapamune©），自2015年以来由 辉瑞制药它是目前的标准治疗，通过抑制mTORC 1发挥作用。西罗莫司有几个临床缺点， 包括相当数量的无应答者，由于其免疫抑制特性而导致的严重不良事件， 和妊娠C类，限制其在育龄妇女中的使用。因此，存在高度未满足的医疗需求， 开发LAM和TS的替代和更安全的治疗方案。 我们已经确定了硝唑尼特（以下简称NTZ; Alinia©），一种FDA批准的安全的抗原虫治疗方法， 治疗1岁或1岁以上患者由贾第鞭毛虫或隐孢子虫引起的腹泻， LAM/TS的潜在新疗法。使用捕获化合物质谱技术，我们发现 NAD（P）H醌氧化还原酶（NQO 1）作为硝唑尼特的人体靶点。此外，我们还表明， NTZ在三种不同的LAM相关小鼠和人细胞中有效抑制mTORC 1相关的细胞信号传导 线解决LAM/TS的潜在病理机制。 我们的目标是这项赠款是调查NTZ的疗效治疗LAM/TS。目标1将是体内证明 在TSC 2-null小鼠模型中比较三种剂量水平的NTZ的概念， Krymskaya在宾夕法尼亚大学。它将评估TSC 2无效病变生长的预防， 在小鼠LAM模型中的免疫组织化学和肺形态学变化和动物存活率。 目标2将涵盖一项体外研究，研究对mTOR和相关信号通路的潜在细胞效应。 体外研究将研究NTZ在LAM衍生的AML（LAMD）细胞和永生化TSC 2-null细胞中的作用。 人源性培养物，随后进行更详细的mTORC 1相互作用研究。除了Moncler NTZ方法，将探索NTZ添加的剂量毒性倾向性雷帕霉素的剂量降低潜力。 该项目旨在为患有LAM/TS毁灭性疾病的患者开发一种新的治疗方法。根据NTZ的 作为一种批准的抗原生动物药物，具有出色的安全性，我们研究中的积极疗效结果将使 根据FDA第505（2）（B）条批准途径和孤儿病认定加速临床开发。