Preclinical development of a novel Nrf2-activator formulation for the treatment of idiopathic pulmonary fibrosis

特发性肺纤维化(IPF)是一种老年性疾病，确诊时的平均年龄为66岁。 尽管有这种强烈的联系，导致衰老倾向于纤维化的细胞/分子机制 疾病还没有被阐明。我们实验室之前的研究已经证明，肌成纤维细胞，关键 效应细胞在纤维化形成中表现出在衰老过程中维持氧化还原动态平衡的能力减弱；这 在一定程度上是由抗氧化反应转录因子Nrf2的缺陷诱导所调节。人类 IPF受试者表现出Nrf2在肌成纤维细胞灶中的表达减少，支持这种细胞氧化还原 人类纤维性疾病中的不平衡。我们的初步数据支持Nrf2氧化还原失衡在一种新的衰老中 小鼠未消退肝纤维化模型。据我们所知，这是第一个衰老的纤维化模型 概括了人类IPF的非解析性。NRF2激活剂富马酸二甲酯(DMF)是FDA- 经批准的通过口服途径治疗多发性硬化症的药物。我们已经开发出一种 使用纳米技术和FDA批准的赋形剂的新型DMF微粒/纳米微粒配方， 它可以作为干粉吸入器(DPI)局部给药到肺部。我们将使用FDA批准的 人体吸入器设备，FDA批准的赋形剂，以及FDA批准的NRF2激活剂，以提供这种新型的NRF2- 活化剂通过吸入途径在体内的配方。实验将在FDA/USP的条件下进行 使用FDA/USP规定的要求的体外试验，包括体外2D细胞培养(即液体覆盖培养 和模拟空气-液体肺界面的空气界面培养)，体外3D细胞培养(即，空气界面培养)， 体内药代动力学/药效学研究。这些研究将评估影响 细胞活力与药物剂量、颗粒-细胞膜相互作用、颗粒细胞摄取、 膜通透性、药物细胞转运和细胞内Nrf2的激活。最后，我们将评估 这种新型NRF2激活剂制剂和给药方法对已建立的衰老小鼠模型的疗效 纤维化症。本申请中拟议的研究旨在：(1)确定DMF的有效性，FDA- 批准的药物(多发性硬化症)，用于新的适应症(肺纤维化)；(2)测试一种新型DMF的疗效 配方，作为DPI；(3)研究IPF口服和吸入(局部)抗氧化剂策略； 以及(4)评估可直接导致IPF临床试验的治疗方法的安全性和有效性。

Idiopathic pulmonary fibrosis (IPF) is a disease of aging, with a mean age of 66 years at the time of diagnosis. Despite this strong association, cellular/molecular mechanisms that account for the aging predilection to fibrotic disease have not been elucidated. Previous studies in our lab have demonstrated that myofibroblasts, key effector cells in fibrogenesis, demonstrate a diminished capacity to maintain redox homeostasis in aging; this was in part regulated by a deficient induction of the antioxidant response transcription factor, Nrf2. Human subjects with IPF exhibit decreased Nrf2 expression in myofibroblastic foci, supporting this cellular redox imbalance in a human fibrotic disease. Our preliminary data support this Nrf2 redox imbalance in a novel aging model of non-resolving fibrosis in mice. This represents, to our knowledge, the first aging model of fibrosis that recapitulates the non-resolving nature of human IPF. The Nrf2-activator, dimethyl fumarate (DMF), is an FDA- approved drug for the treatment of multiple sclerosis via the oral route of administration. We have developed a novel DMF microparticulate/nanoparticulate formulation using nanotechnology and FDA-approved excipients, which can be administered locally to the lungs as a Dry Powder Inhaler (DPI). We will utilize FDA-approved human inhaler devices, FDA-approved excipients, and an FDA-approved Nrf2 activator to deliver this novel Nrf2- activator formulation in vivo via an inhaled route. Experiments will be conducted under FDA/USP conditions using required in vitro tests specified by the FDA/USP, including in vitro 2D cell culture (i.e. liquid-covered culture and air-interface culture mimicking the air-liquid lung interface), in vitro 3D cell culture (i.e. air-interface culture), and in vivo pharmacokinetics/pharmacodynamics studies. These studies will evaluate mechanisms influencing cell viability as a function of drug dose, particle-cellular membrane interactions, particle cellular uptake, membrane permeability, drug cellular transport, and activation of cellular Nrf2. Finally, we will evaluate the efficacy of this novel Nrf2-activator formulation and delivery method in an aged mouse model of established fibrosis. The proposed studies in this application were designed to: (1) Determine the efficacy of DMF, an FDA- approved drug (multiple sclerosis), for a new indication (pulmonary fibrosis); (2) Test the efficacy of a novel DMF formulation, as a DPI; (3) Investigate oral versus inhaled (local) administration of an antioxidant strategy for IPF; and (4) evaluate safety and efficacy profiles of therapeutics that could lead directly to clinical trials for IPF.