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

特发性肺纤维化（IPF）是一种衰老性疾病，诊断时平均年龄为66岁。 尽管有这种强烈的关联，但导致纤维化的衰老倾向的细胞/分子机制仍然存在。 疾病尚未得到阐明。我们实验室以前的研究表明，肌成纤维细胞，关键 纤维化中的效应细胞，表现出在衰老中维持氧化还原稳态的能力减弱;这 部分由抗氧化反应转录因子Nrf 2的诱导缺陷调节。人类 IPF受试者在肌纤维母细胞灶中表现出Nrf 2表达降低，支持这种细胞氧化还原 在人类纤维化疾病中的不平衡。我们的初步数据支持这种Nrf 2氧化还原失衡在一种新的老化 小鼠中的非消退性纤维化模型。据我们所知，这代表了第一个衰老的纤维化模型， 概括了人IPF的非消退性质。Nrf 2激活剂富马酸二甲酯（DMF）是FDA- 用于通过口服给药途径治疗多发性硬化症的批准药物。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/纳米颗粒制剂， 其可以作为干粉吸入剂（DPI）局部给药于肺。我们将使用FDA批准的 人吸入器装置、FDA批准的赋形剂和FDA批准的Nrf 2激活剂，以递送这种新型Nrf 2- 活化剂制剂在体内通过吸入途径。实验将在FDA/USP条件下进行 使用FDA/USP规定的体外试验，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）， 和体内药代动力学/药效学研究。这些研究将评估影响 作为药物剂量、颗粒-细胞膜相互作用、颗粒细胞摄取 膜渗透性、药物细胞转运和细胞Nrf 2活化。最后，我们将评估 这种新Nrf 2激活剂制剂和递送方法在已建立的衰老小鼠模型中的功效 纤维化本申请中拟定的研究旨在：（1）确定DMF（一种FDA- 获批药物（多发性硬化症），用于新适应症（肺纤维化）;（2）测试新型DMF的疗效 （3）研究口服与吸入（局部）抗氧化剂治疗IPF的策略; 和（4）评价可能直接导致IPF临床试验的治疗剂的安全性和有效性。

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.