Ubiquitin pathway inhibitors for treatment of asthma

描述（由申请人提供）：哮喘是一种慢性疾病，发病率相当高，无法治愈。对症治疗是有效的，但产生显着的副作用。因此，需要改进的哮喘靶向治疗。当T细胞（CD 4+）分化为效应T细胞时，哮喘发生，效应T细胞分泌细胞因子，其有助于各种先天免疫细胞的募集和增殖进入组织。Th 2和Th 17细胞通过分泌炎性细胞因子参与发病机制，所述炎性细胞因子诱导中性粒细胞和嗜酸性粒细胞的流入，导致气道炎症。因此，靶向抑制促炎性细胞因子和趋化因子是减弱哮喘过敏反应的有吸引力的方法。在这里提出，通过利用泛素蛋白酶体途径来实现这一治疗目的。蛋白质泛素化是先天性和适应性免疫系统的关键调节机制;连接酶如Itch、Nedd 4（HECT）、Cbl-b和MID 1（RING）调节哮喘中的免疫应答。包括Itch在内的Nedd 4连接酶负调节过敏性免疫应答。值得注意的是，小鼠或人类瘙痒的遗传破坏会导致多系统免疫紊乱和肺部炎症。几种Nedd 4家族E3连接酶以自抑制状态存在; Itch通过N-末端自抑制C2结构域保持失活并被激活 通过衔接蛋白如Nedd 4家族相互作用蛋白1（Ndfip 1），其减轻自抑制。Ndfip 1促进多种E3连接酶的活性，从而限制T细胞中的Th 2分化和IL-4/IL-5 /IL-13产生以及Th 17细胞的致病性，并且Ndfip 1-/-小鼠在肺中发展由产生Th 2细胞因子的T细胞诱导的特征性炎症。此外，发现Ndfip 1中的SNP与哮喘和特应性皮炎相关。因此，Ndfip 1是T细胞调节和过敏性炎症反应的关键负调节剂，Ndfip 1的治疗模拟物应选择性激活泛素化级联反应，以限制Th 2和Th 17细胞因子的产生，从而减少过敏性炎症。这些模拟物将在小分子文库的高通量筛选中使用适于鉴定瘙痒激活剂的Progenra的E3连接酶测定技术来鉴定。细胞概念验证试验将测量选定的小分子激动剂在2型辅助T细胞和相关过敏反应模型中的影响。在II期，将使用最佳激动剂进行先导物优化和额外的临床前研究，以确定其调节瘙痒功能以及减轻相关小鼠模型中炎症的能力。

DESCRIPTION (provided by applicant): Asthma is a chronic disease with considerable morbidity and no cures. Symptomatic treatment is efficacious but produces significant adverse effects. Thus, improved, targeted therapies for asthma are needed. Asthma develops when T cells (CD4+) differentiate to effector T cells, which secrete cytokines that assist in the recruitment and proliferation of various innate immune cells into tissues. Th2 and Th17 cells contribute to pathogenesis by secreting inflammatory cytokines that induce the influx of neutrophils and eosinophils, leading to airway inflammation. Targeted suppression of pro-inflammatory cytokines and chemokines are thus attractive approaches to attenuate allergic responses in asthma. It is proposed here to achieve this therapeutic aim by exploiting the ubiquitin proteasome pathway. Protein ubiquitylation is a key regulatory mechanism of innate and adaptive immune systems; ligases such as Itch, Nedd4 (HECT), Cbl-b, and MID1 (RING) regulate immune responses in asthma. Nedd4 ligases including Itch negatively regulate allergic immune responses. Notably, genetic disruption of Itch in mice or humans causes multi-system immune disorders and lung inflammation. Several Nedd4 family E3 ligases exist in an auto-inhibited state; Itch is kept inactive by the N-terminal auto-inhibitory C2 domain and is activated by adapter proteins such as the Nedd4- family interacting protein 1 (Ndfip1), which relieves the auto-inhibition. Ndfip1 promotes activity of multiple E3 ligases, thereby limiting both Th2 differentiation and IL-4/IL-5 /IL-13 production in T cells as well as the pathogenicity of Th17 cells, and Ndfip1-/- mice develop inflammation in the lungs characteristic of that induced by Th2 cytokine producing T cells. Moreover, SNPs in Ndfip1 are found associated with asthma and atopic dermatitis. Thus, Ndfip1 is a key negative modulator of T cell regulation and allergic inflammatory responses, and therapeutic mimetics of Ndfip1 should selectively activate ubiquitylation cascades to limit Th2 and Th17 cytokine production and thereby diminish allergic inflammation. Such mimetics will be identified in high throughput screening of small molecule libraries with the use of Progenra's E3 ligase assay technology adapted to identify activators of Itch. Cell proof of concept assays will measure the impact of selected small molecule agonists in Type 2 helper T cells and relevant allergy response models. In Phase II, lead optimization and additional preclinical studies will be performed with the best agonists to ascertain their ability o modulate Itch functions as well as to attenuate inflammation in relevant mouse models.