Development of Novel Second Generation Anti-inflammatory Substrate-selective p38 MAP Kinase Inhibitors as Therapy for Acute Respiratory Distress Syndrome

急性呼吸窘迫综合征（ARDS）每年影响约190，000名患者，导致约75，000人死亡 在美国，ARDS的死亡率约为40%，并在幸存者中引起显著的发病率。呼吸 病毒，包括流感和现在的严重急性呼吸道综合征冠状病毒-2（SARS-CoV-2）， ARDS的重要原因，治疗选择有限。VA患者人群有许多风险 ARDS预后不良的因素包括老年、肥胖、高血压、糖尿病和慢性 肺和心脏病。目前尚无有效的药理学治疗方法， ARDS。为了满足这一需求，我们开发了一类新型的底物和功能选择性p38丝裂原， 具有抗炎和内皮屏障稳定活性的活化蛋白激酶（MAPK）抑制剂。 与传统的p38催化抑制剂相比， 炎症通路，我们的新化合物靶向底物对接位点之一，因此仅 阻止某些依赖p38的过程。与我的合作研究者，夏皮罗博士，麦克雷尔博士， 弗莱彻，我们使用计算机辅助药物设计（CADD），以确定一个先导化合物，UM 101，结合 p38 a上的艾德底物对接位点附近的口袋，并表现出独特的生物活性谱。我们 开发了第二代类似物SF 7044，具有更大的溶解度和更好的内皮屏障- 稳定、抗炎和肺保护活性。我们目前正在与Gen 1 e Life合作 许可这些化合物专利的科学有限责任公司完成了临床前测试， SF 7044的临床试验。在我们目前的优异奖，我们确定和筛选了200个额外的 使用改进的CADD策略将化合物靶向与UM 101相同的底物对接位点。我们 鉴定了四种新的先导化合物，具有上级内皮屏障稳定活性和p38 a结合活性 与UM 101和SF 7044相比，具有明显的抗炎作用，但溶解性差。体内 在小鼠模型中的测试将需要重新配制或化学修饰这些新的先导化合物， 改善溶解性（如UM 101）。此次更新的总体目标是使用与 开发SF 7044以设计、合成和表征四种新的 具有改进的活性和药物样性质的先导化合物。由于新的先导化合物具有更大的 内皮稳定活性和p38 a结合比UM 101或SF 7044，我们预计开发第二个， 产生具有显著改善的肺保护活性的类似物。我们将利用CADD和药物 化学原理、蛋白质结合试验以及ALI的人细胞和小鼠模型： 1.设计并合成4种新发现的先导化合物的至少20种类似物， p38 a结合和药物样性质。 2.分析第二代类似物的靶标结合和生物活性。 3.在小鼠模型中分析最活跃的第二代类似物的毒性和有效性 ARDS的渗出期（LPS/高热）和纤维增生期（博来霉素）。 4.评估新化合物的脱靶效应 在这项工作结束时，我们将开发新的非催化的第二代类似物。 p38α抑制剂在小鼠肺损伤模型中具有改善的疗效和安全性特征，以提供一个管道 这些新化合物在用于ARDS的临床试验之前已准备好进行高级临床前试验。

Acute Respiratory Distress Syndrome (ARDS) affects ~190,000 patients and causes ~75,000 deaths per year in the U.S.A. ARDS has a mortality rate of ~40% and causes significant morbidity in survivors. Respiratory viruses, including influenza and now Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), are important causes of ARDS with limited therapeutic options. The VA patient population has many of the risk factors for having poor outcomes with ARDS, including older age, obesity, hypertension, diabetes, and chronic lung and heart disease. There are no currently available pharmacologic therapies proven to be effective in ARDS. To address this need, we developed a novel class of substrate- and function-selective p38 mitogen- activated protein kinases (MAPK) inhibitors with anti-inflammatory and endothelial-barrier stabilizing activity. Compared with conventional p38 catalytic inhibitors that block all downstream signaling including anti- inflammatory pathways, our novel compounds target one of the substrate docking sites and therefore only block certain p38-dependent processes. Together with my co-investigators, Drs. Shapiro, MacKerell, and Fletcher, we used computer-aided drug design (CADD) to identify a lead compound, UM101, that binds a pocket near the ED substrate docking site on p38a and exhibits a unique profile of biological activities. We developed a second-generation analog, SF7044, with greater solubility and improved endothelial barrier- stabilizing, anti-inflammatory, and lung-protective activities. We are currently collaborating with Gen1e Life Sciences, LLC, which licensed the patents for these compounds, completed preclinical testing and began clinical testing of SF7044. During our current Merit Award, we identified and screened 200 additional compounds targeted to the same substrate docking site as UM101 using a refined CADD strategy. We identified four new lead compounds with superior endothelial barrier stabilizing activity and p38a-binding compared with UM101 and SF7044 and distinct anti-inflammatory effect profiles, but poor solubility. In vivo testing in a mouse model will require reformulation or chemical modification of these new lead compounds to improve solubility (like UM101). The overall objective of this renewal is to use the same strategy as used for development of SF7044 to design, synthesize, and characterize second-generation analogs of the four new lead compounds with improved activity and drug-like properties. Since the new lead compounds have greater endothelial-stabilizing activity and p38a-binding than either UM101 or SF7044 we expect to develop second- generation analogs with substantially improved lung-protective activity. We will utilize CADD and medicinal chemistry principles, protein binding assays and human cell and mouse models of ALI: 1. Design and synthesize at least 20 analogs of each the 4 newly discovered lead compounds to improve p38a-binding and drug-like properties. 2. Analyze the second-generation analogs for target binding and biological activities. 3. Analyze toxicity and effectiveness of the most active second-generation analogs in mouse models of the exudative (LPS/hyperthermia) and fibroproliferative (bleomycin) phases of ARDS. 4. Evaluate new compounds for off-target effects At the conclusion of this work, we will have developed second generation analogs of novel non-catalytic p38α inhibitors with improved efficacy and safety profiles in mouse lung injury models to provide a pipeline of new compounds that are ready for advanced preclinical testing prior to clinical testing in ARDS.