Optimization of Novel Molecular Target-based Drugs for Arsenical Skin Injury

砷皮肤损伤新型分子靶点药物的优化

基本信息

批准号：
10023318
负责人：
Mohammad Athar
金额：
$ 74.1万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-10 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10023318
关键词：
Accidents Acids Acute Animal Model Anti-Inflammatory Agents Antidotes Antioxidants Arsenic Arsenicals Asia Attenuated Award Biological Biological Assay British Bulla Cell Death ChIP-seq Characteristics Chelating Agents Chemical Warfare Chemical Weapons Chemicals China Clinic Clinical Complex Cutaneous Cysteine Dangerousness Data Dermatology Development Dose Drug Formulations Event Exposure to FDA approved Family suidae Formulation Funding Generations Germany Health protection Human Human Rights Inflammation Inflammatory Response Injury Intervention Investigation Italy Japan Laboratories Lesion Maximum Tolerated Dose Mediating Miniature Swine Modeling Molecular Molecular Chaperones Molecular Target Mus Mustard Gas Outcome Oxides Pain Pathogenesis Pathway interactions Pharmaceutical Preparations Phosphorylation Poison Population Protective Agents Proteins Publishing Reactive Oxygen Species Records Reporting Research Risk Role Safety Scientist Security Seminal Signal Transduction Skin injury Syria System Therapeutic Therapeutic Agents Tissues Topical application Toxic effect Toxicology Translations USSR United States National Institutes of Health Validation Vesicants World War I World War II base chemical threat clinical candidate clinical translation design effective therapy endoplasmic reticulum stress experience inhibitor/antagonist lead optimization lewisite mouse model novel pre-clinical programs propellant protein folding protein misfolding response skin lesion small molecule therapeutic lead compound transcription factor transcriptome sequencing treatment strategy validation studies weapons web site

项目摘要

Arsenicals such as lewisite, diethylchloroarsine, diphenylchlorarsine, and diphenylcyanoarsine are extremely toxic chemicals that have been used in chemical warfare since World War I and continue to remain a threat to humans, who may be exposed through accidental or intentional mass population exposure. Topical exposure to these agents results in severe cutaneous blistering, inflammation and pain, and therapeutic strategies that safely and effectively attenuate this damage remain urgently needed. Such a strategy has long remained elusive in large part because the molecular mechanisms that underlie the cutaneous damage caused by arsenicals had not been identified. With our previous award, we developed murine and porcine models that, upon topical arsenical exposure, develop cutaneous lesions nearly identical to those that occur in humans. Employing these animal models, we identified a master regulatory signaling cascade underpinning the complex pathobiology of arsenicals. Mechanistically, the inflammatory responses, cell death, tissue disruption, and pain pathways induced by cutaneous arsenicals exposure are mediated by the induction of endoplasmic reticulum (ER) stress and reactive oxygen species generation and subsequent activation of unfolded protein response (UPR) signaling, particularly that involving the ATF4-eIF2α axis. Phosphorylated eIF2α, which we found to be upregulated after arsenicals exposure, blocks translation of most nascent proteins but upregulates the translation of the ATF4 transcription factor. RNA-Seq and CHIP-Seq data confirmed an unbiased role of ATF4 in the pathogenesis of the skin lesions and identified a unified role of ATF4-regulated proteins in this injury. Therefore, we investigated the therapeutic potential of the chemical chaperone 4-phenylbutyric acid (4-PBA), which has been shown to enhance protein folding and reduce ER stress; the antioxidant N-acetyl cysteine (NAC); and the inhibitor of eIF2α phosphorylation ISRIB. Each of these drugs was highly effective in restoring protein translation and diminishing inflammation, tissue disruption, and pain in our mouse model. Thus, we have validated the mechanism-based efficacy of these small molecule agents against cutaneous toxicity induced by arsenicals. 4-BPA and NAC are FDA approved, thus we propose to advance these findings through the lead optimization of 4-PBA and NAC delivered by topical administration after arsenicals exposure in our murine and porcine models. Specifically, we propose to determine the efficacy of the maximum tolerated dose, the window of efficacy, and the durability of response for these drugs, alone and in combination, in treating arsenicals-mediated cutaneous injury in mice (Aim 1); to develop various topical formulations of these drugs and assess the efficacy thereof against arsenicals-mediated cutaneous injury in mice (Aim 2); and to confirm the efficacy of the identified novel outstanding formulation in our porcine model of arsenicals-mediated cutaneous injury (Aim 3). These studies will drive the clinical translation of an antidote for the cutaneous toxicity of arsenicals, which may be further expedited as these drugs are already FDA approved.

砷化合物如路易斯矿、二乙基氯胂、二苯基氯胂和二苯基氰基胂是极其危险的。自第一次世界大战以来一直在化学战中使用的有毒化学品，并继续对人类构成威胁人类，可能通过意外或故意的大规模人群暴露而暴露。这些药物会导致严重的皮肤起泡、炎症和疼痛，治疗策略长期以来，我们仍然迫切需要安全有效地减轻这种损害。很大程度上是难以捉摸的，因为造成皮肤损伤的分子机制凭借我们之前的奖项，我们开发了小鼠和猪模型，局部接触砷后，会出现与人类几乎相同的皮肤损伤。利用这些动物模型，我们确定了支撑该复合物的主要调控信号级联砷的病理学，从机制上讲，炎症反应、细胞死亡、组织破坏和疼痛。皮肤砷暴露诱导的途径是通过内质网的诱导介导的 (ER) 应激和活性氧的产生以及随后激活未折叠蛋白反应 (UPR) 信号传导，特别是涉及磷酸化 eIF2α 轴的信号传导，我们发现它是磷酸化的 eIF2α 轴。砷暴露后上调，阻止大多数新生蛋白质的翻译，但上调 ATF4 转录因子的 RNA-Seq 和 CHIP-Seq 数据的翻译证实了 ATF4 的公正作用。并确定了 ATF4 调节蛋白在这种损伤中的统一作用。因此，我们研究了化学伴侣 4-苯基丁酸 (4-PBA) 的治疗潜力，它已被证明可以增强蛋白质折叠并减少内质网应激；抗氧化剂 N-乙酰半胱氨酸； (NAC)；以及 eIF2α 磷酸化抑制剂 ISRIB，这些药物在恢复方面都非常有效。在我们的小鼠模型中，蛋白质翻译并减少炎症、组织破坏和疼痛。已经验证了这些小分子药物针对皮肤毒性的基于机制的功效 4-BPA 和 NAC 已获得 FDA 批准，因此我们建议通过以下途径推进这些发现。在我们的研究中砷暴露后通过局部给药对 4-PBA 和 NAC 进行先导优化具体来说，我们建议确定最大耐受剂量的功效，这些药物单独或联合使用的疗效窗口和反应的持久性砷介导的小鼠皮肤损伤（目标 1）；开发这些药物的各种外用制剂；并评估其对小鼠砷介导的皮肤损伤的功效（目标2）；已确定的新型出色制剂在砷介导的猪模型中的功效皮肤损伤（目标 3）。这些研究将推动皮肤解毒剂的临床转化。砷的毒性，由于这些药物已获得 FDA 批准，因此可能会进一步加快。