Novel therapeutics for vesicants and toxic inhaled chemicals (U54)

项目概述（见说明）：丹佛的抵消计划旨在开发硫芥（SM）对肺和皮肤的毒性模型，了解损伤发病机制，并评估新型抗氧化剂以拯救SM介导的损伤。SM类似物CEES（2-氯乙基乙基硫醚）在组织中引起炎症、DNA损伤（8-羟基脱氧鸟苷积累；彗星试验；ATM/ATR激活和下游信号传导）和脂质过氧化（4-羟基烯醛），以及炎症。在肺中，CEES引起支气管血管损伤/泄漏。吸入CEES或SM后形成气道阻塞性含纤维蛋白的铸型，氯气（400ppm）后出现类似病变。催化抗氧化剂AEOL10150清除超氧化物、过氧化氢、脂质过氧化物和过氧亚硝酸盐，挽救了：a) CEES造成的上（鼻）下气道损伤；b) SM下气道损伤；C)氯引起的氧化应激、炎症和甲胆碱反应性；d) CEES引起的皮肤损伤，如炎症和中性粒细胞浸润。天然膳食补充剂水飞蓟宾也拯救皮肤损伤的CEES。新的数据显示：a) AEOL10150有效限制CEES和SM暴露升级的损伤，以及更高（250 ppm）氯浓度引起的急性肺损伤，b) CEES吸入和氯模型中的凝血激活，以及c) tPA（组织型纤溶酶原激活剂）在晚期抢救中的作用，从吸入CEES后4小时开始。下一个周期，丹佛中和将：a)建立早期抗凝和/或晚期纤维蛋白溶解剂对CEES、SM和氯气道阻塞的疗效，b)评估AEOL10150对这些药物（单独使用和联合抗凝和/或纤维蛋白溶解方法）造成的深度气道损伤，以及c)评估AEOL10150和黄酮类水飞蓟binin对CEES、氮芥（NM）和SM造成的皮肤损伤的抢救效果。将交叉检查有效的抗凝和/或纤溶药物，以确认皮肤损伤没有恶化。将评估氧化应激、炎症、通透性增加以及肺部气道阻塞的终点。发现对CEES/NM有效的药物将在SM皮肤损伤中进行评估。这些方法将为有毒气体造成的皮肤和肺部损伤确定有希望的救援剂。

PROJECT SUMMARY (See instructions): Denver's CounterACT program was formed to develop models of sulfur mustard (SM) toxicity to lung and skin, understand injury pathogenesis, and evaluate novel antioxidants to rescue SM-mediated injuries. SM analog CEES (2-chloroethyl ethylsulfide) caused inflammation, DNA damage (8-hydroxydeoxy-guanosine accumulation; comet assay; ATM/ATR activation and downstream signaling) and lipid peroxidation (4- hydroxynonenal) in tissues, as well as inflammation. In lung, CEES caused bronchial vascular injury/leak. Airway obstructive fibrin-containing casts formed after inhalation exposures to CEES or SM, and similar lesions were apparent after chlorine (400 ppm). Catalytic antioxidant AEOL10150, which scavenges superoxide, hydrogen peroxide, lipid peroxides, and peroxynitrite, rescued: a) upper (nasal) and lower airway injuries by CEES; b) lower airway injury by SM; c) oxidative stress, inflammation and methacholine reactivity due to chlorine; and d) skin injuries by CEES such as inflammation and neutrophil infiltration. A natural dietary supplement silibinin also rescued skin injuries by CEES. New data showed: a) AEOL10150 efficacy to limit injury by escalated CEES and SM exposures, as well as acute lung injury by higher (250 ppm) chlorine concentrations, b) coagulation activation in the CEES inhalation and chlorine models, and c) efficacy of tPA (tissue-type plasminogen activator) in late rescue, starting 4h after CEES inhalation. Next cycle, Denver CounterACT will: a) establish early anticoagulant and/or late fibrinolytic agent efficacies in airways obstruction by CEES, SM and chlorine, b) evaluate AEOL10150 in profound airway injuries by these agents (both alone and combined with anticoagulant and/or fibrinolytic approaches), and c) evaluate AEOL10150 and flavonoid silibinin to rescue skin injuries by CEES, nitrogen mustard (NM), and SM. Effective anticoagulant and/or fibrinolytic agents will be cross-checked to verify lack of exacerbation of skin injuries. Endpoints of oxidative stress, inflammation, increased permeability, and, for lungs, airway obstruction, will be evaluated. Agent(s) found effective for CEES/NM would be evaluated in skin injury by SM. These approaches will identify promising rescue agents for skin and lung injury by toxic gases.