Novel therapeutics for vesicants and toxic inhaled chemicals (U54)

项目总结（见说明）：丹佛的CounterACT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挽救SM介导的损伤。SM类似物CEES（2-氯乙基乙基硫醚）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）和组织中的脂质过氧化（4-羟基壬烯醛）以及炎症。在肺中，CEES引起支气管血管损伤/泄漏。吸入暴露于CEES或SM后形成了气道阻塞性含纤维蛋白的管型，氯（400 ppm）后也出现了类似的病变。清除超氧化物、过氧化氢、脂质过氧化物和过氧亚硝酸盐的催化抗氧化剂AEOL 10150拯救：a）CEES引起的上（鼻）和下气道损伤; B）SM引起的下气道损伤; c）氯引起的氧化应激、炎症和乙酰甲胆碱反应性;以及d）CEES引起的皮肤损伤，如炎症和中性粒细胞浸润。一种天然的膳食补充剂水飞蓟宾也挽救了CEES造成的皮肤损伤。新数据显示：a）AEOL 10150限制CEES和SM暴露增加所致损伤以及较高（250 ppm）氯浓度所致急性肺损伤的有效性，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）在晚期抢救中的有效性，从CEES吸入后4小时开始。下一个周期，丹佛反ACT将：a）建立早期抗凝剂和/或晚期纤维蛋白溶解剂在CEES、SM和氯引起的气道阻塞中的功效，B）评估AEOL 10150在这些试剂引起的严重气道损伤中的作用（单独和与抗凝剂和/或纤维蛋白溶解方法组合），和c）评估AEOL 10150和类黄酮水飞蓟宾挽救CEES、氮芥（NM）和SM引起的皮肤损伤。将交叉检查有效的抗凝剂和/或纤维蛋白溶解剂，以验证皮肤损伤没有恶化。将评价氧化应激、炎症、通透性增加和肺气道阻塞的终点。发现对CEES/NM有效的药物将通过SM在皮肤损伤中进行评价。这些方法将确定有前途的救援剂的皮肤和肺损伤的有毒气体。

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.