Novel therapeutics for vesicants and toxic inhaled chemicals (U54)

项目摘要（参见说明）：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）引起组织中的炎症、DNA 损伤（8-羟基脱氧鸟苷积累；彗星测定；ATM/ATR 激活和下游信号传导）和脂质过氧化（4-羟基壬烯醛）以及炎症。在肺部，CEES 导致支气管血管损伤/渗漏。吸入暴露于 CEES 或 SM 后会形成气道阻塞性含纤维蛋白管型，并且在氯 (400 ppm) 后也出现类似的病变。催化抗氧化剂 AEOL10150 可清除超氧化物、过氧化氢、脂质过氧化物和过氧亚硝酸盐，可挽救： a) CEES 造成的上（鼻）和下呼吸道损伤； b) SM 造成的下气道损伤； c) 氯引起的氧化应激、炎症和乙酰甲胆碱反应性； d) CEES 造成的皮肤损伤，例如炎症和中性粒细胞浸润。天然膳食补充剂水飞蓟宾也能挽救 CEES 造成的皮肤损伤。新数据显示：a) AEOL10150 可有效限制 CEES 和 SM 暴露增加造成的损伤，以及较高氯浓度 (250 ppm) 造成的急性肺损伤，b) CEES 吸入和氯模型中的凝血激活作用，以及 c) tPA（组织型纤溶酶原激活剂）在晚期救援中的功效（从 CEES 吸入后 4 小时开始）。下一个周期，Denver CounterACT 将：a) 确定早期抗凝剂和/或晚期纤溶剂对 CEES、SM 和氯气道阻塞的疗效，b) 评估 AEOL10150 在这些药物造成的严重气道损伤中的作用（单独使用以及与抗凝剂和/或纤溶剂方法联合使用），以及 c) 评估 AEOL10150 和黄酮类水飞蓟宾以挽救 CEES、氮气造成的皮肤损伤芥末 (NM) 和 SM。将交叉检查有效的抗凝剂和/或纤溶剂，以验证皮肤损伤不会加剧。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。 SM 将评估对 CEES/NM 有效的药剂的皮肤损伤情况。这些方法将确定有前景的有毒气体皮肤和肺部损伤救援剂。

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.