Novel therapeutics for vesicants and toxic inhaled chemicals (U54)

项目摘要(参见说明)：丹佛的中和计划是为了建立硫磺芥末(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-氯乙基乙硫醚)可引起组织炎症、DNA损伤(8-羟基脱氧鸟苷蓄积；彗星试验；ATM/ATR激活和下游信号转导)和脂质过氧化(4-羟基壬烯醛)，以及炎症。在肺部，CEES可引起支气管血管损伤/渗漏。吸入CEES或SM后形成含有纤维蛋白的呼吸道阻塞管型，氯(400ppm)后也有类似的损害。催化抗氧化剂AEOL10150可清除超氧化物、过氧化氢、过氧化脂质和过氧亚硝酸盐，可挽救：a)CEES造成的上(鼻)和下呼吸道损伤；b)SM造成的下呼吸道损伤；c)氯引起的氧化应激、炎症和乙酰甲胆碱反应；以及d)CEES造成的皮肤损伤，如炎症和中性粒细胞渗透。一种天然的膳食补充剂水飞蓟宾也拯救了CEES的皮肤损伤。新的数据显示：a)AEOL10150有效地限制了CEES和SM暴露的升级造成的损伤，以及较高的氯浓度(250ppm)所致的急性肺损伤，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)在晚期救援中的效果，从CEES吸入后4小时开始。下一周期，丹佛中和将：a)建立早期抗凝剂和/或晚期纤溶剂对CEES、SM和氯造成的呼吸道阻塞的疗效，b)评估AEOL10150在这些药物(单独或联合抗凝剂和/或纤溶方法)治疗严重呼吸道损伤中的疗效，以及c)评估AEOL10150和黄酮类水飞蓟宾在CEES、氮芥末(NM)和SM中拯救皮肤损伤的效果。将对有效的抗凝剂和/或纤溶剂进行交叉检查，以证实皮肤损伤没有恶化。将评估氧化应激、炎症、通透性增加的终点，对于肺部，还将评估呼吸道阻塞。发现对CEES/NM有效的S试剂将在皮肤损伤中进行SM评估。这些方法将为有毒气体造成的皮肤和肺损伤确定有前途的救援剂。

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.