Translation of novel and repurposed drugs to address the acute and late effects of mustard exposure

芥子气去污和中和的对策协议可以保护暴露的 暴露后立即给药时，皮肤、眼睛和肺部表面的毒性。然而，这可能不会 足以解决骨髓抑制延迟发作和造血功能破坏相关的 暴露在芥子气中目前建议的影响是提供一个全面的对策方案 这解决了急性皮肤炎介导的芥子气暴露的急性和延迟的有害影响， 炎症和先天免疫系统，特别是巨噬细胞过度活化。 NM介导的皮肤组织损伤导致快速表皮破裂和血管渗漏 导致水肿和疼痛的炎症。在最初的损伤之后， 单核细胞和巨噬细胞。巨噬细胞介导的可溶性信号分子的释放促进了前 炎症状态，其放大其他细胞毒性活性，包括活性氮物质的释放 iNOS、活性氧和蛋白水解酶的诱导。此外，这些激活的iNOS- 产生巨噬细胞在介导和加重全身毒性中起关键作用。 我们建议使用多学科翻译方法来调节皮肤炎症， 在小鼠和人类研究中使用体内技术测定巨噬细胞活性。我们将研究 螺内酯和双膦酸盐（BPs）与维生素D联合使用，以抑制前 炎症因子，包括皮肤和活化巨噬细胞中的TNF α和iNOS。这些药物具有 已在临床上使用多年，具有已知的安全性和毒性。安体舒通是一种钾- 保留利尿剂（水丸），已安全用于治疗原发性高血压，液体 充血性心力衰竭和其他肾脏病变患者的超负荷和水肿。虽然大部分 已经积累了关于螺内酯对盐皮质激素受体的生理病理学的信息 (MR)和肾上腺，MR配体对非经典靶点如皮肤的作用已经发现了新的 和令人兴奋的潜在应用。我们在此提出，螺内酯作为一种弱利尿剂， 能够减少水肿性皮肤损伤的含水量，作为加速伤口愈合的一种方式。使用 安体舒通可在限制水肿和疼痛性皮肤炎症方面具有益处。BP是指 已经使用了40多年。除了骨骼，这些药物存款在皮肤，肝脏， 脾和骨髓，这与我们的研究兴趣一致，因为它们代表器官和组织， 可以是活化的巨噬细胞的储库和/或靶。此外，BP存款并具有持久的 组织中的影响。我们建议将BP与高剂量维生素D3结合使用作为对策， NM暴露。最后，作为一种针对毒性最大的暴露的全身治疗，我们建议使用一种新的免疫疗法， 改性微粒（IMP），其衍生自临床聚合物。在IMP结合和吞噬后， 巨噬细胞被隔离在肝脏和脾脏中降解。IMP策略代表了 这项拨款的新方面是针对严重的全身毒性。为了验证和确定翻译 BPs和维生素D3在抑制芥子气暴露后皮肤炎症中的意义，我们设计了 一项使用紫外线（太阳光谱辐射）作为健康人群芥子气暴露替代品的人体临床研究 人类受试者，以加快未来的转化研究。众所周知，晒伤暴露会导致 发红、肿胀、水疱和皮肤中巨噬细胞的活化，包括iNOS的上调， TNF α。这项人类临床研究的信息将提供信息，并证实我们的 提出了对策。 我们的团队包括来自大学内外不同中心的共同研究者 还有医院共同调查员和顾问小组将监督项目，帮助确定 新的机会和互补的方法，从专业知识中吸取：皮肤药理学; 炎症和巨噬细胞;维生素D;皮肤和组织病理学;临床试验实施和 监管合规性;统计和流行病学-所有这些对于将本研究转化为实际的 干预 这是一种新颖的、高度转化的方法，可以开发潜在的对策来解决关键问题。 发病率和死亡率的因素。螺内酯、双膦酸盐和维生素D是一种 广泛可用且价格低廉，如果证明有益，可能会对世界各地的公共卫生产生广泛影响。 化学武器暴露事件。

Countermeasure protocols of mustard decontamination and neutralization may protect exposed surfaces of the skin, eyes, and lungs when administered immediately after exposure. However, this may not be adequate to address the delayed onset of myelosuppression and disrupted hematopoiesis associated with mustard exposure. The impact of the current proposal is to provide a comprehensive countermeasure regimen that addresses both the acute and delayed detrimental effects of mustard exposure mediated by acute skin inflammation and the innate immune system, specifically macrophage hyper-activation. NM-mediated tissue injury of the skin results in rapid epidermal disruption and vascular leakage resulting edema and painful inflammation. Following the initial injury is an influx of activated inflammatory monocytes and macrophages. Release of macrophage-mediated soluble signaling molecules promotes a pro- inflammatory state which amplifies other cytotoxic activities including release of reactive nitrogen species iNOS, reactive oxygen species, and induction of proteolytic enzymes. Furthermore, these activated iNOS- producing macrophages are critically involved in mediating and exacerbating systemic toxicities. We propose to use a multidisciplinary translational approach to modulate skin inflammation and macrophage activity using in vivo techniques in mouse and human studies. We will study the effect of spironolactone and bisphosphonates (BPs) in combination with vitamin D to inhibit the production of pro- inflammatory factors including; TNFα, and iNOS in the skin and in activated macrophages. These drugs have been in clinical use for many years with known safety and toxicity profiles. Spironolactone is a potassium- sparing diuretic (water pill) that has been safely used for five decades to treat essential hypertension, fluid overload and edema in patients with congestive heart failure and other kidney pathologies. While much information has been accumulated on the physiopathology of spironolactone on the mineralocorticoid receptor (MR) and the adrenal glands, the effect of MR ligands on non-classical targets such as the skin has found new and exciting potential applications. We hereby propose that spironolactone being a weak diuretic has sufficient capacity to reduce water content of edematous skin lesions as a way to accelerate wound healing. The use of spironolactone may have benefits in limiting edema and painful skin inflammation. BPs are medications that has been used for over 40 years. In addition to the skeletal bones, these drugs deposit in the skin, liver, spleen, and bone marrow which aligns with our research interest as they represent organs and tissues that may be reservoirs and/or targets for activated macrophages. Furthermore, BPs deposit and have long-lasting effects in the tissue. We propose to use BPs in conjunction with high dose vitamin D3 as countermeasures to NM exposure. Lastly, as a systemic treatment for the most toxic exposure, we propose to use a novel immune modifying microparticle (IMPs) which are derived from clinical polymers. Upon binding and engulfment of IMPs, macrophages are sequestered for degradation in the liver and spleen. The strategy with IMPs represents a new aspect of this grant to target severe systemic toxicity. To validate and determine the translational significance of BPs and vitamin D3 in suppressing skin inflammation following mustard exposure, we designed a human clinical study using UV (solar spectrum radiation) as a surrogate for mustard exposure in healthy human subjects to expedite future translational studies. It is well known that sunburn exposure leads to redness, swelling, vesication, and activation of macrophages in the skin including upregulation of iNOS and TNFα. Information from this human clinical study will be informative and confirmatory of the use of our proposed countermeasures. Our assembled team involve co-investigators from different centers within and outside the university and hospital. The team of co-investigators and consultants will have oversight of the project, helping to identify new opportunities and complementary approaches drawing from expertise in: dermatopharmacology; inflammation and macrophages; vitamin D; skin and tissue pathology; clinical trials implementation and regulatory compliance; statistics and epidemiology - all critical to the translation of this study into a practical intervention. This is a novel and highly translational approach to develop potential countermeasure to address key morbidity and mortality factors from mustard exposure. Spironolactone, bisphosphonates, and vitamin D are a widely available and inexpensive and, if shown to be beneficial, could have a broad public health impact in the event of chemical weapon exposure.