Therapy for ocular mustard gas exposure using engineered FGF derivatives

 描述（由申请人提供）：芥子气（硫芥子气，SM）和相关的起疱剂已被用作战争武器，并被美国政府认为是对平民的潜在恐怖威胁。这些制剂对皮肤、肺和眼睛造成迅速、严重和使人衰弱的伤害。眼睛，特别是角膜是对辐射最敏感的组织。SM暴露会导致微泡形成和角膜表面去上皮化，并导致显著疼痛和视力下降。目前尚无有效的治疗水疱性损伤的方法。治疗，可以减少角膜病变和加速愈合的角膜表面将是非常有价值的攻击后。本研究的目的是通过使用天然生长因子的工程化版本来产生有效的治疗SM角膜损伤的药物。这些物质，包括成纤维细胞生长因子（FGF）家族的许多成员，是自然伤口愈合反应的一部分，可以加速各种角膜伤口的愈合。在SM损伤的情况下，内源性产生的和外源性应用的生长因子的功效可能受到生长因子对SM灭活的敏感性、损伤后诱导的蛋白酶降解和/或生长因子从眼睛表面和伤口床扩散的限制。我们已经生产了FGF-1的工程版本，FGF-1是一种参与愈合过程的关键生长因子，它应该克服内源性蛋白质的许多限制，并促进其作为芥子气暴露的眼部治疗的使用。该研究项目将首先在兔角膜器官培养模型中测试工程FGF-1（eFGF-1）分子的功效，以证明eFGF-1可以加速水疱性损伤愈合的假设。将评价两种eFGF-1分子，其1）缺乏天然FGF-1的任何游离巯基; 2）具有额外的稳定性增强突变，包括内部二硫键;和3）不需要肝素辅因子。这些衍生物不应被任何残留的SM修饰，对蛋白水解的敏感性大大降低，并且应通过与细胞外基质的硫酸化蛋白聚糖的相互作用保留在伤口部位。将显示这些分子加速眼表面的再生和增殖以及抑制伤口相关标志物的表达的能力。最后，将证明eFGF-1在体内暴露于SM后加速兔眼愈合的能力。如果这项工作成功，这些eFGF-1将进入SM损伤的临床开发。

 DESCRIPTION (provided by applicant): Mustard gas (sulfur mustard, SM) and related vesicants have been used as weapons of war and are considered by the US government to be a potential terrorist threat to the civilian population. These agents cause rapid, significant and debilitating injuries to the skin, lungs and eyes. The eye and particularly the cornea is the tissu most sensitive to exposure. SM exposure drives microvesication with Deepithelialization of the corneal surface and results in significant pain and degradation of vision. No effective treatment for vesicant injury is known. Treatments that could reduce the corneal lesions and accelerate healing of the corneal surface would be highly valuable subsequent to an attack. The goal of this research is to produce effective therapeutics for SM corneal injury by using engineered versions of naturally occurring growth factors. These substances, including many members of the fibroblast growth factor (FGF) family, are part of the natural wound healing response and can accelerate the healing of a wide variety of corneal wounds. The efficacy of both endogenously produced and exogenously applied growth factors in the context of SM injury may be limited by the sensitivity of growth factors to inactivation by SM, degradation by proteases induced following injury and/or by diffusion of the growth factors from the surface of the eye and the wound bed. We have produced engineered versions of FGF-1, a key growth factor involved in the healing process, that should overcome many of the limitations of the endogenous protein and facilitate its use as an ocular therapy for mustard gas exposure. This research project will first test engineered FGF-1 (eFGF-1) molecules for efficacy in the rabbit corneal organ culture model to prove the hypothesis that the eFGF-1s can accelerate healing of vesicant injury. Two eFGF-1 molecules that 1) lack any of the free thiols of native FGF-1; 2) have additional stability enhancing mutations including an internal disulfide bond; and 3) do not require heparin co-factors will be evaluated. These derivatives should not be modified by any residual SM, be considerably less susceptible to proteolysis and should be retained in the wound site through interactions with sulfated proteoglycans of the extracellular matrix. The ability of these molecules to accelerate the regeneration and proliferation of the ocular surface and inhibit the expression of wound related markers will be shown. Finally, the ability of the eFGF-1s to accelerate healing in the rabbit eye following SM exposure in vivo will be demonstrated. If this work is successful, these eFGF-1s will be advanced into clinical development for SM injury.