Therapy for ocular mustard gas exposure using engineered FGF derivatives

 描述(申请人提供)：芥子气(硫磺芥末，SM)和相关发泡剂已被用作战争武器，并被美国政府视为对平民人口的潜在恐怖威胁。这些毒剂会对皮肤、肺部和眼睛造成迅速、严重和令人衰弱的损伤。眼睛，尤其是角膜，是对暴露最敏感的组织。接触SM会导致角膜表面去上皮化的微泡形成，并导致严重的疼痛和视力下降。目前尚无治疗发泡性损伤的有效方法。可以减少角膜损伤并加速角膜表面愈合的治疗方法在疾病发作后将具有极高的价值。这项研究的目标是通过使用自然产生的生长因子的工程化版本来生产有效的治疗SM角膜损伤的方法。这些物质，包括成纤维细胞生长因子(成纤维细胞生长因子)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损伤的背景下，内源性和外源性生长因子的有效性都可能受到生长因子对SM的失活、损伤后引起的蛋白酶降解和/或生长因子从眼表面和伤口床扩散的敏感性的限制。我们已经生产出了工程化的成纤维细胞生长因子-1，这是一种参与愈合过程的关键生长因子，应该可以克服内源性蛋白的许多限制，并有助于将其用作芥子气暴露的眼部治疗。本研究项目将首先在兔角膜器官培养模型中测试工程化成纤维细胞生长因子-1(eFGF-1)分子的有效性，以证明eFGF-1能促进发泡性损伤愈合的假说。1)缺少天然成纤维细胞生长因子-1的任何游离硫醇；2)有额外的稳定性增强突变，包括内部二硫键；以及3)不需要肝素辅助因子的两个eFGF-1分子将被评估。这些衍生物不应被任何残留的SM修饰，对蛋白降解的敏感性大大降低，并应通过与细胞外基质的硫酸蛋白多糖相互作用保留在伤口部位。这些分子能够加速眼表的再生和增殖，并抑制创伤相关标志物的表达。最后，在活体实验中，将证实eFGF-1s在SM暴露后促进兔眼愈合的能力。如果这项工作成功，这些eFGF-1将被推进到SM损伤的临床开发中。

 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.