Multivalent Conjugates of Sonic Hedgehog to Accelerate Diabetic Wound Healing

描述：2010年，超过875,000名美国糖尿病患者被诊断为腿部溃疡。这些伤口未能愈合，每年导致73500多人截肢。Valitor公司正在开发蛋白质-聚合物疗法，以克服糖尿病伤口愈合的微血管后果。我们设计了我们的治疗方法，以补充糖尿病溃疡治疗的标准实践，并减少清创和临床观察这些缓慢愈合的伤口所需的昂贵的系列努力。我们的专利技术是一种化学系链过程，使我们能够控制在受体附近传递的生长因子的价态。利用这项技术，我们将重要的血管生成因子Sonic Hedgehog(Shh)连接到透明质酸(HYA)的线性链上，通过改变Shh：HYA的比例，我们可以调节它激活Shh途径的能力。将Shh连接到大分子上也可以防止其被蛋白水解酶失活，并增强其在靶组织中的分子稳定性。我们将开发这些多价Shh(MvShh)结合物作为一种治疗方法，以加速糖尿病下肢溃疡的伤口愈合，因为Shh先前已被证明能刺激新生血管。Valitor治疗Shh的方法将能够增强Shh的体内生物活性，将我们的治疗药物输送到伤口床可以以较低的治疗成本显著改善患者的长期结果。我们在第一阶段的总体目标是开发mvShh结合物，在协调皮肤伤口愈合的细胞类型中诱导增强的Shh途径激活和下游血管生成基因的表达。在具体目标1中，我们将使用从db/db小鼠获得的真皮成纤维细胞来鉴定体外产生最大Shh途径激活的mvShh配方，db/db小鼠是一种表现出伤口愈合受损和血管生成基因表达减少的糖尿病模型动物。然后我们将确定Shh价在体外对微血管内皮细胞增殖和迁移的影响，因为这是血管生成所必需的基本机制。在具体目标2中，我们将使用全层切除创伤模型证明mvShh偶联物将在db/db小鼠中启动Shh信号和下游血管生成基因的表达。我们的发现将为我们的治疗方法的进一步临床前开发提供有价值的概念验证数据，这是我们为该项目的第二阶段计划的。 公共卫生相关性：2010年，超过875,000名美国糖尿病患者被诊断出患有慢性下肢溃疡，这些患者每年产生超过300亿美元的相关医疗费用。Valitor公司正在开发一种先进的治疗方法，以加速糖尿病伤口的愈合，通过鼓励新生血管和改善伤口床的血液供应来抵消高昂的治疗成本。我们第一阶段项目的总体目标是将我们治疗的治疗参数与血管形成的细胞机制联系起来，并展示我们的技术作为临床治疗的概念验证。

DESCRIPTION: In 2010, more than 875,000 Americans with diabetes were diagnosed with a lower extremity ulcer. Failed closure of these wounds results in more than 73,500 lower extremity amputations annually. Valitor, Inc. is developing protein-polymer therapeutics to overcome the microvascular consequences of diabetic wound healing. We designed our therapy to complement the standard practice of diabetic ulcer management and to reduce the costly serial effort required for debridement and clinical observation of these slowly healing wounds. Our patented technology is a chemical tethering process that enables us control the valency of growth factors that are delivered within the vicinity of a receptor. With this technology, we have conjugated Sonic hedgehog (Shh), an important angiogenic factor, to linear chains of hyaluronic acid (HyA), and by varying the ratio of Shh: HyA, we can modulate its ability to activate the Shh pathway. Conjugating Shh to a large macromolecule may also prevent its deactivation by proteolytic enzymes and enhance its molecular stability in the target tissues. We will develop these multivalent Shh (mvShh) conjugates as a treatment to accelerate wound healing in diabetic lower extremity ulcers, as Shh has previously been shown to stimulate neovascularization. The Valitor approach to Shh therapy will enable enhanced Shh bioactivity in vivo, and delivery of our therapeutic into the wound bed could provide a significant improvement in the patient's long-term outcome at a low treatment cost. Our overall goal during Phase I is to develop mvShh conjugates that elicit enhanced Shh pathway activation and downstream angiogenic gene expression in the cell types that coordinate cutaneous wound healing. In Specific Aim 1, we will identify the mvShh formulations that yield maximal Shh pathway activation in vitro using dermal fibroblasts harvested from db/db mice, a diabetic model animal that exhibits impaired wound healing and diminished angiogenic gene expression. We will then determine the effect of Shh valency on microvascular endothelial cell proliferation and migration in vitro, as these are essential mechanisms required for angiogenesis. In Specific Aim 2, we will demonstrate proof-of-principle that the mvShh conjugates will initiate Shh signaling and downstream angiogenic gene expression in db/db mice using a full-thickness excisional wound model. Our findings will provide valuable proof-of-concept data for further pre-clinical development of our therapeutic, which we have planned for Phase II of this project. PUBLIC HEALTH RELEVANCE: Relevance In 2010, more than 875,000 Americans with diabetes were diagnosed with a chronic lower extremity ulcer, and these patients generate over $30 billion per year in related health care costs. Valitor, Inc. is developing an advanced therapeutic to accelerate healing in diabetic wounds, offsetting the high cost of treatment by encouraging neovascularization and improving blood supply in the wound bed. The overall goal of our Phase I project is to relate the treatment parameters of our therapeutic to the cellular mechanism of blood vessel formation and demonstrate proof-of-concept for our technology as a clinical therapy.