Phase II: Anti-inflammatory and Anti-scarring Actions of Amniotic Membrane Extra

描述（由申请人提供）：在过去的十年中，人们对羊膜移植（AMT）用于眼表重建的兴趣激增。许多研究表明，使用由Bio-Tissue，Inc.制造的冷冻保存的AM（AmnioGraft（R））的AMT，TissueTech，Inc.的子公司，有效促进上皮伤口愈合和减少基质炎症、瘢痕形成和不需要的新血管的形成。为了方便患者护理并降低整体医疗成本，我们获得了SBIR第I和第II阶段赠款（R43/R44 EY 014768），以开发“免缝合”AM移植物，以便AMT可以在医生办公室或重症监护和烧伤病房的床边进行，而无需将患者带到手术室。因此，我们成功开发了ProKera（R），该产品已获得FDA的510（k）许可，被列为II类医疗器械。2008年，我们在美国市场销售了4，000多个单位，用于治疗患有许多威胁视力的角膜表面疾病的患者，这些疾病的特征是伤口愈合不良，不必要的炎症和过度疤痕。我们设想，AM的固有抗炎和抗瘢痕形成作用可以进一步用于治疗AmnioGraft（R）或ProKera（R）无法治疗的眼表疾病，以及用于炎症和瘢痕形成受到极大关注的任何其他身体部位。我们进一步推测，AmnioGraft（R）或ProKera（R）的物理限制的一种解决方案是开发基于AM提取物（AME）的凝胶制剂。通过SBIR I期资助（R43 EY 017497），我们成功验证了生产AME及其冻干粉（AMP）的关键生产步骤，并比较了使用胶原蛋白或透明质酸（HA）作为载体来提供抗炎和抗瘢痕作用的相对效力（目标1）。此外，我们已经生物化学纯化和表征了一种含有HA和α-间抑制剂（IHA）重链（HC）的共价复合物（HC 7 HA），作为AME中的活性组分，负责AM基质中观察到的大多数（如果不是全部）抗炎和抗瘢痕形成作用（目的2）。因此，在II期，我们建议验证纯化的HC 7 HA复合物在发挥体外抗炎和抗瘢痕形成作用方面的效力，验证生产HC 7 HA复合物在其生化组成和纯度方面的一致性，并优化含有HC 7 HA复合物的HA凝胶制剂（目的1）。此外，我们提出证明上述含有HC 7 HA复合物的凝胶制剂在HSV 1诱导的坏死性基质角膜炎的鼠模型中发挥抗炎作用和在准分子激光辅助的光折射性角膜切除术（PRK）的兔模型中发挥抗瘢痕形成作用的临床功效（目的2）。成功完成上述两个目标将使我们能够收集足够的相关数据，以便向FDA提交IDE/IND，从而使我们可以开始将首个眼用局部制剂商业化，并继续建立基于AME或纯化HC 7 HA复合物的新疗法的管道，从而使我们不仅可以扩大眼科的市场空间，还可以扩大其他医学和外科亚专业的市场空间。 公共卫生相关性：本申请提出开发一种基于羊膜提取物及其纯化的含有透明质酸和间-α-抑制剂重链的共价复合物的抗炎和抗血管生成作用的新型治疗剂管道。成功完成我们拟定的II期研究将使我们能够收集充足的相关数据，以便向FDA提交IDE/IND，从而我们可以开始将首个眼用局部制剂商业化，并扩大我们的市场空间，不仅在眼科，而且在其他内科和外科亚专科。

DESCRIPTION (provided by applicant): Over the past decade, there has been a surge of interest in amniotic membrane transplantation (AMT) for ocular surface reconstruction. A number of studies have shown that AMT using cryopreserved AM (AmnioGraft(R)) manufactured by Bio-Tissue, Inc., a subsidiary of TissueTech, Inc., is effective in facilitating epithelial wound healing and reducing stromal inflammation, scarring and formation of unwanted new blood vessels. To facilitate the ease of patient care and reduce the overall medical cost, we have obtained SBIR Phase I and II grants (R43/R44 EY014768) to develop a "sutureless" AM graft so that AMT can be performed in a physician's office or at the bedside of an Intensive Care and Burn Unit without bringing the patient to the operating room. As a result, we have successfully developed ProKera(R), which has received the FDA's 510(k) clearance as a type II medical device. In 2008, we have distributed more than 4,000 units in the U.S. market for treating patients inflicted with a number of sight-threatening corneal surface diseases characterized by poor wound healing, unwanted inflammation, and excessive scarring. We envisioned that AM's inherent anti-inflammatory and anti- scarring actions can further be deployed to treat ocular surface diseases unmanageable by AmnioGraft(R) or ProKera(R), and to any other body sites where inflammation and scarring are of great concern. We further speculated that one solution to the physical constraint of AmnioGraft(R) or ProKera(R) is to develop a gel formulation based on AM extracts (AME). Through SBIR Phase I grant support (R43 EY017497), we have successfully validated the key manufacturing steps of generating AME and its lyophilized powder (AMP), and compared the relative potency of using collagen or hyaluronic acid (HA) as a vehicle to deliver their anti-inflammatory and anti-scarring actions (Aim 1). In addition, we have biochemically purified and characterized one covalent complex (HC7HA) containing HA and heavy chains (HC) of inter-a-inhibitor (IaI) as the active component in AME responsible for most, if not all, anti-inflammatory and anti-scarring effects observed in AM stroma (Aim 2). In Phase II, we thus propose to validate the potency of purified HC7HA complex in exerting in vitro anti-inflammatory and anti-scarring actions, verify the consistency of manufacturing HC7HA complex regarding its biochemical composition and purity, and optimize the HA gel formulation containing HC7HA complex (Aim 1). In addition, we propose to demonstrate clinical efficacies of the aforementioned gel formulation containing HC7HA complex in exerting an anti-inflammatory action in a murine model of HSV1-induced necrotizing stromal keratitis, and an anti-scarring action in a rabbit model of excimer laser-assisted photorefractive keratectomy (PRK) (Aim 2). Successful completion of the above two Aims will allow us to gather sufficient and pertinent data for IDE/IND submission to the FDA so that we may begin to commercialize the first ophthalmic topical formulation and continue to build a pipeline of new therapeutics based on AME or purified HC7HA complex so that we may expand our market spaces not only in ophthalmology, but also in other medical and surgical subspecialties. PUBLIC HEALTH RELEVANCE: This application proposes to develop a novel pipeline of therapeutics based on anti-inflammatory and anti-angiogenic actions of amniotic membrane extract as well as its purified covalent complex containing hyaluronan and heavy chains of inter-a-inhibitor. Successful completion of our proposed studies in Phase II will allow us to gather sufficient and pertinent data for IDE/IND submission to the FDA so that we may begin to commercialize the first ophthalmic topical formulation and expand our market spaces not only in ophthalmology, but also in other medical and surgical subspecialties.