Development of wound care formulations for PVP/NO: A novel nitric oxide releasing platform with potent antimicrobial and anti-biofilm properties

这项SBIR研究工作的中心目标是推进和商业化一种新型生物材料， 用于伤口护理市场的基于氧化物（NO）的输送系统。第一阶段的提案将建立在iFyber的 该生物材料作为有效的抗微生物和抗生物膜平台的强有力的初步数据。具体地说， 通过过去的体外研究，iFyber已经证明该系统具有出色的抗菌和抗生物膜性能 通过体外和离体生物膜模型证明， 用于革兰氏阳性/阴性细菌、酵母和霉菌的市售伤口护理敷料。结果 初步的动物研究已经显示了该系统在体内的活性，但强调了制剂 在进一步推进技术之前，将发展作为一个关键的重点领域。如果得到资助，iFyber的目标是利用这一点， 第一阶段项目，推进有效和伤口相关的局部制剂，提供稳定和持续的 不释放。这些制剂将在II期计划的一系列完整动物研究中进一步推进。 一氧化氮释放技术为标准抗菌治疗提供了一种令人信服的替代方案， 目前用于对抗伤口感染的抗生素。我们目前的数据集清楚地表明， PVP/NO在解决一般微生物感染，更重要的是，预防和 治疗困扰慢性皮肤伤口的微生物生物膜。第一期工程的一个重要里程碑 将通过开发基于PVP/NO的候选局部制剂来扩展PVP/NO技术 其将用作针对微生物感染的非抗生素预防性治疗， 对于慢性感染的伤口-伤口护理领域内的两个未满足的需求。 完成第一阶段的工作将产生一个令人信服的数据集，概述疗效并定义 局部应用方面对产品配方的最低要求。这些研究将有助于 过渡到更广泛的II期研究，旨在支持PVP/NO产品进入FDA的 监管过程。iFyber将解决新开发的NO释放制剂的功效， 生物材料通过第一阶段的目标和任务概述如下： AIM 1.通过筛选活动确定先导和应急局部制剂 任务1.筛选基于PVP/NO的局部制剂以符合目标产品特征（TPP） 要求.过去的研发工作已经确定，如果没有足够的时间，则从PVP/NO中释放NO是快速的。 防止同时暴露于H+/水。这在体内可能难以控制， 需要持续释放。这可以通过控制PVP/NO暴露于H+/水来完成，因为我们 最近通过平衡的亲水性-亲油性有机酸“BHLOA”方法显示， 控制这些系统中的pH变化。在第一阶段，我们将在这些初步研究结果的基础上， 推进BHLOA配方方法，以提供先导和后备PVP/NO配方， 根据NO释放和离体模型研究（任务2）的试验结果进行选择， 初步体内研究（任务3和4）。 任务2.验证抗生物膜功效并选择先导和应急制剂。这个目标 任务是评价来自任务1的候选制剂在离体猪真皮中的功效 生物膜模型，以确定用于伤口的合适的抗菌和抗生物膜半固体敷料。主 本任务的目标是筛选用于中试动物研究的制剂（任务3和4）。 AIM 2.通过临床前试点研究指导PVP/NO原型开发 任务3.对选定的制剂进行体内试验。通过以下途径确定的有希望的配方： 将在Steven Davis教授实验室进行的两项初步体内研究中评价任务1-2 在迈阿密大学。这些研究将确定生物膜预防的功效（研究1）， 减少现有的生物膜相关感染（研究2），这些研究的结果将有助于 进一步向下选择PVP/NO制剂用于未来的II期开发。 任务4.确定PVP/NO的基线体内生物相容性特征。 感染预防和抗生物膜功效，候选PVP/NO制剂对非生物膜的影响， 还将评估感染伤口的伤口愈合过程（研究3）;具体地， 炎症反应、红斑、再上皮化、肉芽组织形成和初始分子 筛选愈合和炎症过程。

The central goal of this SBIR research effort is to advance and commercialize a novel biomaterial with a nitric oxide (NO)-based delivery system for the wound care market. This Phase I proposal will build on iFyber’s strong preliminary data for this biomaterial as a potent antimicrobial and anti-biofilm platform. Specifically, through past in vitro studies, iFyber has shown this system to exhibit excellent antibacterial and anti-biofilm properties as evidenced by both in vitro and ex vivo biofilm models where it significantly outperforms commercially available wound care dressings for Gram positive/negative bacteria, yeast, and mold. Results form pilot animal studies have shown activity of this system, in vivo, but have highlighted formulation development as a key focus area prior to advancing the technology further. If funded, iFyber aims to use this Phase I project to advance effective and wound-relevant topical formulations that provide stable and sustained NO-release. These formulations will be further advanced in a full battery of animal studies planned for Phase II. Nitric oxide-releasing technologies offer a compelling alternative to standard antimicrobial treatments and antibiotics currently used to combat infection in wounds. Our current data set clearly demonstrate the potential clinical utility of PVP/NO in addressing microbial infection in general, and more importantly, prevention and treatment of microbial biofilms that plague chronic skin wounds. An important milestone for this Phase I project will be to expand on the PVP/NO technology by developing candidate PVP/NO-based topical formulations that will serve as a non-antibiotic prophylactic treatment against microbial infection and as a treatment for chronically infected wounds – two unmet needs within the wound care field. Completion of Phase I efforts will result in a compelling data set that outlines the efficacy and defines the minimal requirements for product formulation in terms of topical application. Together, these studies will help in the transition to broader Phase II studies aimed to support entry of a PVP/NO-based product into the FDA's regulatory process. iFyber will address the efficacy of the newly developed formulations of the NO-releasing biomaterial through the Phase I aims and tasks outlined below: AIM 1. Identify lead and contingency topical formulations through a screening campaign Task 1. Screen PVP/NO-based topical formulations for adherence to target product profile (TPP) requirements. Past R&D efforts have established that NO release from the PVP/NO is rapid if not protected from simultaneous exposure to H+/water. This can be difficult to control, in vivo, where sustained release is desired. This can be done by controlling the exposure of PVP/NO to H+/water as we have recently shown through a balanced hydrophilicity-lipophilicity organic acid “BHLOA” approach to controlling pH change in these systems. In Phase I we will build on these preliminary findings by further advancing the BHLOA formulation approach to provide lead and back up PVP/NO formulations that will be selected based on test results from the NO-release and ex vivo model studies (Task 2) followed by pilot in vivo studies (Tasks 3 and 4). Task 2. Verify anti-biofilm efficacy and select lead and contingency formulations. The goal of this task is to evaluate the efficacy of the candidate formulations from Task 1 in an ex vivo porcine dermal model of biofilm to define suitable antimicrobial and anti-biofilm semi-solid dressings for wounds. A main goal in this task will be to down-select formulations for pilot animal studies (Tasks 3 and 4). AIM 2. Guide PVP/NO prototype development through pre-clinical pilot studies Task 3. Conduct in vivo testing on selected formulations. Promising formulations identified through Tasks 1-2 will be evaluated in two pilot in vivo studies conducted in the laboratory of Prof. Steven Davis at the University of Miami. These studies will establish efficacy in biofilm prevention (study 1) and reduction in an existing biofilm-associated infection (study 2), and results of these studies will help to further down-select PVP/NO formulations for future Phase II development. Task 4. Determine the baseline in vivo biocompatibility profile of PVP/NO. In addition to assessing infection prevention and anti-biofilm efficacy, the effects of the candidate PVP/NO formulations on non- infected wounds will also be assessed with respect to the wound healing process (study 3); specifically, inflammatory response, erythema, re-epithelialization, granulation tissue formation, and initial molecular screens for healing and inflammatory processes.