MiniPlasmid vector platform for non-viral gene therapy

描述（由申请人提供）：为了使非病毒基因药物商业化，关键是要提高载体效力（即治疗性转基因表达水平）和治疗效果的持续时间。有效的剂量节省延长持续时间的基因疗法将具有成本和疗效的竞争优势，超过替代技术。在这项I期概念验证研究中，我们将创建一种新型的无抗生素MiniPlasmid基因治疗平台，用于延长持续时间的基因治疗。该载体将提高转基因表达水平的联合收割机瞬时表达增强子与新的270个碱基对复制起点-无抗生素选择盒组合，我们假设该选择盒将在载体递送至身体后促进长持续时间的基因表达。MiniPlasmid平台将用于创建伤口愈合基因治疗产品，以治疗糖尿病神经性足部溃疡。在具体目标1和2中， 创建MiniPlasmid发酵生产平台。在Specific Aim 3中，与常规质粒相比，MiniPlasmid载体平台在体内验证了延长的表达持续时间。利用延长的半衰期耐氧高活性HIF-1突变体（CA 5-HIF-1）开发了用于糖尿病足溃疡治疗的基于缺氧诱导因子1）的基因药物。具体目标3是与伤口愈合基因治疗专家约翰哈蒙博士在约翰霍普金斯大学合作进行。MiniPlasmid载体平台旨在提高转基因表达水平和持续时间，以实现需要延长持续表达时间的多种应用的基因药物开发。第一阶段开发的MiniPlasmid载体将通过出版物、贸易展览和自然技术公司（NTC）网站向研究人员销售，用于各种基因治疗应用。在II期，HIF-1 <$MiniPlasmid基因治疗剂将在III期临床开发之前进行治疗糖尿病足溃疡的临床前安全性和有效性评估。

DESCRIPTION (provided by applicant): To commercialize non-viral gene medicines, it is critical that both vector potency (i.e. therapeutic transgene expression levels) and the duration of the therapeutic effect be improved. Potent dose-sparing extended duration gene therapies will have a cost and efficacy competitive advantage over alternative technologies. In this Phase I proof of concept study, we will create a novel antibiotic-free MiniPlasmid gene therapy platform for extended duration gene therapy. The vectors combine transient expression enhancers that improve transgene expression levels with a novel 270 base pair replication origin-antibiotic free selection cassette that we hypothesize will promote long duration gene expression after vector delivery to the body. The MiniPlasmid platform will be applied to create a wound healing gene therapy product to treat diabetic neuropathic foot ulcers. In Specific Aims 1 and 2 a high yielding MiniPlasmid fermentation manufacturing platform is created. In Specific Aim 3 the MiniPlasmid vector platform is validated in vivo for extended duration expression compared to conventional plasmids. A hypoxia- inducible factor 1¿ (HIF-1 ¿) based gene medicine for diabetic foot ulcer treatment is developed utilizing an extended half-life oxygen resistant highly active HIF-1 ¿ mutant (CA5-HIF-1 ¿). Specific Aim 3 is performed in collaboration with wound healing gene therapy expert Dr. John Harmon at Johns Hopkins University. The MiniPlasmid vector platform is designed to improve transgene expression levels and duration to enable gene medicine development for multiple applications requiring extended duration expression. MiniPlasmid vectors developed in Phase I will be marketed to investigators for a variety of gene therapy applications through publications, trade shows, and the Nature Technology Corporation (NTC) website. In Phase II the HIF-1 ¿ MiniPlasmid gene therapeutic will undergo preclinical safety and efficacy evaluations for treatment of diabetic foot ulcers prior to clinical development in Phase III.