MiniPlasmid vector platform for non-viral gene therapy

描述(由申请人提供)：为了将非病毒基因药物商业化，关键是提高载体效力(即治疗性转基因表达水平)和治疗效果的持续时间。与替代技术相比，有效的节省剂量的延长持续时间的基因疗法将具有成本和疗效竞争优势。在这项第一阶段的概念验证研究中，我们将创建一个新的不含抗生素的微型质粒基因治疗平台，用于延长持续时间的基因治疗。这些载体将提高转基因表达水平的瞬时表达增强剂与一种新型的270碱基对复制来源-无抗生素选择盒结合在一起，我们假设这种盒将在载体传递到体内后促进长期基因表达。微型塑料平台将被应用于创造一种伤口愈合基因治疗产品，用于治疗糖尿病神经性足部溃疡。在具体目标1和2中，高产 创建了微型塑料发酵生产平台。在特定的目的3中，微型质粒载体平台在体内进行了验证，与传统的质粒相比，它的持续表达时间更长。利用耐氧半衰期延长的高活性HIF-1突变体(CA5-HIF-1)，开发了一种以低氧诱导因子-1(HIF-1)为基础的基因药物治疗糖尿病足溃疡。《特殊目的3》是与约翰·霍普金斯大学的创伤修复基因治疗专家约翰·哈蒙博士合作完成的。微型质粒载体平台旨在提高转基因表达水平和持续时间，以使基因药物开发适用于需要延长持续表达时间的多种应用。在第一阶段开发的微型质粒载体将通过出版物、贸易展和自然技术公司(NTC)的网站向研究人员销售各种基因治疗应用。在第二阶段，HIF-1迷你质粒基因疗法将在第三阶段的临床开发之前，接受治疗糖尿病足部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.