VACCINATION BY TOPICAL APPLICATION OF CONDENSED DNA

局部应用DNA疫苗可提供一种安全、简便、无需针头和具有成本效益的免疫方法，并具有改善世界健康的巨大潜力。最近的数据表明，在局部应用裸DNA后，可以转染各种皮肤细胞。已经观察到对dna编码抗原的特异性免疫，尽管皮肤给药后的免疫反应不如IM疫苗接种方案或重组蛋白疫苗强。因此，该技术的商业化受到基因转移效率的技术限制。哥白尼治疗公司开发了一种非病毒基因传递技术，该技术将单个质粒DNA分子压缩成20-25纳米的颗粒，称为PLASmin（TM）复合物。我们的配方简单，可重复，并且PLASmin（TM）复合物容易转染非分裂，有丝分裂后的人类细胞。我们的配方在生理盐水和血清中是稳定的，并且高度抵抗核酸酶消化。纤溶酶（TM）复合物稳定数月，可浓缩至约7mg /ml而不失去稳定性或完整性。为了确定是否PLASmin（TM）复合物可以改善基因转移结果，将几种配方应用于小鼠皮肤，并评估对编码抗原的抗体反应。如果成功，优化后的PLASmin（TM）复合物将用于II期SBIR应用，为人类接种肝炎疫苗。拟议的商业应用：尽管裸DNA应用于皮肤有可能引发免疫反应，但基因表达的限制可能会阻碍商业发展。哥白尼开发了一种新的非病毒技术，将DNA的单分子与多阳离子蛋白凝聚在一起。这种配方大大提高了体外和临床前模型的基因转移结果。如果我们的配方在局部应用后可以在小鼠中引起强大的免疫，那么这个DNA疫苗平台可以用于开发许多针对传染病和癌症适应症的产品。

Topically applied DNA vaccines may provide a safe, easy, needle-less, and cost-effective means of eliciting immunity, and has significant potential to improve world health. Recent data indicate that various skin cells can be transfected following a topical application of naked DNA. Specific immunity to DNA-encoded antigens has been observed, although the immune response following cutaneous administration has been less robust than either IM vaccination protocols or recombinant protein vaccines. Hence, commercialization of this technology is limited by technical constraints in gene transfer efficiency. Copernicus Therapeutics has developed a non-viral gene delivery technology that compacts single plasmid DNA molecules into 20-25 nm particles, termed PLASmin(TM) complexes. Our formulation is simple and reproducible, and PLASmin(TM) complexes readily transfect non-dividing, post-mitotic human cells. Our formulations are stable in normal saline and serum, and highly resistant to nuclease digestion. PLASmin(TM) complexes are stable for months and can be concentrated to approximately 7 mg/ml without loss of stability or integrity. To determine if PLASmin(TM) complexes can improve gene transfer results, several formulations will be applied to murine skin, and antibody responses to encoded antigens will be assessed. If successful, optimized PLASmin(TM) complexes will be used in a Phase II SBIR application to vaccinate humans against hepatitis. PROPOSED COMMERCIAL APPLICATION: Although naked DNA applied to skin has the potential to elicit an immune response, limitations in gene expression may hinder commercial development. Copernicus has developed a novel, non-viral technology in which single molecules of DNA are condensed with polycationic proteins. This formulation greatly improves gene transfer results in vitro and in preclinical models. If our formulation can elicit robust immunity in mice following topical application, then this DNA vaccine platform could be used to develop many products for infectious disease and cancer indications.