SBIR Phase I: Development Of An Orally Administered Gene Delivery Platform For Induced Protein Secretion Via The Gastrointestinal Tract

这个SBIR一期项目将进一步开发一种称为隐窝到达颗粒（CRP）的口服基因递送载体，它能够克服粘液的物理障碍，将基于dna的药物递送到胃肠道细胞。然后，治疗剂将在胃肠道局部以蛋白质的形式表达或分泌到血液中以治疗全身。这种运载工具的开发将带来关于DNA传递的新颖科学见解，这将有利于基础科学和工程领域。成功的商业化将使制药公司受益，因为它为批准的药物和新药提供了一个口服给药平台；使美国患者受益，因为它减少了对注射药物的需求，从而提高了依从性和生活质量；使付款人受益，因为它降低了与注射药物相关的成本，并通过口服给药提高了患者的依从性，从而改善了结果。与药物无关的特性在快速扩大的生物制剂市场中提供了广泛和多样化的市场机会，以及其他治疗药物，从而创造了大量的就业机会。例如，在四个领域（即提供TNF抑制剂，蛋白质替代，肽激素和血液因子）的潜在潜在市场每年超过900亿美元。本项目旨在制备一种稳定的口服隐窝到达颗粒（CRP）制剂，这是一种新型药物不可知的药物递送载体，并证明它可以在体外和体内递送功能性DNA货物，用于肠上皮细胞的表达和分泌。该项目侧重于与保留DNA货物功能相关的技术挑战，作为难以复制的具有挑战性的药物递送场景的一个例子。创新之处在于这种载体的设计可以通过模拟肠道脊髓灰质炎病毒来穿透黏液并转染下层上皮细胞。这将为首次口服基因治疗奠定基础。简而言之，该项目将解决三个目标。首先，通过冻干制备用于口服给药的CRP配方，并分析评估配方和DNA稳定性随时间、温度和ph的变化。其次，通过测量细胞和分泌表达，测试口服配方CRP将编码高斯荧光素酶的功能性DNA质粒传递到体外Caco-2肠上皮细胞的能力。最后，在Fischer 344大鼠体内测试口服和给药CRP将功能性高斯荧光素酶质粒递送到肠上皮细胞的能力，以评估其表达和分泌。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This SBIR Phase I project will further the development of an orally administered gene delivery vehicle called the Crypt-Reaching Particle (CRP) that is able to overcome the physical barriers of the mucus to deliver DNA-based drugs to the cells of the gastrointestinal tract. The therapeutic agent will then be expressed as a protein locally in the gastrointestinal tract or be secreted into the bloodstream to treat the full body. Development of this vehicle will lead to novel scientific insights on DNA delivery that will benefit and educate the field of fundamental science and engineering. Successful commercialization would benefit pharmaceutical companies by providing an oral drug delivery platform for approved and novel drugs, U.S. patients by reducing the need for injectable drugs and thereby increasing compliance and quality of life, and payers by reducing costs associated with injectable drugs and improving outcomes by increasing patient compliance via oral administration. The drug-agnostic feature provides broad and diverse market opportunities within the rapidly expanding market for biologics, among other therapeutic agents leading to substantial job creation. As an example, a potential addressable market in four areas (i.e., delivering TNF inhibitors, protein replacement, peptide hormones, and blood factors) exceeds $90 billion per year.This project is designed to prepare a stable oral formulation of the Crypt-Reaching Particle (CRP), a novel drug-agnostic drug delivery vehicle, and demonstrate that it can deliver a functional DNA cargo for expression and secretion in intestinal epithelial cells in vitro and in vivo. This project focuses on the technical challenges associated with preserving the functionality of a DNA cargo as an example of a challenging drug delivery scenario that is difficult to replicate. The innovation is the design of the vehicle to penetrate through the mucus and transfect underlying epithelial cells by mimicking the enteric poliovirus. This will lay the foundation for the first oral gene therapy. Briefly, the project will address three objectives. First, preparation of a formulation for the CRP for oral administration via lyophilization with analyses to assess formulation and DNA stability as a function of time, temperature, and pH. Second, testing the ability of the orally formulated CRP to deliver a functional DNA plasmid encoding Gaussia luciferase to Caco-2 intestinal epithelial cells in vitro by measuring both cellular and secreted expression. Finally, testing the ability of the orally formulated and administered CRP to deliver a functional Gaussia luciferase plasmid to intestinal epithelial cells in vivo in Fischer 344 rats to assess expression and secretion.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.