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

该SBIR第一阶段项目将进一步开发一种口服基因递送载体，称为隐窝到达颗粒（CRP），能够克服粘液的物理屏障，将基于DNA的药物递送到胃肠道细胞。然后，治疗剂将在胃肠道中局部表达为蛋白质或分泌到血流中以治疗全身。这种载体的开发将导致对DNA递送的新的科学见解，这将有利于基础科学和工程领域的教育。成功的商业化将使制药公司受益，为批准的新药提供口服药物递送平台，美国患者通过减少对注射药物的需求，从而提高依从性和生活质量，支付者通过降低与注射药物相关的成本，并通过口服增加患者依从性来改善结果。药物不可知性的特点在迅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，以及其他治疗剂，从而创造了大量就业机会。例如，四个领域的潜在目标市场（即，递送TNF抑制剂、蛋白质替代物、肽类激素和血液因子）每年超过900亿美元。该项目旨在制备一种稳定的隐窝到达颗粒（CRP）口服制剂，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。该项目的重点是与保留DNA货物的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方案的一个例子。创新之处在于设计了一种载体，通过模仿肠道脊髓灰质炎病毒，穿透粘液并感染底层上皮细胞。这将为首次口服基因治疗奠定基础。简而言之，该项目将实现三个目标。首先，通过冻干制备用于口服给药的CRP制剂，分析以评估制剂和DNA稳定性作为时间、温度和pH的函数。其次，通过测量细胞和分泌表达来测试口服配制的CRP在体外将编码Gaussia荧光素酶的功能性DNA质粒递送至Caco-2肠上皮细胞的能力。最后，测试口服配制和管理CRP的能力，以提供一个功能性高斯荧光素酶质粒肠上皮细胞在体内Fischer 344大鼠，以评估表达和分泌。这个奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

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.