SBIR Phase I: Improving in vitro prediction of oral drug permeability and metabolism using a novel 3D canine organoid model

该小型企业创新研究（SBIR）I期项目的更广泛影响/商业潜力是为制药行业提供改进的口服药物吸收临床前筛选试验。目前用于表征胃肠道吸收的体外方法是基于不可靠的测定，所有开发的药物中有90%最终未能进入市场。实际上，这些限制延缓了急需的治疗药物的开发，并大大增加了药品价格和医疗保健成本。因此，迫切需要开发更具预测性的体外测试测定，以允许早期选择最有希望的候选药物，以减少活体动物研究的数量及其相关成本，同时加速从临床前研究到早期药物开发的过渡。该技术基于成年犬肠道干细胞系的发现，基本特征和生物存档，称为三维（3D）犬肠道类器官。这些微肠比目前可用的方法更接近地模拟功能性肠的生理学，并且具有提供优于目前使用的测定的上级药物筛选的潜力。该SBIR I期是一项建议，旨在确定使用犬肠道类器官与标准2D体外试验（如Caco 2和MDCK细胞系）相比，可以改善口服药物吸收的体外可预测性。在目的1中，目标是确定与常规体外模型相比，治疗药物的肠吸收和渗透性作为疾病和肠段的函数。这将通过量化药物的被动和主动渗透性以及药物转运蛋白在3D犬类器官与常规细胞系统中的表达和功能来实现。在目的2中，目标是确定与标准体外模型相比，治疗药物在犬类器官中的肠代谢作为疾病和肠段的函数。最终，通过这些实验产生的定量数据将被导入到商业软件中，以模拟预定义的候选药物的处置动力学。模型预测的性能将通过比较模拟与观察到的药物动力学等离子体数据从文献中进行验证purpose.This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。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I Project is to provide the pharmaceutical industry with improved pre-clinical screening assay for oral drug absorption. Current in vitro methods for characterizing gastrointestinal absorption are based on unreliable assays, with 90% of all drugs developed ultimately failing to enter the market. Practically, these limitations delay the development of critically needed therapeutic drugs and dramatically increase drug prices and health care costs. There is, therefore, a critical need to develop more predictive in vitro testing assays that will allow for early selection of the most promising drug candidates to reduce the number of live animal studies and their associated costs, while accelerating transition from pre-clinical research to early drug development. The technology is based on the discovery, fundamental characterization and bioarchiving of adult canine intestinal stem cell lines, called 3-dimensional (3D) canine intestinal organoids. These miniguts emulate the physiology of the functional intestine much more closely than currently available methods and have the potential to provide superior drug screening over currently used assays. This SBIR Phase I is a proposal to establish that in vitro predictability of oral drug absorption can be improved using canine intestinal organoids vs. standard 2D in vitro assays, such as Caco2 and MDCK cell lines. In Aim 1, the goal is to determine intestinal absorption and permeability of therapeutic drugs as a function of disease and intestinal segment as compared with conventional in vitro models. This will be achieved by quantifying passive and active permeability of drugs, as well as drug transporter expression and function in 3D canine organoids vs. conventional cell systems. In Aim 2, the goal is to determine intestinal metabolism of therapeutic drugs as a function of disease and intestinal segment in canine organoids compared to standard in vitro models. Ultimately, quantitative data generated through these experiments will be imported into a commercial software to simulate the disposition kinetics of a predefined set of candidate drugs. Performances of the model predictions will be evaluated by comparing simulated vs. observed drug kinetic plasma data from the literature for validation purposes.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.