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

这项小企业创新研究（SBIR）一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分析，90%的开发药物最终未能进入市场。实际上，这些限制延缓了急需治疗药物的开发，并大大提高了药品价格和保健费用。因此，迫切需要开发更具预测性的体外测试方法，以便尽早选择最有希望的候选药物，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向早期药物开发的过渡。这项技术是基于发现、基本特征和成年犬肠道干细胞系的生物存档，称为三维（3D）犬肠道类器官。这些微小的模拟功能肠的生理比目前可用的方法更接近，并有可能提供比目前使用的测定更优越的药物筛选。这项SBIR I期研究旨在建立犬类肠道器官与标准2D体外检测（如Caco2和MDCK细胞系）相比，可以提高口服药物吸收的体外可预测性。在Aim 1中，与常规体外模型相比，目标是确定治疗药物的肠道吸收和通透性作为疾病和肠道段的功能。这将通过量化药物的被动和主动通透性，以及药物转运体在3D犬类器官与传统细胞系统中的表达和功能来实现。在Aim 2中，目的是与标准体外模型相比，确定治疗药物的肠道代谢作为犬类器官疾病和肠道段的功能。最终，通过这些实验产生的定量数据将导入商业软件，以模拟预定义的候选药物集的处置动力学。为了验证目的，模型预测的性能将通过比较文献中模拟和观察到的药物动力学血浆数据来评估。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。

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.