Transporter-Enzyme Interplay Evaluation via Microfluidiic HTS Cell Culture Device

描述（由申请人提供）： 由于过去的许多失败，许多制药科学家认为动物模型在预测人类DMPK（药物代谢和药代动力学）和毒理学方面没有用处。这促进了尽快将NME（新分子实体）推进到人类身上的倡议，而不再强调动物工作。尽管在药物开发早期评估人体NME可能具有显著优势，但我们不认为这是从许多潜在候选化合物中选择DMPK最佳分子的最有效或最有效的方法。我们假设，高代谢化合物（约占市场上药物的70%）的动物模型的可预测性差是由于我们对如何将转运蛋白（吸收和外排）纳入药物代谢的预测模型的初步理解，这种缺乏对转运蛋白-酶相互作用的理解使得传统的药物处置理论简单化和不充分，这是可预测性差的原因。 然而，最重要的是，我们缺乏一个简单的高通量临床前工具来表征转运蛋白酶的相互作用，方便地允许人与动物的比较。在本申请中，我们描述了一种新的临床前工具，由Hurel公司开发的微流控细胞培养生物芯片，并提出研究来测试该系统的适用性，以解决10个具体目标中列出的关注问题。我们将表征Hurel系统中肝脏和肠上皮细胞转运蛋白-酶的相互作用，最初分别使用培养的大鼠和人肝细胞和肠上皮细胞，然后合并使用1类和2类高代谢化合物。将这些数据与主要在本申请外部进行的大鼠肝脏和肠道灌注、大鼠和人肝细胞/肠细胞和微粒体研究以及全动物静脉给药药代动力学研究进行比较。工作的一个主要重点将是测试Hurel装置结果与更费力的动物和人类研究的一致性，并开发模型，允许使用Hurel微流控细胞培养生物芯片预测动物临床前DMPK，以及解决我们目前的药物消除模型的缺陷，这些模型没有充分考虑药物处置中的转运蛋白-酶相互作用。

DESCRIPTION (provided by applicant): Because of numerous past failures, it is the belief of many pharmaceutical scientists that animal models are not useful in predicting human DMPK (drug metabolism and pharmacokinetics) and toxicology. This has promoted initiatives to advance NMEs (new molecular entities) into man as soon as possible with a deemphasis of animal work. Although there can be marked advantages to evaluation of NMEs in humans early in drug development, we do not believe that this is the most efficient or most effective way to select the DMPK-optimal molecule from many potential candidate compounds. We hypothesize that the poor predictability of animal models for highly metabolized compounds (approximately 70% of drugs on the market) is due to our nascent understanding of how to incorporate transporters, both absorptive and efflux, into predictive models of drug metabolism, and this lack of understanding of transporter-enzyme interplay renders traditional drug disposition theory simplistic and inadequate, which accounts for the poor predictability. However, most importantly we lack a simple high-throughput preclinical tool to characterize transporter enzyme interplay that conveniently allows human-animal comparisons. In this application, we describe a novel preclinical tool, the microfluidic cell culture biochip in development by the Hurel Corporation and propose studies to test the applicability of this system to address the issues of concern as outlined in 10 specific aims. We will characterize hepatic and enterocyte transporter-enzyme interplay in the Hurel system, initially separately and then combined, utilizing cultured rat and human hepatocytes and enterocytes for Class 1 and Class 2 highly metabolized compounds. These data will be compared with rat liver and intestinal perfusion, rat and human hepatocyte/enterocyte and microsome studies and whole animal iv dosing pharmacokinetics studies carried out predominantly external to this application. A major focus of the work will be to test the concordance of the Hurel device results with the more laborious animal and human studies and to develop models that will allow prediction of animal preclinical DMPK using the Hurel microfluidic cell culture biochips, as well as address the deficiency of our present models of drug elimination that do not adequately consider transporter-enzyme interplay in drug disposition.