A physiologically based pharmacokinetic model of human airway epithelia

摘要 吸入性药物是治疗肺部疾病的一种重要、有效、但通常昂贵的治疗选择 如哮喘或慢性阻塞性肺病（COPD）。开发品牌的通用等同物 吸入药物的名称可以减少与这些药物相关的经济负担，但受到困难的限制 对呼吸道上皮细胞进行药代动力学（PK）生物等效性研究， 剂.尽管基于生理学的药代动力学（PBPK）模型可以用作直接分析的替代方案，但是， 尽管目前的模型不能解释药物在气道上皮内的作用， 可能影响吸入药物有效浓度的转运蛋白和代谢酶。到 针对这一空白，我们召集了一组气道上皮细胞培养，定量蛋白质组学， PBPK建模和雾化药物的PK测试，以系统地测量相关蛋白质的蛋白质水平。 药物转运蛋白和代谢酶在人类气道上皮细胞。我们的建议利用了 我们机构强大的组织采购和细胞培养核心，可以为我们提供初级 用于培养的人气道上皮细胞代表了广泛的人口统计学因素和解剖学因素， 位置以及用于测试的体内组织样本。我们的计划是制定一个全面的评估， 基于文献综述，气道上皮中存在的药物转运蛋白和代谢酶， 分析我们自己现有的RNA测序数据库，然后利用这些数据开发定量靶向 绝对蛋白质组学（QTAP）方法用于测量相关蛋白质。这种QTAP方法将 应用于代表广泛的人口统计学特征，解剖学位置， 和炎症状态以及体内获得的组织。最终，我们将利用这些数据来开发 增强的PBPK模型，然后我们将通过测试药物和药物的质谱分析进行测试， 在雾化到气道上皮上之后，在顶端、细胞内和基底外侧隔室中的代谢物。 总体而言，本提案中的研究将对药物转运蛋白水平进行全面评价 和代谢酶，以及增强的PBPK建模，可以支持更快速的 开发吸入药物的仿制药。

Abstract Inhaled drugs represent an important, effective, but often expensive therapeutic option for lung diseases such as asthma or chronic obstructive pulmonary disease (COPD). Development of generic equivalents to brand name inhaled drugs could reduce the financial burden associated with these agents but is limited by difficulties performing pharmacokinetic (PK) bioequivalence studies on airway epithelia, the primary target of inhaled agents. Although physiologically based pharmacokinetic (PBPK) models can serve as an alternative to direct measurement of drug PK within airway epithelia, current models fail to account for the actions of drug transporters and metabolizing enzymes that may influence the effective concentrations of inhaled drugs. To address this gap, we have assembled a group of experts on airway epithelial cell culture, quantitative proteomics, PBPK modeling, and PK testing of aerosolized drugs to systematically measure the protein levels of relevant drug transporters and metabolizing enzymes within human airway epithelia. Our proposal takes advantage of the robust Tissue Procurement and Cell Culture Core present at our institution that can provide us with primary human airway epithelial cells for culture representing a wide range of demographic factors and anatomic locations as well as in vivo tissue samples for testing. Our plan is to develop a comprehensive assessment of the drug transporters and metabolizing enzymes present in airway epithelia based on literature review and analysis of our own existing RNA sequencing databases, then utilize this data to develop quantitative targeted absolute proteomic (QTAP) methods for measurement of the relevant proteins. This QTAP methodology will then be applied to human airway epithelia representing a wide range of demographic features, anatomic locations, and inflammatory states as well as tissues obtained in vivo. Ultimately, we will utilize this data to develop enhanced PBPK models, which we will then test be tested via mass spectrometric analysis of test drug and drug metabolites in apical, intracellular, and basolateral compartments after aerosolization onto airway epithelia. Overall, the studies in this proposal will generate a comprehensive evaluation of the levels of drug transporters and metabolizing enzymes in human airway epithelia and enhanced PBPK modeling that can support more rapid development of generic versions of inhaled drugs.