Aim 3: Measurement and multi-scale modeling of pharmaco-kinetics and pharmaco-dyn

项目总结 吸收、分布、代谢和消除(统称为药代动力学，PK) 靶器官中的时间依赖性药物作用(药效学，PD)在疗效和 所有药物的毒性。我们将开发和实施新的方法来在多个尺度上模拟PK-PD 在细胞和亚细胞水平上测量PK的新方法，以及新的细胞培养 更好地模拟肿瘤环境的系统，从而增强了我们预测患者反应的能力 细胞培养数据。我们的翻译目标是创造新的单细胞拆分方法来整合分子 用靶点接触和诱导药物反应的方法了解药物与靶点的相互作用 在组织和生物体中。 Aim 3.1涉及荧光法测量亚细胞分辨率pk的技术开发 伴生成像药物(CID)、标记有荧光团的小分子或蛋白质药物 以保留母体化合物的生物活性和药代动力学的方式进行成像。这些属性 将优化CID的范围(目标3.1.1)，用于药物分布和体内成像的化合物 在活体小鼠中的反应(目标3.1.2)。CID将被用来直接检测单细胞中药物与靶点的相互作用 荧光相关显微镜(AIM 3.1.3)。 Aim 3.2将开发一种新的定量、多重质谱学方法用于分析 基于靶分子共价修饰的细胞和亚细胞水平的构效关系 蛋白质。这将涉及创建新的化学类药物探针(AIM 3.2.1)，然后将对其进行 系统的化学修饰，探索对物化性质的影响，如cLogP，PKA等。 (目标3.2.2)。 目标3.3将涉及开发整合途径水平知识和生物标记物的方法 (预测性和响应性)转化为常规用于翻译的PK-PD模型 工业中的药理和临床试验设计。 Aim 3.4将尝试在培养中重建肿瘤微环境的关键特征，以增加 细胞培养模型的预测性。这将涉及复制时变药物暴露，如在 动物和病人(目标3.4.1)，可溶环境的系统变化(目标3.4.2)，操作 通过改变底物弹性(目标3.4.3)和直接评估 在所有收集的数据中，培养中的药物反应与患者之间的关系目标1-3。

PROJECT SUMMARY Absorption, distribution, metabolism and elimination (collectively called pharmacokinetics, PK), and time-dependent drug actions in target organs (pharmacodynamics, PD) play critical roles in efficacy and toxicity of all drugs. We will develop and implement new methods for modeling PK-PD at multiple scales from cells to patients, new methods for measuring PK at the cellular and subcellular levels, and new cell culture systems that better mimic the tumor environment, thus increasing our ability to predict patient responses from cell culture data. Our translational goal is to create new single-cell resolution methods to integrate a molecular understanding of drug-target interaction with measures of target engagement and induction of drug response in tissues and organisms. Aim 3.1 involves technology development for sub-cellular resolution PK measurement by fluorescence imaging of Companion Imaging Drugs (CIDs), small molecule or protein drugs tagged with a fluorophore for imaging in a manner that retains the bioactivity and pharmacokinetics of the parent compound. The properties of CIDs will be optimized (Aim 3.1.1) and the compounds used for intravital imaging of drug distribution and response in living mice (Aim 3.1.2). CIDs will be used to directly assay drug-target interaction in single cells by fluorescence correlation microscopy (Aim 3.1.3). Aim 3.2 will develop a novel quantitative, multiplexed mass spectrometry method for assaying structure-activity relationships at a cellular and sub-cellular level based on covalent modification of target proteins. This will involve creation of novel chemical drug-like probes (Aim 3.2.1) that will then be subjected to systematic chemical modification to explore the impact of physic0-chemical properties such as cLogP, pKa etc. (Aim 3.2.2). Aim 3.3 will involve development of methods for integrating pathway-level knowledge and biomarkers (both predictive and response) into the kind of PK-PD models that are routinely used for translational pharmacology and clinical trial design in industry. Aim 3.4 will attempt to recreate key features of the tumor microenvironment in culture to increase the predictivity of cell culture models. This will involve reproducing time-varying drug exposure as observed in animals and patients (Aim 3.4.1), systematic variation of the soluble environment (Aim 3.4.2), manipulation of the physical environment through changes in substrate elasticity (Aim 3.4.3) and direct assessment of the relationship between drug response in culture and patients across all the data collected Aims 1-3.