Cytochrome P450-mediated drug interactions at the human blood-brain barrier

细胞色素 P450 介导的人血脑屏障药物相互作用

• 批准号: 10080758
• 负责人: Chaitali Ghosh
• 金额: $ 34.52万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10080758
• 关键词: ABCB1 gene; Affect; Antidepressive Agents; Antiepileptic Agents; Astrocytes; Biological Assay; Biological Availability; Blood - brain barrier anatomy; Brain; Brain Diseases; Brain region; CYP3A4 gene; Carbamazepine; Carrier Proteins; Cells; Clinical; Cortical Dysplasia; Cytochrome P450; Cytochrome a; Disease model; Drug Interactions; Drug Kinetics; Drug Prescriptions; Drug resistance; Drug usage; Electrodes; Electrophysiology (science); Endothelial Cells; Enzymes; Epilepsy; Excision; Extrahepatic; Free Radicals; Gene Expression; Generations; Goals; High Pressure Liquid Chromatography; Histologic; Human; In Situ; In Vitro; Intractable Epilepsy; Ketoconazole; Levetiracetam; Liver; Measurement; Mediating; Metabolic; Metabolism; Mitochondria; Modeling; Molecular; Neurons; Nuclear Receptors; Operative Surgical Procedures; Oxidative Stress; Patients; Pharmaceutical Preparations; Pharmacodynamics; Pharmacogenomics; Pharmacotherapy; Play; Process; Production; Quinolinic Acid; Regulation; Reporting; Research Personnel; Resected; Role; Sampling; Seizures; Sertraline; Serum; Temporal Lobe Epilepsy; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Transfection; base; brain cell; brain endothelial cell; brain metabolism; cell injury; cytotoxicity; differential expression; drug development; drug distribution; drug metabolism; experience; gabapentin; human tissue; in vitro Model; individual patient; inhibitor/antagonist; innovation; lamotrigine; multi drug transporter; multidisciplinary; neurotoxic; neurotoxicity; neurovascular; neurovascular unit; next generation; novel; novel therapeutics; overexpression; oxcarbazepine; patient population; personalized medicine; pregabalin; prevent; side effect; tool; topiramate

ABSTRACT The mechanisms involved in brain drug interactions and reactive metabolites distribution in the brain during pharmacotherapy is largely unknown. Cytochrome P450 (CYP) drug-metabolizing enzymes are known to be functionally active at the blood-brain barrier (BBB). Multidrug transporter proteins (e.g., MDR1) at the BBB were once thought to be the main mechanism of failed antiepileptic drug (AED) therapy in patient suffering from drug-resistant epilepsy (DRE). However, we reported that the BBB participates in this process via CYP-mediated mechanism through the conversion of a therapeutic agent into a neurotoxic molecule. Increased CYP-MDR1 co-expression was found in epileptic brain endothelial cells (EPI-ECs) and neurons. We also found neurotoxic interactions between carbamazepine, (CBZ, an AED) and sertraline (antidepressant) – a consequence of CYP-mediated production of reactive CBZ metabolites and oxidative stress. The extent, relevance and topography of co-prescribed drug metabolism and potential neurotoxicity in the epileptic brain remain unexplored. Some of these drugs are induced and/or inhibited by CYP; we must devise ways to prevent subsequent neuronal changes. Our multidisciplinary team will test the central hypothesis that two co-prescribed AEDs will competitively interact in human BBB endothelial cells to regulate neuronal function and a CYP-MDR mechanism affects drug- metabolite levels in DRE brain. We will study a homogeneous patient population (with focal DRE due to cortical dysplasia, FCD) co-prescribed combination of CYP- and/or non-CYP-regulated AEDs. The Specific Aims will be achieved by using these multifaceted approaches to: (1) assess CYP’s contribution to neurotoxicity and metabolism of AEDs in a humanized in vitro neurovascular unit (NVU); (2) determine the regulatory factors and the extent to which CYP metabolize AEDs by using (a) EPI-ECs derived from brain resections of DRE with FCD to form a NVU and (b) epileptic (dysplastic) and nonepileptic (nondysplastic) tissue resected from the individual patient (ex vivo); and (3) study in situ CYP/MDR1 expression correlated to AED tissue bioavailability, formation of reactive metabolites, and extent of cellular damage in epileptic vs. nonepileptic resected DRE brain regions. The human tissue will be characterized histologically and electrophysiologically through invasive SEEG or subdural grid recording. A novel humanized NVU with primary control or patient-derived DRE brain cells will be used. Drug interactions at the BBB and real-time neuronal effects will be evaluated as a “personalized medicine” tool for disease modeling. Further measurement includes drug/metabolites by HPLC-MS (pharmacokinetics); neurotoxicity; free radical assay; and mitochondrial (pharmacodynamics) and gene expression changes (pharmacogenomics). Molecular manipulations of brain cells (by transfection/inhibitors) will validate CYP-mediated AED interactions in normal, drug-resistant BBB and CYP-MDR1 regulation in DRE brain (epileptic/nonepileptic). Together, there will be ample information to minimize CYP-dependent neurotoxicity and prevent side-effects of a polytherapy.

摘要 脑内药物相互作用机制及反应性代谢产物在脑内的分布 药物治疗在很大程度上是未知的。已知细胞色素P450（CYP 1A 1）药物代谢酶在功能上是 在血脑屏障（BBB）中起作用。多药物转运蛋白（例如，血脑屏障中的MDR 1）曾经被认为是 抗癫痫药物（AED）治疗耐药性癫痫（DRE）失败的主要机制。 然而，我们报道了血脑屏障通过CYP介导的机制参与了这一过程。 将治疗剂转化为神经毒性分子。CYP-MDR 1共表达增加， 癫痫脑内皮细胞（EPI-ECs）和神经元。我们还发现了神经毒性的相互作用， 卡马西平（CBZ，一种AED）和舍曲林（抗抑郁药）-CYP介导的 反应性CBZ代谢物和氧化应激。共同处方药物的程度、相关性和分布 癫痫脑中的代谢和潜在神经毒性尚未探索。有些药物是诱导性的 和/或被神经元抑制;我们必须设计出防止随后神经元变化的方法。 我们的多学科团队将测试中心假设，即两种共同处方的AED将竞争性地 在人血脑屏障内皮细胞中相互作用以调节神经元功能，CYP-MDR机制影响药物- DRE脑中的代谢物水平。我们将研究一个同质的患者人群（由于皮质类固醇导致局灶性DRE）， 发育不良，FCD）共同处方的AED和/或非AED调节的AED的组合。具体目标将是 通过使用这些多方面的方法来实现：（1）评估β-内酰胺酶对神经毒性和代谢的贡献， AED在人源化体外神经血管单位（NVU）中的作用;（2）确定调节因子以及 通过使用（a）衍生自具有FCD的DRE的脑切除的EPI-EC以形成NVU， (b)从个体患者（离体）切除的癫痫（发育不良）和非癫痫（非发育不良）组织;和 (3)原位研究AED/MDR 1表达与AED组织生物利用度、反应性代谢物的形成 癫痫与非癫痫切除的DRE脑区域中的细胞损伤程度。人体组织将被 通过侵入性SEEG或硬膜下网格记录进行组织学和电生理学表征。 将使用具有初级对照或患者来源的DRE脑细胞的新型人源化NVU。药物 BBB的相互作用和实时神经元效应将作为“个性化医学”工具进行评估， 疾病建模进一步测量包括通过HPLC-MS（药代动力学）的药物/代谢物;神经毒性;游离 自由基测定;和线粒体（药效学）和基因表达变化（药物基因组学）。分子 脑细胞的操作（通过转染/抑制剂）将验证CYP介导的AED在正常， DRE脑（癫痫/非癫痫）中的耐药BBB和CYP-MDR 1调节。在一起，将有充足的 最大限度地减少CYP依赖性神经毒性和预防多药治疗副作用的信息。

项目成果

Drug Delivery Challenges in Brain Disorders across the Blood-Brain Barrier: Novel Methods and Future Considerations for Improved Therapy.

• DOI: 10.3390/biomedicines9121834
• 发表时间: 2021-12-04
• 期刊: Biomedicines
• 影响因子: 4.7
• 作者: Achar A;Myers R;Ghosh C
• 通讯作者: Ghosh C

Multiple hurdle mechanism and blood-brain barrier in epilepsy: glucocorticoid receptor-heat shock proteins on drug regulation.

• DOI: 10.4103/1673-5374.313046
• 发表时间: 2021-12
• 期刊: Neural regeneration research
• 影响因子: 6.1
• 作者: Achar A;Ghosh C
• 通讯作者: Ghosh C

Overexpression of pregnane X and glucocorticoid receptors and the regulation of cytochrome P450 in human epileptic brain endothelial cells.

• DOI: 10.1111/epi.13703
• 发表时间: 2017-04
• 期刊: Epilepsia
• 影响因子: 5.6
• 作者: Ghosh C;Hossain M;Solanki J;Najm IM;Marchi N;Janigro D
• 通讯作者: Janigro D

Detection of brain-directed autoantibodies in the serum of non-small cell lung cancer patients.

• DOI: 10.1371/journal.pone.0181409
• 发表时间: 2017
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Banjara M;Ghosh C;Dadas A;Mazzone P;Janigro D
• 通讯作者: Janigro D

Modulation of glucocorticoid receptor in human epileptic endothelial cells impacts drug biotransformation in an in vitro blood-brain barrier model.

• DOI: 10.1111/epi.14567
• 发表时间: 2018-11
• 期刊: Epilepsia
• 影响因子: 5.6
• 作者: Ghosh C;Hossain M;Mishra S;Khan S;Gonzalez-Martinez J;Marchi N;Janigro D;Bingaman W;Najm I
• 通讯作者: Najm I