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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的参与大脑药物相互作用和反应性代谢物在大脑中分布的机制已知细胞色素 P450 (CYP) 药物代谢酶的功能尚不清楚。曾被认为在血脑屏障 (BBB) 上具有活性的多药转运蛋白（例如 MDR1）。耐药性癫痫（DRE）患者抗癫痫药物（AED）治疗失败的主要机制。然而，我们报道 BBB 通过 CYP 介导的机制参与这一过程发现治疗剂转化为神经毒性分子的 CYP-MDR1 共表达增加。我们还发现癫痫脑内皮细胞（EPI-EC）和神经元之间存在神经毒性相互作用。卡马西平（CBZ，一种 AED）和舍曲林（抗抑郁药）——CYP 介导的产生的结果反应性 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的神经毒性仍未被探索，其中一些药物是诱导的。和/或被 CYP 抑制；我们必须想办法防止随后的神经变化。我们的多学科团队将测试中心假设，即两种共同处方的 AED 会具有竞争性与人 BBB 内皮细胞相互作用以调节神经元功能，CYP-MDR 机制影响药物- DRE 大脑中的代谢水平我们将研究同质的患者群体（由于皮质而具有局灶性 DRE）。 CYP 和/或非 CYP 调节的 AED 的共同处方组合将是。通过使用这些多方面的方法来实现：（1）评估 CYP 对神经毒性和代谢的贡献人源化体外神经血管单元 (NVU) 中的 AED (2) 确定调节因素及其程度； CYP 通过使用 (a) 来自 DRE 脑切除和 FCD 的 EPI-EC 来代谢 AED，形成 NVU 和 (b) 从个体患者身上切除的癫痫（发育异常）和非癫痫（非发育异常）组织（离体）； (3) 研究与 AED 组织生物利用度、反应性代谢物形成相关的原位 CYP/MDR1 表达，癫痫与非癫痫切除 DRE 大脑区域的细胞损伤程度。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。将使用具有主要对照或患者来源的 DRE 脑细胞的新型人源化 NVU 药物。 BBB 的相互作用和实时神经效应将被评估为“个性化医疗”工具进一步测量药物/代谢物的 HPLC-MS（药代动力学）；自由基测定；以及线粒体（药效学）和基因表达变化（分子药物基因组学）。对脑细胞的操作（通过转染/抑制剂）将验证正常情况下 CYP 介导的 AED 相互作用， DRE 大脑中的耐药 BBB 和 CYP-MDR1 调节（癫痫/非癫痫）在一起，将会有足够的结果。最大限度地减少 CYP 依赖性神经毒性并预防多种疗法副作用的信息。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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