Targeted drug delivery system to overcome blood-brain barrier and therapeutic resistance to current standard of care in Glioblastoma

靶向药物输送系统可克服血脑屏障和对胶质母细胞瘤现行护理标准的治疗耐药性

• 批准号: 10659749
• 负责人: Meser M. Ali
• 金额: $ 49.95万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659749
• 关键词: Address; Affinity; Alcohols; Alkylating Agents; Alkylation; Amino Acids; Animals; Apoptotic; Binding; Biodistribution; Biological Availability; Biological Markers; Blood; Blood - brain barrier anatomy; Brain; Brain Neoplasms; Carbon; Cell Culture Techniques; Cells; Chemicals; Chemoresistance; Chlorotoxin; Clinical Research; Combined Modality Therapy; Combretastatin A-4; Complex; Copper; Coupled; Curcumin; Cysteine; DNA; DNA Adduction; DNA Adducts; DNA Double Strand Break; DNA Repair; DNA Repair Enzymes; Development; Diagnosis; Disulfides; Disulfiram; Ditiocarb; Dose; Drug Delivery Systems; Drug Targeting; Effectiveness; Encapsulated; Engineering; Environment; Enzymes; FDA approved; Formulation; Fractionated radiotherapy; Glioblastoma; Glioma; Goals; Hematology; Heterogeneity; Immunohistochemistry; In Vitro; Intravenous; Ligands; MAPK8 gene; MGMT gene; Magnetic Resonance Imaging; Malignant Glioma; Mediating; Methods; Methyltransferase; Modeling; Molecular; Monitor; Morphology; Oral; Oral Administration; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Phase II Clinical Trials; Physiological; Play; Polymers; Prognosis; Prognostic Factor; Protein Methyltransferases; Proteins; Radiation therapy; Recurrent disease; Reporting; Research; Research Project Grants; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Safety; Serum Proteins; Solubility; Specificity; Stomach; System; Testing; Tissues; Toxic effect; Treatment Efficacy; Tumor Burden; Ubiquitin; Water; Western Blotting; Work; Xenograft Model; Xenograft procedure; cancer cell; cancer therapy; chelation; clinical application; cytotoxic; cytotoxicity; efficacy evaluation; extracellular; fractionated radiation; hydroxypropyl-beta-cyclodextrin; improved; in vivo; inhibitor; multicatalytic endopeptidase complex; nanoparticle delivery; neoplastic cell; novel; overexpression; phase II trial; pre-clinical; preclinical study; radiation resistance; real time monitoring; receptor; repaired; standard of care; success; targeted agent; targeted delivery; temozolomide; therapy resistant; treatment response; tumor; tumor heterogeneity; water solubility

Project Summary/Abstract: The uniform lethality of glioblastoma (GBM) with a survival of less than 2 years despite best available therapy is attributed to treatment resistance due to DNA repair mechanisms that drive disease relapse and tumor heterogeneity. One prognostic factor identified as a reliable biomarker for GBM sensitivity to temozolomide (TMZ) and radiotherapy (RT) is the overexpression of O6-methylguanine-methyl- transferase (MGMT) enzyme. Patients with active MGMT were found to receive little benefit from TMZ and RT and represent a group of great unmet need with no treatment options that significantly improve survival. Recently, several preclinical and clinical studies suggest that alcohol aversion drug, disulfiram (DSF), inhibited MGMT and improved the efficacy of TMZ in GBM when combined with copper (Cu). However, phase II trial showed that there was no survival benefit from oral Cu/DSF. Nevertheless, the major limitation of oral Cu/DSF has been delivery of fragile DSF within the in vivo system. We have developed 2-hydroxypropyl beta cyclodextrin (HPßCD) encapsulating Cu complex of DSF metabolite, diethyldithiocarbamic acid (DDC), Cu(DDC)2 delivery system that addresses major drawbacks of the Cu(DDC)2: easy degradation in the blood and non-specific interactions with cells and serum proteins and lack of tissue specific delivery. HPßCD providing stability of Cu(DDC)2 is identified. In vitro cell culture study revealed that HPßCD-Cu(DDC)2 inhibited MGMT through the ubiquitin-proteasome pathway. Inhibition of MGMT activity in cell cultures vastly increased the alkylation-induced DNA double-strand breaks, cytotoxicity, and the levels of apoptotic markers like -H2AX, JNK-P and cleavage of PARP-1. Preliminary intravenous delivery of HPßCD- Cu(DDC)2 in combination with TMZ in an MGMT-positive patient derived orthotopic xenograft (PDOX) model demonstrated tumor size regression with prolonged survival. HPßCD-Cu(DDC)2 targets MGMT-145-cysteine and its unique cytotoxic mechanism circumvents MGMT-mediated chemo- and radiation resistance. The present work aimed at the development and application of HPßCD-Cu(DDC)2 for targeted delivery of drug to GBM. Chlorotoxin (CTX), a peptide reported to bind selectively to glioma cells while showing no affinity for non- neoplastic cells, will covalently be coupled to functionalized HPßCD encapsulating Cu(DDC)2. We have shown that PDOXs without MGMT expression are sensitive to RT. Therefore, we hypothesize that the combination of GBM targeting CTX-HPßCD-Cu(DDC)2 with TMZ and RT will overcome TMZ+RT resistance and show synergistic cytotoxic effect in PDOXs which will be monitored by MRI studies. Our objectives of the proposed research are A) To incorporate targeting ligand CTX with HPßCD-Cu(DDC)2 to obtain an active brain drug delivery system, B) To determine the efficacy and safety of oral DSF/Cu versus intravenous CTX-HPßCD- Cu(DDC)2 alone or in combination with TMZ in PDOX, and C) To determine the efficacy and safety of CTX- HPßCD-Cu(DDC)2 in combination with TMZ and fractionated radiation in MGMT upregulated PDOX models.

项目概要/摘要：存活率低于 2 的胶质母细胞瘤 (GBM) 的均匀致死率 尽管有最好的可用治疗方法，但由于 DNA 修复机制导致治疗耐药， 驱动疾病复发和肿瘤异质性的一个预后因素被确定为 GBM 的可靠生物标志物。 对替莫唑胺 (TMZ) 和放射治疗 (RT) 的敏感性是 O6-甲基鸟嘌呤-甲基-的过度表达 发现 MGMT 活性的患者从 TMZ 和 RT 中获益甚微。 代表了一组未满足的巨大需求，并且最近没有显着提高生存率的治疗方案。 多项临床前和临床研究表明，酒精厌恶药物双硫仑 (DSF) 可抑制 MGMT 和 与铜 (Cu) 联合使用可提高 TMZ 在 GBM 中的疗效。然而，II 期试验表明， 口服 Cu/DSF 没有带来生存获益，但口服 Cu/DSF 的主要局限性是。 在体内系统内传递脆弱的 DSF。 我们开发了 2-羟丙基 β 环糊精 (HPßCD) 封装 DSF 代谢物的铜复合物， 二乙基二硫代氨基甲酸 (DDC)、Cu(DDC)2 输送系统可解决 Cu(DDC)2 的主要缺点： 在血液中容易降解，与细胞和血清蛋白发生非特异性相互作用，且缺乏组织 体外细胞培养研究表明，HPßCD 可提供 Cu(DDC)2 的稳定性。 HPßCD-Cu(DDC)2 通过泛素蛋白酶体途径抑制 MGMT 活性。 培养物大大增加了烷基化诱导的 DNA 双链断裂、细胞毒性和 细胞凋亡标记物，如 -H2AX、JNK-P 和 PARP-1 的裂解 初步静脉注射 HPßCD-。 Cu(DDC)2 与 TMZ 联合用于 MGMT 阳性患者衍生的原位异种移植 (PDOX) 模型 HPßCD-Cu(DDC)2 靶向 MGMT-145-半胱氨酸，肿瘤大小消退。 其独特的细胞毒性机制规避了 MGMT 介导的化疗和放射耐药性。 该工作旨在开发和应用 HPßCD-Cu(DDC)2 将药物靶向递送至 GBM。 Chlorotoxin (CTX)，一种据报道可以选择性地与神经胶质瘤细胞结合的肽，同时对非胶质瘤细胞没有亲和力。 我们已经证明，肿瘤细胞将与封装 Cu(DDC)2 的功能化 HPßCD 共价偶联。 没有 MGMT 表达的 PDOX 对 RT 敏感，因此，我们认为两者的组合。 使用 TMZ 和 RT 靶向 CTX-HPßCD-Cu(DDC)2 的 GBM 将克服 TMZ+RT 耐药性并显示 PDOX 的协同细胞毒性作用将通过 MRI 研究进行监测。 研究方向 A) 将靶向配体 CTX 与 HPßCD-Cu(DDC)2 结合以获得活性脑药物 给药系统，B) 确定口服 DSF/Cu 与静脉注射 CTX-HPßCD- 的有效性和安全性 Cu(DDC)2 单独或与 TMZ 联合用于 PDOX，以及 C) 确定 CTX- 的有效性和安全性 HPßCD-Cu(DDC)2 与 TMZ 和分段放射组合在 MGMT 上调 PDOX 模型中。