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.

项目摘要/摘要：胶质母细胞瘤（GBM）的一致致死率，生存率低于2 尽管有最好的治疗方法，但仍持续了10年，这归因于DNA修复机制导致的治疗耐药性， 导致疾病复发和肿瘤异质性。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）和放疗（RT）的敏感性是O 6-甲基鸟嘌呤-甲基- 转移酶（MGMT）。活动性MGMT患者从TMZ和RT中获益甚微 并且代表了一组没有显著提高生存率的治疗选择的巨大未满足的需求。最近， 几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， 当与铜（Cu）组合时，改善TMZ在GBM中的功效。然而，第二阶段试验表明， 口服Cu/DSF没有生存益处。然而，口服Cu/DSF的主要局限性是 脆性DSF在体内系统内的递送。 我们开发了2-羟丙基β环糊精（HP β CD）包封DSF代谢物的Cu络合物， 二乙基二硫代氨基甲酸（DDC），Cu（DDC）2递送系统，其解决了Cu（DDC）2： 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， 具体交付。确定了HP β CD提供Cu（DDC）2的稳定性。体外细胞培养研究表明， HP β CD-Cu（DDC）2通过泛素-蛋白酶体途径抑制MGMT。细胞中MGMT活性的抑制 培养物大大增加了烷基化诱导的DNA双链断裂，细胞毒性和 凋亡标记物如JNK-H2 AX、JNK-P和PARP-1的裂解。HP β CD的初步静脉内递送- Cu（DDC）2与TMZ组合在MGMT阳性患者来源的原位异种移植物（PDOX）模型中 证实了肿瘤大小的消退和延长的生存期。HP β CD-Cu（DDC）2靶向MGMT-145-半胱氨酸 并且其独特的细胞毒性机制避免了MGMT介导的化疗和放射抗性。本 工作旨在开发和应用HP β CD-Cu（DDC）2，用于将药物靶向递送至GBM。 氯毒素（CTX），一种被报道选择性结合胶质瘤细胞而对非胶质瘤细胞没有亲和力的肽。 将共价偶联到包封Cu（DDC）2的官能化HP β CD上。我们已经表明 没有MGMT表达的PDOX对RT敏感。因此，我们假设 用TMZ和RT靶向CTX-HP β CD-Cu（DDC）2的GBM将克服TMZ+RT耐药性，并显示 在PDOX中的协同细胞毒性作用将通过MRI研究监测。我们建议的目标 A）将靶向配体CTX与HP β CD-Cu（DDC）2结合以获得活性脑药物 递送系统，B）为了确定口服DSF/Cu相对于静脉内CTX-HP β CD的功效和安全性， Cu（DDC）2单独或与TMZ组合在PDOX中的作用，和C）为了确定CTX-2的功效和安全性， HP β CD-Cu（DDC）2联合TMZ和分次放射在MGMT中上调PDOX模型。