Lipid nanomedicine targeting multiple signaling pathways of medulloblastoma

靶向髓母细胞瘤多种信号通路的脂质纳米药物

基本信息

批准号：
10663377
负责人：
Ram I. Mahato
金额：
$ 47.56万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-15 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10663377
关键词：
Abbreviations Apoptosis Apoptosis Inhibitor BRD2 gene Binding Biodistribution Biological Assay Blood - brain barrier anatomy Brain Brain Neoplasms Cell Cycle Cell Line Cell Proliferation Cells Cerebellum Chemoresistance Childhood Brain Neoplasm Cholesterol Clinical Combined Modality Therapy Complex Cyclin D1 Data Development Dioxins Dose Drug Transport Encapsulated Erinaceidae Escherichia coli Exhibits Film Goals Growth Human Hydration status Hydrophobicity Impairment In Vitro Individual Lipids MB03 MDM2 gene MYC gene MYCN gene Mutate Mutation N-Myc Protein Names Neoplasm Metastasis Neurocognitive Deficit Neurons Organ Outcome PIK3CG gene Parents Particle Size Pathway interactions Patients Peptides Pharmaceutical Preparations Phosphorylcholine Plasmids Play Polyethylene Glycols Proliferating Proteins Proto-Oncogene Proteins c-akt Recombinant Proteins Recombinants Resistance Roentgen Rays Role SHH gene Series Signal Pathway Signal Transduction Structure Surface TP53 gene Toxic effect Transgenic Organisms Treatment Efficacy Tumor Suppressor Proteins X-Ray Crystallography analog anti-cancer blood-brain barrier penetration cancer stem cell cell growth cell killing chemotherapy design efficacy evaluation in vitro activity in vivo inhibitor inhibitor-of-apoptosis protein innovation lipid nanoparticle medulloblastoma medulloblastoma cell line meter migration mouse model multiple drug use mutant nanomedicine neoplastic cell nerve stem cell neurotoxicity novel phosphoethanolamine protein purification rabies virus glycoprotein G small molecule smoothened signaling pathway stem cell proliferation stem cells synergism systemic toxicity technology platform treatment strategy tumor tumor growth tumorigenesis tumorigenic wound healing x-linked inhibitor of apoptosis protein

项目摘要

PROJECT SUMMARY Medulloblastoma (MB) is the most common childhood brain tumor arising from the cerebellum. Many factors influence the proliferation, differentiation, and migration of cerebellar granular neuronal precursor (GNP). Among them, MDM2 is a major nexus between tumor suppressor TP53 and hedgehog (Hh) signaling in GNPs and promotes MB tumor growth and metastasis. In addition, PI3K and BRD4 signaling also play key roles in MB cell growth, cancer stem cell (CSC) proliferation, and tumorigenesis. Further, MB treatment is challenging due to the development of chemoresistance, inefficient drug transport across the blood brain barrier (BBB) and drug induced neurotoxicity. Hh inhibitors are effective initially to treat SHH-MB, but their repeated use develops chemoresistance due to mutations in smoothened (SMO) but can be overcome by modulating GLI, which is downstream of SMO. In our preliminary studies, we synthesized a series of potent BRD4/PI3K dual inhibitors by modifying structure of parent compound SF2523. One of the compounds 8-(2,3-dihydrobenzo[b][1,4]dioxin- 6-yl)-2-morpholino-4H-chromen-4one (abbreviated as MDP5) was found highly potent. We then determined X- ray crystal structures of the recombinant BD1 and BD2 domains from BRD2 in complex with MDP5. While MDP5 showed higher potency in DOAY cells compared to SF2523 (12.6 µM), IC50 values for MDP5 and SF2523 were similar potency on HD-MB03 MB (MYC amplified) cells. MDP5 decreased the target downstream proteins like p-AKT, MYCN, Cyclin D1, and increased the degradation of MYCN protein indicated by p-MYCN (ser 54). We also discovered a small molecule JW-475A which is a potent dual MDM2 and XIAP inhibitor. MDP5 and JW- 475A (a dual MDM2 and XIAP inhibitor) effectively inhibited the proliferation of MB cells in a dose dependent manner, with significantly higher cell killing when these drugs were used in combination. Treatment of MB cells with the combination of these two drugs significantly decreased the colony formation capacity compared to individual drugs. We prepared PEG-DSPE based lipid nanoparticles (LNPs) with 4.9±0.1% and 4.8±0.1% loading for MDP5 and JW-475A. BBB penetrating targeted LNPs were prepared by surface decorating with rabies virus glycoprotein (RVG) peptide-peptide. Our hypothesis is that inhibition of BRD4/PI3K and MDM2/XIAP simultaneously using MDP5 and JW-475A represents a promising strategy to inhibit MB tumor in vivo. Further, we will use RVG-PEG-DSPE LNPs to encapsulate MDP5 and JW-475A, which have poor drug transport across the BBB. Our specific aims are to i) Synthesize novel MDP5 derivatives as dual function BRD4/PI3K inhibitors and characterize in vitro activity; ii) Evaluate anti-cancer efficacy of JW-475A in combination with MDP5 in vitro.; iii) Formulate MDP5 and JW-475A into LNPs decorated with RVG peptide and determine their biodistribution, therapeutic efficacy, and systemic/organ toxicity in in SHH and MYC driven cells and PDX-based orthotopic and transgenic SmoA1 MB mouse models. Long-term significance. Successful completion of this project will provide a platform technology for treating brain tumors using this innovative LNP-based combination therapy.

项目概要髓母细胞瘤（MB）是最常见的儿童脑肿瘤，由多种因素引起。影响小脑颗粒神经元前体（GNP）的增殖、分化和迁移。他们认为，MDM2 是肿瘤抑制因子 TP53 和 GNP 中的刺猬 (Hh) 信号之间的主要联系，促进MB肿瘤生长和转移此外，PI3K和BRD4信号在MB细胞中也发挥着关键作用。此外，由于MB治疗具有挑战性。化学耐药性的发展、穿过血脑屏障（BBB）的药物转运效率低下以及药物 Hh 抑制剂最初对治疗 SHH-MB 有效，但重复使用会产生神经毒性。由于平滑 (SMO) 突变而产生化学耐药性，但可以通过调节 GLI 来克服，GLI 是在我们的初步研究中，我们合成了一系列有效的 BRD4/PI3K 双重抑制剂。通过修改母体化合物 SF2523 的结构，其中一种化合物8-(2,3-二氢苯并[b][1,4]二恶英-。 6-yl)-2-morpholino-4H-chromen-4one（缩写为 MDP5）被发现具有高效能。 BRD2 与 MDP5 复合物的重组 BD1 和 BD2 结构域的射线晶体结构。与 SF2523 (12.6 µM) 相比，在 DOAY 细胞中显示出更高的效力，MDP5 和 SF2523 的 IC50 值为 MDP5 对 HD-MB03 MB（MYC 扩增）细胞具有相似的功效，可降低目标下游蛋白，如 p-AKT、MYCN、Cyclin D1，并增加 p-MYCN（第 54 号序列）表明的 MYCN 蛋白的降解。还发现了一种小分子 JW-475A，它是一种有效的 MDM2 和 XIAP 双重抑制剂。 475A（一种 MDM2 和 XIAP 双重抑制剂）以剂量依赖性方式有效抑制 MB 细胞的增殖当这些药物联合使用治疗MB细胞时，具有更显着的细胞杀伤作用。与相比，这两种药物的组合显着降低了集落形成能力我们制备了基于 PEG-DSPE 的脂质纳米颗粒 (LNP)，负载量为 4.9±0.1% 和 4.8±0.1%。 MDP5 和 JW-475A 通过用狂犬病病毒进行表面修饰来制备 BBB 穿透靶向 LNP。我们的假设是抑制 BRD4/PI3K 和 MDM2/XIAP。同时使用 MDP5 和 JW-475A 代表了一种有前途的体内抑制 MB 肿瘤的策略。我们将使用 RVG-PEG-DSPE LNP 来封装 MDP5 和 JW-475A，它们的药物转运能力较差我们的具体目标是 i) 合成新型 MDP5 衍生物作为双功能 BRD4/PI3K 抑制剂并表征体外活性；ii) 评估 JW-475A 与 MDP5 组合的体外抗癌功效。 iii) 将MDP5和JW-475A配制为用RVG肽修饰的LNP并测定其生物分布， SHH 和 MYC 驱动的细胞以及基于 PDX 的原位和转基因SmoA1 MB小鼠模型的成功完成将具有长期意义。提供使用这种基于 LNP 的创新联合疗法治疗脑肿瘤的平台技术。