Nanomedicine of Hedgehog and AKT/ERK Dual Inhibitors for Pancreatic Cancer

Hedgehog和AKT/ERK双重抑制剂治疗胰腺癌的纳米药物

基本信息

批准号：
10555244
负责人：
Ram I. Mahato
金额：
$ 43.33万
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：
2022-01-25 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10555244
关键词：
Abraxane Adopted Apoptosis Apoptosis Inhibitor BCL2 gene Binding Biodistribution Biological Blood Vessels Carbonates Cell Death Cells Chemoresistance Clinical Clinical Trials Combined Modality Therapy Cyclin D1 Desmoplastic Development Dodecanol Dose Limiting Down-Regulation Drug Delivery Systems Drug Formulations Drug Kinetics Encapsulated Epidermal Growth Factor Receptor Epithelium Erinaceidae Extracellular Matrix Extracellular Signal Regulated Kinases FRAP1 gene Failure Family member Feedback Formulation Genes Glutathione Glycolysis Goals Hepatotoxicity In Vitro Incubated Inhibition of Cell Proliferation Intervention KPC model KRAS2 gene Laboratories Ligands MEK inhibition MEKs Malignant Neoplasms Malignant neoplasm of pancreas Mediating Mesenchymal Metabolism Mus Mutate Mutation Neoplasm Metastasis Oncogenic Organ Oxidation-Reduction PI3K/AKT PIK3CG gene Pancreatic Ductal Adenocarcinoma Pathway interactions Peptides Pharmaceutical Preparations Phosphatidylinositols Phosphorylation Phosphotransferases Play Polymers Proliferating Proteins Proto-Oncogene Proteins c-akt Ras/Raf Reporting Resistance Role SHH gene Signal Transduction Site Surface Survival Rate TNF-related apoptosis-inducing ligand TNFRSF10A gene TNFRSF10B gene Therapeutic Toxic effect Treatment Efficacy Tumor Burden Tumor Promotion biological adaptation to stress cancer cell cancer stem cell cancer therapy carcinogenesis chemotherapy combinatorial cost density ethylene glycol gemcitabine improved in vivo inhibitor innovation mouse model mutant nanomedicine nanoparticulate novel novel strategies novel therapeutics overexpression pancreatic cancer cells pancreatic neoplasm pancreatic stellate cell propylene resistance mechanism side effect stem cell biomarkers stem cell proliferation synergism therapeutic evaluation therapy development treatment strategy tumor tumor growth uptake

项目摘要

PROJECT SUMMARY Gemcitabine (GEM), a frontline drug, shows limited efficacy due to its rapid metabolism and inefficient delivery to the desmoplastic pancreatic tumor site. Hedgehog (Hh) signaling activates pancreatic stellate cells (PSCs) and plays a critical role in the formation of desmoplasia and proliferation of cancer stem cells (CSCs). KRAS is predominantly mutated in pancreatic cancer (PC), yet KRAS remains a difficult target. Since inhibition of mTORC1/2 increases ERK phosphorylation, we propose combination therapy of GEM with ONC201, which is an AKT/ERK dual inhibitor to effectively treat PC. ONC201 inhibits cell proliferation and induces TNF-related apoptosis inducing ligand (TRAIL)-mediated apoptosis. Further, we have adopted a stroma depletion strategy by sequentially administrating Hh inhibitor MDB5 for reducing physical barrier of drug delivery to the tumor site. While sonic hedgehog (Shh)-deficient tumors have reduced stromal content, such tumors are aggressive with increased vascularity and metastatic potential. Therefore, reduction of desmoplasia by inhibiting Hh pathway will allow efficient delivery of ONC201 and GEM loaded into EGFR targeted NPs to the pancreatic tumor site. We have identified an effective combinatorial treatment strategy using clinically viable inhibitors, which can be applied to PDAC tumors with different KRAS mutations. In our preliminary studies, (i) compared to free GEM, mPEG-co-PCC-g-GEM-g-DC NPs increased GEM accumulation in orthotopic tumor by 2.5-fold. To control GEM release into the tumor, we synthesized mPEG-co-P(Asp)-g-DC-S-S-GEM with GEM payload of 14% w/w. There was 90% GEM release from the polymer upon incubation with L-glutathione (GSH). Combination of GEM with ONC201 showed synergy in killing resistant PC cells in vitro and reduced tumor growth in vivo more effectively than their monotherapies. We also synthesized 2-chloro-N1-[4-chloro-3-(2-pyridinyl) phenyl]-N4, N4-bis(2- pyridinylmethyl)-1,4-benzenedicarboxamide (MDB5), which inhibited Hh ligands and CSC markers more efficiently than vismodegib. Targeted NPs were prepared and optimized by decorating their surface with EGFR binding peptide GE11 at different ligand density. Systemic administration of MDB5 loaded GE11-NPs into PC tumor bearing mice resulted in higher drug concentration in the tumor at 4h post administration compared to non-targeted NPs. Therefore, we hypothesize that sequential administration of MDB5 loaded NPs will increase GEM and ONC201 delivery to the tumor and result in synergistic inhibition of PC by reversing resistance induced by desmoplasia and CSC proliferation more efficiently. Our specific aims are to i) assess the effects of ONC201 and GEM combination in GEM resistant PC cells in vitro and in vivo, ii) development of targeted redox sensitive nanomedicine of MDB5, ONC201 and GEM, and iii) nanoparticulate delivery of MDB5, ONC201, and GEM combination in orthotopic, PDX and spontaneous KPC mouse models. Long-term impact is to develop novel strategies to reduce desmoplasia-induced chemoresistance in PC using multifunctional nanomedicine of MDB5, GEM and ONC201.

项目概要吉西他滨 (GEM) 是一种一线药物，由于其快速代谢和低效递送而显示出有限的疗效到促纤维增生性胰腺肿瘤部位。 Hedgehog (Hh) 信号激活胰腺星状细胞 (PSC) 并在结缔组织增生的形成和癌症干细胞（CSC）的增殖中发挥关键作用。 KRAS 是主要在胰腺癌 (PC) 中发生突变，但 KRAS 仍然是一个困难的目标。由于抑制 mTORC1/2 增加 ERK 磷酸化，我们建议 GEM 与 ONC201 联合治疗，即一种有效治疗 PC 的 AKT/ERK 双重抑制剂。 ONC201抑制细胞增殖并诱导TNF相关凋亡诱导配体（TRAIL）介导的细胞凋亡。此外，我们采用了基质消耗策略通过顺序施用 Hh 抑制剂 MDB5 来减少药物递送到肿瘤部位的物理屏障。虽然音刺猬 (Shh) 缺陷型肿瘤的基质含量减少，但此类肿瘤具有侵袭性增加血管分布和转移潜力。因此，通过抑制 Hh 途径来减少结缔组织增生将允许将装载到 EGFR 靶向纳米颗粒中的 ONC201 和 GEM 有效递送至胰腺肿瘤部位。我们已经确定了一种使用临床上可行的抑制剂的有效组合治疗策略，该策略可以应用于具有不同KRAS突变的PDAC肿瘤。在我们的初步研究中，(i) 与免费 GEM 相比， mPEG-co-PCC-g-GEM-g-DC NPs 使原位肿瘤中 GEM 的积累增加了 2.5 倍。控制创业板释放到肿瘤中，我们合成了 mPEG-co-P(Asp)-g-DC-S-S-GEM，GEM 有效负载为 14% w/w。那里与 L-谷胱甘肽 (GSH) 一起孵育后，90% 的 GEM 从聚合物中释放。创业板与 ONC201在体外表现出协同杀死耐药PC细胞的作用，并更有效地减少体内肿瘤的生长比他们的单一疗法。我们还合成了2-氯-N1-[4-氯-3-(2-吡啶基)苯基]-N4, N4-bis(2- 吡啶基甲基)-1,4-苯二甲酰胺 (MDB5)，更能抑制 Hh 配体和 CSC 标记物比 vismodegib 更有效。通过用 EGFR 修饰其表面来制备和优化靶向纳米颗粒不同配体密度下的结合肽GE11。将 MDB5 加载的 GE11-NP 系统管理到 PC 中与荷瘤小鼠相比，给药后 4 小时肿瘤中药物浓度更高非靶向 NP。因此，我们假设装载 MDB5 的 NP 的顺序给药会增加 GEM 和 ONC201 递送至肿瘤并通过逆转诱导的耐药性协同抑制 PC 更有效地促进结缔组织增生和 CSC 增殖。我们的具体目标是 i) 评估 ONC201 的效果和 GEM 组合在体外和体内的 GEM 抗性 PC 细胞中，ii) 开发靶向氧化还原敏感的 MDB5、ONC201 和 GEM 的纳米药物，以及 iii) MDB5、ONC201 和 GEM 的纳米颗粒递送在原位、PDX 和自发 KPC 小鼠模型中进行组合。长期影响是开发新颖的使用 MDB5 多功能纳米药物减少 PC 中结缔组织增生诱导的化疗耐药性的策略， GEM 和 ONC201。