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

• 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
• 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

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.

项目摘要 吉西他滨（GEM）是一种前线药物，由于其快速代谢和效率递送而显示出有限的功效 到脱肿瘤胰腺肿瘤部位。刺猬（HH）信号传导激活胰腺星状细胞（PSC） 并在癌细胞（CSC）形成和癌细胞增殖的形成中起着至关重要的作用。克拉斯是 在胰腺癌（PC）中主要突变，但KRAS仍然是一个困难的目标。由于抑制 MTORC1/2增加了ERK磷酸化，我们提出了GEM与Onc201的组合疗法，即 AKT/ERK双重抑制剂可有效治疗PC。 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，我们采用了基质耗竭策略 通过依次管理HH抑制剂MDB5来减少药物输送到肿瘤部位的物理障碍。 虽然缺乏的Sonic刺猬（SHH）肿瘤减少了基质含量，但这种肿瘤具有侵略性 增加血管性和转移潜力。因此，通过抑制HH途径减少脱木质 使ONC201的有效输送和将GEM递送到EGFR靶向NP中，以供胰腺肿瘤部位。我们 已经确定了使用临床上可行抑制剂的有效组合治疗策略，这可以是 应用于具有不同KRAS突变的PDAC肿瘤。在我们的初步研究中，（i）与自由宝石相比 MPEG-CO-PCC-GEM-GEM-GEM-G-DC NPS增加了原位肿瘤中的GEM积累2.5倍。控制宝石 释放到肿瘤中，我们合成了MPEG-CO-P（ASP）-G-DC-S-S-GEM，GEM有效载荷为14％w/w。那里 与L-Glutathione（GSH）孵育后，从聚合物中释放了90％的宝石。宝石与 ONC201显示在体外杀死抗性PC细胞的协同作用，并在体内降低肿瘤生长 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-双（2-）（2- 吡啶基甲基）-1,4-苯甲酰胺（MDB5），它抑制HH配体和CSC标记更多 比Vismodegib有效。通过用EGFR装饰其表面来制备目标NP 在不同的配体密度下结合肽GE11。将MDB5的全身性给药加载到PC中 肿瘤轴承小鼠在给药后4H导致肿瘤的药物浓度较高，而 非目标的NP。因此，我们假设MDB5负载的NP的顺序给药会增加 GEM和ONC201递送到肿瘤，并通过逆转电阻引起的抑制PC协同抑制 通过Desmoplasia和CSC增殖更有效。我们的具体目的是i）评估ONC201的影响 在体外和体内耐宝石抗PC细胞中的宝石组合，ii）靶向氧化还原敏感的发展 MDB5，ONC201和GEM的纳米医学，以及III）纳米明确的MDB5，ONC201和GEM 原位，PDX和自发KPC小鼠模型的组合。长期影响是发展新颖 使用MDB5的多功能纳米医学降低PC中脱木质诱导的化学抗性的策略， 宝石和ONC201。