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: 10346555
• 负责人: Ram I. Mahato
• 金额: $ 45.47万
• 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/10346555
• 关键词: Abraxane; Adopted; Apoptosis; Apoptosis Inhibitor; BCL2 gene; Binding; Biodistribution; Biological; Blood Vessels; Carbonates; Cell Death; Cell Proliferation; 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; Epithelial; Erinaceidae; Extracellular Matrix; Extracellular Signal Regulated Kinases; FRAP1 gene; Failure; Family member; Feedback; Formulation; Genes; Glutathione; Glycolysis; Goals; Hepatotoxicity; In Vitro; 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; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Peptides; Pharmaceutical Preparations; Phosphatidylinositols; Phosphorylation; Phosphotransferases; Play; Polymers; 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; Toxic effect; Treatment Efficacy; Tumor Burden; base; biological adaptation to stress; cancer cell; cancer stem cell; cancer therapy; 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; smoothened signaling pathway; 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 胰腺结缔组织增生性肿瘤部位Hedgehog（Hh）信号激活胰腺星状细胞（PSC） 并在结缔组织形成和癌症干细胞（CSC）增殖中起关键作用。KRAS是 KRAS在胰腺癌（PC）中主要突变，但KRAS仍然是一个困难的靶点。由于抑制 mTORC 1/2增加ERK磷酸化，我们建议GEM与ONC 201联合治疗， AKT/ERK双重抑制剂，有效治疗PC。ONC 201抑制细胞增殖并诱导TNF相关的凋亡。 凋亡诱导配体（TRAIL）介导的凋亡。此外，我们还采用了基质耗竭策略， 通过顺序施用Hh抑制剂MDB 5以减少药物递送至肿瘤部位的物理屏障。 虽然音刺猬（Shh）缺陷型肿瘤具有减少的基质含量，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通路减少结缔组织增生将 允许将装载到EGFR靶向NP中的ONC 201和GEM有效递送到胰腺肿瘤部位。我们 已经确定了一种使用临床上可行的抑制剂的有效的组合治疗策略， 应用于具有不同KRAS突变的PDAC肿瘤。在我们的初步研究中，（i）与游离GEM相比， mPEG-共-PCC-g-GEM-g-DC NP使原位肿瘤中的GEM积累增加2.5倍。控制GEM 为了释放到肿瘤中，我们合成了具有14%w/w GEM有效负载的mPEG-co-P（Asp）-g-DC-S-S-GEM。那里 当与L-谷胱甘肽（GSH）孵育时，从聚合物释放90%的GEM。GEM与 ONC 201在体外杀伤耐药PC细胞方面显示出协同作用，并且在体内更有效地减少肿瘤生长 而不是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-吡啶基）苯基 对Hh配体和CSC标记物的抑制作用更强 比Vismodegib更有效。通过用EGFR修饰它们的表面来制备和优化靶向NPs 在不同配体密度下结合肽GE 11。将负载MDB 5的GE 11-NP全身施用至PC中 荷瘤小鼠在给药后4小时肿瘤中的药物浓度高于 非目标NP。因此，我们假设序贯施用负载MDB 5的NP将增加 GEM和ONC 201递送至肿瘤并通过逆转诱导的耐药性导致PC的协同抑制 通过结缔组织增生和CSC增殖更有效。我们的具体目标是i）评估ONC 201的作用， ii）开发靶向氧化还原敏感性靶向细胞， iii）MDB 5、ONC 201和GEM的纳米颗粒递送 在原位、PDX和自发KPC小鼠模型中，长远的影响是开发新颖的 使用MDB 5的多功能纳米药物减少PC中结缔组织诱导的化学抗性的策略， GEM和ONC 201。