Lipid-dendrimer micellar nanocarriers for siRNA/drug co-delivery in MDR cancer

用于 MDR 癌症中 siRNA/药物共递送的脂质树枝状聚合物胶束纳米载体

• 批准号: 9005996
• 负责人: Vladimir P Torchilin
• 金额: $ 34.07万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-07 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9005996
• 关键词: Antibodies; Antineoplastic Agents; Apoptosis; Binding; Biological Preservation; Blood Circulation; Breast Adenocarcinoma; Caspase; Cells; Charge; Clinical; Complex; DU145; Defense Mechanisms; Dendrimers; Down-Regulation; Doxorubicin; Ethanolamines; Family; Formulation; Gene Targeting; Genes; Human; In Vitro; Individual; Ligands; Lipids; MCF7 cell; MDA MB 231; Malignant Neoplasms; Malignant neoplasm of ovary; Micelles; Modeling; Multi-Drug Resistance; Mus; Nucleosomes; P-Glycoprotein; PC3 cell line; Paclitaxel; Penetration; Permeability; Pharmaceutical Preparations; Polymers; Preparation; Prostate carcinoma; Proteins; Research; Resistance; Small Interfering RNA; Surface; System; Therapeutic; Toxic effect; Transfection; base; cancer cell; cancer therapy; chemotherapy; cytotoxicity; effective therapy; ethylene glycol; immunogenicity; in vivo Model; inhibitor/antagonist; malignant breast neoplasm; nanocarrier; novel; public health relevance; self assembly; survivin; tumor; tumor growth; vector; zeta potential

 DESCRIPTION (provided by applicant): The effective therapy of multidrug resistant (MDR) tumors remains an important unsolved problem in cancer therapy. Certain proteins over-expressed in MDR cells, such as P-glycoprotein (Pgp) or survivin, are involved in the resistance of these cells towards traditional chemotherapy. Specific down-regulation of such proteins in cancer cells can be achieved by applying the corresponding siRNA. The combination of this approach and traditional chemotherapy could provide a promising strategy against MDR tumors. The clinical use of siRNA is limited by problems with its stabilization and delivery. We have recently suggested a poly(ethylene glycol)-phosphatidyl ethanolamine (PEG-PE)-modified G(4)-PAMAM nanocarrier for siRNA. We have also developed a unique dendrimer-based mixed micellar system from this new G(4)- PAMAM-PEG-PE and PEG-PE constructs. Such micelles can complex siRNA via the dendrimer part and entrap poorly soluble anti-cancer drugs within their lipid core. Such a system has high micellization efficiency with low CMC and higher drug loading; stabilizes and protects siRNA against enzymatic degradation; provides enhanced cell binding, penetration, and endosomal escape because of its positive charge; and shows less cytotoxicity due to PEGylation. It will allow a co-delivery of siRNA and drug in one nanopreparation, including the targeted one if the dendrimer-based micelles are surface-modified with the targeting ligand. Our general hypothesis is that PEG-lipid-modified dendrimer-based polymeric mixed micelles will complex siRNA via the dendrimer part and load poorly soluble anticancer drugs into the core resulting in siRNA/drug-co-loaded nanopreparations, which can be additionally targeted to tumor by surface- attached anti-cancer antibodies (mAb 2C5 developed by us earlier), and will serve as an effective mean against MDR tumors. The following specific aims will be pursued: (1) To prepare, characterize, and evaluate PAMAM-PEG-PE/siRNA (P-gp and survivin siRNAs) complexes and their mixed micellar nanopreparations with PEG-PE and anti-cancer drugs (doxorubicin or paclitaxel) modified or non-modified with mAb 2C5; (2) To study the interaction of non-targeted or Ab-targeted siRNA/drug-co-loaded dendrimer-based nanopreparations from the Aim 1 with various MDR cancer cells in vitro using non-MDR cells as controls: (a) To follow cell association and intracellular accumulation of siRNA and drugs; (b) Investigate the efficiency of the nanopreparations in down-regulation of target genes in MDR cells; (c) Evaluate their cytotoxicity and MDR reversal in MDR cells using scrambled siRNA as control; (3) To demonstrate the possibility of efficient siRNA/drug co-delivery to MDR tumors and enhanced tumor therapy using the developed systems in two different in vivo models and treatment agents selected after the Aim 2: (a) To investigate tumor accumulation of chosen non-targeted and targeted siRNA/drug-co-loaded nanopreparations in tumor-bearing mice; (b) To evaluate their activity in tumor growth inhibition using scrambled siRNA as a control.

 描述（由申请人提供）：多药耐药（MDR）肿瘤的有效治疗仍然是癌症治疗中尚未解决的重要问题。 MDR 细胞中过度表达的某些蛋白质，例如 P-糖蛋白 (Pgp) 或生存素，与这些细胞对传统化疗的抵抗有关。通过应用相应的 siRNA 可以实现癌细胞中此类蛋白质的特异性下调。这种方法与传统化疗的结合可以提供一种有前景的针对 MDR 肿瘤的策略。 siRNA 的临床使用因其稳定性和递送问题而受到限制。我们最近提出了一种用于 siRNA 的聚（乙二醇）-磷脂酰乙醇胺（PEG-PE）修饰的 G（4）-PAMAM 纳米载体。我们还利用这种新的 G(4)-PAMAM-PEG-PE 和 PEG-PE 结构开发了一种独特的基于树枝状聚合物的混合胶束系统。这种胶束可以通过树枝状聚合物部分复合siRNA，并将难溶性抗癌药物捕获在其脂质核心内。该体系胶束化效率高，CMC低，载药量高；稳定并保护 siRNA 免受酶促降解；由于其正电荷，提供增强的细胞结合、渗透和内体逃逸；并且由于聚乙二醇化而表现出较低的细胞毒性。它将允许在一种纳米制剂中共同递送 siRNA 和药物，如果基于树枝状聚合物的胶束用靶向配体进行表面修饰，则包括靶向纳米制剂。 我们的总体假设是，PEG-脂质修饰的基于树枝状聚合物的聚合混合胶束将通过树枝状聚合物部分与siRNA复合，并将难溶性抗癌药物负载到核心中，形成siRNA/药物共负载的纳米制剂，该纳米制剂可以通过表面附着的抗癌抗体（我们之前开发的mAb 2C5）另外靶向肿瘤，并将作为对抗肿瘤的有效手段。 耐多药肿瘤。将追求以下具体目标：（1）制备、表征和评估 PAMAM-PEG-PE/siRNA（P-gp 和生存素 siRNA）复合物及其与 PEG-PE 和经 mAb 2C5 修饰或未修饰的抗癌药物（阿霉素或紫杉醇）的混合胶束纳米制剂； (2) 使用非 MDR 细胞作为对照，研究来自 Aim 1 的非靶向或 Ab 靶向 siRNA/药物共载树枝状大分子纳米制剂与各种 MDR 癌细胞在体外的相互作用： (a) 跟踪细胞关联以及 siRNA 和药物的细胞内积累； (b) 研究纳米制剂下调 MDR 细胞中靶基因的效率； (c) 使用乱序 siRNA 作为对照，评估其在 MDR 细胞中的细胞毒性和 MDR 逆转； (3) 证明使用开发的系统在两种不同的体内模型和目标 2 之后选择的治疗剂中有效地将 siRNA/药物共同递送至 MDR 肿瘤并增强肿瘤治疗的可能性： (a) 研究选定的非靶向和靶向 siRNA/药物共载纳米制剂在荷瘤小鼠中的肿瘤积累； (b) 使用乱序 siRNA 作为对照来评估它们在肿瘤生长抑制中的活性。