Multifunctional matrix metalloprotease-2-sensitive anti-cancer nanopreparations

多功能基质金属蛋白酶2敏感抗癌纳米制剂

• 批准号: 8701689
• 负责人: Vladimir P Torchilin
• 金额: $ 16.91万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-07 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8701689
• 关键词: A549; Antineoplastic Agents; Apoptosis; Binding; Biodistribution; Blood; Camptothecin; Cancer cell line; Cell Line; Cells; Charge; Clinical; Curcumin; Data; Drug Delivery Systems; Ensure; Environment; Enzymes; Ethanolamines; Experimental Neoplasms; Extracellular Matrix; Feedback; Hospitals; Human; In Vitro; Israel; Lead; Longevity; Malignant Neoplasms; Mediating; Medical; Metalloproteases; Micelles; Modeling; Mus; Non-Small-Cell Lung Carcinoma; Paclitaxel; Penetration; Peptides; Permeability; Pharmaceutical Preparations; Polyethylene Glycol 2000; Polyethylene Glycols; Polymers; Preparation; Process; Prodrugs; Research; Risk; Route; Software Design; Solid; Specificity; Stimulus; Structure; System; Testing; Toxic effect; aqueous; base; cell killing; chemical property; cytotoxicity; design; drug mechanism; fibrosarcoma; improved; in vivo; innovation; medical schools; meetings; mouse model; nanomedicine; nanoparticle; neoplastic cell; novel; premature; programs; public health relevance; response; self assembly; success; tumor; tumor growth; uptake

DESCRIPTION (provided by applicant): This study is based on two challenges: (1) The need to develop an effective system for tumor delivery of poorly-soluble anti-cancer drugs, and (2) The need to achieve enhanced drug delivery into tumor cells in response to local tumoral stimuli. We have chosen paclitaxel (PTX) as a poorly soluble drug and extracellular matrix metalloprotease 2 (MMP2) that is up-regulated in many tumors as a local tumor stimulus, to which a drug delivery system for PTX should react. An easy-to-prepare self-assembling micellar system was chosen as a general platform in this research. We have designed a novel amphiphilic polyethylene glycol (PEG) 2000-PTX conjugate where PEG and PTX are conjugated via a MMP2-sensitive peptide (PEG2000- peptide-PTX, pro-drug), which can self-assemble into a "core-shell" structure and respond to up-regulated MMP2, resulting in tumor-specific drug release. This conjugate with two other amphiphilic conjugates - PEG1000-phosphatidyl ethanolamine (PEG1000-PE) for additional micelle stabilization and TATp-PEG1000- PE to add a cell-penetrating function - self-assembles into mixed micelles, where drug and TATp are "shielded" by the longer PEG2000 chains when en route to a tumor, but become exposed after MMP2-mediated detachment of the PEG2000 blocks in the tumor and allow for enhanced intracellular delivery of the active drug. Such a preparation ensures: (i) higher PTX loading efficiency, (ii) low premature PTX release, (iii) blood longevity and better tumor accumulation via the enhanced permeability and retention (EPR) effect, and (iv) enhanced drug internalization by tumor cells due to the presence of TATp moieties, which become exposed inside the tumor after the MMP2 action. Using this micellar system, promising in vitro and early in vivo data have been obtained in terms of its tumor cell-specificity and cytotoxicity. We hypothesize that this novel MMP2-sensitive system will significantly improve the delivery of the drug into tumor cells, resulting in an enhanced antitumor effect and decreased off-target toxicity. The following specific aims will be pursued to test this hypothesis: (1) To prepare and optimize the MMP2- responsive paclitaxel(PTX)-loaded self-assembly micellar nanoparticles using TATp-PEG1000-DOPE; PEG1000-PE; and MMP2-sensitive PEG2000-peptide-PTX (pro-drug) building blocks; (2) To investigate cellular uptake and cytotoxicity of PTX-containing, MMP2-sensitive nanopreparation in vitro using A549 human non-small cell lung cancer and HT-1080 human fibrosarcoma cell lines with up-regulated extracellular matrix MMP2; (3) To perform experimental tumor therapy in vivo in mice using a cancer cell line selected after Aim 2 based on the highest response (maximum level of cell killing) to the MMP2-sensitive nanopreparation. This study will introduce a novel type of stimuli-sensitive preparations of poorly-soluble anticancer drugs with increased efficacy, and become the basis for the subsequent R01 proposal to show the applicability of the approach for other poorly-soluble drugs and the broad variety of tumors with up-regulated MMP2.

描述（由申请人提供）：本研究基于两个挑战：（1）需要开发一种有效的系统来向肿瘤递送难溶性抗癌药物，以及（2）需要增强药物递送到肿瘤细胞中以响应局部肿瘤刺激。我们选择紫杉醇 (PTX) 作为难溶性药物，并选择在许多肿瘤中上调的细胞外基质金属蛋白酶 2 (MMP2) 作为局部肿瘤刺激物，PTX 的药物递送系统应对其做出反应。本研究选择一种易于制备的自组装胶束系统作为通用平台。我们设计了一种新型两亲性聚乙二醇（PEG）2000-PTX缀合物，其中PEG和PTX通过MMP2敏感肽（PEG2000-肽-PTX，前药）缀合，它可以自组装成“核-壳”结构并响应上调的MMP2，从而导致肿瘤特异性药物释放。这种缀合物与另外两种两亲性缀合物 - PEG1000-磷脂酰乙醇胺 (PEG1000-PE) 提供额外的胶束稳定性，TATp-PEG1000-PE 增加细胞穿透功能 - 自组装成混合胶束，其中药物和 TATp 在到达肿瘤的途中被较长的 PEG2000 链“屏蔽”，但在到达肿瘤后会暴露出来。 MMP2 介导的 PEG2000 分离会阻断肿瘤并增强活性药物的细胞内递送。这样的制剂确保：(i)更高的PTX负载效率，(ii)较低的过早PTX释放，(iii)通过增强的渗透性和保留(EPR)效应实现血液寿命和更好的肿瘤积累，以及(iv)由于TATp部分的存在而增强肿瘤细胞的药物内化，TATp部分在MMP2作用后暴露在肿瘤内部。使用这种胶束系统，在其肿瘤细胞特异性和细胞毒性方面获得了有希望的体外和早期体内数据。 我们假设这种新型 MMP2 敏感系统将显着改善药物向肿瘤细胞的输送，从而增强抗肿瘤作用并降低脱靶毒性。将追求以下具体目标来检验这一假设： (1) 利用TATp-PEG1000-DOPE制备并优化MMP2响应的紫杉醇(PTX)自组装胶束纳米粒子； PEG1000-PE；和 MMP2 敏感的 PEG2000-肽-PTX（前药）构建模块； (2) 利用细胞外基质 MMP2 上调的 A549 人非小细胞肺癌细胞系和 HT-1080 人纤维肉瘤细胞系，体外研究含 PTX 的 MMP2 敏感纳米制剂的细胞摄取和细胞毒性； (3) 使用目标 2 后基于对 MMP2 敏感纳米制剂的最高反应（最大细胞杀伤水平）选择的癌细胞系，在小鼠体内进行实验性肿瘤治疗。 这项研究将推出一种新型的难溶性抗癌药物刺激敏感制剂，其疗效更高，并成为后续R01提案的基础，以展示该方法对其他难溶性药物和MMP2上调的广泛肿瘤的适用性。