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）需要开发一种用于肿瘤递送难溶性抗癌药物的有效系统，以及（2）需要响应局部肿瘤刺激实现增强的药物递送至肿瘤细胞中。我们选择紫杉醇（PTX）作为一种难溶性药物，并选择在许多肿瘤中上调的细胞外基质金属蛋白酶2（MMP 2）作为局部肿瘤刺激物，PTX的药物递送系统应与之反应。本研究选择了一种易于制备的自组装胶束体系作为研究的通用平台。我们设计了一种新型的两亲性聚乙二醇（PEG）2000-PTX缀合物，其中PEG和PTX通过MMP 2敏感肽（PEG 2000- peptide-PTX，前药）缀合，其可以自组装成“核-壳”结构并响应于上调的MMP 2，导致肿瘤特异性药物释放。该缀合物与另外两种两亲性缀合物-PEG 1000-磷脂酰乙醇胺（PEG 1000-PE）用于额外的胶束稳定，TATp-PEG 1000- PE用于增加细胞穿透功能-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PEG 2000链“屏蔽”，但在肿瘤中MMP 2介导的PEG 2000阻滞剂脱离后暴露，并允许活性药物的增强的细胞内递送。这种准备确保：（i）更高的PTX负载效率，（ii）低的过早PTX释放，（iii）通过增强的渗透性和滞留（EPR）效应的血液寿命和更好的肿瘤积累，和（iv）由于存在TATp部分而增强的肿瘤细胞的药物内化，所述TATp部分在MMP 2作用后暴露在肿瘤内。使用这种胶束系统，在肿瘤细胞特异性和细胞毒性方面，已经获得了有希望的体外和早期体内数据。 我们假设这种新型MMP 2敏感系统将显著改善药物向肿瘤细胞的递送，从而增强抗肿瘤作用并降低脱靶毒性。为了检验这一假设，将追求以下具体目标： (1)采用TATp-PEG 1000-DOPE、PEG 1000-PE和MMP 2敏感的PEG 2000-peptide-PTX制备并优化MMP 2响应性紫杉醇（PTX）自组装胶束纳米粒（前药）构件;（2）研究含PTX的细胞摄取和细胞毒性，MMP-2敏感性纳米制剂的体外实验研究（3）使用在目标2之后基于对MMP 2敏感性纳米制剂的最高响应（最大水平的细胞杀伤）选择的癌细胞系在小鼠中进行体内实验性肿瘤治疗。 这项研究将介绍一种新型的难溶性抗癌药物的刺激敏感制剂，其疗效增加，并成为随后R 01提案的基础，以显示该方法对其他难溶性药物和MMP 2上调的各种肿瘤的适用性。