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: 8833261
• 负责人: Vladimir P Torchilin
• 金额: $ 20.29万
• 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 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8833261
• 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; antitumor 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）：这项研究基于两个挑战：（1）需要开发有效的系统来递送肿瘤可溶性不佳的抗癌药物，以及（2）需要将增强的药物递送到肿瘤中细胞响应局部肿瘤刺激。我们选择了紫杉醇（PTX）作为一种可溶的药物和细胞外基质金属蛋白酶2（MMP2），在许多肿瘤中被上调为局部肿瘤刺激，PTX的药物输送系统应反应。在这项研究中，选择了一个易于培训的自组装胶束系统作为一般平台。我们已经设计了一种新型的两亲聚乙二醇（PEG）2000-PTX结合物，其中PEG和PTX通过MMP2敏感肽（PEG2000-肽-PTX，Pro-Drug）结合，可以自组成“ Core-Shell” “结构并响应上调的MMP2，导致肿瘤特异性药物释放。这与另外两个两亲性结合物 - PEG1000-磷脂酰乙醇胺（PEG1000-PE）进行额外的胶束稳定和TATP-PEG1000-PE，以增加细胞渗透功能 - 自我组成型 - 在药物和TATP中的混合胶束中自我组成术，到达肿瘤的途中，较长的PEG2000链会在MMP2介导的肿瘤中PEG2000块的脱离后暴露，并允许增强活性药物的细胞内递送。这样的准备工作可确保：（i）较高的PTX加载效率，（ii）低过早的PTX释放，（iii）血液寿命和通过增强的渗透性和保留率（EPR）效应（iv）增强肿瘤的药物内在化，通过增强的肿瘤积累。由于存在TATP部分，细胞在MMP2作用后暴露在肿瘤内。使用该胶束系统，已经根据其肿瘤细胞特异性和细胞毒性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该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。将追求以下具体目标来检验这一假设： （1）使用tatp-peg1000-dope准备和优化MMP2响应式紫杉醇（PTX）的自组装胶束纳米颗粒； PEG1000-PE;和对MMP2敏感的PEG2000肽-PTX（Pro-Pro-Proug）构建块； （2）使用A549人类非小细胞肺癌和HT-1080人类纤维肉瘤细胞系在体外研究含PTX的细胞摄取和细胞毒性，并在体外进行了纳米毒性，并具有上心的细胞外基质基质MMP2； （3）基于对MMP2敏感性纳米反应的最高反应（最大细胞杀伤水平），使用AIM 2选择后选择的癌细胞系在小鼠中进行实验性肿瘤治疗。 这项研究将引入一种新型类型的刺激敏感性制剂，以增加疗效的溶质抗癌药物不良药物，并成为随后的R01提议的基础，以显示该方法适用于其他溶质溶剂差的药物的适用性和各种各样的肿瘤。具有上调的MMP2。

项目成果

Modification of Nanoparticles with Transferrin for Targeting Brain Tissues.

用转铁蛋白修饰纳米颗粒以靶向脑组织。

• DOI: 10.1007/978-1-0716-1617-8_5
• 发表时间: 2021
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Attia,SaraAly;Li,Xiang;Filipczak,Nina;Costa,DanielF;Torchilin,VladimirP
• 通讯作者: Torchilin,VladimirP

Synthesis of Doxorubicin and miRNA Stimuli-Sensitive Conjugates for Combination Therapy.

用于联合治疗的阿霉素和 miRNA 刺激敏感缀合物的合成。

• DOI: 10.1007/978-1-4939-9220-1_8
• 发表时间: 2019
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Costa,DanielF;Sarisozen,Can;Torchilin,VladimirP
• 通讯作者: Torchilin,VladimirP

Mixed Nanosized Polymeric Micelles as Promoter of Doxorubicin and miRNA-34a Co-Delivery Triggered by Dual Stimuli in Tumor Tissue.

• DOI: 10.1002/smll.201600925
• 发表时间: 2016-09
• 期刊: Small (Weinheim an der Bergstrasse, Germany)
• 作者: Salzano G;Costa DF;Sarisozen C;Luther E;Mattheolabakis G;Dhargalkar PP;Torchilin VP
• 通讯作者: Torchilin VP