PTD-Mediated Protein or Drug Delivery for Cancer Therapy

PTD 介导的蛋白质或药物递送用于癌症治疗

基本信息

批准号：
8185708
负责人：
VICTOR C YANG
金额：
$ 25.77万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-05-04 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8185708
关键词：
Accounting Adopted Adsorption Adverse effects Affect Animals Antibodies Antidotes Antigens Antineoplastic Agents Apoptosis Behavior Binding Carcinoembryonic Antigen Cell physiology Cell surface Cells Charge Clinical Colonic Neoplasms Colorectal Cancer Colorectal Neoplasms Complex Coupled Coupling Cytosol Diagnosis Disease Doctor of Philosophy Dose Drug Delivery Systems Drug Kinetics Engineering Exhibits Future Gelonium multiflorum GEL protein Generic Drugs Glioma Grant Heparin Heparin Binding Human Hydrolysis In Vitro Institutes Investigation Large Intestine Carcinoma Link Malignant Neoplasms Manuscripts Masks Mediating Methods Monitor Mus Neoplasm Metastasis Normal tissue morphology Oxidoreductase Paper Pathway interactions Peptide Hydrolases Peptides Pharmaceutical Preparations Pharmacotherapy Phase Plasma Prodrugs Protamine Sulfate Protamines Proteins Publications Reporting Research Small Interfering RNA Solutions Surface Surface Antigens System Technology Time Toxic effect Toxin Translations Treatment Efficacy Xenograft procedure abstracting anticancer research asparaginylendopeptidase cancer therapy cancer type chemotherapy clinical efficacy cytotoxic design improved in vivo innovation leukemia malignant colon tumor mouse model neoplastic cell pre-clinical prevent success tumor tumor growth uptake

项目摘要

DESCRIPTION (provided by applicant): Drug therapy has thus far attained little success in managing cancers, primarily due to a lack clinical efficacy and presence of drug-induced toxicity. In the previous project, we instituted principles to overcome such obstacles in achieving an ideal drug therapy of maximum therapeutic efficacy with minimal drug-induced toxicity. This drug delivery system (DDS) is comprised of two components: [1] a targeting part composed of heparin linked to a mAb, and [2] a drug part, comprised of a drug coupled with a cell penetrating peptide (CPP). These two components can self-assemble into a tight complex via charge interaction between heparin and CPP. Binding CPP with heparin would mask its cell-penetrating function due to inhibition of its adsorption to the cell surfaces. The drug complex provides a prodrug behavior and prevents CPP-mediated uptake by normal tissues, alleviating drug-induced side effects. Following tumor targeting via the mAb, protamine will be administered to unmask heparin inhibition and restore the activity of the released CPP-Drug, inducing apoptosis to only tumor cells. Exceptional progress was made during the past grant period, with publication of 36 papers and graduation of 12 PhD/Postdoctoral trainees. All aims were accomplished, and utility/applicability of this DDS was confirmed. Despite promise, two crucial factors - efficacy of antibody targeting and extent of prodrug protection - were found in need of improvement in order to realize a clinically enabled drug therapy. Recent advancement in colorectal cancer research seems to offer a solution to these two issues. First, carcinoembryonic antigen (CEA), a surface antigen known to associate with tumor growth/metastasis, was shown to be overly expressed in colonic cancers but limited in normal tissues. Secondly, we discovered that >85% of the injected T84.66, a mouse mAb against human CEA, accumulated at the colorectal tumor target via an antigen-mediated elimination pathway. Thirdly, the protease legumain was recently reported to express profoundly on the surface of colorectal tumors but is almost undetectable in normal tissues. Using this information, we further engineered a highly improved DDS to cross the afore-cited barriers. Briefly, a potent toxin (gelonin) or siRNA drug will be covalently linked with LMWP, a proven but non-toxic CPP, via S-S bond. This LMWP-Drug will be further linked to T84.66 through a peptide linker containing the legumain-degradable specific AANL sequence. The drug complex would then exhibit a prodrug behavior during tumor targeting, since LMWP is embedded between the antibody and drug molecule, thus unable to transverse the drug through tumor cells in exerting cytotoxic effects. Following tumor accumulation via T84.66 targeting, release of LMWP-Drug from T84.66 will take place on tumor surface via cleavage of the AANL linker by surface-expressed legumain. LMWP-Drug will then internalize into tumor cells via LMWP-mediated cell entry action, initiating tumor apoptosis. The specific aims are to validate the pre-clinical capability of this system in managing specifically colorectal cancers, through extensive in vitro and in vivo studies using a well-established LS174T xenograft mouse model; thus paving the road for future clinical translation. PUBLIC HEALTH RELEVANCE: Drug therapy has thus far attained little success in managing cancers, primarily due to a lack of clinical efficacy and presence of drug-induced toxicity. In this project, we propose to develop a drug delivery system, which would take advantage of the tumor-specific expression of legumain as the activator of a prodrug that consists of T84.66 antibody, a protein (or siRNA) toxin, and a cell-penetrating peptide. The construction of the DDS is designed to improve treatment efficacy against colorectal cancers, while also reducing systemic toxicities.

描述（由申请人提供）：迄今为止，药物治疗在管理癌症方面几乎没有成功，这主要是由于缺乏临床功效和药物诱导的毒性的存在。在上一个项目中，我们提出了克服这种障碍，以实现最大程度的治疗疗效的理想药物治疗，并使用最少的药物诱导的毒性。该药物递送系统（DDS）由两个组成部分组成：[1]由与MAB相关的肝素组成的靶向部分，[2]由药物组成的药物组成，该药物由与细胞穿透肽（CPP）结合的药物组成。这两个组件可以通过肝素和CPP之间的电荷相互作用自组装成紧密的复合物。与肝素结合的CPP将由于抑制其吸附到细胞表面而掩盖其细胞穿透功能。该药物复合物提供了前药行为，并防止了CPP介导的正常组织摄取，从而减轻了药物诱导的副作用。通过MAB靶向肿瘤后，精蛋白将被施用以揭示肝素抑制并恢复释放的CPP-CPR-grug的活性，从而诱导仅对肿瘤细胞的细胞凋亡。在过去的赠款期间取得了出色的进步，并发表了36篇论文，毕业12位博士/博士后学员。所有目标均已完成，并确认了该DDS的实用性/适用性。尽管有希望，但发现需要改进的两个关键因素 - 抗体靶向和前药保护程度的功效，以实现临床上启用的药物治疗。结直肠癌研究的最新进展似乎为这两个问题提供了解决方案。首先，已知与肿瘤生长/转移相关的表面抗原（CEA）被证明在结肠癌中表达过多，但在正常组织中有限。其次，我们发现，通过抗原介导的消除途径在结直肠肿瘤靶标积累的注射的T84.66（针对人CEA的小鼠mAb）中> 85％。第三，最近据报道，蛋白酶在大肠肿瘤的表面上表达了深刻的表达，但在正常组织中几乎无法检测到。使用这些信息，我们进一步设计了高度改进的DD，以越过上述障碍。简而言之，通过S-S键，有效的毒素（胶质素）或siRNA药物将与LMWP共价（一种经过验证但无毒的CPP）。该LMWP-grug将通过包含豆科素降解的特定AANL序列的肽接头将进一步链接到T84.66。然后，由于LMWP嵌入抗体和药物分子之间，因此该药物复合物将在肿瘤靶向过程中表现出前药行为，因此无法通过肿瘤细胞横穿药物来发挥细胞毒性作用。通过T84.66靶向肿瘤积累后，将通过表面表达的豆药通过裂解AANL接头在肿瘤表面上释放LMWP-Prug。然后，LMWP-Prug将通过LMWP介导的细胞进入作用内部化为肿瘤细胞，从而启动肿瘤凋亡。具体的目的是通过大量的体外和体内研究验证该系统在管理大肠癌方面的临床前能力，并使用良好的LS174T异种移植小鼠模型来验证该系统的临床能力。因此为未来的临床翻译铺平了道路。公共卫生相关性：迄今为止，药物治疗在管理癌症方面几乎没有成功，这主要是由于缺乏临床功效和药物诱导的毒性的存在。在这个项目中，我们建议开发一种药物输送系统，该系统将利用豆科蛋白酶的肿瘤特异性表达作为由T84.66抗体，蛋白质（或siRNA）毒素和细胞培养肽组成的前药的激活剂。 DDS的构建旨在提高针对结直肠癌的治疗功效，同时还可以降低全身毒性。