ROS-responsive Chemical Modification of Protein and Its Delivery

蛋白质的 ROS 响应性化学修饰及其传递

• 批准号: 10000918
• 负责人: Yamin Li
• 金额: $ 34.82万
• 依托单位: TUFTS UNIVERSITY MEDFORD
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-18 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10000918
• 关键词: 4T1; Antineoplastic Agents; Arsenic Trioxide; Behavior; Binding; Biodistribution; Biological; Biological Process; Breast Cancer Cell; Breast Cancer Model; Cancerous; Carbonates; Cations; Cell Surface Receptors; Cells; Charge; Chemicals; Cleaved cell; Cytoplasm; Development; Disease; Drug Kinetics; Electrostatics; Experimental Designs; Formulation; Genetic Diseases; Goals; Growth Factor; Human; Hydrogen Peroxide; Hydrophobicity; In Vitro; Intravenous; Libraries; Lipids; Malignant Neoplasms; Measurement; Mediating; Methods; Modification; Monitor; Monoclonal Antibodies; Mus; Nature; Neoplasm Metastasis; Normal Cell; Organ; Paclitaxel; Peptides; Pharmaceutical Preparations; Post-Translational Protein Processing; Property; Proteins; Reactive Oxygen Species; Resistance; Scheme; Surface; System; Tail; Testing; Therapeutic Studies; Toxic effect; Treatment Efficacy; base; cancer cell; cancer therapy; cancer type; cytokine; cytotoxic; disulfide bond; improved; in situ imaging; in vivo; malignant breast neoplasm; mouse model; nanocomplexes; nanoformulation; nanoparticle; novel; physical property; protein complex; protein function; public health relevance; side effect; small molecule; success; targeted cancer therapy; targeted delivery; therapeutic protein; triple-negative invasive breast carcinoma; tumor; tumor growth

PROJECT SUMMARY Intracellular protein therapies are tremendously appealing, promising treatments for many heretofore untreatable diseases, including genetic diseases and cancers. However, the safe, efficient intracellular delivery of proteins remains a challenge. Here we propose the development of an effective cytoplasmic delivery strategy for therapeutic proteins for cancer treatment: A combination of reversible chemical modification of proteins and nanocomplexation with cationic lipid-based nanoparticles. The chemical modification will inactivate the cytotoxic proteins, making them effectively nontoxic to normal cells. However, upon encountering increased level of reactive oxygen species (ROS), i.e. inside the cancer cells, the modified chemical moiety will be cleaved and the biological function of the proteins will be restored, resulting in cytotoxic action. Simultaneously, chemical modification reduces the surface charge of proteins, leading to more efficient nanocomplexation and delivery by bioreducible cationic lipid-like molecules. This protein delivery platform may thus serve as an efficient tumor- specific targeting system for cancer treatment. To accomplish the objective of this application, we propose the following three Specific Aims: Specific Aim 1: ROS-responsive modification of saporin and its characterization. To develop an approach for chemical modification of saporin, characterize the modification, and investigate the cleavage of the modified moieties at elevated ROS levels. Specific Aim 2: Intracellular delivery of saporin-NBC using synthetic lipids. To study a new class of synthetic lipid-based nanoparticles for intracellular saporin-NBC delivery in both cancerous and non-cancerous cells, and to study the synergistic effect of intracellular delivery of saporin-NBC and ROS-inducing anticancer drugs in suppressing triple-negative breast cancer cells. Specific Aim 3: Studying the in vivo saporin-NBC delivery in mouse breast cancer model. To study the therapeutic efficacy of bioreducible lipids in delivering proteins to inhibit tumor growth in murine breast cancer mouse models.

项目总结 细胞内蛋白疗法是非常有吸引力的，有望治疗许多迄今无法治愈的疾病。 疾病，包括遗传病和癌症。然而，安全、高效的细胞内蛋白质输送 仍然是一个挑战。在这里，我们建议开发一种有效的细胞质递送策略 用于癌症治疗的治疗性蛋白质：蛋白质的可逆化学修饰和 阳离子脂基纳米粒的纳米络合作用。化学修饰会使细胞毒素失活 蛋白质，使它们对正常细胞有效无毒。然而，在遇到更高水平的 活性氧物种(ROS)，即在癌细胞内，修饰的化学部分将被切割，并 这些蛋白质的生物学功能会被恢复，从而产生细胞毒作用。同时，化学物质 修饰降低了蛋白质的表面电荷，通过以下方式导致更有效的纳米络合和传递 生物可还原的阳离子类脂分子。因此，这种蛋白质输送平台可能会作为一种有效的肿瘤- 癌症治疗的特异性靶向系统。 为了实现本应用程序的目标，我们提出了以下三个具体目标： 具体目标1：皂苷的ROS响应性修饰及其表征。开发一种方法 对于皂苷的化学修饰，表征了修饰，并研究了修饰后的裂解 部分处于升高的ROS水平。 具体目标2：使用合成脂类在细胞内传递皂苷-NBC。学习一个新的班级 合成脂质纳米粒在癌和非癌组织中的细胞内皂苷-NBC传递 细胞内给药，研究皂苷-NBC细胞内给药与ROS诱导抗癌的协同作用 抑制三阴性乳腺癌细胞的药物。 具体目的3：研究Saporin-NBC体内给药对小鼠乳腺癌模型的影响。 生物可还原脂类转运蛋白抑制小鼠肿瘤生长的疗效研究 乳腺癌小鼠模型。