Developing targeted activatable peptides to amplify radiosensitizer delivery

开发靶向可激活肽以增强放射增敏剂的输送

• 批准号: 9379141
• 负责人: Sunil J Advani
• 金额: $ 7.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9379141
• 关键词: Address; Advanced Malignant Neoplasm; Adverse effects; Architecture; Area; Biodistribution; Biosensing Techniques; Cancer Patient; Cells; Charge; Cleaved cell; Clinical; Cytotoxic Chemotherapy; DNA Repair; Data; Dependency; Development; Diagnosis; Diagnostic; Disease; Dose; Drug Delivery Systems; Drug Kinetics; Drug Monitoring; Drug Sensitization; Drug Targeting; Environment; Extracellular Matrix; Fluorescence Resonance Energy Transfer; Functional Imaging; Gelatinase A; Generations; Goals; Head and Neck Cancer; Head and neck structure; In Situ; Ionizing radiation; Irradiated tumor; Kinetics; Label; Link; MMP14 gene; Malignant Neoplasms; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Measures; Modeling; Molecular; Monitor; Morbidity - disease rate; Mus; Neoplasm Metastasis; Normal tissue morphology; Organ Preservation; Patient-Focused Outcomes; Patients; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Pharmacology; Play; Prodrugs; Property; Quality of life; Radiation exposure; Radiation therapy; Radiation-Sensitizing Agents; Radioresistance; Radiosensitization; Resistance; Role; Technology; Testing; Therapeutic; Tissues; Toxic effect; Unresectable; Work; base; chemotherapy; cyanine dye 5; design; effective therapy; extracellular; flexibility; fluorophore; improved; improved outcome; inhibitor/antagonist; innovation; interest; killings; neoplastic cell; non-invasive imaging; overexpression; peptide drug; polypeptide; pre-clinical; ratiometric; response; theories; tool; tumor; tumor growth; tumor microenvironment; tumor progression; uptake

Project Summary/Abstract Tumor resistance to radiotherapy continues to be a significant problem in improving outcomes of patients diagnosed with locally advanced, unresectable cancers including head and neck (HNC). Cellular sensitivity to ionizing radiation (IR) is governed by intracellular and extracellular factors. To help overcome tumor radioresistance, drugs that sensitize tumor to ionizing radiation (IR) are used. In theory, more potent radiosensitizers should increase tumor kill and improve patient outcomes. In practice, the clinical utility of of such drugs is curtailed by side effects that limit doses. To address these issues, we are developing activatable cell penetrating peptide (ACPP) probes to monitor and selectively deliver conjugated radiosensitizers to tumors based on extracellular tumor protease activity. ACPP consist of a drug conjugated polycationic cell penetrating peptide and an autoinhibitory polyanionic peptide separated from each other by a flexible peptide linker. Our initial peptide linker is cleaved by matrix metalloproteinases (MMP) 2/9, abundantly expressed in tumors. Intact, drug conjugated ACPP is “pro-drug” where the the drug linked cell penetrating peptide is neutralized by the polyanionic peptide, blocking intracellular drug uptake. Upon encountering MMP 2/9 within tumors, ACPP is cleaved (i.e. “activated”) and releases the drug conjugated cell-penetrating peptide, which is then taken up by tumor cells. An advantage of our ACPP probes is that they can be designed to non-invasively image extracellular tumor protease activities by synthesizing ratiometric ACPP. Ratiometric ACPP are labeled with Cy5 and Cy7 on the polycationic and polyanionic peptide respectively, acting as a FRET donor/acceptor pair. By using different protease sensitive peptide linker sequences between the two charged polypeptides, ratiometric ACPP Cy5:Cy7 emission ratio non-invasively measures distinct tumor protease activities in situ. A unique property of ACPP is the dependency on protease cleavage begets enzymatic amplification in the area of interest. The goals of our proposal are to develop ACPP probes to selectively radiosensitize tumors. In Aim 1, we will test the pharmacokinetics, biodistribution and efficacy of ACPP conjugated radiosensitizers in murine HNC models. To achieve this, drugs and fluorophores will be conjugated to ACPP and combined with IR. In Aim 2, we will interrogate in situ how IR alters extracellular protease activities in a dose dependent and temporal manner using a panel of ratiometric ACPP with distinct linkers cleaved preferentially by specific proteases. Identifying IR inducible ACPP cleavage will promote IR guided drug delivery development. In summary, ACPP radiosensitizer delivery has several innovative features that warrant pre-clinical inquiry. Diagnostically, ratiometric ACPP allows for non-invasive imaging of intrinsic tumor microenvironment proteases and IR's extrinsic influence. Therapeutically, ACPP's modular architecture allows for permutations of different classes radiosensitizing drugs to be conjugated to ACPPs with distinct protease linkers. Together, our studies will test how ACPPs can be exploited to improve the therapeutic ratio of radiotherapy.

项目摘要/摘要 肿瘤对放射治疗的抵抗仍然是改善患者预后的一个重大问题。 被诊断为局部晚期、不能切除的癌症，包括头部和颈部(HNC)。细胞对 电离辐射(IR)受细胞内和细胞外因素的影响。帮助战胜肿瘤 放射抗性是指使肿瘤对电离辐射(IR)敏感的药物。从理论上讲，更有效力 放射增敏剂应该增加肿瘤的杀伤力，改善患者的预后。在实践中，OF的临床应用 此类药物会受到限制剂量的副作用的限制。为了解决这些问题，我们正在开发Actiatable 细胞穿透肽(ACPP)探针用于监测和选择性地将偶联放射增敏剂输送到肿瘤 基于细胞外肿瘤蛋白水解酶活性。ACPP由药物偶联聚阳离子细胞穿透组成 多肽和通过柔性多肽连接物彼此分离的自抑制多阴离子多肽。我们的 起始肽连接蛋白被基质金属蛋白酶(MMP2/9)切割，在肿瘤中大量表达。 完整的药物结合的ACPP是“前药物”，药物连接的细胞穿透肽被中和 多阴离子多肽，阻断细胞内药物摄取。在肿瘤内遇到MMP2/9后，ACPP 被裂解(即“激活”)并释放药物结合的细胞穿透肽，然后被摄取 通过肿瘤细胞。我们的ACPP探头的一个优势是它们可以被设计成非侵入性成像 比率计量式ACPP合成的胞外肿瘤蛋白酶活性。比率计式ACPP标有 Cy5和Cy7分别位于聚阳离子和聚阴离子多肽上，作为FRET供体/受体对。 通过在两个带电多肽之间使用不同的蛋白水解酶敏感多肽接头序列， 比率ACPP Cy5：Cy7发射比非侵入性地原位测量不同的肿瘤蛋白酶活性。一个 ACPP的独特性质是依赖于蛋白酶的切割在该区域产生酶扩增 感兴趣的人。我们建议的目标是开发ACPP探针来选择性地使肿瘤放射增敏。在AIM 1，我们将测试ACPP偶联放射增敏剂在小鼠体内的药代动力学、生物分布和疗效。 HNC模型。为了实现这一点，药物和荧光团将与ACPP结合并与IR结合。在……里面 目的2，我们将在原位询问IR如何以剂量依赖和 使用一组比率计量型ACPP的时间方式，其中不同的连接子优先被特定的 蛋白酶。识别IR诱导的ACPP裂解将促进IR导向药物传递的发展。在……里面 总结，ACPP放射增敏剂给药有几个创新特征，值得进行临床前调查。 在诊断方面，比率计量式ACPP允许对固有的肿瘤微环境蛋白酶进行无创成像 以及IR的外在影响。在治疗方面，ACPP的模块化架构允许不同的排列 将放射增敏药物与具有不同蛋白酶连接物的ACPP偶联的类别。共同努力，我们的研究 将测试如何利用ACPPS来提高放射治疗的比率。