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 在改善患者的预后方面，对放射疗法的耐肿瘤性仍然是一个重大问题 被诊断出患有局部高级，不可切除的癌症，包括头颈部（HNC）。细胞对细胞的敏感性 电离辐射（IR）受细胞内和细胞外因子的控制。帮助克服肿瘤 使用放射线，使用肿瘤对电离辐射（IR）的药物。从理论上讲，更强大 放射敏剂应增加肿瘤杀伤并改善患者预后。实际上，临床实用程序 这种药物会因限制剂量的副作用而减少。为了解决这些问题，我们正在开发可激活的 细胞穿透性肽（ACPP）问题，以监测并选择性地将共轭辐射效应递送到肿瘤 基于细胞外肿瘤蛋白酶活性。 ACPP由穿透的药物共轭多阳离子细胞组成 肽和自身抑制性的聚苯故肽通过柔性肽接头彼此分离。我们的 初始肽接头被基质金属蛋白酶（MMP）2/9裂解，在肿瘤中绝对表达。 完整的，药物共轭的ACPP是“促药”，其中连接的药物穿透性肽被中和 聚苯故肽，阻断细胞内药物的摄取。在肿瘤中遇到MMP 2/9时，ACPP 被裂解（即“激活”），并释放出药物共轭细胞穿透肽，然后将其吸收 由肿瘤细胞。我们ACPP问题的优势是它们可以设计为非侵入性图像 细胞外肿瘤蛋白酶活性通过合成比率ACPP。比例测量ACPP标记 Cy5和Cy7分别在聚阳离子和聚苯故的胡椒粉上，充当FRET供体/受体对。 通过在两个带电多肽之间使用不同蛋白酶敏感肽接头序列， 比率ACPP CY5：CY7发射比非侵入性测量不同的肿瘤蛋白酶活性。一个 ACPP的独特特性是对蛋白酶裂解的依赖性，该地区的酶扩增 感兴趣的。我们建议的目标是开发ACPP问题，以选择性地放射敏感性肿瘤。目标 1，我们将测试ACPP共轭放射增强剂的药代动力学，生物分布和效率 HNC模型。为了实现这一目标，药物和荧光团将与ACPP结合并与IR结合。在 AIM 2，我们将在原位询问IR如何在剂量依赖性中改变细胞外蛋白酶的活动 使用一组比值计aCPP的临时方式，并以特定的方式更优选地裂解不同的接头 蛋白酶。鉴定IR诱导的ACPP裂解将促进IR引导的药物递送的开发。在 摘要，ACPP放射敏化剂的交付具有一些创新的功能，可以保证临床前查询。 诊断，比例计aCPP允许对内在肿瘤微环境蛋白酶进行非侵入性成像 和IR的外在影响。在治疗上，ACPP的模块化体系结构允许不同 将放射敏化药物​​与具有不同蛋白酶接头的ACPP结合的类别。在一起，我们的研究 将测试如何探索ACPP以提高放射疗法的治疗比率。