Near-IR Ratiometric Fluorescence Probes for Assessing Cargo Delivery to Prostate Tumors

用于评估前列腺肿瘤的货物输送的近红外比率荧光探针

基本信息

批准号：
10328982
负责人：
Clifford Berkman
金额：
$ 17.27万
依托单位：
WASHINGTON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-14 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10328982
关键词：
3-Dimensional Achievement Active Sites Address Albumins Antigen Targeting Binding Biological Markers Castration Cell model Cells Clinic Companions Complex Cytotoxic agent Development Disease Dose Drug Delivery Systems Drug Targeting Drug toxicity Dyes Endosomes Endothelium Enzymes Epithelial FOLH1 gene Fluorescence Goals Human Image In Vitro Lysosomes Malignant neoplasm of prostate Modeling Outcome Performance Pharmaceutical Preparations Physiological Positioning Attribute Positron-Emission Tomography Primary Neoplasm Process Prostatic Prostatic Neoplasms Reporting Resistance Site System Testing Therapeutic Therapeutic Agents Tissues Translations Tumor-Associated Vasculature Urea Vascularization Work Xenograft procedure base bioprinting cell type chemotherapeutic agent design drug candidate drug discovery drug efficacy drug release kinetics efficacy testing expectation imaging agent improved in vivo in vivo imaging individualized medicine mouse model neoplastic cell neovasculature novel novel therapeutics phosphoramidate predictive modeling prostate cancer cell ratiometric scaffold small molecule spatiotemporal targeted agent targeted biomarker targeted delivery trafficking tumor tumor xenograft uptake

项目摘要

PROJECT SUMMARY/ABSTRACT Prostate-specific membrane antigen (PSMA) is the hallmark enzyme-biomarker for prostate cancer because it is expressed in the epithelium of nearly all prostate cancers. In addition to this unique expression in prostate cancer, which has led to targeted delivery of imaging and therapeutic agents, the endothelial expression of PSMA on the neovasculature of both prostatic and non-prostatic tumors offers a broader opportunity to modulate the vascularization and improve drug delivery. We recently demonstrated that the addition of an albumin-binding motif to small-molecule PSMA-targeted agents dramatically slows clearance and allows for tumor uptake of nearly 50% of an injected dose. Our phosphoramidate targeting molecules selectively target the active-site of PSMA, and rapidly penetrate prostate tumor cells, internalizing to endosomes/lysosomes. In addition, our Phos-Am linker system can rapidly release cargo at endosomal pH but is stable at physiologically pH. Based on these key achievements, we are currently positioned to develop the first effective PSMA-targeted SMDC. However, a key question remains for the successful development and optimization of PSMA-targeted drug-conjugates, “is uptake of PSMA-targeted conjugates in tumor and non-target tissues directly correlated with their cargo release?” Our hypothesis is that a PSMA-targeted near-IR ratiometric fluorescence probe (PSMA-NIR-RFP) can report on both uptake and cargo-release in PSMA-positive cells and tumors using dual- channel in vivo imaging. We will test our hypothesis by pursuing the following specific aims. The in vivo performance of the probe will be evaluated using a novel multi-cell type, 3D bioprinted prostatic tumor model accurately represents PSMA(+) primary tumor vasculature and can be employed in xenograft mouse models. Aim #1: Determine the in vitro spatiotemporal subcellular accumulation and release of cargo from a PSMA-NIR-RFP in PSMA(+) prostate tumor cells and tumor-associated vasculature. It is our assertion that a PSMA-NIR-RFP can report on the uptake, trafficking, and cargo release in endosomes and lysosomes of PSMA(+) cells, through their pH-triggered dye release. Aim #2: Determine the extent of cargo-release from PSMA-targeted conjugates in target and non- target tissues in PSMA(+) tumor-xenograft mouse model. It is our expectation that quantification of tumor- specific uptake and controlled cargo-release can be assessed using a PSMA-NIR-RFP in combination with dual-channel in vivo imaging. The most important outcome of the proposed work is that a PSMA-NIR-RFP will be developed and employed to model and report on the in vivo uptake and cargo-release of PSMA-targeted conjugates, which can be subsequently applied to the optimization of PSMA and other biomarker-targeted drug-conjugates.

项目概要/摘要前列腺特异性膜抗原 (PSMA) 是前列腺癌的标志性酶生物标志物因为它在几乎所有前列腺癌的上皮细胞中都有表达。前列腺癌，这导致了成像和治疗剂的靶向递送，内皮细胞 PSMA 在前列腺和非前列腺肿瘤的新血管中的表达提供了更广泛的研究调节血管化和改善药物输送的机会。我们最近证明，在小分子 PSMA 靶向中添加白蛋白结合基序药物显着减慢清除速度，并允许肿瘤吸收注射剂量的近 50%。氨基磷酸酯靶向分子选择性靶向PSMA活性位点，快速渗透前列腺肿瘤细胞，内化到内体/溶酶体此外，我们的 Phos-Am 连接器系统可以快速释放。货物在内体 pH 值下稳定，但在生理 pH 值下稳定。基于这些关键成就，我们目前正在研究。定位于开发第一个有效的 PSMA 目标 SMDC。然而，针对 PSMA 的成功开发和优化仍然存在一个关键问题。药物缀合物，“肿瘤和非靶组织中 PSMA 靶向缀合物的摄取直接相关与他们的货物释放？”我们的假设是，一种针对 PSMA 的近红外比率荧光探针 (PSMA-NIR-RFP) 可以使用双-报告 PSMA 阳性细胞和肿瘤中的摄取和货物释放我们将通过追求以下特定目标来检验我们的假设。将使用新型多细胞类型 3D 生物打印前列腺肿瘤模型来评估探针的性能准确地代表 PSMA(+) 原发性肿瘤脉管系统，可用于异种移植小鼠模型。目标#1：确定体外时空亚细胞积累和释放的货物我们的断言是 PSMA(+) 前列腺肿瘤细胞和肿瘤相关脉管系统中的 PSMA-NIR-RFP。 PSMA-NIR-RFP 可以报告内体和溶酶体中的摄取、运输和货物释放 PSMA(+) 细胞通过 pH 值触发染料释放。目标#2：确定目标和非目标中 PSMA 靶向缀合物的货物释放程度我们期望对 PSMA(+) 肿瘤异种移植小鼠模型中的靶组织进行定量。可以使用 PSMA-NIR-RFP 结合双通道活体成像。拟议工作最重要的成果是将开发 PSMA-NIR-RFP，并用于模拟和报告 PSMA 靶向缀合物的体内摄取和货物释放，随后可应用于 PSMA 和其他生物标志物靶向药物偶联物的优化。