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

• 批准号: 10112677
• 负责人: Clifford Berkman
• 金额: $ 22.48万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-14 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10112677
• 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

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.

项目摘要/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取消器的上皮中表示。除了这种独特的表达 前列腺癌已导致成像和治疗剂的靶向递送（内皮） PSMA在前列腺和非遗传性肿瘤的新生血管中的表达提供了更广泛的表达 调节血管化并改善药物输送的机会。 我们最近证明，在小分子PSMA靶向的小分子中添加了相册的图案 药物会大大减慢清除率，并允许注射剂量的肿瘤吸收近50％。我们的 磷脂酰胺靶向分子有选择地靶向PSMA的活动位置，并迅速穿透前列腺 肿瘤细胞，内化到内体/溶酶体。此外，我们的Phos-AM链接系统可以迅速释放 内体pH时的货物，但在物理上是稳定的。基于这些关键成就，我们目前是 定位于开发第一个有效的PSMA靶向SMDC。 但是，对于成功开发和优化PSMA靶向的关键问题仍然存在 毒品偶联，“是在肿瘤和非目标时间直接相关的PSMA靶向偶体的摄取 他们的货物释放？ （PSMA-NIR-RFP）可以使用双重阳性细胞和肿瘤中的摄取和货物释放 频道在体内成像。我们将通过追求以下特定目标来检验我们的假设。体内 探针的性能将使用新型多细胞类型的3D生物打印前列腺肿瘤模型进行评估 准确代表PSMA（+）原发性肿瘤脉管系统，可以在异种移植小鼠模型中进行。 AIM＃1：确定体外时空亚细胞的积累，并从中释放货物 PSMA（+）前列腺肿瘤细胞和肿瘤相关的脉管系统中的PSMA-NIR-RFP。这是我们的断言 PSMA-NIR-RFP可以在内体和溶酶体中报告摄取，贩运和货物释放 PSMA（+）细胞通过pH触发的染料释放。 AIM＃2：确定目标和非PSMA靶向共轭物的货物释放程度 PSMA（+）肿瘤 - Xenograpt小鼠模型中的目标时间。我们期望肿瘤数量 可以使用PSMA-NIR-RFP与特定的吸收和受控货物释放一起评估 双通道在体内成像。 拟议工作的最重要结果是将开发PSMA-NIR-RFP，并且 用于建模并报告体内吸收和货物的货物释放，这是 随后可以应用于PSMA和其他靶向的药物偶联的PSMA和其他生物标志物的优化。