BIND Chemistry for Imaging Intracellular Targets

用于细胞内靶标成像的 BIND 化学

基本信息

批准号：
8901980
负责人：
RALPH WEISSLEDER, MD, PHD
金额：
$ 10.62万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8901980
关键词：
A Mouse Affect Affinity Alder plant Amino Acids Appearance BODIPY Behavior Binding Binding Proteins Biochemical Biocompatible Biological Biological Models Cancer cell line Cell Culture Techniques Cell Nucleus Cells Centrosome Chemicals Chemistry Collaborations Combined Modality Therapy Coupled Data Detection Development Dose Drug Kinetics Drug resistance Enzymes Fluorescence Generic Drugs Genetic Engineering Goals Gold Housing Human Image Imagery In Vitro Interphase KRAS2 gene Kidney Kinetics Label Lead Libraries Life Ligands Liver Malignant Neoplasms Mass Spectrum Analysis Measures Methods Microscopic Mitosis Mitotic spindle Modeling Mus Organ PLK1 gene Pancreas Pancreatic Ductal Adenocarcinoma Pathway Analysis Patients Pharmaceutical Preparations Phosphotransferases Physicians Poly(ADP-ribose) Polymerases Positron-Emission Tomography Process Protein Binding Proteins Proteome Proteomics Reaction Reagent Recombinant Fusion Proteins Reporter Research Signal Transduction Small Interfering RNA Source Stable Isotope Labeling Technology Testing Therapeutic Toxic effect Treatment Efficacy Tumor Cell Line Work base cancer cell catalyst cellular imaging chemical synthesis comparative cycloaddition cytotoxic gemcitabine human PLK1 protein imaging agent imaging modality imaging probe in vivo in vivo imaging intravital imaging intravital microscopy molecular imaging mouse model novel protein complex research study response scaffold small molecule success therapeutic target tumor validation studies whole body imaging

项目摘要

There is a need to develop more sensitive, specific and robust strategies capable of imaging and quantitating therapeutic targets In cancers in vivo. To date, many biological targets and processes remain largely unobserved due to a) lack of affinity ligands, b) mismatches between target abundance and the amount of imaging agent required for target visualization, c) delivery barriers, d) unfavorable pharmacokinetics, e) high background signals, f) the unavailability of appropriate affinity ligands, and/or g) the sheer numbers of targets. The goal of this project is to use a recently developed bioorthogonal platform technology (known as BIND) for the rapid development of intracellular imaging agents such as polo-lil<e kinase (PLK1) and poly-ADP-ribose-polymerase 1 (PARP1), which are key intracellular cancer targets. BIND involves a {4+2} inverse Diels-Alder cycloaddition between fraA7s-cyclooctene (TCO) and a tetrazine (Tz), either attached to an imaging reporter or to a small molecule affinity ligand. BIND is biocompatible, has one of the most favorable reaction kinetics among bioorthogonal reactions, does not require catalysts, works well with small-molecule therapeutic drugs and can be combined with efficient amplifications strategies. This reaction can also be easily incorporated into development strategies for the vast number of existing small molecule affinity ligands for kinases and other intracellular targets. Cell permeable, small-molecule binders for PLK1 and PARP will be modified using TCO/Tz and imaged using complementary TCO/Tzimaging reporters. We will first validate the approach using cell based microscopic imaging, before performing in vivo experiments in mouse models of pancreatic ductal adenocarcinoma (PDAC). The specific aims are: 1) to develop a library of PLK-1 and PARP1-targeted BIND agents and characterize them biochemically, using live cell imaging and intravital microscopy; 2) to perform target-identification and network analysis using in vivo proteomics (SILAC); and 3) to explore the translational potential using 18F-labeled agents in gemcitabine and PLKHnhibitor/PARPinhibitor treated mice.

有必要制定能够成像和定量体内癌症的治疗靶标。迄今为止，许多生物学目标和过程由于a）缺乏亲和力配体，b）目标丰度不匹配，因此在很大程度上没有观察到目标可视化所需的成像剂量，c）输送障碍，d）不利药代动力学，e）高背景信号，f）适当亲和力配体的不可用和/或 g）目标数量的数量。该项目的目的是使用最近开发的生物正交平台技术（称为结合），以快速发展细胞内成像剂，例如polo-lil <e激酶（PLK1）和多-ADP-核糖 - 聚合酶1（PARP1），它是关键的细胞内癌靶标。绑定涉及a {4+2} Fraa7s-Cyclooclene（TCO）和四嗪（TZ）之间的逆Diels-Alder Cyclotition 附着在成像记者或小分子亲和力配体上。绑定是生物相容性的，具有生物正交反应中最有利的反应动力学，不需要催化剂，作品与小分子的治疗药物一起很好，可以与有效的放大策略结合使用。该反应也可以轻松地纳入大量现有的发展策略中激酶和其他细胞内靶标的小分子亲和力。细胞渗透，小分子 PLK1和PARP的粘合剂将使用TCO/TZ进行修改，并使用互补的TCO/Tzimaging成像记者。在进行体内之前，我们将首先使用基于细胞的微观成像验证该方法胰腺导管腺癌（PDAC）的小鼠模型中的实验。具体目的是：1）开发PLK-1和PARP1靶向绑定剂的库，并使用生物化学表征它们活细胞成像和插入式显微镜； 2）使用体内蛋白质组学（SILAC）； 3）使用18F标记的代理探索转化电位吉西他滨和Plkhnhibitor/parpin抑制剂治疗的小鼠。