Novel Extended Red-Shifted Rhodamine Dyes

描述（由申请人提供）：在电磁波谱的长可见/近红外（NIR）区域工作的荧光染料是分子探针、荧光配体、酶底物、成像造影剂和一般示踪剂的优良报告组分。在常用的短波长和中波长染料如香豆素、荧光素和罗丹明的现有应用中，测定或组织组分可严重干扰含有这些染料的示踪剂的荧光性质。例如，在细胞色素P450（CYP 450）测定中，酶本身可以完全淬灭这些染料的报告信号。较长波长/ NIR染料目前供应非常短缺，并且那些商业上可获得的染料非常昂贵或仅在限制性许可协议下可获得。这项工作的主要重点是发现新的，长波长的近红外染料，设计用于高通量筛选（HTS）荧光偏振（FP）和荧光共振能量转移（FRET）生物测定新的药物和农业化学品发现研究。该染料还可用于生物医学体内成像、免疫荧光显微术和荧光激活细胞分选和计数。将开发新的合成化学途径，这将允许高纯度染料的多克规模合成，这将导致新产品以比通常具有设计不良的化学合成途径的现有染料更低的成本更广泛地提供给研究和科学界。第一阶段有五个具体目标： 1. 72种新型扩展磺酰罗丹明染料的液相化合物库的设计与合成2.薄层色谱平行纯化及新型染料的表征3.新染料的光物理表征4.在模型荧光偏振（FP）研究中评估新染料的性能5.在荧光共振能量转移（FRET）研究中评估新染料的性能。 一个成功的第一阶段研究将随后进行第二阶段研究，涉及规模扩大的合成和纯化的染料与商业潜力。这些染料也将作为体内光学成像的组分进行测试，并将组装含有新染料的各种FP和FRET HTS检测试剂盒。

DESCRIPTION (provided by applicant): Fluorescent dyes that operate in the long visible / near infra-red (NIR) region of the electromagnetic spectrum are excellent reporter components of molecular probes, fluorescent ligands, enzyme substrates, imaging contrast agents and general tracers. In existing applications of commonly used short and medium wavelength dyes such as coumarins, fluoresceins and rhodamines, assay or tissue components can severely interfere with the fluorescence properties of tracers containing these dyes. For example, in cytochrome P450 (CYP) assays, the enzyme itself can completely quench the reporting signal of such dyes. Longer wavelength / NIR dyes are in very short supply at the present time and those that are commercially available are very expensive or are only available under restrictive license agreements. The main focus of this work is to discover novel, long wavelength near-infrared dyes that are designed for use in high throughput screening (HTS) fluorescent polarization (FP) and fluorescence resonance energy transfer (FRET) biological assays for new drug and agrochemical discovery research. The dyes should also find uses in biomedical in vivo imaging, immunofluorescence microscopy, and fluorescence activated cell sorting and counting. Novel synthetic chemistry pathways will be developed that will allow multi-gram scale synthesis of the dyes in high purity that will result in new products that will be more widely available to the research and scientific community at a lower cost than existing dyes which generally have poorly designed chemical synthetic pathways. There are five specific aims in phase I: 1. Design and Synthesis of a Solution Phase Compound Library of 72 Novel Extended Sulforhodamine Dyes 2. Parallel Purification by Thin Layer Chromatography and Characterization of the Novel Dyes 3. Photophysical Characterization of the New Dyes 4. Assessment of the Performance of the New Dyes in a Model Fluorescence Polarization (FP) Study 5. Assessment of the Performance of the New Dyes in a Fluorescence Resonance Energy Transfer (FRET) Study. A successful phase I study will be followed by a phase II study that involves scale up syntheses and purification of dyes with commercial potential. The dyes will also be tested as components for in vivo optical imaging and a variety of FP and FRET HTS assay kits will be assembled containing the new dyes.