Preparation of New Long Wavelength Fluorescent Dyes

描述(由申请人提供)：在电磁光谱的长可见/近红外(NIR)区域工作的新型荧光染料已被证明是这种靶向分子探针和示踪剂的优秀报告成分。这类染料目前供不应求，那些商业上可以获得的染料非常昂贵，或者只有在严格限制的许可协议下才能获得。许多现有的染料具有不受欢迎的性质，例如相对较高的非特异性结合和聚集的倾向，并且部分由于未优化的合成方法而昂贵得令人望而却步。这一应用的主要焦点是通过新的组合化学技术发现新型的长波长近红外扩展方酸染料，这些组合化学技术旨在用于高通量筛选、荧光偏振、生物医学成像和其他应用技术，如免疫荧光显微镜和荧光激活细胞分类和计数。将开发新的固相合成路径，使高纯度染料能够进行克级合成，并将产生更广泛地供研究和科学界使用的产品。 具体目标有8个：具体目标1：利用第一阶段开发的合成方法，进一步合成240种新型长波长/近红外扩展方块因染料。具体目标2：新染料的小规模提纯和表征。具体目标3：从目标1和2放大合成20种精选的菁染料。目标4：荧光偏振(FP)研究。具体目标5：评估新染料在荧光共振能量转移(FRET)研究中的性能。具体目标6：新染料的免疫组织化学研究。具体目标7：染料稳定性研究。具体目标8：蛋白质标记效率研究。

DESCRIPTION (provided by applicant): Novel fluorescent dyes that operate in the long visible / near infra-red (NIR) region of the electromagnetic spectrum have been shown to act as excellent reporter components of such targeting molecular probes and tracers. Such dyes are in very short supply at the present time and those that are commercially available are very expensive or are only available under severely restrictive license agreements. Many of the available dyes have undesirable properties such as relatively high non-specific binding and a tendency to aggregate and are prohibitively expensive due in part to unoptimized synthetic methods. The main focus of this application is to discover novel, long wavelength near-infrared extended squaraine dyes by novel combinatorial chemistry techniques that are designed for use in high throughput screening, fluorescence polarization, biomedical imaging and other applicable techniques such as immunofluorescence microscopy and fluorescence activated cell sorting and counting. Novel solid phase synthetic pathways will be developed that will allow gram-scale synthesis of the dyes in high purity and will result in products that will be more widely available to the research and scientific community. There are 8 specific aims: Specific Aim 1: To Synthesize a Further 240 Novel Longer Wavelength / Near-Infrared Extended Squaraine Dyes By Using The Synthetic Methodologies Developed In Phase I. Specific Aim 2: Small Scale Purification and Characterization of the New Dyes. Specific Aim 3: Scale up Synthesis of 20 Selected Cyanine Dyes from Aims 1 and 2. Aim 4: Fluorescence Polarization (FP) Studies. Specific Aim 5: To Assess the Performance of the New Dyes in Fluorescence Resonance Energy Transfer (FRET) Studies. Specific Aim 6: Immunohistochemical Studies with the New Dyes. Specific Aim 7: Dye Stability Studies. Specific Aim 8: Protein Labeling Efficiency Studies.