A New Class of Water Soluble Cyanine Dyes

描述（由申请人提供）：长波长和近红外（NIR）染料已被证明在靶向分子探针和示踪剂中充当优异的报告组分。许多目前可用的荧光染料和探针具有不期望的性质，例如相对高的非特异性蛋白质结合和聚集倾向，并且部分由于未优化的合成方法而过于昂贵。这项工作的主要重点是发现新的，水溶性的，长波长的近红外菁染料，设计用于高通量筛选，成像和其他适用的技术，如免疫荧光显微镜，荧光共振能量转移和荧光激活细胞分选和计数。将开发新的合成溶液和固相化学途径，这将允许以克级合成高纯度的染料，并将产生更好的产品，这些产品将以更低的成本更广泛地提供给研究和科学界。本研究的主要目的有10个：（1）利用第一阶段的合成方法，制备120个新的苯并冠醚长波长菁染料的组合化学库。(2)利用第一阶段开发的合成方法制备180种新的聚乙二醇长波长菁染料的组合化学库。(3)新染料的纯化和表征。(4)染料稳定性和溶解度研究。(5)非特异性结合研究。(6)15种菁染料的放大合成。(7)荧光偏振（FP）研究。(8)评价新染料在荧光共振能量转移（FRET）研究中的性能。(9)新染料的免疫组织化学研究。(10)替代反应性手柄的合成。

DESCRIPTION (provided by applicant): Long wavelength and near infrared (NIR) dyes have been shown to act as excellent reporter components in targeting molecular probes and tracers. Many of the currently available fluorescent dyes and probes have undesirable properties such as relatively high non-specific protein binding and a tendency to aggregate and are prohibitively expensive due in part to unoptimized synthetic methods. The main focus of this work is to discover novel, water soluble, long wavelength near-infrared cyanine dyes that are designed for use in high throughput screening, imaging and other applicable techniques such as immunofluorescence microscopy, fluorescence resonance energy transfer and fluorescence activated cell sorting and counting. Novel synthetic solution and solid phase chemistry pathways will be developed that will allow gram-scale synthesis of the dyes in high purity and will result in better products that will be more widely available at lower cost to the research and scientific community. There are 10 specific aims: (1) To Prepare a Combinatorial Chemistry Library of 120 New Benzocrown-ether Long Wavelength Cyanine Dyes Using the Synthetic Methodologies Developed in Phase I. (2) To Prepare a Combinatorial Chemistry Library of 180 New Polyethylene Glycol Long Wavelength Cyanine Dyes Using the Synthetic Methodologies Developed in Phase I. (3) Purification and Characterization of the New Dyes. (4) Dye Stability and Solubility Studies. (5) Non-Specific Binding Studies. (6) Scale up Synthesis of 15 Selected Cyanine Dyes. (7) Fluorescence Polarization (FP) Studies. (8) To Assess the Performance of the New Dyes in Fluorescence Resonance Energy Transfer (FRET) Studies. (9) Immunohistochemical Studies with the New Dyes. (10) Synthesis of Alternate Reactive Handles.