Synthesis of Novel Macrocycle-Containing Polymers as Protease Sensors

作为蛋白酶传感器的新型含大环聚合物的合成

• 批准号: 7677963
• 负责人: Mindy Levine
• 金额: $ 4.07万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7677963
• 关键词: Biological Assay; Biomedical Research; Cancer Detection; Cleaved cell; Detection; Development; Diagnosis; Disease; Dyes; Encapsulated; Energy Transfer; Fluorescence; Fluorescence Spectroscopy; Funding; Goals; Lead; Measures; Mediating; Methods; Monitor; Oligopeptides; Pathology; Peptide Hydrolases; Peptides; Polymers; Research; Residual state; System; United States National Institutes of Health; Vertebral column; base; copolymer; disease diagnosis; in vivo; monomer; novel; public health relevance; sensor; squaraine; tool

DESCRIPTION (provided by applicant): Amplifying fluorescent polymers developed in the Swager research group have shown a remarkable ability to act as sensitive and specific sensors for various analytes. This proposal seeks to harness this sensing ability of amplifying fluorescent polymers for the development of new assays for protease detection. To do so, the synthesis of an amplifying mixed co-polymer with some percentage of macrocyclic units is proposed. These macrocycles will encapsulate squaraine dyes in close proximity to the polymer backbone, and will contain oligopeptide linkers that can be cleaved by a target protease. In the absence of the target protease, the dye will be held in the macrocycle and efficient Dexter-type energy transfer will occur from the polymer backbone to the squaraine dye. Excitation of the polymer will result in selective amplified emission from the squaraine dye. Protease-mediated cleavage of the macrocycle will release the squaraine dye, leading to a quenching of the Dexter energy transfer between the polymer and the dye. The use of Dexter-type energy transfer will increase the sensitivity of the system, as the emission of the squaraine dye will occur on a completely dark background, without residual polymer fluorescence. The efficiency of energy transfer will be monitored using fluorescence spectroscopy to detect the emission of the squaraine dye, which occurs in the near-infrared spectral region. Fluorescence spectroscopy will further be used to measure the rate at which the target protease cleaves the macrocycle, as this will result in a quenching of the Dexter energy transfer between the polymer and a dye and a silencing of the dye's emission. PUBLIC HEALTH RELEVANCE This proposal describes the synthesis of a new class of amplifying fluorescent polymers that can be utilized for ultrasensitive protease detection. Protease activity has been implicated in a variety of diseases, including cancer; the detection of proteases in vivo is therefore critical both for the diagnosis of multiple diseases and for the development of new disease treatments. The proposed macrocyclic polymer can easily be adapted to detect virtually any target protease, as well as other biologically relevant targets.

描述(申请人提供)：Swager研究小组开发的放大荧光聚合物显示了作为各种分析物的敏感和特定传感器的非凡能力。这项提议试图利用放大荧光聚合物的这种传感能力来开发新的蛋白酶检测方法。为此，建议合成一种具有一定比例的大环单元的放大混合共聚物。这些大环将方酸染料包裹在聚合物主干附近，并将包含可被目标蛋白酶切割的寡肽连接物。在没有目标酶的情况下，染料将被保持在大环中，高效的右旋糖苷酸型能量将从聚合物主链转移到方酸染料。聚合物的激发将导致方酸染料的选择性放大发射。蛋白水解酶介导的大环裂解将释放方酸染料，导致聚合物和染料之间的右旋能量传递猝灭。由于方酸染料的发射将在完全黑暗的背景下发生，没有残留的聚合物荧光，因此使用右旋转移型能量转移将提高系统的灵敏度。将使用荧光光谱来监测能量转移的效率，以检测方块因染料的发射，该发射发生在近红外光谱区域。荧光光谱学将进一步用于测量目标蛋白酶裂解大环的速度，因为这将导致聚合物和染料之间的Dexter能量传递猝灭，并使染料的发射沉默。公共卫生相关性本提案描述了一种新型放大荧光聚合物的合成，该聚合物可用于超灵敏的蛋白酶检测。蛋白水解酶活性与多种疾病有关，包括癌症；因此，体内蛋白水解酶的检测对于多种疾病的诊断和新的疾病治疗方法的开发都是至关重要的。所提出的大环聚合物可以很容易地被改装成检测几乎任何目标蛋白酶，以及其他生物相关的目标。