Energy transfer dyes comprising donor amp; acceptor dye moieties linked via an aryl/heteroaryl containing bridge, amp; their use as labels in biological systems

摘要

Energy transfer dyes, their preparation, and their use as labels in biological systems is disclosed. The dyes are preferably in the form of cassettes which enable their attachment to a variety of biological materials. The dyes and the reagents that can be made from them offer a wide variety of fluorescent labels with large Stokes' shifts enabling their use in a variety of fluorescence applications over a wide range of the visible spectrum. The energy transfer dyes may be of the formula:BR BR DSB1/SB- LSB1/SB-A-LSB2/SB- DSB2/SBBR BR whereinBR SL LIDSB1/SB is a first dye moiety suitable as an acceptor or donor in an energy transfer arrangement; LIDSB2/SB is a second dye moiety suitable as a donor or acceptor in an energy transfer arrangement with the first dye; /SL ```A comprises 5-20 linked atoms, wherein the linked atoms contain at least one aryl or heteroaryl group and wherein the atoms are independently selected from the group consisting of carbon, nitrogen, sulfur, oxygen, phosphorus;BR ```LSB1/SB and LSB2/SB comprise a group of linked atoms and LSB2/SB is adapted for attaching to a biological target. The following are specifically claimed 5-FAM-Phe-5-REG-11-ddUTP, 5-FAM-Phe-5-REG-11-ddCTP, 5-FAM-Phe-5-REG-11-ddATP, 5-FAM-Phe-5-REG-11-ddGTP, 5-FAM-Phe-5-ROX-11-ddCTP, 5-FAM-Phe-5-ROX-11-ddUTP, 5-FAM-Phe-5-ROX-11-ddATP, 5-FAM-Phe-5-ROX-11-ddGTP, 5-FAM-Phe-5-TAMARA-11-ddGTP, 5-FAM-Phe-5-TAMARA-11-ddATP, 5-FAM-Phe-5-TAMARA-11-ddCTP, 5-FAM-Phe-5-TAMARA-11-ddUTP, 5-FAM-Phe-5-R110-11-ddATP, 5-FAM-Phe-5-R110-11-ddUTP, 5-FAM-Phe-5-R110-11-ddGTP, and 5-FAM-Phe-5-R110-11-ddCTP. An energy transfer cassette may be of the formula: EMI ID=1.1 HE=19 WI=43 LX=1009 LY=1917 TI=CF PC```wherein FSP1/SP is a first dye suitable as an acceptor or donor in an energy transfer arrangement;BR ```FSP2/SP is a second dye that is suitable as a donor or acceptor in an energy transfer arrangement with the first dye;BR ```B is one or more alkyl groups containing from 1 to 30 atoms, wherein said atoms comprise one or more -NH-, -O-, -CH=CH-, -C I C- and phenylenyl groups, and wherein said atoms are selected from the group consisting of CSB1-4/SB linear or branched alkyl, hydroxyl, halo, or phenyl;BR ```MSP3/SP comprises hydrogen or a reactive or functional group adapted for attaching the energy transfer dye to a target material; andBR ```D comprises an atom or group adapted for attaching FSP2/SP to the linker group B.