4,7-dichlororhodamine dyes useful as molecular probes

C - Chemistry – Metallurgy – 09 – B

Patent

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C09B 11/24 (2006.01) C07D 221/12 (2006.01) C07D 491/147 (2006.01) C07F 9/6558 (2006.01) C07H 21/00 (2006.01) C09B 11/28 (2006.01) C09K 11/06 (2006.01) C12Q 1/68 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2367868

A set of 4,7-dichlororhodamine compounds useful as fluorescent dyes are disclosed having the structures (I) and (VI) wherein R1-R6 are hydrogen, fluorine, chlorine, lower alkyl, lower alkene, lower alkyne, sulfonate, sulfone, amino, amido, nitrile, lower alkoxy, linking group, or, when taken together, R1 and R6 is benzo, or, when taken together, R4 and R5 is benzo; R7-R10, R12-R16 and R18 may be hydrogen, fluorine, chlorine, lower alkyl, lower alkene, lower alkyne, sulfonate, sulfone, amino, amido, nitrile, lower alkoxy, linking group; R11 and R17 may be hydrogen, lower alkyl, lower alkene, lower alkyne, phenyl, aryl, linking group; Y1-Y4 are hydrogen, lower alkyl, or cycloalkyl, or, when taken together, Y1 and R2, Y2 and R1, Y3 and R3, and/or Y4 and R4 is propano, ethano, or substituted forms thereof; and X1-X3 taken separately are hydrogen, chlorine, fluorine, lower alkyl, carboxylate, sulfonate, hydroxymethyl, and linking group, or any combinations thereof. In another aspect, the invention includes reagents labeled with the 4,7-dichlororhodamine dye compounds, including deoxynucleotides, dideoxynucleotides, and polynucleotides. In an additional aspect, the invention includes methods utilizing such dye compounds and reagents including dideoxy polynucleotide sequencing and fragment analysis methods.

L'invention porte sur un ensemble de composés 4,7-dichlororhodamine utilisés comme colorants fluorescents et ayant les structures (I) et (VI) dans lesquelles R1-R6 représentent hydrogène, fluor, chlore, alkyle inférieur, alcène inférieur, alkyne inférieur, sulfonate, sulfone, amino, amido, nitrile, alkoxy inférieur, un groupe de liaison ou, réunis, R1 et R6 représentent benzo, ou, réunis, R4 et R5 représentent benzo; R7-R10, R12-R16 et R18 peuvent être hydrogène, fluor, chlore, alkyle inférieur, alcène inférieur, alkyne inférieur, sulfonate, sulfone, amino, amido, nitrile, alcoxy inférieur, un groupe de liaison; R11 et R17 peuvent être hydrogène, alkyle inférieur, alcène inférieur, alkyne inférieur, phényle, aryle, un groupe de liaison; Y1-Y4 représentent hydrogène, alkyle inférieur ou cycloalkyle ou, réunis, Y1 et R2, Y2 et R1 Y3 and R3, et/ou Y4 et R4 représentent propano, éthano, ou des formes substituées de ceux-ci; et X1-X3, pris séparément, représentent hydrogène, chlore, fluor, alkyle inférieur, carboxylate, sulfonate, hydroxyméthyle, et un groupe de liaison, ou des combinations quelconques de ceux-ci. En variante, cette invention porte sur des réactifs marqués avec les composés colorants 4,7-dichlororhodamine, incluant des désoxynucléotides, didésoxynucléotides, et des polynucléotides. Dans une autre variante, l'invention porte sur des procédés utilisant ces composés colorants et ces réactifs incluant le séquençage des didésoxypolynucléotides et sur des procédés d'analyse de leurs fragments.

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