公开(公告)号：US20040225119A1

公开(公告)日：2004-11-11

申请号：US10656826

申请日：2003-09-05

Applicant: Applera Corporation

Inventor： Scott C. Benson ,  Steven M. Menchen ,  Peter D. Theisen ,  Kevin M. Hennessey ,  Vergine C. Furniss ,  Joan D. Hauser

IPC: C12Q001/68 ,  C07H021/04 ,  C07D311/78

CPC classification number: C07H21/00 ,  C07C39/38 ,  C09B11/00 ,  C09B11/08 ,  C12Q1/6869

Abstract: A class of asymmetric monobenzoxanthene compounds useful as fluorescent dyes are disclosed having the structure 1 wherein Y1 and Y2 are individually hydroxyl, amino, imminium or oxygen, R1-R8 are hydrogen, fluorine, chlorine, alkyl alkene, alkyne, sulfonate, amino, amido, nitrile, alkoxy, linking group, and combinations thereof and R9 is acetylene, alkane, alkene, cyano, substituted phenyl and combinations thereof. The invention further includes novel intermediate compounds useful for the synthesis of asymmetric benzoxanthene compounds having the general structure 2 where substituents R3-R7 correspond to like-referenced substituents in the structure of described above, and Y2 is hydroxyl or amine. In another aspect, the invention includes methods for synthesizing the above dye compounds and intermediates. In yet another aspect, the present invention includes reagents labeled with the asymmetric benzoxanthene dye compounds, including deoxynucleotides, dideoxynucleotides, phosphoramidites, and polynucleotides. In an additional aspect, the invention includes methods utilizing such dye compounds and reagents including dideoxy polynucleotide sequencing and fragment analysis methods.

Abstract translation: 公开了一类用作荧光染料的不对称单苯并氧蒽化合物，其结构是其中Y 1和Y 2分别是羟基，氨基，亚氨基或氧，R 1 -R 8是氢，氟，氯，烷基烯烃，炔烃，磺酸酯，氨基，酰胺基， 腈，烷氧基，连接基团及其组合，R 9为乙炔，烷烃，烯烃，氰基，取代的苯基及其组合。 本发明还包括可用于合成具有通式结构的不对称苯并氧杂蒽化合物的新型中间体化合物，其中取代基R 3〜R 7对应于上述结构中的相同取代基，Y 2为羟基或胺。 另一方面，本发明包括合成上述染料化合物和中间体的方法。 在另一方面，本发明包括用不对称苯并氧杂蒽染料化合物标记的试剂，包括脱氧核苷酸，双脱氧核苷酸，亚磷酰胺和多核苷酸。 在另一方面，本发明包括利用这种染料化合物的方法和包括双脱氧多核苷酸测序和片段分析方法的试剂。