Nucleotide compounds including a rigid linker

C - Chemistry – Metallurgy – 07 – H

Patent

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C07H 21/00 (2006.01) C07H 19/06 (2006.01) C07H 19/16 (2006.01) C07H 19/20 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2344643

A nucleoside/tide compound having the structure: NUC-L-S-LB/LG, is described wherein NUC is a nucleoside/tide having a nucleobase portion B, L is a rigid linkage, S is a spacer; and LB/LG is a member of a linkage pair or a label. NUC is attached to L through B such that when B is a purine, L is attached to the 8-position of the purine, when B is 7-deazapurine, L is attached to the 7-position of the 7-deazapurine, and when B is pyrimidine, L is attached to the 5-position of the pyrimidine. In an important aspect of the invention, L has the structure (a) wherein each of n, o and p are integers ranging from 0 to 3, and the sum of n, o and p is at least 2, and each of W, X, Y and Z is selected from the group consisting of carbon and nitrogen. The invention further includes polynucleotide compounds comprising the nucleoside/tide, and primer extension methods utilizing the nucleoside/tide, particularly when used in combination with certain mutant polymerase enzymes.

L'invention concerne un composé nucléosidique/tidique ayant la structure ci-après: NUC-L-S-LB/LG, dans laquelle NUC représente un nucléoside/tide comportant une partie nucléobase B, L représente une liaison rigide, S représente un espaceur, et LB/LG représente un élément d'une paire de liaison ou d'un marqueur. NUC est attaché à L par B de manière que lorsque B représente une purine, L est attaché à la position 8 de la purine, que lorsque B représente une 7-déazapurine, L est attaché à la position 7 de la 7-déazapurine, et que lorsque B représente une pyrimidine, L est attaché à la position 5 de la pyrimidine. Selon un aspect important de l'invention, L a la structure (a) où n, o et p sont chacun des entiers compris entre 0 et 3, la somme de n, o et p étant au moins égale à 2, et W, X, Y et Z sont chacun sélectionnés dans le groupe constitué du carbone et de l'azote. L'invention concerne également des composés polynucléotidiques comprenant ledit nucléoside/tide, ainsi que des méthodes d'extension des amorces utilisant ce nucléoside/tide, notamment lorsqu'il est utilisé en association avec certaines polymérases mutantes.

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