US 7,888,136 B2 · Unsymmetrical cyanine dimer compounds and their application

Abstract

Embodiments of the present invention provide methods and nucleic acid reporter molecules for the detection of nucleic acid in a sample. The nucleic acid reporter molecule comprises two unsymmetrical cyanine monomer moieties, which may be the same or different, that are covalently attached by a linker comprising at least one aromatic, heteroaromatic, cyclic or heterocyclic moiety comprising 3-20 non-hydrogen atoms selected from the group consisting of O, N, S, P and C. The linker may be rigid, relatively flexible or some degree thereof. The unsymmetrical cyanine monomer moieties comprise a substituted or unsubstituted benzazolium moiety and a substituted or unsubstituted pyridinium or quinolinium moiety that is connected by a methine bridge that is monomethine, trimethine or pentamethine. The linkers form the cyanine dimer compounds by attaching to the pyridinium or quinolinium moiety of the monomer moieties. The present nucleic acid reporter molecules find utility in forming a nucleic acid-reporter molecule complex and detecting the nucleic acid. In particular, present nucleic acid reporter molecules with a rigid linker and monomer moieties with a monomethine bridge find utility in detecting RNA in the presence of DNA.

Claims35

Dependent claims

A nucleic acid reporter molecule that has a RNA/DNA ratio of fluorescence enhancement greater than about one wherein the nucleic acid reporter molecule comprises a first monomethine unsymmetrical cyanine monomer moiety, a second monomethine unsymmetrical cyanine monomer moiety and a linker that is

wherein R8 is a substituted heteroatom or an unsubstituted heteroatom;
R9 is a substituted heteroatom or an unsubstituted heteroatom;
R12 is a substituted heteroatom or an unsubstituted heteroatom;
each Z is —(CH2)g wherein g is 0 or 1;
and the first unsymmetrical cyanine monomer moiety is covalently attached to the linker;
and the second unsymmetrical cyanine monomer moiety is covalently attached to the linker.

3 more elements

A kit for detecting nucleic acid in a sample, wherein the kit comprises:

a nucleic acid reporter molecule that is according to the formula
wherein each R3 is independently hydrogen, unsubstituted C1-C6 alkyl, substituted C1-C6 alkyl, unsubstituted C1-C6 alkoxy, substituted C1-C6 alkoxy, unsubstituted fused benzene, substituted fused benzene, unsubstituted trifluoromethyl, substituted trifluoromethyl, unsubstituted halogen, substituted halogen, unsubstituted reactive group, substituted reactive group, unsubstituted carrier molecule, substituted carrier molecule, unsubstituted solid support or substituted solid support;
each R4 is independently a unsubstituted C1-C6 alkyl substituted C1-C6 alkyl unsubstituted reactive group, substituted reactive group, unsubstituted carrier molecule, substituted carrier molecule, unsubstituted solid support or substituted solid support;
X is O, S or CR6R7 wherein each R6 and R7 are independently a unsubstituted C1-C6 alkyl, substituted C1-C6 alkyl unsubstituted reactive group substituted reactive group unsubstituted carrier molecule, substituted carrier molecule, unsubstituted solid support, substituted solid support or R6 and R7 taken together form a 5- or 6-membered saturated ring;
j is 0, 1, or 2;
linker is a series of stable covalent bonds comprising 1-30 nonhydrogen atoms selected from the group consisting of C, N, O, S and P;
and,
K is substituted pyridinium, unsubstituted pyridinium, substituted quinolinium, or unsubstituted quinolinium;
wherein the linker comprises at least one aromatic, heteroaromatic, cyclic or heterocyclic moiety comprising 3-20 non-hydrogen atoms selected from the group consisting of O, N, S, P and C.

6 more elements

Dependencies are inferred from the claim text.

Unsymmetrical cyanine dimer compounds and their application

Life of this patent

Unsymmetrical cyanine dimer compounds and their application

Life of this patent

Abstract

Claims35