Dimers of unsymmetrical cyanine dyes containing pyridinium moieties

Claim 2

A compound of the formula: α--(CH═CH).sub.n --β--BRIDGE--γ--(CH═CH).sub.n' --δ where each of α and δ is independently a benzazolium ring moiety that is a benzoxazolium, benzothiazolium, benzimidazolium, 3,3-dialkylindolium heterocycle or a naphthazolium ring moiety that is a naphthoxazolium, naphthothiazolium, napthimidazolium or benzo-3,3-dialkylindolium heterocycle, each of the nitrogen atoms of which is substituted by alkyl with 1 to 6 carbons, which benzazolium ring moiety is linked to --(CH═CH) n -- or --(CH═CH) n' -- respectively through a 2-position of the ring moiety, wherein the remaining substituents of the ring moiety are independently hydrogen, halogen, trifluoromethyl, alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; n and n' are independently 0, 1 or 2; β is a first 2-methine-substituted pyridinium: ##STR17## or a first 4-methine-substituted pyridinium: ##STR18## that is linked to --(CH═CH) n -- through an exocyclic methine residue of β, which pyridinium is optionally substituted by hydrogen, halogen, trifluoromethyl, alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; BRIDGE is an aliphatic chain containing a backbone of 4 to 19 methylene groups (--CH 2 --), which is optionally interspersed at one or more intervals with a heteroatom, each of which is independently N, O or S, wherein each N heteroatom is additionally substituted by H, alkyl or two alkyl groups with 1 to 6 carbons, which alkyl substituents may be the same or different, provided that any heteroatom is separated from another heteroatom by at least 2 methylene groups, wherein one methylene terminus of BRIDGE is attached to a nitrogen atom of β and another methylene terminus of BRIDGE is attached to a nitrogen atom of γ; γ is a second 2-methine-substituted pyridinium or a second 4-methine-substituted pyridinium or a 2-methine-substituted quinolinium: ##STR19## or a 4-methine-substituted quinolinium: ##STR20## that is linked to --(CH═CH) n' -- through an exocyclic methine residue of γ, which pyridinium or quinolinium is optionally substituted by hydrogen, halogen, trifluoromethyl, alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; and Ψ is a counterion or counterions that neutralize positive charges on the compound.

Claim 3

A compound, as claimed in claim 1, where α and δ are each benzothiazolium or benzoxazolium moieties.

Claim 4

A compound, as claimed in claim 2, where β and γ are each pyridinium moieties.

Claim 5

A compound, as claimed in claim 2, in which n and n' are both 0.

Claim 6

A compound, as claimed in claim 2, in which n and n' are both 1.

Claim 7

A compound, as claimed in claim 1, where β and γ are each pyridinium moieties.

Claim 8

A compound, as claimed in claim 6, where α and δ are each benzoxazolium moieties.

Claim 9

A compound, as claimed in claim 6, where α and δ are each benzothiazolium moieties.

Claim 10

A compound, as claimed in claim 1, where BRIDGE has the formula: --(CH.sub.2).sub.p --J--(CH.sub.2).sub.q --K--(CH.sub.2).sub.r --L--(CH.sub.2).sub.s -- where s is 2-12 and p, q and r are independently 0 or 2-6, provided that if any of p, q or r is 0 then the adjacent moiety J, K or L is absent; and wherein J, K and L are independently O, S, imino (--NH--), immonium (--NH 2 + --), alkylimino (--NR 1 --), alkylimmonium (--NHR 1 + --) or dialkylimmonium (--NR 1 R 2+ --), wherein R 1 and R 2 independently have 1 to 6 carbons.

Claim 11

A compound, as claimed in claim 9, where α and δ are each benzothiazolium or benzoxazolium moieties.

Claim 12

A compound, as claimed in claim 10, where β and γ are each pyridinium moieties.

Claim 13

A compound, as claimed in claim 11, in which n and n' are both 0.

Claim 14

A compound, as claimed in claim 11, in which n and n' are both 1.

Claim 15

A compound, as claimed in claim 9, where β and γ are each pyridinium moieties.

Claim 16

A compound, as claimed in claim 14, where α and δ are each benzoxazolium moieties.

Claim 17

A compound, as claimed in claim 14, where α and δ are each benzothiazolium moieties.

Claim 18

A compound, as claimed in claim 9, where at least one of J, K and L is imino (--NH--), immonium (--NH 2 + --), alkylimino (--NR 1 --), alkylimmonium (--NHR 1 + --) or dialkylimmonium (--NR 1 R 2+ --).

Claim 19

A compound, as claimed in claim 9, where at least two of J, K and L are dimethylammonium.

Claim 20

A compound of the formula: ##STR21## where X is S or O; R 3 is C 1 to C 6 alkyl; Ψ is a counterion or counterions that neutralize positive charges on the compound; and wherein n and n' are independently 0, 1 or 2.

Claim 21

A compound of the formula: ##STR21## where X is S or O; R 3 is C 1 to C 6 alkyl; Ψ is a counterion or counterions that neutralize positive charges on the compound; and wherein n and n' are independently 0, 1 or 2.

Claim 22

A compound, as claimed in claim 19, wherein X is S or O, R 3 is methyl and n and n' are both 0.

Claim 23

A compound, as claimed in claim 19, wherein X is S or O, R 3 is methyl and n and n' are both 1.

Claim 24

A compound comprising a nucleic acid polymer bound to one or more dye molecules of the formula: α--(CH═CH).sub.n --β--BRIDGE--γ--(CH═CH).sub.n' --δ where each of α and δ is independently a benzazolium ring moiety that is a benzoxazolium, benzothiazolium, benzimidazolium, 3,3-dialkylindolium heterocycle or a naphthazolium ring moiety that is a naphthoxazolium, naphthothiazolium, napthimidazolium or benzo-3,3-dialkylindolium heterocycle, each of the nitrogen atoms of which is substituted by alkyl with 1 to 6 carbons, which benzazolium ring moiety is linked to --(CH═CH) n -- or --(CH═CH) n' -- respectively through a 2-position of the ring moiety, wherein the remaining substituents of the ring moiety are independently hydrogen, halogen, trifluoromethyl; alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; n and n' are independently 0, 1 or 2; β is a first 2-methine-substituted pyridinium: ##STR22## or a first 4-methine-substituted pyridinium: ##STR23## that is linked to --(CH═CH) n -- through an exocyclic methine residue of β, which pyridinium is optionally substituted by hydrogen, halogen, trifluoromethyl, alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; BRIDGE is an aliphatic chain containing a backbone of 4 to 19 methylene groups (--CH 2 --), which is optionally interspersed at one or more intervals with a heteroatom, each of which is independently N, O or S, wherein each N heteroatom is additionally substituted by H, alkyl or two alkyl groups with 1 to 6 carbons, which alkyl substituents may be the same or different, provided that any heteroatom is separated from another heteroatom by at least 2 methylene groups, wherein one methylene terminus of BRIDGE is attached to a nitrogen atom of β and another methylene terminus of BRIDGE is attached to a nitrogen atom of γ; γ is a second 2-methine-substituted pyridinium or a second 4-methine-substituted pyridinium or a 2-methine-substituted quinolinium: ##STR24## or a 4-methine-substituted quinolinium: ##STR25## that is linked to --(CH═CH) n' -- through an exocyclic methine residue of γ, which pyridinium or quinolinium is optionally substituted by hydrogen, halogen, trifluoromethyl, alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; and Ψ is a counterion or counterions that neutralize positive charges on the compound such that the fluorescence of said dye molecules when bound to the nucleic acid polymer is enhanced.

Claim 25

A compound comprising a nucleic acid polymer bound to one or more dye molecules of the formula: α--(CH═CH).sub.n --β--BRIDGE--γ--(CH═CH).sub.n' --δ where each of α and δ is independently a benzazolium ring moiety that is a benzoxazolium, benzothiazolium, benzimidazolium, 3,3-dialkylindolium heterocycle or a naphthazolium ring moiety that is a naphthoxazolium, naphthothiazolium, napthimidazolium or benzo-3,3-dialkylindolium heterocycle, each of the nitrogen atoms of which is substituted by alkyl with 1 to 6 carbons, which benzazolium ring moiety is linked to --(CH═CH) n -- or --(CH═CH) n' -- respectively through a 2-position of the ring moiety, wherein the remaining substituents of the ring moiety are independently hydrogen, halogen, trifluoromethyl; alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; n and n' are independently 0, 1 or 2; β is a first 2-methine-substituted pyridinium: ##STR22## or a first 4-methine-substituted pyridinium: ##STR23## that is linked to --(CH═CH) n -- through an exocyclic methine residue of β, which pyridinium is optionally substituted by hydrogen, halogen, trifluoromethyl, alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; BRIDGE is an aliphatic chain containing a backbone of 4 to 19 methylene groups (--CH 2 --), which is optionally interspersed at one or more intervals with a heteroatom, each of which is independently N, O or S, wherein each N heteroatom is additionally substituted by H, alkyl or two alkyl groups with 1 to 6 carbons, which alkyl substituents may be the same or different, provided that any heteroatom is separated from another heteroatom by at least 2 methylene groups, wherein one methylene terminus of BRIDGE is attached to a nitrogen atom of β and another methylene terminus of BRIDGE is attached to a nitrogen atom of γ; γ is a second 2-methine-substituted pyridinium or a second 4-methine-substituted pyridinium or a 2-methine-substituted quinolinium: ##STR24## or a 4-methine-substituted quinolinium: ##STR25## that is linked to --(CH═CH) n' -- through an exocyclic methine residue of γ, which pyridinium or quinolinium is optionally substituted by hydrogen, halogen, trifluoromethyl, alkyl, alkoxy, amino, mono- or dialkylamino, the alkyl portions of which contain 1 to 6 carbons; and Ψ is a counterion or counterions that neutralize positive charges on the compound such that the fluorescence of said dye molecules when bound to the nucleic acid polymer is enhanced.

Claim 26

A compound, as claimed in claim 22, wherein BRIDGE has the formula: --(CH.sub.2).sub.p --J--(CH.sub.2).sub.q --K--(CH.sub.2).sub.r --L--(CH.sub.2).sub.s -- where s is 2-12, and p, q and r are independently 0 or 2-6, provided that if any of p, q or r is 0 then the adjacent moiety J, K or L is absent; and wherein J, K and L are independently O, S, imino (--NH--), immonium (--NH 2 + --), alkylimino (--NR 1 --), alkylimmonium (--NHR 1 + --) or dialkylimmonium (--NR 1 R 2+ --), wherein R 1 and R 2 independently have 1 to 6 carbons.

Claim 27

A compound, as claimed in claim 22, wherein the dye has the formula: ##STR26## where X is S or O; R 3 is C 1 to C 6 alkyl; and wherein n and n' are independently 0, 1 or 2.

Claim 28

A compound, as claimed in claim 22, where the nucleic acid polymer is DNA.

Claim 29

A compound, as claimed in claim 22, where the nucleic acid polymer is RNA.

Claim 30

A compound, as claimed in claim 22, where the nucleic acid polymer is intracellular.

Claim 31

A compound, as claimed in claim 22, where the fluorescence enhancement of the dye molecules when bound to the nucleic acid is greater than about 50-fold.

Claim 32

A compound, as claimed in claim 22, containing at least two different dye molecules having different spectral properties, where a first dye molecule has an emission maximum that substantially overlaps an excitation maximum of a second dye molecule.

Claim 33

A compound, as claimed in claim 29, where one of the different dye molecules are selected from the group consisting of: ##STR27## where X is S or O; R 3 is C 1 to C 6 alkyl; and wherein n and n' are independently 0, 1 or 2.