US5582977A
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Dimers of unsymmetrical cyanine dyes
Assignee: Molecular Probes Inc
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Filing: 1994-01-06
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Granted: 1996-12-10
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Expires: 2014-01-06
Abstract
Abstract The invention relates to dimers of unsymmetrical cyanine dyes, typically dimers of benzthiazole or benzoxazole derivatives, that exhibit enhanced fluorescence on binding with DNA or RNA, The dimers generally have the formula: ##STR1## where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-6 carbons; X is O, S, or N--R 3 , where R 3 is H or an alkyl group having 1-6 carbons; Z is O, S, or N--R 4 , where R 4 is H or an alkyl group having 1-6 carbons; n and s, which may be the same or different, =0, 1, or 2; Y is HC═CH; and p, m, q, and r=0 or 1, such that p+m=1 and q+r=1; and where -BRIDGE- has the general formula: --(CH.sub.2).sub.α --[A.sup.1 --(CH.sub.2).sub.β --].sub.I [A 2 --(CH 2 ).sub.γ --] II A 3 --(CH 2 ).sub.δ -- where α, β, γ, and δ, which may be the same or different, are integers greater than 1 and less than 5; I and II, which may be the same or different, =0 or 1; and A 1 , A 2 , and A 3 , which may be the same or different, are independently O; S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or--(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons.
Inventors
- 👤 Stephen T. Yue
- 👤 Iain D. Johnson
- 👤 Richard P. Haugland
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 15:29
CPC Classifications
C; C09; C09B; C09B23/00; C09B23/02; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/52
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
278 words
A compound of the formula: ##STR8## where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-6 carbons; X is O, S, or N--R 3 , where R 3 is H or an alkyl group having 1-6 carbons; Z is O, S, or N--R 4 , where R 4 is H or an alkyl group having 1-6 carbons; n and s, which may be the same or different, =0, 1, or 2; Y is HC═CH; and p, m, q, and r=0 or 1, such that p+m=1 and q+r=1; and where -BRIDGE- has the general formula: --(CH.sub.2).sub.α --[A.sup.1 --(CH.sub.2).sub.β --].sub.I [A.sup.2 --(CH.sub.2).sub.γ --].sub.II A.sup.3 --(CH.sub.2).sub.δ -- where α, β, γ, and δ, which may be the same or different, are integers greater than 1 and less than 5; I and II, which may be the same or different, =0 or 1; A 1 and A 2 , which may be the same or different, are independently O; S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons, and A 3 is S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons.
Claim 2
Dependent Claim
278 words
A compound of the formula: ##STR8## where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-6 carbons; X is O, S, or N--R 3 , where R 3 is H or an alkyl group having 1-6 carbons; Z is O, S, or N--R 4 , where R 4 is H or an alkyl group having 1-6 carbons; n and s, which may be the same or different, =0, 1, or 2; Y is HC═CH; and p, m, q, and r=0 or 1, such that p+m=1 and q+r=1; and where -BRIDGE- has the general formula: --(CH.sub.2).sub.α --[A.sup.1 --(CH.sub.2).sub.β --].sub.I [A.sup.2 --(CH.sub.2).sub.γ --].sub.II A.sup.3 --(CH.sub.2).sub.δ -- where α, β, γ, and δ, which may be the same or different, are integers greater than 1 and less than 5; I and II, which may be the same or different, =0 or 1; A 1 and A 2 , which may be the same or different, are independently O; S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons, and A 3 is S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons.
Claim 3
Dependent Claim
40 words
A compound, as claimed in claim 1, where A 1 , A 2 and A 3 , which may be the same or different, are independently (CH 2 ).sub.μ, --(NR 5 )--, or --N + R 6 R 7 )--.
Claim 4
Dependent Claim
25 words
A compound, as claimed in claim 1, where X and Z are the same; R 1 and R 2 are the same; n=s and m=r.
Claim 5
Dependent Claim
14 words
A compound, as claimed in claim 1, where m and r are each 1.
Claim 6
Dependent Claim
22 words
A compound, as claimed in claim 1, where X and Z, which may be the same or different, are O or S.
Claim 7
Dependent Claim
14 words
A compound, as claimed in claim 1, where n is not equal to s.
Claim 8
Dependent Claim
72 words
A compound, as claimed in claim 1, where A 1 and A 3 , which may be the same or different, are independently --(NR 5 )-- where R 5 is H or an alkyl group having 1-2 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-2 carbons; and II=0.
Claim 9
Dependent Claim
134 words
A compound, as claimed in claim 1, where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-2 carbons; X and Z, which may be the same or different are O or S; m and r=1; in the BRIDGE formula α, β, γ, and δ, which may be the same or different, are 2 or 3; A 1 , A 2 , and A 3 , which may be the same or different, are (CH 2 ).sub.μ where μ=0; --(NR 5 )-- where R 5 is H or an alkyl group having 1-2 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-2 carbons.
Claim 10
Dependent Claim
119 words
A compound, as claimed in claim 3, where R 1 and R 2 are alkyl groups having 1-2 carbons; X and Z are O or S; m and r=1; in the BRIDGE formula α, β, γ, and δ, which may be the same or different, are 2 or 3; A 1 , A 2 , and A 3 , which may be the same or different, are (CH 2 ).sub.μ where μ=0; --(NR 5 )-- where R 5 is H or an alkyl group having 1-2 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-2 carbons.
Claim 11
Dependent Claim
235 words
A fluorescent compound comprising a nucleic acid polymer bound to a dye of the general formula: ##STR9## where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-6 carbons; X is O, S, or N--R 3 , where R 3 is H or an alkyl group having 1-6 carbons; Z is O, S, or N--R 4 , where R 4 is H or an alkyl group having 1-6 carbons; n and s, which may be the same or different, =0, 1, or 2; Y is HC═CH; and p, m, q, and r=0 or 1, such that p+m=1 and q+r=1; and where -BRIDGE- has the general formula: --(CH.sub.2).sub.α --[A.sup.1 --(CH.sub.2).sub.β --].sub.I [A.sup.2 --(CH.sub.2).sub.γ --].sub.II A.sup.3 --(CH.sub.2).sub.δ -- where α, β, γ, and δ, which may be the same or different, are integers greater than 1 and less than 5; I and II, which may be the same or different, =0 or 1; and A 1 , A 2 , and A 3 , which may be the same or different, are independently O; S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons.
Claim 12
Dependent Claim
235 words
A fluorescent compound comprising a nucleic acid polymer bound to a dye of the general formula: ##STR9## where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-6 carbons; X is O, S, or N--R 3 , where R 3 is H or an alkyl group having 1-6 carbons; Z is O, S, or N--R 4 , where R 4 is H or an alkyl group having 1-6 carbons; n and s, which may be the same or different, =0, 1, or 2; Y is HC═CH; and p, m, q, and r=0 or 1, such that p+m=1 and q+r=1; and where -BRIDGE- has the general formula: --(CH.sub.2).sub.α --[A.sup.1 --(CH.sub.2).sub.β --].sub.I [A.sup.2 --(CH.sub.2).sub.γ --].sub.II A.sup.3 --(CH.sub.2).sub.δ -- where α, β, γ, and δ, which may be the same or different, are integers greater than 1 and less than 5; I and II, which may be the same or different, =0 or 1; and A 1 , A 2 , and A 3 , which may be the same or different, are independently O; S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons.
Claim 13
Dependent Claim
40 words
A compound, as claimed in claim 10, where A 1 , A 2 and A 3 , which may be the same or different, are independently (CH 2 ).sub.μ, --(NR 5 )--, or --(N + R 6 R 7 )--.
Claim 14
Dependent Claim
72 words
A compound, as claimed in claim 10, where A 1 and A 3 , which may be the same or different, are independently --(NR 5 )-- where R 5 is H or an alkyl group having 1-2 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-2 carbons; and II=0.
Claim 15
Dependent Claim
134 words
A compound, as claimed in claim 10, where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-2 carbons; X and Z, which may be the same or different are O or S; m and r=1; in the BRIDGE formula α, β, γ, and δ, which may be the same or different, are 2 or 3; A 1 , A 2 , and A 3 , which may be the same or different, are (CH 2 ).sub.μ where μ=0; --(NR 5 )-- where R 5 is H or an alkyl group having 1-2 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-2 carbons.
Claim 16
Dependent Claim
25 words
A compound, as claimed in claim 11, where X and Z are the same; R 1 and R 2 are the same; n=s and m=r.
Claim 17
Dependent Claim
30 words
A compound, as claimed in claim 11, where m and r are each 1; and where X and Z, which may be the same or different, are O or S.
Claim 18
Dependent Claim
14 words
A compound, as claimed in claim 11, where n is not equal to s.
Claim 19
Dependent Claim
14 words
A compound as claimed in claim 11, where the nucleic acid polymer is DNA.
Claim 20
Dependent Claim
14 words
A compound as claimed in claim 11, where the nucleic acid polymer is RNA.
Claim 21
Dependent Claim
119 words
A compound, as claimed in claim 14, where R 1 and R 2 are alkyl groups having 1-2 carbons; X and Z are O or S; m and r=1; in the BRIDGE formula α, β, γ, and δ, which may be the same or different, are 2 or 3; A 1 , A 2 , and A 3 , which may be the same or different, are (CH 2 ).sub.μ where μ=0; --(NR 5 )-- where R 5 is H or an alkyl group having 1-2 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-2 carbons.
Claim 22
Independent Claim
274 words
A method of forming a fluorescent nucleic acid complex, comprising: a) combining a sample thought to contain nucleic acid polymers with a dye of the general formula: ##STR10## where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-6 carbons; X is O, S, or N--R 3 , where R 3 is H or an alkyl group having 1-6 carbons; Z is O, S, or N--R 4 , where R 4 is H or an alkyl group having 1-6 carbons; n and s, which may be the same or different, =0, 1, or 2; Y is HC═CH; and p, m, q, and r=0 or 1, such that p+m=1 and q+r=1; and where -BRIDGE- has the general formula: --(CH.sub.2).sub.α --[A.sup.1 --(CH.sub.2).sub.β --].sub.I [A.sup.2 --(CH.sub.2).sub.γ --].sub.II A.sup.3 --(CH.sub.2).sub.δ -- where α, β, γ, and δ, which may be the same or different, are integers greater than 1 and less than 5; I and II, which may be the same or different, =0 or 1; and A 1 , A 2 , and A 3 , which may be the same or different, are independently O; S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons; b) after the dye has been allowed to combine with any nucleic acid polymers in said sample, detecting fluorescence resulting from the dye combining with nucleic acid polymers.
Claim 23
Independent Claim
274 words
A method of forming a fluorescent nucleic acid complex, comprising: a) combining a sample thought to contain nucleic acid polymers with a dye of the general formula: ##STR10## where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-6 carbons; X is O, S, or N--R 3 , where R 3 is H or an alkyl group having 1-6 carbons; Z is O, S, or N--R 4 , where R 4 is H or an alkyl group having 1-6 carbons; n and s, which may be the same or different, =0, 1, or 2; Y is HC═CH; and p, m, q, and r=0 or 1, such that p+m=1 and q+r=1; and where -BRIDGE- has the general formula: --(CH.sub.2).sub.α --[A.sup.1 --(CH.sub.2).sub.β --].sub.I [A.sup.2 --(CH.sub.2).sub.γ --].sub.II A.sup.3 --(CH.sub.2).sub.δ -- where α, β, γ, and δ, which may be the same or different, are integers greater than 1 and less than 5; I and II, which may be the same or different, =0 or 1; and A 1 , A 2 , and A 3 , which may be the same or different, are independently O; S; (CH 2 ).sub.μ where μ=0 or 1; --(NR 5 )-- where R 5 is H or an alkyl group having 1-6 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-6 carbons; b) after the dye has been allowed to combine with any nucleic acid polymers in said sample, detecting fluorescence resulting from the dye combining with nucleic acid polymers.
Claim 24
Dependent Claim
18 words
A method, as in claim 20, wherein the nucleic acid polymers in the sample are contained in cells.
Claim 25
Dependent Claim
15 words
A method, as in claim 20, wherein the sample contains nucleic acid polymers in solution.
Claim 26
Dependent Claim
132 words
A method, as in claim 20, where R 1 and R 2 , which may be the same or different, are alkyl groups having 1-2 carbons; X and Z, which may be the same or different are O or S; m and r=1; in the BRIDGE formula α, β, γ, and δ, which may be the same or different, are 2 or 3; A 1 , A 2 , and A 3 , which may be the same or different, are (CH 2 ).sub.μ ; --(NR 5 )-- where R 5 is H or an alkyl group having 1-2 carbons; or --(N + R 6 R 7 )-- where R 6 and R 7 , which may be the same or different, are independently hydrogen or an alkyl group having 1-2 carbons.
Claim 27
Dependent Claim
15 words
A method, as in claim 21, wherein said fluorescence indicates loss of cellular membrane integrity.
References Cited (Backward Citations)
US4304908A
Cited
US4883867A
Cited
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