Method for labeling and detecting materials employing arylsulfonate cyanine dyes

• US 5,268,486 A
• Filed: 05/15/1992
• Issued: 12/07/1993
• Est. Priority Date: 04/18/1986
• Status: Expired due to Term

• Current Assignee: Carnegie Mellon University
• Original Assignee: Carnegie Mellon University
• Inventors: Mujumdar, Ratnakar B., Waggoner, Alan S., Ernst, Lauren A.
• Primary Examiner(s): Shah, Mukund J.
• Assistant Examiner(s): Grumbling, Matthew V.
• Application Number:
  US07/884,636
• Time in Patent Office: 571 Days
• Field of Search: 544/212, 544/328, 546/272, 546/273, 548/427, 548/455
• US Class Current: 548/427
• CPC Class Codes: C09B 23/0008 substituted on the polymethine chain
• CPC Class Codes: C09B 23/02 the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
• CPC Class Codes: C09B 23/10 The polymethine chain containing an even number of >CH- groups styryl dyes C09B23/14, C09B23/14 takes precedence
• CPC Class Codes: C09B 23/145 the ethylene chain carrying an heterocyclic residue, e.g. heterocycle-CH=CH-C6H5
• CPC Class Codes: C09B 62/0025 Specific dyes not provided for in groups C09B62/004 - C09B62/018
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

A luminescent cyanine dye having generally the following structure ##STR1## wherein the dotted lines represent one to three rings having 5 to 6 atoms in each ring. R₃, R₄, R₈ and R₉ groups are attached to the rings. At least one of the R₈ and R₉ groups is a sulfonic acid or sulfonate group and at least one of the R₁, R₂, R₃, R₄ and R₇ groups is a reactive moiety that reacts with amino, hydroxy or sulfhydryl nucleophiles.

Claims

1. A water soluble luminescent dye consisting of:

a cyanine having the structure ##STR12## wherein;

the dotted lines each represent carbon atoms necessary for the formation of one ring to three fused rings having 5 to 6 atoms in each ring and said R₃, R₄, R₈ and R₉ groups are attached to said rings;

X and Y are each CH₃ --C--CH₃ ;

m is an integer selected from the grouping consisting of 1, 2 and 3;

at least one of said R₁, R₂, R₃, R₄ and R₇ groups reacts with amino, hydroxy or sulfhydryl nucleophiles and is a reactive moiety selected from the group consisting of ##STR13## wherein Q is Br or Cl, and n=0, 1, 2, 3, 4, 5, 6, 7 or 8 for R₃, R₄ and R₇, and n=1, 2, 3, 4, 5, 6, 7 or 8 for R₁ and R₂ ;

and when any one of said R₃, R₄, and R₇ groups is not selected from one said reactive moiety, said remaining R₃, R₄, and R₇ are hydrogen or E--F, a polar group wherein E is a spacer group having the structure --(CH₂)_n -- and n of said spacer group=0, 1, 2, 3, or 4, and F is hydroxy, sulfonate, sulfate, carboxylate, or a lower alkyl substituted amino;

and when any one of said R₁ and R₂ groups is not selected from one said reactive moiety, said remaining R₁ or R₂ are a lower alkyl or said E--F and n of said spacer group=1, 2, 3, or 4;

at least one said R₈ and R₉ groups is selected from the group consisting of a sulfonic acid and a sulfonate moiety attached directly to said ring for conferring improved luminescence to said dye when in use as compared to a dye which does not have said at least one sulfonic acid or sulfonate moiety;

and when one of said R₈ or R₉ group is not a sulfonic acid or a sulfonate, said remaining R₈ or R₉ group is a hydrogen.

2. The luminescent dye of claim 1 wherein said cyanine has the structure:

##STR14##

3. The luminescent dye recited in claim 1 wherein the cyanine has the structure:

##STR15## wherein R₁₀ is selected from the group consisting of said reactive moiety wherein n=0, 1, 2, 3, 4, 5, 6, 7 or 8, hydrogen and sulfonic acid or sulfonate moiety.

4. A water soluble luminescent dye consisting of a cyanine having the structure ##STR16## wherein:

X and Y are CH₃ --C--CH₃ ;

m is an integer selected from the group consisting of 1, 2 and 3;

at least one of said R₁, R₂, R₃, R₄, R₇, R₁₀ and R₁₁ groups reacts with amino, hydroxy or sulfhydryl nucleophiles and is a reactive moiety selected from the group consisting of;

##STR17## wherein Q is Br or Cl, and n=0, 1, 2, 3, 4, 5, 6, 7 or 8 for R₃, R₄, R₇, R₁₀ and R₁₁, and n=1, 2, 3, 4, 5, 6, 7 or 8 for R₁ and R₂ ;

and when any one of said R₃, R₄, and R₇ groups is not selected from one said reactive moiety, said remaining R₃, R₄, and R₇ are hydrogen or E--F, a polar group wherein E is a spacer group having the structure --(CH₂)_n -- and n=0, 1, 2, 3, or 4, and F is hydroxy, sulfonate, sulfate, carboxylate, or lower alkyl substituted amino;

and when any one of said R₁ and R₂ groups is not selected from one said reactive group, said remaining R₁ and R₂ are lower alkyl or said E--F and n=1, 2, 3, or 4;

said R₈ and R₉, and said R₁₀ and R₁₁ groups when said R₁₀ and R₁₁ groups are not said reactive moiety, are selected from the group consisting of hydrogen and a sulfonic acid or sulfonate moiety, wherein at least one of said R₈, R₉, R₁₀ and R₁₁ groups is one of said sulfonic acid or said sulfonate moiety attached directly to a ring of said dye for conferring improved luminescence to said dye when in use as compared to a dye which does not have said at least one sulfonic acid or sulfonate moiety.

5. A water soluble luminescent dye consisting of:

##STR18## wherein X and Y are CH₃ --C--CH₃ ;

m is an integer selected from the group consisting of 1, 2 and 3;

at least one of said R₁ and R₂ is ##STR19## and the other is selected from the group consisting of ##STR20## --CH₂ CH₂ OH, --(CH₂)_k SO₃ where n=1, 2, 3, 4, 5, 6, 7 or 8 and k=2, 3, 4 or 5; and

at least one of said R₈ and R₉ groups is a sulfonate moiety attached directly to a ring of said dye for conferring improved luminescence to said dye when in use as compared to a dye which does not have at least one sulfonic acid or sulfonate moiety;

and when one of said R₈ and R₉ group is not said sulfonate moiety, said remaining R₈ or R₉ group is a hydrogen.

6. A water soluble luminescent dye consisting of:

##STR21## wherein X and Y are CH₃ --C--CH₃ ;

m is an integer selected from the group consisting 1, 2 and 3;

at least one of said R₁ and R₂ is ##STR22## and the other is selected from the group consisting of ##STR23## --CH₂ CH₂ OH, --(CH₂)_k SO₃ where n=1, 2, 3, 4, 5, 6, 7, or and k=2, 3, 4, or 5;

at least two of said R₈, R₉,R₁₀ and R₁₁ groups are a sulfonate moiety attached directly to a ring of said dye for conferring improved luminescence to said dye when in use as compared to a dye which does not have at least one sulfonic acid or sulfonate moiety;

and when any one of said R₈, R₉, R₁₀ and R₁₁ groups is not said sulfonate moiety, it is a hydrogen.

7. A water soluble luminescent dye consisting of:

##STR24## wherein X and Y are CH₃ --C--CH₃ ;

m is an integer selected from the group consisting of 1, 2, and 3;

at least one of said R₁ and R₂ is ##STR25## and the other is selected from the group consisting of ##STR26## where n=1, 2, 3, 4, 5, 6, 7 or 8 and k=2, 3, 4 or 5;

at least two of said R₈, R₉ and R₁₀ groups are a sulfonate moiety attached directly to a ring of said dye for conferring improved luminescence to said dye when in use as compared to a dye which does not have at least one sulfonic acid or sulfonate moiety;

and when any one of said R₈, R₉ or R₁₀ is not said sulfonate moiety, it is a hydrogen.

8. A water soluble luminescent dye consisting of:

##STR27## wherein m is an integer selected from the group consisting of 1 and 2 and n=5.