Chemically reactive pyrenyloxy sulfonic acid dyes

• US 5,132,432 A
• Filed: 09/22/1989
• Issued: 07/21/1992
• Est. Priority Date: 09/22/1989
• Status: Expired due to Term

• Current Assignee: Molecular Probes Incorporated (Thermo Fisher Scientific Incorporated)
• Original Assignee: Molecular Probes Incorporated (Thermo Fisher Scientific Incorporated)
• Inventors: Haugland, Richard P., Whitaker, James E.
• Primary Examiner(s): Shah, Mukund J.
• Assistant Examiner(s): Ward, Edward C.
• Application Number:
  US07/410,837
• Time in Patent Office: 1,033 Days
• Field of Search: 562/45, 562/46, 562/47, 562/55, 548/518, 548/543, 548/528, 560/10, 552/6, 558/46, 558/6, 558/17, 436/172, 8/648
• US Class Current: 548/518
• CPC Class Codes: C07C 2603/50 Pyrenes; Hydrogenated pyrenes
• CPC Class Codes: C07C 309/43 having at least one of the sulfo groups bound to a carbon atom of a six-membered aromatic ring being part of a condensed ring system
• CPC Class Codes: C07D 207/16 Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
• CPC Class Codes: C07D 207/404 with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms, e.g. succinimide
• CPC Class Codes: C09B 57/001 Pyrene dyes

Abstract

The invention relates to novel fluorescent dyes based on the following pyrenyloxy sulfonic acid structure: ##STR1## wherein at least one of R₁, R₂, R₃, and R₄ is --OCH₂ C(O)OCH₃, --OCH₂ CON(O)H, --OCH₂ CON₃, --OCH₂ CONHNH₂, ##STR2## or --CH₂ CONH(CH₂)_n NH₂ where n is an integer from 1 to 10; at least one of R₁, R₂, R₃, and R₄ is sulfonic acid or alkali salts of sulfonic acid; and all other of R₁, R₂, R₃, and R₄ are hydrogen or a hydroxyl group. The new reagents contain functional groups typically found in biomolecules or polymers. The spectral properties of the fluorescent dyes are sufficiently different in wavelengths and intensity from fluorescein as to permit simultaneous use of both dyes with minimum interference. The dyes have narrower spectral band-widths and smaller Stokes'"'"' shifts than other reactive dyes with similar spectral properties, do not show appreciable sensitivity to pH, have higher solubilities in aqueous solvents and have good quantum yields and photostability.

Claims

1. A fluorescent dye having the structure:

##STR9## wherein (a) at least one of R₁, R₂, R₃, and R₄ has the structure --O--R₅ wherein R₅ includes a chemically reactive functional group selected from the group consisting of --CH₂ C(O)OCH₃, --CH₂ C(O)OH, --CH₂ CON₃, --CH₂ CONHNH₂, ##STR10## and --CH₂ CONH(CH₂)_n NH₂ where n is an integer from 1 to 10;

(b) at least one of R₁, R₂, R₃, and R₄ is selected from the group consisting of sulfonic acid and alkali salts of sulfonic acid; and

(c) all other of R₁, R₂, R₃, and R₄ are selected from the group consisting of a hydrogen and a hydroxyl group.

2. A fluorescent dye as recited in claim 1 wherein R₁ has the structure --OCH₂ C(O)OCH₃ and R₂, R₃, and R₄ have the structure --SO₃ X, wherein X is an alkali metal.

3. A fluorescent dye as recited in claim 1 wherein R₁ has the structure --OCH₂ C(O)OH and R₂, R₃, and R₄ have the structure --SO₃ X, wherein X is an alkali metal.

4. A fluorescent dye as recited in claim 1 wherein R₁ has the structure --OCH₂ CON₃ and R₂, R₃, and R₄ have the structure --SO₃ X, wherein X is an alkali metal.

5. A fluorescent dye as recited in claim 1 wherein R₁ has the structure --OCH₂ CONHNH₂ and R₂, R₃, and R₄ have the structure --SO₃ X, wherein X is an alkali metal.

6. A fluorescent dye as recited in claim 1 wherein R₁ has the structure ##STR11## and R₂, R₃, and R₄ have the structure --SO₃ X, wherein X is an alkali metal.

7. A fluorescent dye as recited in claim 1 wherein R₁ has the structure --OCH₂ CONH(CH₂)_n NH₂, where n is an integer from 1 to 10, and R₂, R₃, and R₄ have the structure --SO₃ X, wherein X is an alkali metal.

8. A fluorescent dye as recited in claim 1 wherein R₁ has the structure --OCH₂ C(O)OH, R₂ is a hydroxyl group, and R₃ and R₄ have the structure --SO₃ X, wherein X is an alkali metal.

9. A fluorescent dye as recited in claim 1 wherein R₁ and R₂ have the structure --OCH₂ C(O)OH and R₃ and R₄ have the structure --SO₃ X, wherein X is an alkali metal.