Derivatives of 1,2-dihydro-7-hydroxyquinolines containing fused rings

• US 7,816,519 B2
• Filed: 11/16/2006
• Issued: 10/19/2010
• Est. Priority Date: 08/04/2000
• Status: Expired due to Fees

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Liu, Jixiang, Diwu, Zhenjun, Gee, Kyle
• Primary Examiner(s): POWERS, FIONA
• Application Number:
  US11/560,579
• Publication Number:
  US 20070093652A1
• Time in Patent Office: 1,433 Days
• Field of Search: None
• US Class Current: 544/99
• CPC Class Codes: C07D 471/04 Ortho-condensed systems
• CPC Class Codes: C07D 491/04 Ortho-condensed systems
• CPC Class Codes: C07D 491/044 with only one oxygen atom as ring hetero atom in the oxygen-containing ring
• CPC Class Codes: C07D 491/14 Ortho-condensed systems
• CPC Class Codes: C07D 491/147 the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom
• CPC Class Codes: C07D 491/22 in which the condensed system contains four or more hetero rings
• CPC Class Codes: C07D 495/04 Ortho-condensed systems
• CPC Class Codes: C07D 495/14 Ortho-condensed systems
• CPC Class Codes: C07D 495/22 in which the condensed system contains four or more hetero rings
• CPC Class Codes: C07D 498/14 Ortho-condensed systems
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10T 436/141111 Diverse hetero atoms in same or different rings [e.g., alkaloids, opiates, etc.]
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/145555 Hetero-N

Abstract

The present invention describes novel dyes, including coumarins, rhodamines, and rhodols that incorporate additional fused aromatic rings. The dyes of the invention absorb at a longer wavelength than structurally similar dyes that do not possess the fused aromatic rings. Many of the dyes of the invention are useful fluorescent dyes. The invention includes chemically reactive dyes, dye-conjugates, and the use of such dyes in staining samples and detecting ligands or other analytes.

Claims

1. A compound of the formula:

3. The compound according to claim 1, wherein G and W are 5-membered heteroaromatic rings.

4. The compound according to claim 1, wherein G and W are 6-membered heteroaromatic rings.

5. The compound according to claim 1, wherein R²⁸ has the formula

6. The compound according to claim 1, wherein said R_X is independently selected from the group consisting of an acrylamide, an activated ester of a carboxylic acid, an acyl azide, an acyl nitrile, an aldehyde, an alkyl halide, an amine, an anhydride, an aniline, an aryl halide, an azide, an aziridine, a boronate, a carboxylic acid, a diazoalkane, a haloacetamide, a halotriazine, a hydrazine, an imido ester, an isocyanate, an isothiocyanate, a maleimide, a phosphoramidite, a reactive platinum complex, a sulfonyl halide, and a thiol group.

7. The compound according to claim 1, wherein said S_C is independently selected from the group consisting of an amino acid, a peptide, a protein, a tyramine, a carbohydrate, a metal chelating moiety, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid, a hapten, a psoralen, a drug, a hormone, a lipid, a lipid assembly, a polymer, a polymeric microparticle, a biological cell, and a virus.

8. The compound according to claim 1, wherein G or W is furan, thiophene, pyrrol, benzofuran, benzothiophene, indole, pyridine, quinoline, isoquinoline, oxazole, benzoxazole, thiazole, benzothiazole, isoxazole, imidazole, benzimidazole, naphthofuran, naphthothiophene, benzindole, benzoquinoline, or acridine.

9. The compound according to claim 1, wherein one of said R⁵, R⁶, R³⁰, R³¹, R³², R³³, and R³⁴ is -L-R_X or -L-S_C.

10. The compound according to claim 1, wherein

said R³ and R⁴ are each methyl;

and

R⁶ is H.

11. The compound according to claim 1, wherein Q is CR²⁸ and R²⁸ has the formula

2. A compound of the formula:

12. A kit for staining a sample, wherein the kit comprises a solution comprising a buffer and a compound having formula:

13. The kit according to claim 12, wherein the kit further comprises an additional detection reagent, a purification medium, or standards.

14. The kit according to claim 12, wherein at least one of R³, R⁴, R⁵, R⁶, R²⁸, R³⁰, R³¹, R³², R³³, R³⁴, R⁴³, R⁴⁴, R⁴⁵ and R⁴⁶ is L-R_x wherein said R_x is independently selected from the group consisting of a carboxylic acid, an activated ester of a carboxylic acid, an amine, an azide, a hydrazine, a haloacetamide, an alkyl halide, an isothiocyanate, and a maleimide group.

15. The kit according to claim 14, wherein at least one of R³¹, R³² or R³⁴ is L-R_x and R³⁰ is carboxylic acid or sulfonic acid.

16. The kit according to claim 12, wherein at least one of R³, R⁴, R⁵, R⁶, R²⁸, R³⁰, R³¹, R³², R³³, R³⁴, R⁴³, R⁴⁴, R⁴⁵ and R⁴⁶ is L-Sc, wherein said Sc is independently selected from the group consisting of an amino acid, a peptide, a protein, an antibody, an antibody fragment, a carbohydrate, a nucleotide, a nucleoside, an oligonucleotide, a nucleic acid polymer, an ion-complexing moiety, a lipid, a non-biological organic polymer and polymeric microparticle.

17. The kit according to claim 16, wherein the Sc is an antibody or fragment thereof.

18. A method of staining a sample, the method comprising steps:

a) combining a solution with the sample, wherein the solution comprises a compound of the formula

19. The method according to claim 18, wherein the method further comprises combining the sample with an additional detection reagent.

20. The method according to claim 18, wherein said sample comprises cells, growth medium, tissue, proteins, peptides, or biological fluids.

21. The method according to claim 18, wherein said sample is immobilized in or on a solid or semi-solid matrix that is a membrane, an electrophoretic gel, a silicon chip, a glass slide, a microwell plate, or a microfluidic chip.

22. The method according to claim 18, wherein at least one of R³⁰, R³¹, R³², R³³, and R³⁴ is -L-R_x or -L-S_C;

R_x is selected from the group consisting of a carboxylic acid, an activated ester of a carboxylic acid, an amine, an azide, a hydrazine, a haloacetamide, an alkyl halide, an isothiocyanate, and a maleimide group; and

S_c is selected from the group consisting of an amino acid, a peptide, a protein, a

polysaccharide, a nucleotide, a nucleoside, an oligonucleotide, a nucleic acid polymer, an ion-complexing moiety, a lipid, or a non-biological organic polymer or polymeric microparticle, wherein said Sc is optionally bound to one or more additional fluorophores.

23. The method according to claim 18, wherein at least one of R³⁰, R³¹, R³², R³³, and R³⁴ is -L-S_C, and said S_C is a nucleoside, a nucleotide, an oligonucleotide, or a nucleic acid polymer.