Fluorescent dyes (AIDA) for solid phase and solution phase screening

• US 20050227299A1
• Filed: 05/27/2005
• Published: 10/13/2005
• Est. Priority Date: 12/21/1998
• Status: Abandoned Application

• Current Assignee: Hubert Gstach, Manfred Auer
• Original Assignee: Hubert Gstach, Manfred Auer
• Inventors: Auer, Manfred, Gstach, Hubert
• Application Number:
  US11/139,753
• Publication Number:
  US 20050227299A1
• Time in Patent Office: Days
• US Class Current: 435/7.500
• CPC Class Codes: C07D 231/56 Benzopyrazoles; Hydrogenated benzopyrazoles
• CPC Class Codes: C07D 403/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07D 495/04 Ortho-condensed systems
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

The invention relates to new fluorescent dyes of formula (I) which can be used in high throughput screening both, on the solid phase as well as in homogeneous solution.

Claims

1-11. -11. (canceled)

12. Compounds comprising structures of the formulae II-III

A-B-D-C-D′

-  

 

(Formula (II))
A-B-D- and -D′

-C  

 

(Formula (III))

wherein

A is selected from the group consisting of functionalized polystyrene based resins, polyacrylamide based polymers, polystyrene/polydimethylacrylamide composites, PEGA resins, polystyrene-polyoxyethylene based supports, Tentagel resins, PEG-polystyrene graft polymeric supports, glass surfaces, functionalized surfaces, materials grafted with functionalized surfaces, and polyethylenglycol;

B is a linker allowing cleavage of fluorescent conjugates of formula (II-III) for liberation of the D and C containing fragments;

C is a compound selected from formula (I)

wherein

one of the radicals R¹ or R² and one of the radicals R³ or R⁴ is hydrogen and the other is independently —

COOH, —

COOR⁷, —

CONH₂, —

CONH(CH₂)_nOH, —

CONR⁸R⁹, —

CH₂OH, —

CH₂NH₂, —

NO₂, —

NR¹⁰R¹¹, —

NHCOR¹², Cl, Br, F, —

CF₃, —

N═

C═

O, —

N═

C═

S, —

SO₃H, —

SO₂NH(CH₂)_nNH₂, (C₁-C₄) alkyl, (C₁-C₁₆)-alkyl substituted at the terminal carbon with —

COOH, —

COOR⁷, —

CONH₂, —

CONR⁸R⁹, —

CONH(CH₂)_nOH, —

CH₂OH, —

CH₂NH₂, —

N═

C═

O, —

N═

C═

S, —

SO₃H, —

SO₂NH(CH₂)_nNH₂, —

CONH(CH₂)_nNH₂, and the —

NH₂ group could also be substituted by (C₁-C₄) alkyl or a commonly used amino protecting group;

and one of the radicals R⁵ or R⁶ is hydrogen and the other is hydrogen, halogen, —

NO₂, —

NR¹⁰R¹¹, —

NHCOR¹², (C₁-C₄) alkyl, (C₁-C₁₆)-alkyl substituted at the terminal carbon with —

COOH, —

COOR⁷, —

CONH₂, —

CONR⁸R⁹, —

CONH(CH₂)_nOH, —

CH₂OH, —

CH₂NH₂, —

N═

C═

O, —

N═

C═

S, —

SO₃H, —

SO₂NH(CH₂)_nNH₂, —

CONH(CH₂)_nNH₂, wherein and the —

NH₂ group could also be substituted by (C₁-C₄) alkyl or a commonly used amino protecting group;

n is 2-8;

with the proviso that only one of R¹-R⁶ is nitro;

R⁷ is a commonly used carboxyl protecting or carboxyl activating group;

R⁸ or R⁹ is hydrogen and the other is lower alkyl (C₁-C₄), phenyl, benzyl, or R⁸ and R⁹ are part of a 5 or 6 membered ring;

R¹⁰ and R¹¹ are independently hydrogen or (C₁-C₄)alkyl; and

R¹² is (C₁-C₁₀)alkyl, phenyl, which both can be substituted by (C₁-C₄) alkyl, protected amino group or halogen; and

D and D′

are independently a bond or a spacer selected from α

,ω

-diamino-alkanes, diaminocyclohexyl, bis-(aminomethyl)-substituted phenyl, α

-amino-ω

-hydroxy-alkanes, alkylamines, cyclic alkylamines or cyclic alkyldiamines or amino acids without or with additional functionality in the side chain.

13. Compounds of claim 12, wherein

B is selected from benzyl, benzhydryl, benzhydryliden, trityl, xanthenyl, benzoin, silicon, or allyl based linkers.

14. Compounds of claim 12 of the following structures:

15. Compounds of claim 12 wherein the amino protecting group is tert-butyloxycarbonyl, 9-fluorenylmethoxycarbonyl, phthalimido, trifluoroacetamido, methoxycarbonyl, ethoxycarbonyl, benzyloxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, or 2-(trimethylsilyl)ethoxy-carbonyl.

16. Compounds of claim 12 wherein C is of the following structures: