Phosphonate fluorescent dyes and conjugates

• US 7,671,218 B2
• Filed: 08/12/2005
• Issued: 03/02/2010
• Est. Priority Date: 08/13/2004
• Status: Active Grant

• Current Assignee: Elitechgroup MDX LLC
• Original Assignee: Elitech Holding B. V.
• Inventors: Lukhtanov, Eugeny, Vorobiev, Alexei
• Primary Examiner(s): Sullivan; Daniel M
• Assistant Examiner(s): Katakam; Sudhakar
• Application Number:
  US11/202,635
• Publication Number:
  US 20070172832A1
• Time in Patent Office: 1,663 Days
• Field of Search: 549/223, 549/224, 435/6, 436/172, 536/26.6
• US Class Current: 549/223
• CPC Class Codes: C07F 9/1411 with hydroxyalkyl compounds with further substituents on alkyl
• CPC Class Codes: C07F 9/5728 condensed with carbocyclic rings or carbocyclic ring systems
• CPC Class Codes: C07F 9/6533 Six-membered rings
• CPC Class Codes: C07F 9/6547 condensed with carbocyclic rings or carbocyclic ring systems
• CPC Class Codes: C07F 9/65522 condensed with carbocyclic rings or carbocyclic ring systems
• CPC Class Codes: C07F 9/65583 each of the hetero rings containing nitrogen as ring hetero atom
• CPC Class Codes: C07F 9/65586 at least one of the hetero rings does not contain nitrogen as ring hetero atom
• CPC Class Codes: C07F 9/6561 containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
• CPC Class Codes: C07F 9/65842 Cyclic amide derivatives of acids of phosphorus, in which one nitrogen atom belongs to the ring
• CPC Class Codes: C09B 11/08 Phthaleins; Phenolphthaleins; Fluorescein
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C09B 23/083 five >CH- groups

Abstract

Reagents are provided for the introduction of phosphonate groups into fluorescent dyes. Methods are also provided for preparing dye conjugates.

Claims

1. A fluorescent dye reagent having the formula:

2. A fluorescent dye reagent of claim 1, wherein said fluorescent dye component is selected from the group consisting of coumarin dyes, resorufin dyes, dipyrrometheneboron difluoride dyes, ruthenium bipyridyl dyes, energy transfer dyes, thiazole orange dyes and N-aryl-1,8-naphthalimide dyes.

3. A fluorescent dye reagent of claim 1, wherein L is C₂-C₈ alkylene.

4. A fluorescent dye reagent of claim 1, wherein N^a is an ammonium ion group having the formula —

N⁺(R^D)₃ wherein each R^D is independently selected from H, C₁-C₈ alkyl, aryl and arylC₁-C₄ alkyl.

5. A fluorescent dye reagent of claim 4, wherein each R^D is H or C₁-C₈alkyl.

6. A fluorescent dye reagent of claim 1, wherein m is 0 and n is 1.

7. A fluorescent dye reagent of claim 1, wherein m is 0 and n is 2.

8. A fluorescent dye reagent of claim 1, wherein m is 1 and n is 1.

9. A fluorescent dye reagent of claim 2, wherein said fluorescent dye component is a xanthene or benzoxanthene dye selected from the group consisting of fluorescein dyes, rhodamine dyes and rhodol dyes.

10. A fluorescent dye reagent of claim 9, having formula I(a) or I(b):

11. A fluorescent dye reagent of claim 1, selected from compound:

12. A fluorescent dye reagent having the formula:

Fl-P¹

wherein

Fl is a fluorescent dye component selected from the group consisting of coumarins, benzocoumarins, xanthenes, benzo[a]xanthenes, benzo[b]xanthenes, benzo[c]xanthenes, cyanines, acridines, dipyrrometheneboron difluorides, phenoxazines, benzo[a]phenoxazines, benzo[b]phenoxazines and benzo[c]phenoxazines; and

P¹ is functionalized phosphonate group having the formula;

13. A fluorescent dye reagent of claim 12, wherein said fluorescent dye component is a xanthene or benzoxanthene dye selected from the group consisting of fluorescein dyes, rhodamine dyes and rhodol dyes.