Fluorescence dyes and their use as fluorescence markers

• US 6,552,199 B1
• Filed: 05/11/2000
• Issued: 04/22/2003
• Est. Priority Date: 05/20/1999
• Status: Expired due to Fees

• Current Assignee: Roche Diagnostics GmbH (Roche Holding AG)
• Original Assignee: Roche Diagnostics GmbH (Roche Holding AG)
• Inventors: Reiss, Alexander, Daltrozzo, Ewald
• Primary Examiner(s): Huang, Evelyn Mei
• Application Number:
  US09/568,679
• Time in Patent Office: 1,076 Days
• Field of Search: 546/198, 546/174, 546/152, 544/135, 548/156, 548/546, 548/547, 549/223
• US Class Current: 546/174
• 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: G01N 33/582 with fluorescent label

Abstract

The subject matter of the invention are new xanthene derivatives of the formula I,

wherein R1,R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, and X, Y, are as defined herein. The compounds according to the invention provide molecules that are—due to their spectral properties (absorption maxima in the range of approx. 650 nm and above as well as emission maxima above 670 nm)—very suitable for the use as dyes and in particular as fluorescence dyes. The compounds of the formula I according to the invention are used for the production of fluorescence conjugates, for their application in immunoassays, for DNA analytics, for in-vivo diagnostics or as a laser dye.

Claims

1. A compound of the formula:

wherein R1, R2, R3, R4 and R5 are selected from the group consisting of hydrogen, alkyl, aryl, heteroaryl, alkoxy, cyano, hydroxy, halogen, isocyanate, isothiocyanate, carboxylic acid, sulfonic acid, and derivatives of carboxylic acid or sulfonic acid selected from the group consisting of alkoxycarbonyl, aryloxycarbonyl, N-hydroxysuccinimide esters, acid halogenides, unsubstituted amino and substituted amino selected from the group consisting of ω

-amino alcohols, ω

-aminocarboxylic acids and ω

-aminohalogen alkanes, with the proviso that at least one of R1, R2, R3, R4 and R5 is a carboxylic acid or derivative thereof,

R6, R7, R8, R9, R10 and R11 are selected from the group consisting of hydrogen, alkyl, substituted alkyl, and halogen, and

X is selected from the group consisting of amines, substituted amines, heteroarylidenecyanomethine residues, and malonic acid derivatives selected from the group consisting of 2-propyldinitril residues and cyanoacetic acid esters;

Y is selected from the group consisting of heteroarylidenecyanomethine residues, and malonic acid derivatives selected from the group consisting of 2-propyldinitril residues and cyanoacetic acid esters.

2. The compound of claim 1, wherein said heteroarylidenecyanomethine residues are selected from the group consisting of 2-benzthiazolylidenecyanomethine and 2- or 4-quinolinylidenecyanomethine.

3. The compound of claim 1, wherein X and Y are identical.

4. The compound of claim 1, wherein X and Y are different.

5. The compound 3′

,6′

-bis-(2-benzthiazolylidene-cyanomethine)-fluoran.

6. The compound 5,6,2′

,7′

-tetrachloro-3′

,6′

-bis-(2-propyldinitrile)-fluoran.

7. The compound 3′

,6′

-bis-(cyanoacetic acid ethyl ester-2-yl)-fluoran.

8. The compound 3′

-(2-benzthiazolylidene-cyanomethine)-3′

-(2-quinolinylidene-cyanomethine)-4,5,6,7-tetrachlorofluoran.

9. The compound 3′

-(2-benzthiazolylidene-cyanomethylene)-6′

-chlorofluoran.

10. The compound 3′

,6′

-bis-(2-benzthiazolylidene-cyanomethine)-5(6)-carboxyfluoran.

11. The compound 3′

,6′

-bis-(2-benzthiazolylidene-cyanomethine)-5(6)-sulfofluoran.

12. The compound 3-(2-benzthiazolyl-cyanomethine)-6-(2-benzthiazolylidene-cyanomethine)-9-[2-phenylcarboxylic acid-(2-chloroethyl)-ester]-xanthene-3-ylidene.

13. The compound 3-(2-benzthiazolyl-cyanomethine)-6-(2-benzthiazolylidene-cyanomethine)-9-[2-phenylcarboxylic acid-(N-hydroxysuccinimidyl)-ester]-xanthene-3-ylidene.

14. The compound 3-(2-benzthiazolyl-cyanomethine)-6-(2-benzthiazolylidene-cyanomethine)-9-{phenyl-2,4,(5)-bis[carboxylic acid-(N-hydroxysuccinimidyl)-ester]}-xanthene3-ylidene.

15. The compound 5(6)-carboxy-3′

,6′

-bis-(2-propyldinitril)-fluoran.

16. The compound 2,7-dichloro-3,6-bis-(2-propyldinitril)-9-(2,4(5)-phenyl-dicarboxylic acid-di-(N-hydroxysuccinimidyl ester)-xanthene-3-ylidene.

17. The compound 3,6bis-(cyanoacetic acid ethyl ester-2-yl)-9-[2-phenyl-carboxylic acid-(N-hydroxysuccinimidyl)-ester]-xanthene-3-ylidene.

18. The compound 3,6-bis-(cyanoacetic acid ethyl ester-2-yl)-9-[4,5-dichloro-2-phenyl-carboxylic acid-N-hydroxysuccinimidyl)ester]-xanthene-3-ylidene.

19. The compound 3,6-bis-(cyanoacetic acid ethyl ester-2-yl)-9-(2,4(5)-phenyl-dicarboxylic acid-di-(N-hydroxysuccinimidyl ester)-xanthene-3-ylidene.