Pentacyclic compounds and their use as absorption or fluorescent dyes

• US 5,750,409 A
• Filed: 05/30/1995
• Issued: 05/12/1998
• Est. Priority Date: 11/18/1991
• Status: Expired due to Term

• Current Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Original Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Inventors: Arden-Jacob, Jutta, Herrmann, Rupert, Josel, Hans-Peter, Drexhage, Karl-Heinz
• Primary Examiner(s): ACHUTAMURTHY, PONNATHAPURA N
• Application Number:
  US08/454,440
• Time in Patent Office: 1,078 Days
• Field of Search: 436/512, 436/513, 436/517, 436/536, 436/164, 436/166, 436/172, 436/800, 436/815, 435/7.1, 546/36, 546/48, 546/42
• US Class Current: 436/517
• 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 11/28 Pyronines ; Xanthon, thioxanthon, selenoxanthan, telluroxanthon dyes
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

Pentacyclic derivatives having the general formulae (Ia), (Ib) and (Ic) ##STR1## denote: hydrogen, alkyl with 1 to 20 carbon atoms polyoxyhydrocarbyl, phenyl, phenylalkyl with 1 to 3 carbon atoms in the alkyl chain, wherein the alkyl residues or/and phenyl residues can be substituted by one or several hydroxy, halogen, sulfo, carboxy or alkoxycarbonyl groups in which alkoxy can have 1 to 4 carbon atoms;

R⁷ denotes an alkyl group with 1 to 7 carbon atoms, substituted by at least one halogen, or denotes a carboxyalkyl group or a phenyl group which is substituted by a carboxy or alkoxycarbonyl group located at the o-position relative to the carbon atom bound to the pentacyclic ring system and by at least one halogen, wherein alkoxy can have 1 to 4 carbon atoms, or a carboxymethylene-oxy-alkyloxy group; the residues R¹⁴ and R¹⁵ as well as the positions on the rings in the formulae that are not marked by specific symbols denote any desired substituents which may optionally be linked together;

the two bonds marked by a broken line mean that the two carbon atoms which are joined by the bond marked by a broken line can be linked together by a single or double bond;

and X^- is a counterion; as well as their derivatives activated at least at one of the residues R¹, R⁷ or/and R¹³.

The compounds can be used as absorption or fluorescent dyes, in particular as markers for hapten/antibody/protein conjugates, as laser dyes and as an absorption medium for solar collectors.

Claims

1. A compound of the formula ##STR10## wherein R¹ and R¹³ independently of each other represent a hydrogen, an alkyl residue having 1 to 20 carbon atoms, a polyoxyhydrocarbyl, a phenyl group, or a phenylalkyl residue having 1 to 3 atoms in the alkyl chain;

R⁷ is (i) an alkyl group of 1 to 20 carbon atoms, substituted by at least one halogen,

(ii) a phenyl group, substituted by a carboxy or an alkoxycarbonyl group having 1 to 4 carbon atoms, wherein said carboxy or alkoxycarbonyl group is located in the ortho position of the carbon atom bound to a pentacyclic ring system of said compound and by at least one halogen,

(iii) a carboxy group,

(iv) a carboxyalkyl group having 1 to 10 carbon atoms in the alkylene chain, or

(v) a carboxymethylene-oxy-alkyloxy group;

X.sup.⊖

is a counterion;

R², R³, R⁴, R⁵, R⁶, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹⁴, R¹⁵ R¹⁶, R¹⁷, R¹⁸, and R¹⁹ independently of each other represent a hydrogen, an alkyl or a substituted alkyl having 1 to 20 carbons and two of R², R³, R⁴, R⁵, R⁶, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, and R¹⁹ are optionally linked together;

----- represents a single or double bond;

wherein said compound can be activated at at least one of R¹, R⁷ and R¹³.

2. A compound of claim 1, wherein at least one of R¹ or R¹³, is an alkyl or phenyl group substituted by a hydroxy, a halogen, a sulfo group, a carboxy group or an alkoxycarbonyl group having 1 to 4 carbon atoms.

3. A compound of claim 1, wherein at least one of R², R³, R⁴, R⁵, R⁶, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹⁴, R¹⁵ , R¹⁶, R¹⁷, R¹⁸, and R¹⁹ is a substituted alkyl.

4. A compound of claim 1, wherein at least two of R², R³, R⁴, R⁵, R⁶, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, and R¹⁹, are linked together.

5. A compound of claim 1, wherein at least one of R¹ and R¹³, is a hydrogen, an optionally substituted group of 1 to 4 carbon atoms, or a benzyl group.

6. A compound of claim 5, wherein said alkyl is substituted by a hydroxy, a halogen, a carboxy group, an alkoxycarbonyl group, or a sulfo group.

7. A compound of claim 1, wherein at least one of R¹ and R¹³ is a polyoxyhydrocarbyl.

8. A compound of claim 7, wherein said polyoxyhydrocarbyl is a polyether, a polyol, a water soluble carbohydrate, or a water soluble carbohydrate substituted by amino or acid groups.

9. A compound of claim 1, wherein R⁷ is a perhalogenated alkyl group having 1 to 3 carbon atoms.

10. A compound of claim 9, wherein R⁷ is a perfluorinated alkyl group having 1 to 3 carbon atoms.

11. A compound of claim 1, wherein R⁷ is a carboxyalkyl group having 1-10 carbon atoms in the alkylene chain, a carboxy group or a carboxymethylene-oxy-alkyloxy group.

12. A compound of claim 1, having the formula, ##STR11## wherein R¹ is not linked with R¹⁵, and R¹³ is not linked with R¹⁴.

13. A compound of claim 1, wherein at least one of said ---- is a double bond.

14. A compound of claim 1, wherein X.sup.⊖

is a perchlorate, a heptafluorobutyrate, COO-- or SO₃.

15. A compound of claim 1, wherein R⁷ is an alkyl group having 1 to 7 carbon atoms.

16. A compound of claim 1, wherein R¹ and R¹³, independently of each other, represent a hydrogen, a methyl, a ethyl, a 3-sulfopropyl, a 4-sulfobutyl, a 3-carboxypropyl, a 4-carboxybutyl, a 3-methoxycarbonylpropyl, a 3-ethoxycarbonylpropyl, a methoxy-ethoxy-ethyl, a hydroxyethoxy-ethyl, or a benzyl group.

17. A compound of claim 1, wherein R⁷ is selected from the group consisting of trifluoromethyl, pentafluoroethyl, heptafluoropropyl, a 3,4,5,6-halo-2-carboxyphenyl, a 3,4,5,6-halo-2-ethoxy-carboxyl-phenyl and a carboxyethyl group.

19. A method for determining a first immunologically bindable substance in a sample, comprising:

contacting a conjugate of a compound of claim 1 and a second immunologically bindable substance which is bindable with said first immunologically bindable substance, with said sample containing said first immunologically bindable substance, and

determining the change in absorption or fluorescence of the compound caused by immunological binding which is specific for the first immunologically bindable substance as a measure of the amount of said first immunologically bindable substance in said sample.

20. A method for determining a first immunologically bindable substance in a sample, comprising:

contacting a conjugate of the compound of claim 1 and a second immunologically bindable substance, with a known amount of a binding partner which binds to said first immunologically bindable substance and to said second immunologically bindable substance, and with said sample containing said first immunologically bindable substance, and

determining the change in absorption or fluorescence of the compound caused by immunological binding which is specific for the first immunologically bindable substance in said sample as a measure of the amount of said first immunologically bindable substance.

18. A compound of the formula:

##STR12## wherein R¹ and R¹³ independently of each other represent a hydrogen, an alkyl residue having 1 to 20 carbon atoms, a polyoxyhydrocarbyl, a phenyl residue, or a phenylalkyl residue having 1 to 3 atoms in the alkyl chain;

R⁷ is (i) an alkyl group of 1 to 20 carbon atoms, substituted by at least one halogen,

(ii) a phenyl group, substituted by a carboxy or an alkoxycarbonyl group having 1 to 4 carbon atoms, wherein said carboxy or alkoxycarbonyl group is located in the ortho position of the carbon atom bound to a pentacyclic ring system of said compound and by at least one halogen;

(iii) a carboxy group;

(iv) a carboxyalkyl group having 1 to 10 carbon atoms in the alkylene chain;

or

(v) a carboxymethylene-oxy-alkyloxy group;

X.sup.⊖

is a counterion;

R², R³, R⁴, R⁵, R⁶, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, and R¹⁹ independently of each other represent a hydrogen, an alkyl or a substituted alkyl having 1 to 20 carbons and two of R², R³, R⁴, R⁵, R⁶, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, and R¹⁹ are optionally linked together;

---- represents a single or double bond;

wherein said compound can be activated at at least one of R¹, R⁷ and R¹³ ; and

wherein there is an N-hydroxy-succinimide ester or an organic acid chloride at at least one of R¹, R⁷, and R¹³.