Novel alkyl substituted fluorescent compounds and polyamino acid conjugates

• US 4,351,760 A
• Filed: 09/07/1979
• Issued: 09/28/1982
• Est. Priority Date: 09/07/1979
• Status: Expired due to Term

• Current Assignee: Dade Behring Marburg GmbH (Siemens AG)
• Original Assignee: Syva Co. (Siemens AG)
• Inventors: Ullman, Edwin F., Khanna, Pyare
• Primary Examiner(s): Schain, Howard E.
• Application Number:
  US06/073,158
• Time in Patent Office: 1,117 Days
• Field of Search: 260/112 R, 260/112 B, 260/121, 424/85, 424/88, 435/188, 525/420
• US Class Current: 530/395
• CPC Class Codes: C07D 311/82 Xanthenes
• CPC Class Codes: C07J 41/0055 the 17-beta position being substituted by an uninterrupted chain of at least three carbon atoms which may or may not be branched, e.g. cholane or cholestane derivatives, optionally cyclised, e.g. 17-beta-phenyl or 17-beta-furyl derivatives
• CPC Class Codes: C07J 41/0088 containing unsubstituted amino radicals
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: Y10S 435/961 including a step of forming, releasing, or exposing the antigen or forming the hapten-immunogenic carrier complex or the antigen per se
• CPC Class Codes: Y10S 435/968 High energy substrates, e.g. fluorescent, chemiluminescent, radioactive
• CPC Class Codes: Y10S 530/806 Antigenic peptides or proteins

Abstract

Fluorescent antigen conjugates are provided comprising antigens covalently bonded to at least one 2,7-dialiphatic substituted-9-phenyl-6-hydroxy-3H-xanthen-3-one, wherein the 1- and 8-positions are unsubstituted. Also provided are novel fluorescent compounds absorbing at wavelengths in excess of 500 nm, having active functionalities for linking to the antigen. Finally, methods are provided for analyzing antigens in serum, whereby serum interference is avoided.

Claims

1. A compound of the formula:

##STR9## wherein;

E^b is hydrogen or chloro;

E² is chloro;

Z² is carboxy;

R² is alkylene of from 1 to 3 carbon atoms;

D² is hydrogen or carboxy;

W² is a bond, alkylene, carboxamidoalkylene, or poly(carboxamidoalkylene), wherein alkylene is of from 1 to 2 carbon atoms;

Y² is non-oxo-carbonyl, carbamoyl, thiocarbamoyl, amino, methylene, oxy or thio;

A² is a poly(amino acid) ligand or receptor;

m² is 0 to 3;

x² is 0 to 4; and

n² is 1 to the molecular weight of A² divided by 500.

2. A compound according to claim 1, wherein A² has a molecular weight of from about 2,000 to 600,000.

3. A compound according to claim 2, wherein E^b is chloro.

4. A compound according to any one of claims 1 to 3, wherein W² is a bond and Y² is non-oxo-carbonyl.

5. A composition of matter consisting of a conjugate bonded to a Support, and of the formula:

##STR10## wherein;

n³ is 1 to the molecular weight of A² divided by 500, with the proviso that when q is 0, n³ is 1;

q is 0 or 1;

p is at least 1 and up to the molecular weight of Support divided by 500;

Support is a molecule of at least about 10,000 molecular weight having a plurality of functionalities for linking;

the group in the brackets is bonded by any convenient functionality by a bond or linking group to the Support;

E^b is hydrogen or chloro;

E² is chloro;

Z² is carboxy;

R² is alkylene of from 1 to 3 carbon atoms;

D² is hydrogen or carboxy;

W² is a bond, alkylene, carboxyamidoalkylene or poly(carboxyamidoalkylene), wherein alkylene is of from 1 to 2 carbon atoms;

Y² is non-oxo-carbonyl, carbamoyl, thiocarbamoyl, amino, methylene, oxy or thio;

A² is a poly(amino acid) ligand or receptor bonded to Y² ;

m² is 0 to 3.

x² is 0 to 4.

6. A composition of matter according to claim 5, wherein said support is a polysaccharide.

7. A composition of matter according to any of claims 5 and 6, wherein A² is a poly(amino acid) of from about 2,000 to 600,000 molecular weight and Y² is non-oxo-carbonyl.