Chemiluminescent reagents

• US 7,256,299 B2
• Filed: 01/23/2004
• Issued: 08/14/2007
• Est. Priority Date: 03/12/2002
• Status: Expired due to Term

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Stavrianopoulos, Jannis G., Rabbani, Elazar
• Primary Examiner(s): Riley; Jezia
• Application Number:
  US10/764,388
• Publication Number:
  US 20050004350A1
• Time in Patent Office: 1,299 Days
• Field of Search: 549/510, 436/172, 252/301.16, 252/186.42
• US Class Current: 549/510
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C09B 23/02 the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
• CPC Class Codes: C09B 47/00 Porphines; Azaporphines non-dyeing compounds C07D487/22
• CPC Class Codes: C09B 57/00 Other synthetic dyes of known constitution
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

This invention provides for labeling reagents, labeled targets and processes for preparing labeling reagents. The labeling reagents can take the form of cyanine dyes, xanthene dyes, porphyrin dyes, coumarin dyes or composite dyes. These labeling reagents are useful for labeling probes or targets, including nucleic acids and proteins. These reagents can be usefully applied to protein and nucleic acid probe based assays. They are also applicable to real-time detection processes.

Claims

1. A chemiluminescent reagent having the structure:

2. The chemiluminescent reagent of claim 1, wherein said Q comprises an adamantyl group.

3. The chemiluminescent reagent of claim 1, wherein said cyclic ring comprises an aromatic ring.

4. The chemiluminescent reagent of claim 1, wherein R₂ comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.

5. The chemiluminescent reagent of claim 4, wherein said substituted aliphatic group comprises halogen or sulfonates.

6. The chemiluminescent reagent of claim 1, wherein enzymatically convertible R₁ comprises glucose, xylose, fucose, amino acids or esters of phosphates, carboxylic acids or fatty acids.

7. The chemiluminescent reagent of claim 1, wherein R₁ is enzymatically converted into R₁* through the action of enzymes comprising an amidases, trypsin or chymotrypsin.

8. The chemiluminescent reagent of claim 1, wherein after said enzymatic conversion of R₁ to R₁* and before said conversion of said dioxetane to the unstable light-emitting dioxetane form, an intermediate five- or six-membered ring is formed comprising a linkage between said G₁ and G_2.

9. A chemiluminescent reagent having the structure:

10. The chemiluminescent reagent of claim 9, wherein said non-cleaving enzymatic process comprises oxidation or reduction.

11. The chemiluminescent reagent of claim 9, wherein said Q comprises an adamantyl group.

12. The chemiluminescent reagent of claim 9, wherein R comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.

13. The chemiluminescent reagent of claim 12, wherein said substituted aliphatic group comprises halogen, or sulfonate.

14. The chemiluminescent reagent of claim 9, wherein said reactive site comprises an oxygen, a nitrogen or a sulfur atom.

15. The chemiluminescent reagent of claim 9, wherein said enzymatic process is carried out by an enzyme comprising an oxidase or reductase.

16. The chemiluminescent reagent of claim 9, wherein after said enzymatic process, said dioxetane is converted to an unstable light-emitting dioxetane form.