Novel chemiluminescent reagents

• US 20050004350A1
• Filed: 01/23/2004
• Published: 01/06/2005
• Est. Priority Date: 03/12/2002
• Status: Active Grant

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Inventors: Stavrianopoulos, Jannis G., Rabbani, Elazar
• Granted Patent: US 7,256,299 B2
• Time in Patent Office: Days
• US Class Current: 530/351
• 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

1-286. -286. (Canceled)

287. A chemiluminescent reagent having the structure:

288. The chemiluminescent reagent of claim 287, wherein said Q comprises an adamantyl group.

289. The chemiluminescent reagent of claim 287, wherein said cyclic ring comprises an aromatic ring.

290. The chemiluminescent reagent of claim 287, wherein R₂ comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.

291. The chemiluminescent reagent of claim 290, wherein said substituted aliphatic group comprises halogen, nitrates, sulfonates or nitrites.

292. The chemiluminescent reagent of claim 287, wherein enzymatically convertible R₁ comprises amides, esters, phosphates, carboxylic acids, fatty acids, glucose, xylose, fucose, or amino acids.

293. The chemiluminescent reagent of claim 287, wherein R₁ is enzymatically converted into R₁* through the action of enzymes comprising amidases, esterases, acetylcholinesterases, acid phosphatases, alkaline phosphatases, decarboxylases, lipases, glucosidases, xylosidases, fucosidases, trypsin or chymotrypsin.

294. The chemiluminescent reagent of claim 287, 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 G₂.

295. A chemiluminescent reagent having the structure:

296. The chemiluminescent reagent of claim 295, wherein said non-cleaving enzymatic process comprises oxidation or reduction.

297. The chemiluminescent reagent of claim 295, wherein said Q comprises an adamantyl group.

298. The chemiluminescent reagent of claim 295, wherein R comprises a substituted or unsubstituted aliphatic group or an unsubstituted aromatic group.

299. The chemiluminescent reagent of claim 298, wherein said substituted aliphatic group comprises halogen, nitrate, sulfonate or nitrite.

300. The chemiluminescent reagent of claim 295, wherein said reactive site comprises an oxygen, a nitrogen or a sulfur atom.

301. The chemiluminescent reagent of claim 295, wherein said enzymatic process is carried out by an enzyme comprising an oxidase or reductase.

302. The chemiluminescent reagent of claim 295, wherein after said enzymatic process, said dioxetane is converted to an unstable light-emitting dioxetane form.