Labeling reagents and labeled targets comprising nonmetallic porphyrins

• US 20060172308A1
• Filed: 01/22/2004
• Published: 08/03/2006
• 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 7,537,751 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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. (canceled)

2-286. -286. (canceled)

287. A labeling reagent comprising a nonmetallic porphyrin, said reagent comprising:

288. The labeling reagent of claim 287, wherein said reactive group R₀ is attached directly or indirectly to any one of the four non-pyrrole positions.

289. The labeling reagent of claim 287, wherein said reactive group R₀ comprises sulfhydryl, hydroxyl, amine, isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonylhalide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imidoester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde.

290. The labeling reagent of claim 287, wherein as a reactive group R₀ is capable of forming a carbon-carbon linkage with a target.

291. The labeling reagent of claim 287, wherein said reactive group R₀ comprises an alkene group, an alkyne group or a halogenated compound.

292. The labeling reagent of claim 287, wherein any of said R₁ through R₈ alkyl groups comprises methyl, ethyl or propyl.

293. The labeling reagent of claim 287, wherein any of said R₁ through R₈ alkyl groups further comprises a polar or charged group.

294. The labeling reagent of claim 287, wherein said reactive group R₀ is attached indirectly to said nonmetallic porphyrin through a linker arm.

295. The labeling reagent of claim 294, wherein said linker arm comprises at least two consecutive peptide bonds.

296. A labeled target comprising a nonmetallic porphyrin, said reagent comprising:

297. The labeled target of claim 296, wherein said reactive group R₀ is attached directly or indirectly to any one of the four non-pyrrole positions.

298. The labeled target of claim 296, wherein said target T comprises a protein, a peptide, a nucleic acid, a nucleotide or a nucleotide analog, a receptor, a natural or synthetic drug, a synthetic oligomer, a synthetic polymer, a hormone, a lymphokine, a cytokine, a toxin, a hapten, an antibody, a carbohydrate, a sugar or an oligo- or polysaccharide.

299. The labeled target of claim 298, wherein said nucleic acid or nucleotide or nucleotide analog is modified.

300. The labeling reagent of claim 296, wherein any of said R₁ through R₈ alkyl groups comprises methyl, ethyl or propyl.

301. The labeling reagent of claim 296, wherein any of said R₁ through R₈ alkyl groups further comprises a polar or charged group.

302. The labeling reagent of claim 296, wherein said target T is attached indirectly to said nonmetallic porphyrin through a linker arm.

303. The labeling reagent of claim 302, wherein said linker arm comprises at least two consecutive peptide bonds.