Porphyrin compounds, their conjugates and assay methods based on the use of said conjugates

• US 6,582,930 B1
• Filed: 04/20/2001
• Issued: 06/24/2003
• Est. Priority Date: 11/09/1998
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Arctic Diagnostics Oy
• Inventors: Meltola, Niko Jarmo Juhani, Soini, Aleksi Elias, Yashunsky, Dmitry Vladimirovich, Ponomarev, Gelii Vasilevich
• Primary Examiner(s): CEPERLEY, MARY
• Application Number:
  US09/807,866
• Time in Patent Office: 795 Days
• Field of Search: 436/544, 436/546, 436/800
• US Class Current: 435/40.5
• CPC Class Codes: C07D 487/22 in which the condensed system contains four or more hetero rings
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

The preparation of phosphorescent metalloporphyrin labelling reagents and their use for preparation of phosphorescent conjugates with biomolecules. The labelling reagents obtainable are water soluble monofunctional derivatives of Pt- and Pd-coproporphyrins, where the term “monofunctional” refers to the number of reactive groups in the porphyrin moiety.

Claims

1. A substantially pure compound of formula (I) or (II):

wherein

M is palladium (II) or platinum (II);

R₁ is —

OH, —

O⁻

X⁺ or —

OR;

where R is an alkyl, an aryl, an alkyaryl group or an aralkyl group and X⁺ is a cation

R₂ is —

Y—

Z, wherein

Y is either a covalent bond, a straight or branched chain alkylene, an arylene, an alkarylene or an aralkylene group, and may contain heteroatoms or side chains containing heteroatoms, or Y is a residue of a polypeptide, polysaccharide, polynucleotide or a polyether, and Z is a functional group selected from the group consisting of isothiocyanato and maleimido.

2. The compound of claim 1 wherein Y is an alkaryl group.

3. The compound of claim 1 wherein Z is a maleimido group.

4. The compound of claim 1 wherein Y is an alkaryl group and Z is an isothiocyanato group.

5. The compound of claim 1 wherein R₁ is —

OH or —

O⁻

X⁺.

6. The compound of claim 1 wherein R₁ is —

OH or —

O⁻

X⁺ and R₂ is —

(CH₂)_n—

Ph—

NCS where Ph is a phenyl and n is an integer from 1 to 10.

7. A labelled conjugate comprising a bioaffinity reactant labelled with one or several phosphorescent labels wherein the phosphorescent label is a compound of claim 1.

8. A conjugate of claim 7 wherein the bioaffinity reactant is a biologically active molecule.

9. A bioaffinity assay method wherein at least one of the biospecific reactants is labelled with the compound of claim 1.

10. The method of claim 9 wherein the analyte is a biologically active molecule.

11. A cytological or histological staining method wherein at least one of the biospecific reactants is labelled with the compound of claim 1.

12. The conjugate of claim 8, wherein said biologically active molecule is a member of the group consisting of a hapten, a biologically active ligand, a drug, a peptide, an oligonucleotide, a nucleotide, a nucleic acid, a polypeptide, a protein, an antibody and an antibody fragment.

13. The method of claim 10, wherein said biologically active molecule is a member of the group consisting of a hapten, a biologically active ligand, a drug, a peptide, an oligonucleotide, a nucleotide, a nucleic acid, a polypeptide, a protein, an antibody and an antibody fragment.