Fluorophore chelated lanthanide luminescent probes with improved quantum efficiency

• US 9,732,378 B2
• Filed: 12/28/2015
• Issued: 08/15/2017
• Est. Priority Date: 01/13/2010
• Status: Active Grant

• Current Assignee: New Jersey Institute of Technology, Rutgers University
• Original Assignee: New Jersey Institute of Technology, Rutgers University
• Inventors: Kozlov, Maxim, Marras, Salvatore, Krasnoperov, Lev, Wirpsza, Laura, Pillai, Shyamala, Mustaev, Arkady
• Primary Examiner(s): Hartley, Michael G
• Assistant Examiner(s): SAMALA, JAGADISHWAR RAO
• Application Number:
  US14/980,342
• Publication Number:
  US 20160194690A1
• Time in Patent Office: 596 Days
• CPC Class Codes: A61K 49/0021 the fluorescent group being a small organic molecule
• CPC Class Codes: A61K 49/0028 Oxazine dyes
• CPC Class Codes: A61K 49/0052 Small organic molecules oligomers, polymers, dendrimers A61K49/0054
• CPC Class Codes: C07D 215/227 only one oxygen atom which is attached in position 2
• CPC Class Codes: C07D 215/233 only one oxygen atom which is attached in position 4
• CPC Class Codes: C07D 215/38 Nitrogen atoms nitro radicals C07D215/18
• CPC Class Codes: C07D 257/02 not condensed with other rings
• 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: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C09B 57/00 Other synthetic dyes of known constitution
• CPC Class Codes: C09B 57/02 Coumarine dyes
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/103 luminescence
• CPC Class Codes: C12Q 2565/1015 labels being on the same oligonucleotide
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

The invention relates to novel luminescent compositions of matter containing a fluorophore, synthetic methods for making the compositions, macromolecular conjugates of the compositions, and the use of the compositions in various methods of detection. The invention also provides kits containing the compositions and their conjugates for use in the methods of detection.

Claims

1. An in vitro method of detecting a target with a probe, the method comprising:

contacting a sample with a luminescent probe composition comprising a targeting-probe moiety having an affinity for a target, the targeting-probe moiety covalently conjugated to a luminescent chelate composition comprising;

(i) a fluorophore of Formula (I) or Formula (II); and

(ii) a chelating moiety covalently joined, optionally through a first linker, to the fluorophore,

wherein Formula (I) is;

3. The method of claim 1, wherein the detection comprises detecting fluorescence emissions from the fluorophore of the luminescent probe composition.

2. An in vivo method of detecting a target with a probe, the method comprising:

administering to a subject a luminescent probe composition comprising a target-probe moiety having an affinity for a target, the targeting-probe moiety covalently conjugated to a luminescent chelate composition comprising;

(i) a fluorophore of Formula (I) or Formula (II); and

(ii) a chelating moiety covalently joined, optionally through a first linker, to the fluorophore,

wherein Formula (I) is;

4. The method of claim 2, wherein the detection comprises detecting fluorescence emissions from the fluorophore of the luminescent probe composition.

5. A method for detecting an analyte in a sample comprising the steps of:

(a) contacting a sample containing the analyte with a luminescent chelate composition to specifically bind the analyte to form a luminescent-analyte complex;

(b) illuminating the sample with excitation radiation; and

(c) detecting emission radiation of at least one luminescent-analyte complex;

wherein the luminescent chelate composition comprises;

(i) a fluorophore of Formula (I) or Formula (II); and

(ii) a chelating moiety covalently joined, optionally through a first linker, to the fluorophore;

wherein Formula (I) is;

6. The method of claim 5, wherein the analyte is contacted with a first luminescent chelate composition and a second luminescent chelate composition, and wherein the first luminescent chelate composition comprises a first lanthanide and the second luminescent chelate composition comprises a second lanthanide, wherein the first and second lanthanides are different.

7. The method of claim 5, wherein the analyte is contacted with a first luminescent chelate composition and a second luminescent chelate composition, and wherein the first and second luminescent chelate compositions have the same emission spectra, but different emission lifetime.

8. The method of claim 7, wherein the R₁ chelating moieties of the first and second luminescent chelate compositions independently comprise at least one modifier selected from the group consisting of hydrazine, diamine, tetraamine, and cyclic tetraamine.

9. The method of claim 5, wherein the analyte is contacted with a first luminescent chelate composition and a second luminescent chelate composition, and wherein the first and second luminescent chelate compositions can be selectively excited at different wavelengths.