Fluorophore chelated lanthanide luminescent probes with improved quantum efficiency

Patent metadata

Field	Value
Publication number	US-9732378-B2
Application number	US-201514980342-A
Country	US
Kind code	B2
Filing date	Dec 28, 2015
Priority date	Jan 13, 2010
Publication date	Aug 15, 2017
Grant date	Aug 15, 2017

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.

First claim

What is claimed is:

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:

and wherein:
R 1 is the site of a covalent attachment, optionally through a linker, of the sensitizer moiety to the chelating moiety;
R 2 is a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear (C 1 -C 20 ) alkoxylene, a branched (C 3 -C 20 ) alkoxylene, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N or O; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene, alkylarene, or alkoxylene moieties are optionally further substituted with from 1-4 halogen atoms;
R 3 is a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear (C 1 -C 20 ) alkoxylene, a branched (C 3 -C 20 ) alkoxylene, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N or O; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene, alkylarene, or alkoxylene moieties are optionally further substituted with from 1-4 halogen atoms;
R 4 is H; a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear (C 1 -C 20 ) alkoxylene, a branched (C 3 -C 20 ) alkoxylene, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N, O, or S; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene, alkylarene, or alkoxylene moieties are optionally further substituted with from 1-4 halogen atoms;
wherein at least one of R 2 , R 3 , or R 4 further contains a crosslinking group selected from the group consisting of nitrogen mustard, epoxide, maleimide, disulfide, activated ester, imidate, azide, acetylene, aldehyde, sulfonyl chloride, acylazide, and acylhydrazide;
and wherein Formula (II) is:

and wherein:
R 1 is the site of a covalent attachment, optionally through a linker, of the sensitizer moiety to the chelating moiety;
R 5 is H; a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N, O, or S; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene, or alkylarene moieties are optionally further substituted with from 1-4 halogen atoms;
R 6 is a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N, O, or S; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene, or alkylarene moieties are optionally further substituted with from 1-4 halogen atoms;
R 7 is H; a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N, O, or S; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene, or alkylarene moieties are optionally further substituted with from 1-4 halogen atoms;
wherein at least one of R 5 , R 6 , or R 7 further contains a crosslinking group selected from the group consisting of nitrogen mustard, epoxide, maleimide, disulfide, activated ester, imidate, azide, acetylene, aldehyde, sulfonyl chloride, acylazide, and acylhydrazide;
wherein the R 1 chelating moiety of Formula (I) or Formula (II) is selected from the group consisting of EDTA, DTPA, TTHA, DOTA, TAGA, DOTP, DTPA-BMA, DO2P, and HP-DO3A; and
wherein the luminescent composition further comprises a lanthanide chelated to the chelating moiety, the lanthanide selected from the group consisting of: Gd(III), Dy(III), Ho(III), Er(III), Eu(III), Tb(III), Sm(III), Ce(III), Pr(III), Yb(III), Tm(III), Nd(III), and Tb(IV); and
detecting a signal produced from 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:

and wherein:
R 1 is the site of a covalent attachment, optionally through a linker, of the sensitizer moiety to the chelating moiety;
R 2 is a linear alky

Assignees

• Univ Rutgers
• New Jersey Inst Technology

Inventors

• Mustaev Arkady
• Kozlov Maxim
• Marras Salvatore
• Krasnoperov Lev
• Wirpsza Laura
• Pillai Shyamala

Classifications

• C07K14/00

  Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof · CPC title

• C12Q2565/1015

  labels being on the same oligonucleotide · CPC title

• C12Q1/6816 Primary

  characterised by the detection means (C12Q1/6804 takes precedence) · CPC title

• A61K49/0021

  the fluorescent group being a small organic molecule · CPC title

• C07H21/00

  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 title