Grafted dinuclear metal complexes, and use thereof as cellular microparticle sensors

Patent metadata

Field	Value
Publication number	US-10180427-B2
Application number	US-201615255704-A
Country	US
Kind code	B2
Filing date	Sep 2, 2016
Priority date	Mar 23, 2011
Publication date	Jan 15, 2019
Grant date	Jan 15, 2019

Abstract

Novel grafted dinuclear metal complexes having formula (I) or of formula (II)

wherein M represents a metal ion,
and also the use thereof as sensors in a method for detecting and/or characterising cellular microparticles.

First claim

The invention claimed is:

1. A method for detection and/or characterisation of cellular microparticles, comprising:
placing in contact of a compound having formula (I) or (II)

wherein
M +i is a metal ion, and i is 1, 2 or 3
L is an exchangeable ligand
X is a —(CH 2 ) m —NH-A group, or a —CH 2 —NHC(O)—R—NH-A group wherein R is a C 2 -C 10 alkyl group, either substituted or unsubstituted, linear or branched, and A is H or a fluorophore group
m=1 to 12
n=1, 2 or 3
Y is (CH 2 ) p —NH 2 wherein p=0 to 12,

when p=0, the pyridine group is not substituted by Y, with a sample of biological fluid that is likely to contain cellular microparticles, and detection and/or characterisation of the microparticles captured by the said compound having formula (I) or (II).

2. The method of claim 1 , wherein M is selected from the group consisting of Zn, Cu, Mn, Co, Ni and Fe.

3. The method of claim 1 , wherein:
M is Zn or Cu
X is —(CH 2 ) m —NH-A, where m=1, A is H, n=1 and p=0; or
X is —(CH 2 ) m —NH-A, where m=1, A is H, n=2 and p=0; or
X is —CH 2 —NHC(O)—R—NH-A, A is H, n=1 and p=0.

4. The method of claim 1 , wherein X is —(CH 2 ) m —NH-A, —CH 2 —NHC(O)—R—NH-A wherein R is a C 2 -C 10 alkyl group, either substituted or unsubstituted, linear or branched, and A is a fluorophore group.

5. The method of claim 1 , wherein the compound having formula (I) is, in a first step, immobilised on a solid support.

6. The method of claim 5 , further comprising the following steps:
immobilisation of a compound having formula (I) or (II) on the surface of the solid support,
placing in contact with a sample of biological fluid that is likely to contain cellular microparticles
capture of the cellular microparticles on the said compound having formula (I) or (II), and
detection and/or characterisation of the captured cellular microparticles.

7. The method of claim 1 , wherein the solid support is activated prior to immobilisation of the compound having formula (I) or (II).

8. The method of claim 7 , further comprising the following steps:
activation of a solid support,
immobilisation of a compound having formula (I) or (II) on a surface of the activated support,
placing in contact with a sample of biological fluid that is likely to contain cellular microparticles
capture of the cellular microparticles on the said compound having formula (I) or (II), and
detection and/or characterisation of the captured cellular microparticles.

9. The method of claim 1 , further comprising the following steps:
immobilisation of a compound having formula (III) on a solid support

in which
Z is NH 2 or a —(CH 2 ) m —NH-A group, or a -CH 2 —NHC(O)—R—NH-A group wherein R is a C 2 -C 10 alkyl group, either substituted or unsubstituted, linear or branched, and A is H or a fluorophore group, provided that when m=2, A is different from H
m=1 to 12
n=1, 2 or 3
Y is (CH 2 ) p -NH 2 where p=0 to 12,
incubation of the support-ligand ensemble in the presence of the metal ion, in a manner such that the dinuclear metal complex having formula (I) or (II) is formed in situ,
the placing in contact of said compound having formula (I) or (II) with a sample of biological fluid that is likely to contain cellular microparticles,
the capture of the cellular microparticles on the said compound having formula (I) or (II), and
the detection and/or characterisation of the captured cellular microparticles.

10. The method of claim 1 , wherein the solid support is a microtiter plate, a sheet, a cone, a tube, a well, a bead, a particle or a strip.

11. The method of claim 1 , wherein the compound having formula (I) or (II) is in a solution.

12. The method of claim 11 , wherein in the compound having formula (I) or (II), X is —(CH 2 ) m —NH-A or CH 2 —NHC(O)—R—NH-A, wherein R is C 2 -C 10 alkyl group, either substituted or unsubstituted, linear or branched, and A is a fluorophore.

13. The method of claim 1 , wherein the captured microparticles are revealed by using specific labelled antibodies that may be detected by means of photometric analysis or using fluorescence measurement.

14. The method of claim 13 , wherein the antibodies are labelled with an enzyme and then detected by means of chromogenic analysis using a suitable substrate.

Assignees

• Centre Nat Rech Scient
• Univ De Caen Basse Normandie
• Univ Joseph Fourier
• Univ Grenoble Alpes

Inventors

• Belle Catherine
• Gellon Gisele
• Plawinski Laurent
• Doeuvre Loic
• Angles Cano Eduardo

Classifications

• C07F1/08

  Copper compounds · CPC title

• C07D401/14

  containing three or more hetero rings · CPC title

• C07D213/38 Primary

  having only hydrogen or hydrocarbon radicals attached to the substituent nitrogen atom · CPC title

• C07F3/06

  Zinc compounds · CPC title

• G01N33/54353 Primary

  with ligand attached to the carrier via a chemical coupling agent (coatings G01N33/54393) · CPC title