Polynitrogen compounds and uses thereof as fluorescent chromophores

Patent metadata

Field	Value
Publication number	US-10954436-B2
Application number	US-201615746386-A
Country	US
Kind code	B2
Filing date	Jul 19, 2016
Priority date	Jul 20, 2015
Publication date	Mar 23, 2021
Grant date	Mar 23, 2021

Abstract

The present invention relates to the use, as a fluorescent chromophore, of a compound with formula (I) wherein: A1 is —N— or —C(Y1)—; A2 is —N— or —C(Y2)—; A3 is —N— or —C(Y3)—; A4 is —N— or —C(Y4)—; at least one of A1, A2, A3 and A4 representing —N—; X1 is —N— or —C(Y5)—; X2 is —N— or —C(Y6)—; X3 is —N— or —C(Y7)—; and Y1, Y2, Y3, Y4, Y5, Y6 and Y7 are in particular chosen independently of one another from the group made up of: H, electron-donor groups and electron-attracting groups.

First claim

What is claimed is:

1. A method of detecting a biological molecule by fluorescence, comprising:
conjugating the biological molecule with a fluorescent chromophore, and
detecting a fluorescent signal from said fluorescent chromophore,
wherein said fluorescent chromophore is a compound of formula (I):

wherein:
A 1 is —N— or —C(Y 1 )—;
A 2 is —N— or —C(Y 2 )—;
A 3 is —N— or —C(Y 3 )—;
A 4 is —N— or —C(Y 4 )—;
at least one of A 1 , A 2 , A 3 and A 4 representing —N—;
Y 1 , Y 2 , Y 3 and Y 4 are selected independently of one another from the group consisting of: H, electron-donor groups and electron-attracting groups,
where Y 1 and Y 2 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 30 atoms,
and/or Y 2 and Y 3 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 30 atoms,
and/or Y 3 and Y 4 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 30 atoms,
X 1 is —N— or —C(Y 5 )—;
X 2 is —N— or —C(Y 6 )—;
X 3 is —N— or —C(Y 7 )—;
Y 5 , Y 6 and Y 7 are selected independently of one another from the group consisting of H, electron-donor groups and electron-attracting groups,
where Y 5 and Y 6 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 10 atoms,
and/or Y 6 and Y 7 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 10 atoms,

said compound with formula (I) being able to be in salt or pure stereoisomer form or in the form of a mixture of enantiomers and/or diastereoisomers, including racemic mixtures, or in the form of tautomers,
with the exception of the compound with formula:

2. The method according to claim 1 , wherein, in formula (I), Y 1 , Y 2 , Y 3 and Y 4 are selected independently of one another from the group consisting of: H, halogen, (C 1 -C 6 )alkyl, (C 6 -C 10 )aryl, heteroaryl comprising from 5 to 10 atoms, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkynyl, CN, hydroxy, (C 1 -C 6 )alcoxy, thiol, (C 1 -C 6 )alkylthio, (CH 2 ) n SO 2 —OR a where n=1-6, CH 2 SO 2 —NR a R b , NO 2 , SO 3 R a , NR a R b , C(O)OR a , C(O)R a , and C(O)NR a R b , R a and R b representing H or a (C 1 -C 6 )alkyl group.

3. The method according to claim 1 , wherein, in formula (I), Y 5 , Y 6 , and Y 7 represent, independently of one another, a hydrogen atom or a halogen atom, or are selected from the group consisting of:
a (C 1 -C 6 )alkyl, (C 3 -C 6 )cycloalkyl or (C 6 -C 10 )aryl group,
said (C 6 -C 10 )aryl group optionally being substituted by at least one substituent chosen from among: halogen, CN, (C 1 -C 6 )alkyl, OH, (C 1 -C 6 )alkoxy, C(O)OR a and NR a R b , R a and R b representing H or a (C 1 -C 6 )alkyl group,
a heteroaryl group comprising from 5 to 10 chain links and containing from 1 to 4 heteroatoms chosen from among O, S or N, said heteroaryl group optionally being substituted by at least one substituent chosen from among: halogen, CN, (C 1 -C 6 )alkyl, OH, (C 1 -C 6 )alkoxy, C(O)OR a and NR a R b , R a and R b representing H or a (C 1 -C 6 )alkyl group,
a heterocycloalkyl group comprising from 4 to 10 chain links and containing from one to three heteroatoms chosen from among O, S or N,
NO 2 ,
CHO,
a C(O)OR a group, R a being as defined above,
a —HC═CH—Ar group, Ar representing a (C 6 -C 10 )aryl group,
SO 3 H,
a NR c R d group, R c and R d representing H or a (C 1 -C 6 )alkyl group, or being able to form a hetero(C 2 -C 5 )cycloalkyl group with the nitrogen atom that carries them;
an OR e group, R e representing H, a (C 1 -C 6 )alkyl group or a (C 6 -C 10 )aryl group; and
a group with the following formula (A):

4. The method according to claim 1 , wherein A 1 and A 4 are —N—, A 2 is —C(Y 2 )— and A 3 is —C(Y 3 )—.

5. A conjugate comprising a biological molecule and a compound with formula (I) according to claim 1 , in which said compound with formula (I) is linked to the biological molecule via a linker, said biological molecule being selected from the group consisting of antibodies, proteins, peptides, carbohydrates, lipids, polysaccharides, fatty acids, amino acids, deoxyribonucleic acids, ribonucleic acids, oligonucleotides, medicinal drugs and ligands.

6. The conjugate according to claim 5 , with the following formula (I″):

wherein:
A 1 is —N— or —C(Y 1 )—;
A 2 is —N— or —C(Y 2 )—;
A 3 is —N— or —C(Y 3 )—;
A 4 is —N— or —C(Y 4 )—;

at least one of A 1 , A 2 , A 3 and A 4 representing —N—;
Y 1 , Y 2 , Y 3 and Y 4 are selected independently of one another from the group consisting of: H, electro-donor groups and electro-attracting groups, or can represent a group with formula -L′-Z′, L′ representing a linker and Z′ representing a biological molecule chosen from the group made up of antibodies, proteins, peptides, carbohydrates, lipids, polysaccharides, fatty acids, amino acids, deoxyribonucleic acids, ribonucleic acids, oligonucleotides, medicinal drugs and ligands,

where Y 1 and Y 2 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 30 atoms,
and/or Y 2 and Y 3 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 30 atoms,
and/or Y 3 and Y 4 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 30 atoms,
X 1 is —N— or —C(Y 5 )—;
X 2 is —N— or —C(Y 6 )—;
X 3 is —N— or —C(Y 7 )—;
Y 5 , Y 6 and Y 7 are selected independently of one another from the group consisting of H, electron-donor groups and electron-attracting groups, or represent a group with formula -L′-Z′, L′ and Z′ being as defined above,

where Y 5 and Y 6 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 10 atoms,
and/or Y 6 and Y 7 together form, with the carbon atoms that carry them, a (hetero)cycloalkyl group or a (hetero)aryl group comprising from 5 to 10 atoms,

said compound with formula (I″) being able to be in salt or pure stereoisomer form or in the form of a mixture of enantiomers and/or diastereoisomers, including racemic mixtures, or in the form of tautomers,
wherein at least one of Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 and Y 7 is a group with formula -L′-Z′.

7. A method for detecting at least one biological molecule in a biological medium, comprising:
introducing the conjugate of claim 6 into said biological medium, said conjugate comprising a fluorescent marker and a biological molecule,
exciting said medium, and
detecting at least one fluorescence signal of said fluorescent marker.

8. A method for detecting a biological molecule by fluorescence, the method comprising contacting a conjugate according to claim 5 with at least one biological molecule.

9. A method f

Assignees

• Univ Orleans
• Centre Nat Rech Scient

Inventors

• Suzenet Franck
• Sirbu Doina
• Guillaumet Gérald
• Bonnet Pascal

Classifications

• C07D471/14

  Ortho-condensed systems · CPC title

• C07F5/025

  Boronic and borinic acid compounds · CPC title

• C07D487/14

  Ortho-condensed systems · CPC title

• C07D487/04 Primary

  Ortho-condensed systems · CPC title

• C07F7/0812

  comprising a heterocyclic ring · CPC title