Macrophage identification agent, and identification method, sorting method, evaluation method, screening method and kit using the macrophage identifier agent

Patent metadata

Field	Value
Publication number	US-10041937-B2
Application number	US-201414766386-A
Country	US
Kind code	B2
Filing date	May 29, 2014
Priority date	May 30, 2013
Publication date	Aug 7, 2018
Grant date	Aug 7, 2018

Abstract

An object of the present invention is to provide an identification method for a macrophage subtype using an organic compound. Another object of the present invention is to provide a macrophage identification agent containing an organic compound in which a spectral characteristic obtained when the organic compound is added is different depending on a macrophage subtype. Still another object of the present invention is to provide an evaluation method for a macrophage subtype using a macrophage identification agent, an analysis method for correlation between a macrophage subtype and a test substance, a screening method for correlation between a macrophage subtype and a test substance, and a kit. An identification method for a macrophage subtype utilizing that a spectral characteristic obtained with an organic compound added is different depending on a macrophage subtype is provided.

First claim

The invention claimed is:

1. A method for identifying macrophage subtype M1 and macrophage subtype M2, wherein the subtype M1 and the subtype M2 are identified based on a spectral characteristic obtained with a dye compound having a molecular weight of 2000 or less added,
wherein the dye compound is at least one compound represented by general formula (1):

wherein, in the general formula (1), R 1 represents a hydrogen atom, an alkyl group, an aralkyl group, an alkenyl group, or an aryl group; R 2 to R 5 each independently represents a hydrogen atom, an alkyl group, an aryl group, a carboxyl group, a carboxylalkyl group, or an alkylcarbonyl group, wherein R 2 and R 4 may be bound to each other to form an aliphatic ring; R 6 represents a hydrogen atom, an alkyl group, an alkoxy group, or a halogen atom; R 7 and R 8 each independently represents a hydrogen atom, an alkyl group, or an aryl group; R 9 represents a dicyanomethylene group, a cyanocarboxymethylene group, or any one of general formulas (2) to (4); Y 1 represents a carbon atom having aryl group, a cyclopentene ring, a cyclohexene ring or a carbon atom; and n represents 0 or 1,

wherein, in the general formula (2), R 10 represents an alkyl group, a carboxylalkyl group, an alkoxycarbonylalkyl group, or an alkylcarbonyloxyalkyl group; R 11 represents a sulfur atom, a 2-thioxothiazolidin-4-one group, a thiazoline-2,4-one group, or general formula (5); and “*” represents a binding site,

wherein, in the general formula (5), R 12 represents an alkyl group, a carboxylalkyl group, an alkoxycarbonylalkyl group, or an alkylcarbonyloxyalkyl group; R 13 and R 14 each represent a hydrogen atom and may be bound to each other to form a benzene ring; Q 1 represents an oxygen atom, a sulfur atom, a N-alkyl nitrogen atom, or —C(R 15 )(R 16 )—, wherein R 15 and R 16 each independently represent an alkyl group and may be bound to each other to form a ring; X 1 − represents an anionic group; and “*” represents a binding site,
wherein, in the general formula (3), R 17 represents an alkyl group, a carboxylalkyl group, an alkoxycarbonylalkyl group, or an alkylcarbonyloxyalkyl group; R 18 and R 19 each represents a hydrogen atom and may be bound to each other to form a benzene ring; Q 2 represents an oxygen atom, a sulfur atom, a N-alkyl nitrogen atom, or —C(R 20 )(R 21 )—, wherein R 20 and R 21 each independently represents an alkyl group and may be bound to each other to form a ring; X 2 − represents an anionic group; and “*” represents a binding site, and
wherein, in the general formula (4), R 22 represents an alkyl group or an aryl group; R 23 represents an alkyl group or a carboxyl group; and “*” represents a binding site.

2. The method according to claim 1 , wherein the ring formed when R 2 and R 4 of the general formula (1) are bound to each other is a cyclopentane ring.

3. The method according to claim 1 , wherein R 9 is a compound represented by the general formula (2).

4. The method according to claim 1 , wherein R 9 is a compound represented by the general formula (3).

5. The method according to claim 1 , wherein R 9 is a compound represented by the general formula (4).

6. A method for identifying macrophage subtype M1 and macrophage subtype M2, wherein the subtype M1 and the subtype M2 are identified based on a spectral characteristic obtained with a dye compound having a molecular weight of 2000 or less added,
wherein the dye compound is at least one compound represented by general formula (6):

wherein, in the general formula (6), R 24 and R 25 each independently represents an alkyl group, a carboxylalkyl group, an alkoxycarbonylalkyl group, or an alkylcarbonyloxyalkyl group; R 26 to R 33 each independently represents a hydrogen atom, an alkyl group, an aryl group, an alkoxy group, a halogen atom, an alkoxysulfonyl group, a N-alkylsulfamoyl group, an alkyloxycarbonyl group, a carbamoyl group, or a N-alkylcarbamoyl group, wherein R 26 and R 27 , R 28 and R 29 , R 30 and R 31 , and R 32 and R 33 may be independently cyclized to form a benzene ring; X 3 − represents an anionic group; Y 2 represents a group containing *1, *2, and *5 and containing an oxygen atom, a sulfur atom, a N-alkyl nitrogen atom or —C(R 34 )(R 35 )—, or a group containing *1, *2, and *5 and represented by *1-C*5-CH═CH-*2 or *1-CH═CH-*5-*2; Y 3 represents a group containing *3, *4, and *6 and containing an oxygen atom, a sulfur atom, a N-alkyl nitrogen atom, or —C(R 34 )(R 35 )—, or a group containing *3, *4, and *6 and represented by *3-Y 3 -*6-*4, *3-C*6-CH═CH-*4, or *3-CH═CH-*6-*4,
wherein R 34 and R 35 each independently represents an alkyl group and may be bound to each other to form a ring; and
A represents a 3-oxocyclobutenolate ring or is represented by general formula (7) or (8):

wherein, in the general formula (7), R 36 to R 38 each independently represents a hydrogen atom, an alkyl group or an aryl group, and n represents a value of 0 to 2,
wherein, in the general formula (8), R 39 represents a hydrogen atom, a phenyl group, a thiol group, an alkoxy group, an aryloxy group, or a halogen atom; and R 40 and R 41 each independently represents a hydrogen atom, an alkyl group, or an alkyloxycarbonyl group.

7. The method according to claim 6 , wherein the compound represented by the general formula (6) is represented by general formula (9):

wherein, in the general formula (9), R 42 and R 43 each independently represents an alkyl group, a carboxylalkyl group, an alkylcarbonyloxyalkyl group, or an alkoxycarbonyl alkyl group; R 44 to R 51 each independently represents a hydrogen atom, an alkyl group, an aryl group, an alkoxy group, a halogen atom, an alkoxysulfonyl group, a N-alkylsulfamoyl group, an alkyloxycarbonyl group, or a N-alkylcarbamoyl group, wherein R 44 and R 45 , R 46 and R 47 , R 48 and R 49 , and R 50 and R 51 may be independently cyclized to form a benzene ring; R 49 to R 51 each independently represents a hydrogen atom, an alkyl group or an aryl group; m represents a value of 0 to 2; X 4 − represents an anionic group; and Y 4 and Y 5 each represents an oxygen atom, a sulfur atom, or an alkylene group, wherein the alkylene group may have a substituent, the substituent used in this case is an alkyl group, and the substituents may be bound to each other to form an aliphatic ring.

8. The method according to claim 6 , wherein A of the general formula (6) is a 3-oxocyclobutenolate ring.

9. A method for identifying macrophage subtype M1 and macrophage subtype M2, wherein the subtype M1 and the subtype M2 are identified based on a spectral characteristic obtained with a dye compound having a molecular weight of 2000 or less added,
wherein the dye compound is at least one compound represented by general formula (10):

wherein, in the general formula (10), R 55 and R 56 each independently represents an alkyl group; R 57 and R 58 each independen

Assignees

• Canon Kk

Inventors

• Koide Fumiyo
• Nomoto Tsuyoshi
• Yamauchi Sachiko
• Watanabe Kohei
• Shintou Taichi
• Miyazaki Takeshi
• Tabata Yasuhiko
• Sanda Eri

Classifications

• C07D403/06

  linked by a carbon chain containing only aliphatic carbon atoms · CPC title

• C07D209/80

  [b, c]- or [b, d]-condensed · CPC title

• C07F5/027

  Organoboranes and organoborohydrides · CPC title

• C07D413/06

  linked by a carbon chain containing only aliphatic carbon atoms · CPC title

• C07D403/08

  linked by a carbon chain containing alicyclic rings · CPC title