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-2016018389-A1
Application number	US-201414766386-A
Country	US
Kind code	A1
Filing date	May 29, 2014
Priority date	May 30, 2013
Publication date	Jan 21, 2016
Grant date	—

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

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 an organic compound added.

2 . The method according to claim 1 , wherein the organic compound is a dye compound having a molecular weight of 2000 or less.

3 . The method according to claim 2 , wherein the organic compound comprises one or more compounds 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 represent 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 represent 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 represent 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 represent 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.

4 . The method according to claim 3 , wherein the ring formed when R 2 and R 4 of the general formula (1) are bound to each other is a cyclopentane ring.

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

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

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

8 . The method according to claim 2 , wherein the organic compound contains at least compound represented by general formula (6):

wherein, in the general formula (6), R 24 and R 25 each independently represent an alkyl group, a carboxylalkyl group, an alkoxycarbonylalkyl group, or an alkylcarbonyloxyalkyl group; R 26 to R 33 each independently represent 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 1 − 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 represent 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 represent 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 represent a hydrogen atom, an alkyl group or an alkyloxycarbonyl group.

9 . The method according to claim 8 , 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 represent an alkyl group, a carboxylalkyl group, an alkylcarbonyloxyalkyl group, or an alkoxycarbonyl alkyl group; R 44 to R 51 each independently represent 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 represent a hydrogen atom, an alkyl group or an aryl group; m represents a value of 0 to 2; X 2 − represents an anionic group; and Y 4 and Y 5 each represent 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.

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

11 . The method according to claim 2 , wherein the organic compound contains one or more compounds represented by general formula (10):

wherein, in the general formula (10), R 55 and R 56 each independently represent an alkyl group; R 57 and R 58 each independently represent a hydrogen atom or an alkyl group, wherein R 55 and R 57 , and R 56 and R 58 may be independently bound to each other to form a ring; R 59 represents a hydrogen atom, an alkyl group, an aryl group, an alkoxy group or a halogen atom; X 5 rep

Assignees

• Canon Kk

Inventors

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

Classifications

• C07D401/06

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

• C07D498/04

  Ortho-condensed systems · CPC title

• C07D209/14

  Radicals substituted by nitrogen atoms, not forming part of a nitro radical · CPC title

• C07D277/64

  with only hydrocarbon or substituted hydrocarbon radicals attached in position 2 · CPC title

• C07D413/08

  linked by a carbon chain containing alicyclic rings · CPC title