Probe for a biological specimen and labelling method and screening method using the probe

Abstract

Provided is a novel probe for a biological specimen for labelling by itself and clearly visualizing one of a specific cell and a specific cell organ in a living body, the probe having excellent spectral characteristics and exhibiting excellent storage stability. The probe for a biological specimen contains, as an active agent, at least one kind of compound represented by a general formula (I).

1 . A probe for a biological specimen, comprising, as an active agent, at least one kind of compound represented by the general formula (I): 
       
         
           
           
               
               
           
         
         where: 
         R 1  represents one of a hydrogen atom, an alkyl group, an aralkyl group, an alkenyl group, an aryl group, a heterocyclic group, and an acyl group; 
         R 2  to R 5  each independently represent one of a hydrogen atom, an alkyl group, an aryl group, a carboxylic acid group, a carboxylic acid ester group, and an acyl group, and R 2  and R 4  may be bonded to each other to form a ring; and 
         R 6  represents one of a hydrogen atom, an alkyl group, an alkoxy group, and a halogen atom, R 7  and R 8  each independently represent one of a hydrogen atom, an alkenyl group, a cyano group, a carboxylic acid group, a carboxylic acid ester group, a sulfonic acid group, an acyl group, and a heterocyclic group, and R 7  and R 8  may be bonded to each other to form a ring. 
       
     
     
         2 . A probe for a biological specimen according to  claim 1 , wherein any one of R 7  and R 8  in the general formula (I) represents one of a cyano group, a carboxylic acid group, and a heterocyclic group. 
     
     
         3 . A probe for a biological specimen according to  claim 1 , wherein any one of R 7  and R 8  in the general formula (I) represents a hydrogen atom, and the other represents one of a heterocyclic group represented by the following general formula (II) and an alkenyl group: 
       
         
           
           
               
               
           
         
         where: 
         R 9  represents one of an alkyl group and an aryl group; 
         R 10  to R 13  each independently represent a hydrogen atom, an alkyl group, an aryl group, an alkoxy group, a carboxylic acid group, a sulfonic acid group, a heterocyclic group, an amino group, and a halogen atom, one of R 10  and R 11 , R 11  and R 12 , and R 12  and R 13  may be bonded to each other to form a ring, and X— represents an anionic group; 
         Q 1  represents one of a sulfur atom, an oxygen atom, —C(R 14 )(R 15 )—, —CH═CH—, and —N(R 16 )—; and 
         R 14  to R 16  each represent one of a hydrogen atom, an alkyl group, and an aryl group. 
       
     
     
         4 . A probe for a biological specimen according to  claim 3 , wherein the heterocyclic group represented by the general formula (II) comprises a benzothiazolyl ring group, a benzoxazolyl ring group, and a dimethylindolenyl ring group. 
     
     
         5 . A probe for a biological specimen according to  claim 1 , wherein R 7  and R 8  represented in the general formula (I) are bonded to each other to form a ring, and the ring comprises a heterocycle formed of one of 5- and 6-membered rings. 
     
     
         6 . A probe for a biological specimen according to  claim 5 , wherein the heterocycle formed of a 5-membered ring is represented by the following general formula (III): 
       
         
           
           
               
               
           
         
         where: 
         R 17  represents one of a hydrogen atom, an alkyl group, an aryl group, and a heterocyclic group; and 
         R 18  represents one of an alkyl group, an aryl group, a carboxylic acid group, a carboxylic acid ester group, a hydroxyl group, and an amino group. 
       
     
     
         7 . A probe for a biological specimen according to  claim 5 , wherein the heterocycle formed of a 5-membered ring is represented by the following general formula (IV): 
       
         
           
           
               
               
           
         
         where: 
         Q 2  represents one of an oxygen atom, a sulfur atom, and —N(R 21 )—; 
         R 19  represents one of a hydrogen atom, an alkyl group, an aryl group, and a heterocyclic group; 
         R 20  represents one of a sulfur atom, an oxygen atom, ═NR 22 , a heterocyclic group, a methylene group replaced by a heterocyclic group, and a dicyanomethylene group; 
         R 21  and R 22  each represent one of a hydrogen atom, an alkyl group, an aryl group, and a heterocyclic group. 
       
     
     
         8 . A probe for a biological specimen according to  claim 7 , wherein Q 2  represents a sulfur atom. 
     
     
         9 . A probe for a biological specimen according to any one of  claim 7 , wherein R 20  in the general formula (IV) represents one of a sulfur atom and 2-thioxothiazolidin-4-one having a substituent at a 3-position. 
     
     
         10 . A probe for a biological specimen according to  claim 1 , wherein R 2  and R 4  represented in the general formula (I) are bonded to each other to form a ring, and the ring comprises an aliphatic ring. 
     
     
         11 . A probe for a biological specimen according to  claim 1 , wherein R 2  and R 4  represented in the general formula (I) are bonded to each other to form a ring, and the ring comprises a cyclopentane ring. 
     
     
         12 . A probe for a biological specimen according to  claim 1 , wherein the compounds represented by the general formulae (I) to (IV) comprise at least one of a carboxylic acid group, a sulfonic acid group, a polyethylene glycol group, a carboxylic acid salt, and a sulfonic acid salt. 
     
     
         13 . A labelling method, wherein one of staining and differential staining is performed by bringing a biological specimen into contact with the probe for a biological specimen according to  claim 1 . 
     
     
         14 . A labelling method according to  claim 13 , wherein the biological specimen is used in vivo, in vitro, and ex vivo. 
     
     
         15 . A screening method, wherein the probe for a biological specimen according to  claim 1  is utilized. 
     
     
         16 . A screening method according to  claim 15 , wherein a small bony fish is used. 
     
     
         17 . A screening method according to  claim 16 , wherein the small bony fish comprises Zebrafish. 
     
     
         18 . A diagnostic composition, comprising, as an active agent, the probe for a biological specimen according to  claim 1 . 
     
     
         19 . A diagnostic device, comprising the probe for a biological specimen according to  claim 1 .