Probe for a hair cell, and labelling method for a hair cell using the probe for a hair cell

Abstract

Provided are a novel probe for a hair cell for clearly identifying various conditions of a hair cell, and a labelling method for a hair cell using the probe for a hair cell, more particularly, a probe for a hair cell containing, as an active agent, at least one kind selected from staining compounds represented by one of the general formulae (I) and (II), and a labelling method for a hair cell using the probe for a hair cell.

1 - 17 . (canceled) 
     
     
         18 . A labelling method for a hair cell comprising administering to a biological specimen a probe for a hair cell which contains as an active agent, a staining compound represented by general formula (I): 
       
         
           
           
               
               
           
         
         wherein: 
         R 1  represents a hydrogen atom, an alkyl group, or an aryl group; 
         R 2  to R 5  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, or a halogen atom, and R 2  and R 3 , R 3  and R 4 , or R 4  and R 5  are optionally bonded to each other to form a ring; 
         R 6  represents a hydrogen atom or an alkyl group; 
         R 7  represents a hydrogen atom, an alkyl group, a carboxylic acid group, or a cyano group; 
         X 1   −  represents an anionic group; 
         Y represents a sulfur atom, an oxygen atom, —N(R 8 )—, or —C(R 9 )(R 10 )—, wherein R 8  to R 10  each independently represent a hydrogen atom, an alkyl group, or an aryl group, and R 9  and R 10  are optionally bonded to each other to form a ring; 
         n represents an integer of 1 to 3; and 
         A represents the general formula (III): 
       
       
         
           
           
               
               
           
         
         wherein: 
         R 18  to R 21  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, or a halogen atom, and R 18  and R 19 , R 19  and R 20 , or R 20  and R 21  are optionally bonded to each other to form a ring; 
         R 22  represents a hydrogen atom, an alkyl group, or an aryl group; and 
         Z represents a sulfur atom, an oxygen atom, or —C(R 23 )(R 24 )—, wherein R 23  and R 24  each represent a hydrogen atom, an alkyl group, or an aryl group, and R 23  and R 24  are optionally bonded to each other to form a ring. 
       
     
     
         19 . The labelling method for a hair cell according to  claim 18 , wherein the compound represented by the general formula (I) comprises at least one of a carboxylic acid group or a sulfonic acid group. 
     
     
         20 . An imaging method for a hair cell, comprising:
 administering to a biological specimen a probe for a hair cell; and   observing fluorescence derived from the probe for a hair cell by irradiating the biological specimen with excitation light;   wherein the probe for a hair cell contains as an active agent a staining compound represented by the general formula (I)   
       
         
           
           
               
               
           
         
         wherein: 
         R 1  represents a hydrogen atom, an alkyl group, or an aryl group; 
         R 2  to R 5  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, or a halogen atom, and R 2  and R 3 , R 3  and R 4 , or R 4  and R 5  are optionally bonded to each other to form a ring; 
         R 6  represents a hydrogen atom or an alkyl group; 
         R 7  represents a hydrogen atom, an alkyl group, a carboxylic acid group, or a cyano group; 
         X 1   −  represents an anionic group; 
         Y represents a sulfur atom, an oxygen atom, —N(R 8 )—, or —C(R 9 )(R 10 )—, wherein R 8  to R 10  each independently represent a hydrogen atom, an alkyl group, or an aryl group, and R 9  and R 10  are optionally bonded to each other to form a ring; 
         n represents an integer of 1 to 3, and 
         A represents the general formula (III): 
       
       
         
           
           
               
               
           
         
         wherein: 
         R 18  to R 21  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, or a halogen atom, and R 18  and R 19 , R 19  and R 20 , or R 20  and R 21  are optionally bonded to each other to form a ring; 
         R 22  represents a hydrogen atom, an alkyl group, or an aryl group; and 
         Z represents a sulfur atom, an oxygen atom, or —C(R 23 )(R 24 )—, wherein R 23  and R 24  each represent a hydrogen atom, an alkyl group, or an aryl group, and R 23  and R 24  are optionally bonded to each other to form a ring. 
       
     
     
         21 . An evaluation method for auditory toxicity of a chemical substance, comprising:
 administering the chemical substance to an organism;   administering to the organism a probe for a hair cell; and   observing fluorescence derived from the probe for a hair cell by irradiating the organism with excitation light;   wherein the probe for a hair cell contains as an active agent a staining compound represented by the general formula (I)   
       
         
           
           
               
               
           
         
         wherein: 
         R 1  represents a hydrogen atom, an alkyl group, or an aryl group; 
         R 2  to R 5  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, or a halogen atom, and R 2  and R 3 , R 3  and R 4 , or R 4  and R 5  are optionally bonded to each other to form a ring; 
         R 6  represents a hydrogen atom or an alkyl group; 
         R 7  represents a hydrogen atom, an alkyl group, a carboxylic acid group, or a cyano group; 
         X 1   −  represents an anionic group; 
         Y represents a sulfur atom, an oxygen atom, —N(R 8 )—, or —C(R 9 )(R 10 )—, wherein R 8  to R 10  each independently represent a hydrogen atom, an alkyl group, or an aryl group, and R 9  and R 10  are optionally bonded to each other to form a ring; 
         n represents an integer of 1 to 3, and 
         A represents the general formula (III): 
       
       
         
           
           
               
               
           
         
         wherein: 
         R 18  to R 21  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, or a halogen atom, and R 18  and R 19 , R 19  and R 20 , or R 20  and R 21  are optionally bonded to each other to form a ring; 
         R 22  represents a hydrogen atom, an alkyl group, or an aryl group; and 
         Z represents a sulfur atom, an oxygen atom, or —C(R 23 )(R 24 )—, wherein R 23  and R 24  each represent a hydrogen atom, an alkyl group, or an aryl group, and R 23  and R 24  are optionally bonded to each other to form a ring. 
       
     
     
         22 . The evaluation method for auditory toxicity of a chemical substance according to  claim 21 , wherein the organism comprises Zebrafish. 
     
     
         23 . A screening method for a therapeutic drug for hearing loss, comprising:
 administering a test substance to a hearing loss model animal;   administering a diagnostic composition for an auditory function to the hearing loss model animal; and   examining a staining condition of the diagnostic composition for an auditory function for a hair cell of the hearing loss model animal;   wherein the diagnostic composition for an auditory function comprises a probe for a hair cell, and   wherein the probe for a hair cell contains as an active agent a staining compound represented by the general formula (I)   
       
         
           
           
               
               
           
         
         wherein: 
         R 1  represents a hydrogen atom, an alkyl group, or an aryl group; 
         R 2  to R 5  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, or a halogen atom, and R 2  and R 3 , R 3  and R 4 , or R 4  and R 5  are optionally bonded to each other to form a ring; 
         R 6  represents a hydrogen atom or an alkyl group; 
         R 7  represents a hydrogen atom, an alkyl group, a carboxylic acid group, or a cyano group; 
         X 1   −  represents an anionic group; 
         Y represents a sulfur atom, an oxygen atom, —N(R 8 )—, or —C(R 9 )(R 10 )—, wherein R 8  to R 10  each independently represent a hydrogen atom, an alkyl group, or an aryl group, and R 9  and R 10  are optionally bonded to each other to form a ring; 
         n represents an integer of 1 to 3, and 
         A represents the general formula (III): 
       
       
         
           
           
               
               
           
         
         wherein: 
         R 18  to R 21  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, or a halogen atom, and R 18  and R 19 , R 19  and R 20 , or R 20  and R 21  are optionally bonded to each other to form a ring; 
         R 22  represents a hydrogen atom, an alkyl group, or an aryl group; and 
         Z represents a sulfur atom, an oxygen atom, or —C(R 23 )(R 24 )—, wherein R 23  and R 24  each represent a hydrogen atom, an alkyl group, or an aryl group, and R 23  and R 24  are optionally bonded to each other to form a ring. 
       
     
     
         24 . The screening method according to  claim 23 , wherein the hearing loss model animal comprises Zebrafish. 
     
     
         25 . An evaluation method for a therapeutic drug for hearing loss, comprising:
 administering a test substance to a hearing loss model animal;   administering a diagnostic composition for an auditory function to the hearing loss model animal; and   examining a staining condition of the diagnostic composition for an auditory function for a hair cell of the hearing loss model animal;   wherein the diagnostic composition for an auditory function comprises a probe for a hair cell, and   wherein the probe for a hair cell contains as an active agent a staining compound represented by the general formula (I)   
       
         
           
           
               
               
           
         
         wherein: 
         R 1  represents a hydrogen atom, an alkyl group, or an aryl group; 
         R 2  to R 5  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, or a halogen atom, and R 2  and R 3 , R 3  and R 4 , or R 4  and R 5  are optionally bonded to each other to form a ring; 
         R 6  represents a hydrogen atom or an alkyl group; 
         R 7  represents a hydrogen atom, an alkyl group, a carboxylic acid group, or a cyano group; 
         X 1   −  represents an anionic group; 
         Y represents a sulfur atom, an oxygen atom, —N(R 8 )—, or —C(R 9 )(R 10 )—, wherein R 8  to R 10  each independently represent a hydrogen atom, an alkyl group, or an aryl group, and R 9  and R 10  are optionally bonded to each other to form a ring; 
         n represents an integer of 1 to 3, and 
         A represents the general formula (III): 
       
       
         
           
           
               
               
           
         
         wherein: 
         R 18  to R 21  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, or a halogen atom, and R 18  and R 19 , R 19  and R 20 , or R 20  and R 21  are optionally bonded to each other to form a ring; 
         R 22  represents a hydrogen atom, an alkyl group, or an aryl group; and 
         Z represents a sulfur atom, an oxygen atom, or —C(R 23 )(R 24 )—, wherein R 23  and R 24  each represent a hydrogen atom, an alkyl group, or an aryl group, and R 23  and R 24  are optionally bonded to each other to form a ring 
       
     
     
         26 . The evaluation method according to  claim 25 , wherein the hearing loss model animal comprises Zebrafish.