Labeling composition for intraocular tissue, labeling method of intraocular tissue, and screening method

Abstract

The invention provides a labeling composition for an intraocular tissue of a living individual, which specifically labels the intraocular tissue without need of an invasive operation such as exposure of an ocular tissue or injection of a staining agent into the ocular tissue or a nerve tissue linking to the ocular tissue, a method of noninvasively labeling an intraocular tissue of a living individual, and a screening method using the labeling composition for the intraocular tissues. The composition contains a compound capable of labeling at least a photoreceptor cell layer of a retina, wherein the compound is a staining compound having a particular structure as a partial structure thereof.

1 . A labeling composition for an intraocular tissue of a living individual, which comprises a compound capable of labeling at least a photoreceptor cell layer of a retina, wherein the compound is a staining compound having a structure represented by the following general formula (I) or general formula (II) as a partial structure thereof, 
       
         
           
           
               
               
           
         
         wherein in the general formula (I), R 1  and R 2  are, independently of each other, a hydrogen atom, an alkyl group or an aryl group, R 1  and R 2  may bond to each other to form a ring, and A is a structure represented by any one of the following general formulae (III) to (IX) and (VIII′) 
       
       
         
           
           
               
               
           
         
         wherein in the general formula (III), R 6  is an aryl group, R 7 , R 8 , R 10  and R 11  are, independently of one another, a hydrogen atom, an alkyl group, an aryl group or a halogen atom, and R 9  is an ammonium salt group having a counter anion, in the general formula (IV), R 12  and R 13  are, independently of each other, a hydrogen atom, an alkyl group, an aryl group or a heterocyclic group, and R 14  to R 16  are, independently of one another, a hydrogen atom, an alkyl group, an aryl group, a heterocyclic group, a halogen atom or an amino group, in the general formula (V), R 17  and R 18  are, independently of each other, a hydrogen atom, an alkyl group or an alkoxy group, R 19  and R 20  are, independently of each other, a hydrogen atom, an alkyl group, an alkoxy group, a hydroxyl group or a cyano group, R 21  is a heterocyclic group or —CH═C(R 22 )(R 23 ), R 22  and R 23  are, independently of each other, a hydrogen atom, a cyano group, a heterocyclic group, a carboxyl group or a carboxylate group, and R 1  and R 17 , R 2  and R 18 , and R 22  and R 23  may, independently of one another, bond to each other to form a ring, in the general formula (VI), R 24  is a hydrogen atom, an alkyl group, an aryl group or a heterocyclic group, in the general formula (VII), R 25  is an oxygen atom, a sulfur atom or N(R 27 ), R 26  is a hydrogen atom, an alkyl group, an alkoxy group or a sulfonic group, and R 27  is a hydrogen atom, an alkyl group or an aryl group, in the general formula (VIII), R 28  is a hydrogen atom, an alkyl group or an aryl group, in the general formula (VIII′), R 28 ′ is a halogen atom, an alkoxy group or an aryloxy group, and in the general formula (IX), R 29  is a hydrogen atom, an alkyl group or an aryl group; and wherein A is also a structure represented by the following general formula (X) 
       
       
         
           
           
               
               
           
         
         wherein in the general formula (X), R 31  and R 32  are individually a sulfonic group or a salt thereof, and R 33  and R 34  are individually a hydrogen atom, an alkyl group, an alkoxy group or a halogen atom, with the proviso that R 1  and R 2  in the general formula (I) bond to each other to form one substituent group ═N—R 30 , and R 30  is an aryl group or a heterocyclic group, 
         further wherein in the general formula (II), R 3  and R 4  are, independently of each other, a hydrogen atom, an alkenyl group, a cyano group, a carboxyl group, a carboxylate group, a sulfonic group, an acyl group or a heterocyclic group, R 3  and R 4  may bond to each other to form a ring, and R 5  is a hydrogen atom, an alkyl group, an aryl group or a heterocyclic group, with the proviso that one of R 3  and R 4  is a hydrogen atom, and the other is a heterocyclic group represented by the following general formula (XI) 
       
       
         
           
           
               
               
           
         
         wherein in the general formula (XI), R 35  is an alkyl group or an aryl group, R 36  to R 39  are, independently of one another, a hydrogen atom, an alkyl group, an aryl group, an alkoxy group, a carboxyl group, a sulfonic group, a heterocyclic group, an amino group or a halogen atom, R 36  and R 37 , R 37  and R 38 , or R 38  and R 39  may bond to each other to form a ring, X −  is an anionic group, and Q 1  is a sulfur atom, oxygen atom, —C(R 40 )(R 41 )—, —CH═CH— or —N(R 42 )—, in which R 40  to R 42  are individually a hydrogen atom, an alkyl group or an aryl group, and R 40  and R 41  may bond to each other to form a ring, and wherein the ring formed by bonding R 3  and R 4  in the general formula (II) to each other is represented by any one of the following general formulae (XII), (XIII) and (XV) 
       
       
         
           
           
               
               
           
         
         wherein in the general formula (XII), R 43  is a hydrogen atom, an alkyl group, an aryl group or a heterocyclic group, and R 44  is an alkyl group, an aryl group, a carboxyl group, a carboxylate group, a hydroxyl group or an amino group, in the general formula (XIII), Q 2  is an oxygen atom, a sulfur atom or —N(R 52 )—, R 45  is a hydrogen atom, an alkyl group, an aryl group or a heterocyclic group, R 46  is a sulfur atom, an oxygen atom, ═NR 53 , a heterocyclic group or a dicyanomethylene group, and R 52  and R 53  are individually a hydrogen atom, an alkyl group, an aryl group or a heterocyclic group, and in the general formula (XV), R 49  and R 50  are, independently of each other, a hydrogen atom, an alkyl group or an aryl group, and R 51  is an oxygen atom or a sulfur atom, and 
         B is a structure represented by any one of the following general formulae (XVI) to (XVIV) and (XVII′) 
       
       
         
           
           
               
               
           
         
         wherein in the general formula (XVI), R 54  is a hydrogen atom, an alkyl group, an aralkyl group, an alkenyl group, an aryl group, a heterocyclic group or an acyl group, R 55  to R 58  are, independently of one another, a hydrogen atom, an alkyl group, an aryl group, a carboxyl group, a carboxylate group or an acyl group, R 55  and R 57  may bond to each other to form a ring, and R 59  is a hydrogen atom, an alkyl group, an alkoxy group or a halogen atom, in the general formula (XVII), R 60  is a hydrogen atom, an alkyl group or an aryl group, R 61  to R 64  are, independently of one another, a hydrogen atom, an alkyl group, an aryl group, an alkoxy group, a carboxyl group, a sulfonic group, a heterocyclic group, an amino group or a halogen atom, R 61  and R 62 , R 62  and R 63 , or R 63  and R 64  may bond to each other to form a ring, and Q is a sulfur atom, an oxygen atom, —C(R 65 )(R 66 )— or —CH═CH—, in which R 65  and R 66  bond to each other to form a ring, in the general formula (XVII′), R 60 ′ is a hydrogen atom, an alkyl group or an aryl group, Q 3 ′ is a sulfur atom or an oxygen atom, and dotted lines in the general formula (XVII′) represent the case where a benzene ring is present or absent, in the general formula (XVIII), R 68  and R 69  are, independently of each other, a hydrogen atom, an alkyl group or an aryl group, and R 70  is an oxygen atom or a sulfur atom, and in the general formula (XVIV), R 71  is an alkyl group, 
         further wherein B and R 5  may bond to each other to form a benzothiophen-3-one ring, and at that time, R 3  and R 4  bond to each other to form a benzothiophen-3-one ring. 
       
     
     
         2 . The labeling composition for the intraocular tissue according to  claim 1 , wherein the intraocular tissue is a retinal tissue. 
     
     
         3 . The labeling composition for the intraocular tissue according to  claim 1 , wherein the compound selectively labels at least one cell selected from the group consisting of rod cells, plural kinds of cone cells different in sensitivity to a wavelength region, and a part of the cell bodies of these cells. 
     
     
         4 . The labeling composition for the intraocular tissue according to  claim 1 , which contains the compound at a concentration of 0.001 ng/mL or more and 100 μg/mL or less. 
     
     
         5 . The labeling composition for the intraocular tissue according to  claim 1 , wherein the compound is a fluorescent compound. 
     
     
         6 . A labeling composition for labeling an intraocular tissue of a living individual, which comprises a compound capable of labeling at least a photoreceptor cell layer of the retina. 
     
     
         7 . The labeling composition for the intraocular tissue according to  claim 1 , wherein the compound is administered without causing a surgical damage to an ocular tissue or a nerve tissue linking to the ocular tissue to label the intraocular tissue. 
     
     
         8 . A staining agent for an intraocular tissue, which comprises the labeling composition for the intraocular tissue according to  claim 1 . 
     
     
         9 . A diagnostic composition comprising, as an active agent, the labeling composition for the intraocular tissue according to  claim 1  or a staining agent for the intraocular tissue, which comprises the labeling composition for the intraocular tissue. 
     
     
         10 . A staining method for an ocular tissue, which comprises a step of bringing the labeling composition for the intraocular tissue according to  claim 1  or a staining agent for the intraocular tissue, which comprises the labeling composition for the intraocular tissue, into contact with the ocular tissue. 
     
     
         11 . A multiple labeling method for an ocular tissue, which comprises using plural kinds of labeling compositions including at least one of the labeling composition for the intraocular tissue according to  claim 1  and a staining agent for the intraocular tissue, which comprises the labeling composition for the intraocular tissue. 
     
     
         12 . The multiple labeling method according to  claim 11 , wherein at least one of the plural kinds of labeling compositions contains at least one antibody. 
     
     
         13 . An imaging method for an intraocular tissue by conducting labeling with the labeling method according to  claim 10 , which comprises a step of irradiating the intraocular tissue with excitation light to observe fluorescence. 
     
     
         14 . An evaluating method for a condition of a retina, which comprises using the imaging method of the intraocular tissue according to  claim 13 . 
     
     
         15 . A screening method comprising using a model animal labeled with the labeling composition for the intraocular tissue according to  claim 1 . 
     
     
         16 . A screening method for a disease curing medicine or disease preventing medicine, which comprises the steps of administering a test substance to a model animal, administering the labeling composition for the intraocular tissue according to  claim 1  to the model animal, and checking a labeled condition caused by the labeling composition for the intraocular tissue with respect to the intraocular tissue of the model animal. 
     
     
         17 . The screening method according to  claim 15 , wherein the model animal is Zebrafish. 
     
     
         18 . The screening method according to  claim 15 , wherein the model animal is mouse or rat.