Fluorescent Probe

Abstract

A fluorescent probe which specifically and efficiently traps nitrogen monoxide, zinc ion etc. to emit fluorescence is provided. A compound represented by the following general formula (I): [wherein R 1 and R 2 represent hydrogen atom, or a group represented by the following formula (A): (wherein X 1 to X 4 represent hydrogen atom, an alkyl group, or a protective group for amino group, and m and n represent 0 or 1); R 3 and R 4 represent hydrogen atom, a C 1-6 alkyl group, or a C 1-6 alkoxy group; R 5 to R 12 represent hydrogen atom, sulfo group, phospho group, a halogen atom, or a C 1-6 alkyl group; R 13 and R 14 represent a C 1-18 alkyl group; Z 1 represents oxygen atom, sulfur atom, or —N(R 15 )— (wherein R 15 represents hydrogen atom, or a C 1-6 alkyl group); Y 1 and Y 2 represent —C(═O)—, —C(═S)—, or —C(R 16 )(R 17 ) (wherein R 16 and R 17 represent a C 1-6 alkyl group); and M − represents a counter ion in a number required for neutralizing the charge].

1 . A compound represented by the following general formula (I):  
       
         
           
           
               
               
           
         
       
       wherein R 1  and R 2  independently represent a hydrogen atom, or a group represented by the following formula (A):  
       
         
           
           
               
               
           
         
       
       (wherein X 1 , X 2 , X 3 , and X 4  independently represent a hydrogen atom, an alkyl group which may have a substituent, or a protective group for amino group, and m and n independently represent 0 or 1, provided that R 1  and R 2  do not simultaneously represent a hydrogen atom; R 3  and R 4  independently represent a hydrogen atom, a C 1-6  alkyl group which may have a substituent, or a C 1-6  alkoxy group which may have a substituent; R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , and R 12  independently represent a hydrogen atom, a sulfo group, a phospho group, a halogen atom, or a C 1-6  alkyl group which may have a substituent; R 13  and R 14  independently represent a C 1-18  alkyl group which may have a substituent; Z 1  represents an oxygen atom, a sulfur atom, or —N(R 15 )— (wherein R 15  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent; Y 1  and Y 2  independently represent —C(═O)—, —C(═S)—, or —C(R 16 )(R 17 ) (wherein R 16  and R 17  independently represent a C 1-6  alkyl group which may have a substituent; and M −  represents a counter ion in a number required for neutralizing the charge  3 .  
     
     
         2 . A fluorescent probe containing the compound represented by the general formula (I) according to  claim 1  except for a compound wherein any one or more of X 1 , X 2 , X 3 , and X 4  represent a protective group for an amino group.  
     
     
         3 . A compound represented by the following general formula (IA):  
       
         
           
           
               
               
           
         
       
       wherein R 21  and R 22  represent amino groups substituting at adjacent positions on the benzene ring, and one of the amino groups may have one alkyl group which may have a substituent; R 23  and R 24  independently represent a hydrogen atom, a C 1-6  alkyl group which may have a substituent, or a C 1-6  alkoxy group which may have a substituent; R 25 , R 26 , R 27 , R 28 , R 29 , R 30 , R 31 , and R 32  independently represent a hydrogen atom, a sulfo group, a phospho group, a halogen atom, or a C 1-6  alkyl group which may have a substituent; R 33  and R 34  independently represent a C 1-18  alkyl group which may have a substituent; Z 21  represents an oxygen atom, a sulfur atom, or —N(R 35 )— wherein R 35  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent); Y 21  and Y 22  independently represent —C(═O)—, —C(═S)—, or —C(R 36 )(R 37 )— wherein R 36  and R 37  independently represent a C 1-6  alkyl group which may have a substituent; and M −  represents a counter ion in a number required for neutralizing the charge.  
     
     
         4 . The compound according to  claim 3 , wherein R 23 , R 24 , R 25 , R 26 , R 27 , R 28 , R 29 , R 30 , R 31 , and R 32  are hydrogen atoms, R 33  and R 34  are C 1-6  alkyl groups substituted with a sulfo group, Z 21  is an oxygen atom, and Y 21  and Y 22  are —C(CH 3 ) 2 —.  
     
     
         5 . A reagent for measuring nitrogen monoxide, which contains the compound represented by the general formula (IA) according to  claim 3 .  
     
     
         6 . A compound represented by the following general formula (IB):  
       
         
           
           
               
               
           
         
       
       wherein R 41  and R 42  combine together to represent a group represented by —N═N—NR 58 — which forms a ring at the adjacent positions on the benzene ring wherein R 58  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent, or R 41  and R 42  represent a combination of an amino group (which may have a C 1-6  alkyl group which may have a substituent, or a protective group for an amino group and a nitro group substituting at adjacent positions on the benzene ring; R 43  and R 44  independently represent a hydrogen atom, a C 1-6  alkyl group which may have a substituent, or a C 1-6  alkoxy group which may have a substituent; R 45 , R 46 , R 47 , R 48 , R 49 , R 50 , R 51 , and R 52  independently represent a hydrogen atom, a sulfo group, a phospho group, a halogen atom, or a C 1-6  alkyl group which may have a substituent; R 13  and R 54  independently represent a C 1-18  alkyl group which may have a substituent; Z 41  represents an oxygen atom, a sulfur atom, or —N(R 55 )— wherein R 55  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent; Y 41  and Y 42  independently represent —C(═O)—, —C(═S)—, or —C(R 56 )(R 57 )— (wherein R 56  and R 57  independently represent a C 1-6  alkyl group which may have a substituent); and M −  represents a counter ion in a number required for neutralizing the charger.  
     
     
         7 . The compound according to  claim 6 , wherein R 43 , R 44 , R 45 , R 46 , R 47 , R 48 , R 49 , R 50 , R 51 , and R 52  are hydrogen atoms, R 53  and R 54  are C 1-6  alkyl groups substituted with a sulfo group, Z 41  is an oxygen atom, and Y 41  and Y 42  are —C(CH 3 ) 2 —.  
     
     
         8 . A method for measuring nitrogen monoxide, which comprises (a) reacting the compound represented by the general formula (IA) with nitrogen monoxide;  
       
         
           
           
               
               
           
         
       
       wherein R 21  and R 22  represent amino groups substituting at adjacent positions on the benzene ring, and one of the amino groups may have one alkyl group which may have a substituent; R 23  and R 24  independently represent a hydrogen atom, a C 6  alkyl group which may have a substituent, or a C 1-6  alkoxy group which may have a substituent; R 25 , R 26 , R 27 , R 28 , R 29 , R 30 , R 31 , and R 32  independently represent a hydrogen atom, a sulfo group, a phospho group, a halogen atom, or a C 1-6  alkyl group which may have a substituent: R 33  and R 34  independently represent a C 1-18  alkyl group which may have a substituent; Z 21  represents an oxygen atom, a sulfur atom, or —N(R 35 )—, wherein R 35  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent; Y 21  and Y 22  independently represent —C(═O)—, —C(═S)—, or —C(R 36 )(R 37 )—, wherein R 36  and R 37  independently represent a C 1-6  alkyl group which may have a substituent; and M −  represents a counter ion in a number required for neutralizing the charge: and (b) the step detecting the compound of the general formula (IB)  
       
         
           
           
               
               
           
         
       
       wherein R 41  and R 42  combine together to represent a group represented by —N═N—NR 58 — which forms a ring at the adjacent positions on the benzene ring, wherein R 58  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent, or R 41  and R 42  represent a combination of an amino group which may have a C 1-6  alkyl group which may have a substituent, or a protective group for an amino group; and a nitro group substituting at adjacent positions on the benzene ring, R 43  and R 44  independently represent a hydrogen atom, a C 1-6  alkyl group which may have a substituent, or a C 1-6  alkoxy group which may have a substituent; R 45 , R 46 , R 47 , R 48 , R 49 , R 50 , R 51 , and R 52  independently represent a hydrogen atom, a sulfo group, a phospho group, a halogen atom, or a C 1-6  alkyl group which may have a substituent; R 53  and R 54  independently represent a C 1-18  alkyl group which may have a substituent; Z 41  represents an oxygen atom, a sulfur atom, or —N(R 55 )—, wherein R 55  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent; Y 41  and Y 42  independently represent —C(═O)—, —C(═S)—, or —C(R 56 )(R 57 )—, wherein R 56  and R 57  independently represent a C 1-6  alkyl group which may have a substituent; and M −  represents a counter ion in a number required for neutralizing the charge: 
 wherein R 41  and R 42  combine together to represent a group represented by —N═N—NR 58 — which forms a ring at the adjacent positions on the benzene ring (wherein R 58  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent produced in (a).  
 
     
     
         9 . A compound represented by the following general  
       
         
           
           
               
               
           
         
       
       wherein R 61  and R 62  independently represent a hydrogen atom, or a group represented by the following formula (B):  
       
         
           
           
               
               
           
         
       
       wherein X 61 , X 62 , X 63 , and X 64  independently represent a hydrogen atom, an alkyl group which may have a substituent, or a protective group for amino group, and p and q independently represent 0 or 1), provided that R 61  and R 62  do not simultaneously represent a hydrogen atom, and when R 61  and R 62  simultaneously represent a group represented by the formula (B), in at least one of the groups represented by the formula (B), either p or q, or both represent 1; R 63  and R 64  independently represent a hydrogen atom, a C 1-6  alkyl group which may have a substituent, or a C 1-6  alkoxy group which may have a substituent; R 65 , R 66 , R 67 , R 68 , R 69 , R 70 , R 71 , and R 72  independently represent a hydrogen atom, a sulfo group, a phospho group, a halogen atom, or a C 1-6  alkyl group which may have a substituent; R 73  and R 74  independently represent a C 1-18  alkyl group which may have a substituent; Z 61  represents an oxygen atom, a sulfur atom, or —N(R 75 )— wherein R 75  represents a hydrogen atom, or a C 1-6  alkyl group which may have a substituent; Y 61  and Y 62  independently represent —C(═O)—, —C(═S)—, or —C(R 76 )(R 77 )— (wherein R 76  and R 77  independently represent a C 1-6  alkyl group which may have a substituent); and M −  represents a counter ion in a number required for neutralizing the charged.  
     
     
         10 . A fluorescent probe for zinc containing the compound represented by the general formula (IC) according to  claim 9  (except for a compound wherein any one or more of X 61 , X 62 , X 63 , and X 64  are protective group for amino group).  
     
     
         11 . A zinc complex formed from the compound represented by the general formula (IC) according to  claim 9  (except for a compound wherein any one or more of X 61 , X 62 , X 63 , and X 64  are protective group for amino group), and a zinc ion.  
     
     
         12 . A method for measuring zinc ions, which comprises (a) reacting the compound represented by the general formula (IC) according to  claim 9  except for a compound wherein any one or more of X 61 , X 62 , X 63 , and X 64  are protective group for amino group) with a zinc ion, and (b) measuring fluorescence intensity of a zinc complex produced in (a).