Fluorescent probes for saccharrides

Abstract

The spectroscopic and photophysical properties of fluorescent probes comprising donor-acceptor derivatives comprising the boric acid group or a derivative of boric acid, B(OH) 3 (or borate ion, BO(OH) 2 −1 ), arsenious acid, H 3 AsO 3 (or arsenite ion, H 2 AsO 3 −1 ), telluric acid, H 6 TeO 6 (or tellurate ion, H 5 TeO 6 −1 ) or germanic acid, Ge(OH) 6 (or germanate ion, GeO(OH) 3 −1 ) are described. Method of using said probes are also provided.

1 . A fluorescent compound which is represented by the formula  
       
         
           
           
               
               
           
         
       
       wherein R 1  represents the boric acid group or a derivative of boric acid, B(OH) 3  (or borate ion, BO(OH) 2   −1 ), arsenious acid, H 3  AsO 3  (or arsenite ion, H 2 AsO 3   −1 ), telluric acid, H 6 TeO 6  (or tellurate ion, H 5  TeO 6   −1 ) or germanic acid, Ge(OH) 6  (or germanate ion, GeO(OH) 3   −1 ); 
 Y represents a compound of the formula  
                     
 (—C═C—)n  
 (C≡C)n  
 or a phenyl group  
 and wherein R″ represents a compound selected from the group consisting of H, N(CH 3 ) 2 , CN and OCH 3  or a substituent capable of donating an electron to or accepting an electron from a the aromatic system of the compound; and n is a whole number from 1 to 5.  
 
     
     
         2 . The fluorescent compound of  claim 1  which is represented by the formula  
       
         
           
           
               
               
           
         
       
       wherein R 2  represents a compound selected from the group consisting of H, N(CH 3 ) 2 , CN and OCH 3 , or a substituent capable of donating an electron to or accepting an electron from a the aromatic system of the compound(C═C and a phenyl linker); and n is a whole number from 1 to 5.  
     
     
         3 . The fluorescent compound of  claim 2  which is represented by a chemical formula selected from the group consisting of  
       
         
           
           
               
               
           
         
       
     
     
         4 . The fluorescent compound of  claim 2  which is represented by the chemical formula  
       
         
           
           
               
               
           
         
       
     
     
         5 . The fluorescent compound of  claim 2  which is represented by the chemical formula  
       
         
           
           
               
               
           
         
       
     
     
         6 . The fluorescent compound of  claim 1  which is represented by the chemical formula  
       
         
           
           
               
               
           
         
       
     
     
         7 . A fluorescent compound represented by the chemical formula  
       
         
           
           
               
               
           
         
       
     
     
         8 . The compound of  claim 1 , wherein the compound is attached to a solid support.  
     
     
         9 . The compound of  claim 1 , wherein the compound is immobilized in a polymer matrix.  
     
     
         10 . The compound of  claim 8 , wherein the compound is covalently attached to the solid support.  
     
     
         11 . The compound of  claim 9 , wherein the compound is covalently attached to the polymer matrix.  
     
     
         12 . The compound of  claim 9 , wherein the compound is embedded within the polymer matrix.  
     
     
         13 . The compound of  claim 9 , wherein the matrix is biocompatable.  
     
     
         14 . A method for detecting the presence or concentration of a diol in a sample which comprises the following steps: 
 a) contacting the sample with a probe which comprises the compound of  claim 1;  and    b)measuring any change in fluorescence emitted by the probe upon binding of the diol to the probe, thereby detecting the presence or concentration of the diol.    
     
     
         15 . The method of  claim 14  wherein the diol is a sugar.  
     
     
         16 . The method of  claim 15  wherein the sugar is selected from the group consisting of glucose, fructose and galactose.  
     
     
         17 . The method of  claim 14  wherein said change is a change in the intensity of fluorescence of said compound.  
     
     
         18 . The method of  claim 14  wherein said change is a change in the lifetime of fluorescence of said compound.  
     
     
         19 . The method of  claim 14  wherein said change is a change in the intensity ratio of said compound.  
     
     
         20 . A method as in  claim 14  which is performed in vivo in an animal.  
     
     
         21 . A method as in  claim 14  wherein said measuring step comprises a measurement of the anisotropy or polarization of the fluorescence emitted by the compound.  
     
     
         22 . A method as in  claim 14  wherein said measuring step comprises a measurement of the anisotropy decay time of the fluorescence emitted by the compound.  
     
     
         23 . A method as in  claim 14  wherein said measuring step comprises a measurement of the anisotropy correlation time of the fluorescence emitted by the compound.  
     
     
         24 . A method as in  claim 14  wherein said measuring step comprises a measurement of the anisotropy differential phase angle of the fluorescence emitted by the compound.  
     
     
         25 . A method as in  claim 14  wherein said measuring step comprises a measurement of the anisotropy modulation ratio of the fluorescence emitted by the compound.  
     
     
         26 . A method as in  claim 14  wherein said measuring step comprises a measurement of the modulated anisotropy of the fluorescence emitted by the compound.  
     
     
         27 . A method for detecting the presence or concentration of a diol in a sample which comprises the following steps: 
 a) contacting the sample with a probe which comprises the compound of  claim 7;  and    b)measuring any change in fluorescence emitted by the probe upon binding of the diol to the probe, thereby detecting the presence or concentration of the diol.    
     
     
         28 . The method of  claim 27  wherein the diol is a sugar.  
     
     
         29 . The method of  claim 28  wherein the sugar is selected from the group consisting of glucose, fructose and galactose.  
     
     
         30 . The method of  claim 27  wherein said change is a change in the intensity of fluorescence of said compound.  
     
     
         31 . The method of  claim 27  wherein said change is a change in the lifetime of fluorescence of said compound.  
     
     
         32 . The method of  claim 27  wherein said change is a change in the intensity ratio of said compound.  
     
     
         33 . A method as in  claim 27  which is performed in vivo in an animal.  
     
     
         34 . A method as in  claim 27  wherein said measuring step comprises a measurement of the anisotropy or polarization of the fluorescence emitted by the compound.  
     
     
         35 . A method as in  claim 27  wherein said measuring step comprises a measurement of the anisotropy decay time of the fluorescence emitted by the compound.  
     
     
         36 . A method as in  claim 27  wherein said measuring step comprises a measurement of the anisotropy correlation time of the fluorescence emitted by the compound.  
     
     
         37 . A method as in  claim 27  wherein said measuring step comprises a measurement of the anisotropy differential phase angle of the fluorescence emitted by the compound.  
     
     
         38 . A method as in  claim 27  wherein said measuring step comprises a measurement of the anisotropy modulation ratio of the fluorescence emitted by the compound.  
     
     
         39 . A method as in  claim 27  wherein said measuring step comprises a measurement of the modulated anisotropy of the fluorescence emitted by the compound.  
     
     
         40 . A method for detecting the presence or concentration of a diol in a sample which comprises the following steps: 
 a) contacting the sample with a probe which comprises a compound selected from the group consisting of                          b) measuring any change in fluorescence lifetime emitted by the probe upon binding of the diol to the probe, thereby detecting the presence or concentration of the diol.    
     
     
         41 . The method of  claim 40  wherein the diol is a sugar.  
     
     
         42 . The method of  claim 41  wherein the sugar is selected from the group consisting of glucose, fructose and galactose.  
     
     
         43 . A method as in  claim 40  which is performed in vivo in an animal.  
     
     
         44 . A method as in  claim 40  wherein said change is a change in the intensity ratio of said compound.  
     
     
         45 . A method as in  claim 40  wherein said measuring step comprises a measurement of the anisotropy or polarization of the fluorescence emitted by the compound.  
     
     
         46 . A method as in  claim 40  wherein said measuring step comprises a measurement of the anisotropy decay time of the fluorescence emitted by the compound.  
     
     
         47 . A method as in  claim 40  wherein said measuring step comprises a measurement of the anisotropy correlation time of the fluorescence emitted by the compound.  
     
     
         48 . A method as in  claim 40  wherein the animal is a human.  
     
     
         49  A kit for detecting the presence of a diol in a sample which comprises a compound as in  claim 1  and packaging material, and optionally labeling material and instructions for using said compound to detect the presence of the diol.  
     
     
         50 . A kit as in  claim 49  wherein the compound is bound to a solid support.  
     
     
         51 . A kit for detecting the presence of a diol in a sample which comprises a compound as in  claim 7  and packaging material, and optionally labeling material and instructions for using said compound to detect the presence of the diol.  
     
     
         52 . A kit as in  claim 51  wherein the compound is bound to a solid support.  
     
     
         53 . A kit for detecting the presence of a diol in a sample which comprises a compound as in  claim 40  and packaging material, and optionally labeling material and instructions for using said compound in a method as in  claim 40 .  
     
     
         54 . A kit as in  claim 53  wherein the compound is bound to a solid support.