Fluorescent compounds

Abstract

Fluorescent dye compounds of formula (I) are disclosed. These compounds are useful as they interact with organic compounds in a manner such that excitation with certain wavelengths of light results in fluorescent emmission. Detection and/or monitoring of the fluorescence provides a means for the detection or quantification of organic compounds when bound to these fluorescent dye compounds. Formula (I), wherein: each of R, R′ and R″ is a hydrogen atom, halogen atom or a straight or branched chain (C 1-20 alkyl, alkenyl or alkynyl group optionally substituted with one or more halogen, hydroxy and/or oxy group: rings A, B and C optionally include one or more double bonds; rings B and C are optionally substituted with one or more halogen atoms.

1 . A compound according to formula (I):  
       
         
           
           
               
               
           
         
       
       wherein: 
 each of R, R′ and R″ is a hydrogen atom, halogen atom or a straight or branched chain C 1-20  alkyl, alkenyl or alkynyl group optionally substituted with one or more halogen, hydroxy and/or oxy group;  
 rings A, B and C optionally include one or more double bonds;  
 rings B and C are optionally substituted with one or more halogen atoms, including tautomers of formula (I), and  
 the compound is capable of interacting with an organic compound and when interacting with the organic compound, the compound emits fluorescence after excitation at a broad range of wavelengths.  
 
     
     
         2 . A compound as claimed in  claim 1  according to the formula (Ia), including tautomers thereof:  
       
         
           
           
               
               
           
         
       
     
     
         3 . A compound as claimed in  claim 2  having the stereochemistry as depicted in formula (Ib), including tautomers thereof:  
       
         
           
           
               
               
           
         
       
     
     
         4 . A compound as claimed in any one of  claims 1  to  3  wherein R is a straight or branched chain C 1-20  conjugated alkenyl group optionally substituted with hydroxy and oxy groups; R′ is a straight or branched chain C 1-20  alkyl group optionally substituted with a hydroxy group and R″ is a straight or branched chain C 1-20  alkyl group, including tautomers thereof.  
     
     
         5 . A compound as claimed in  3  wherein R is —C(OH)CHC(O)(CH) 6 CH 3 , R′ is —CH 2 OH and R′ is ′Me being the compound 5,6-dihydro-3-[(1Z, 4E, 6E, 8E)-1-hydroxy-3 -oxo-1,4,6,8-decatetraenyl]-6-hydroxymethyl-9a-methyl-2H-furo[3,2-g][2]benzopyran-2-9(9aH)-dione according to formula (Ic), including tautomers thereof:  
       
         
           
           
               
               
           
         
       
     
     
         6 . A compound according to formula (I):  
       
         
           
           
               
               
           
         
       
       wherein each of R, R′ and R″ is a hydrogen atom, halogen atom or a straight or branched chain C 1-20  alkyl, alkenyl or alkynyl group optionally substituted with one or more halogen, hydroxy and/or oxy groups; 
 ring A, B and C are optionally substituted with one or more halogen atoms; and  
 rings A, B and C optionally include one or more double bonds; with the proviso that: 
 (i) R≠—C(OH)CHC(O)(CH) 6 C(Et)(Me) when R′=—CH 2 OH, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a;  
 (ii) R≠—C(OH)CHC(O)(CH) 6 C(Et)(Me) when R′=Me, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a;  
 (iii) R≠—C(OH)CHC(O)(CH) 6 C(Et)(Me) when R′=Me, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (iv) R≠—C(O)(CH) 4 Me when R′=-n-propyl, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (v) R≠—C(O)(CH) 6 Me when R′=-n-propyl, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (vi) R≠—C(O)(CH) 5 Me when R′=—(CH) 2 Me, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (vii) R≠—C(O)(CH) 6 Me when R′=—(CH) 2 Me, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (viii) R≠—(CH) 2 C(Me)CHC(Me)(Et) when R′=—Ac, R″=Me, C4=Cl, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (ix) R≠—(CH) 2 C(Me)CHC(Me)(Et) when R′=—Ac, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (x) R≠—(CH) 2 C(Me)CH-i-Pr when R′=—Ac, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (xi) R≠—C(O)(CH) 4 Me when R′=—(CH) 2 Me, R″=Me, ring A includes a double bond between C3 and C3a; ring B includes a double bond between C4 and C4a; and ring C includes a double bond between C8 and C8a and C5 and C6;  
 (xii) R≠—C(O)(CH) 4 Me when R′=-n-Pr, R″=Me, ring A does not include a double bond; and rings B and C include double bonds between C8a and C4a and C5 and C6;  
 (xiii) R≠—C(O)(CH) 4 Me when R′=-n-Pr, R″=Me, ring A, B and C do not include double bonds;  
 (xiv) R≠—C(O)(CH) 6 Me when R′=—(CH) 2 Me, R″=Me, ring A does not include a double bond; and rings B and C include double bonds between C8a and C4a and C5 and C6;  
 (xv) R≠—C(O)(CH) 4 Me when R′=—C(CH 2 )(Me), R″=Me, ring A does not include a double bond; and rings B and C include double bonds between C8a and C4a and C5 and C6;  
 (xvi) R≠—C(O)(CH) 4 Me when R′=—(CH) 2 Me, R″=Me, ring A does not include a double bond; and rings B and C include double bonds between C8a and C4a and C5 and C6, including tautomers thereof.  
 
 
     
     
         7 . A compound as claimed in  claim 6  according to the formula (Ia):  
       
         
           
           
               
               
           
         
       
     
     
         8 . A compound as claimed in  claim 7  having the stereochemistry as depicted in formula (Ib):  
       
         
           
           
               
               
           
         
       
     
     
         9 . A compound as claimed in any one of  claims 1  to  8 , wherein the compound when interacting with an organic compound, emits fluorescence after excitation at blue wavelengths.  
     
     
         10 . A compound as claimed in  claim 9  wherein the compound emits fluorescence when excited by light in the range 300-560 nm.  
     
     
         11 . A compound as claimed in  claim 10  wherein the organic compound is a protein and the compound when interacting with the protein emits fluorescence after excitation at 307 nm.  
     
     
         12 . A compound as claimed in  claim 10  wherein the compound emits fluorescence when excited by light at about 400 nm, 488 nm and 514 nm.  
     
     
         13 . A compound as claimed in  claim 12  wherein compound emits fluorescence at 600 nm after excitation at 488 nm.  
     
     
         14 . A composition including a compound as claimed in any one of  claims 1  to  13 , an analytically acceptable carrier and optionally, a flurochrome.  
     
     
         15 . A process for the preparation of a compound as claimed in any one of  claims 1  to  13  comprising culturing a fungal species under conditions such that the fungal species produces the compound, and separating the compound from the culture.  
     
     
         16 . Use of a compound as claimed in any one of  claims 1  to  13  as a fluorescent dye or marker.  
     
     
         17 . Use as in  claim 16  in a technique for staining, labelling and/or detecting organic molecules.  
     
     
         18 . A method for fluorescent labelling an organic compound, comprising causing a compound as claimed in any one of  claims 1  to  13  to bind to an organic compound in a manner such that the organic compound is fluorescently labelled with the compound.  
     
     
         19 . A compound, method, composition or use as claimed in any one of  claims 1  to  14  and  16  to  18 , wherein the organic compound is a protein, peptide, sugar, nucleic acid, cell surface molecule or detergent, or is included in an antibody or a cell.  
     
     
         20 . A compound method or use as in  claim 19  wherein the compound is bound to the organic compound through a linking molecule.  
     
     
         21 . A method of detecting a organic compound in a sample including the organic compound, the method comprising labelling the organic compound according to the method of  claim 18 , and detecting the organic compound in the sample by monitoring or detecting its fluorescence.  
     
     
         22 . A method as claimed in  claim 21  wherein the labelled organic compound is detected when the sample is exposed to light from a wide range of wavelengths, preferably wavelengths in the range of 300-560 nm, more preferably blue wavelengths.  
     
     
         23 . A method as claimed in  claim 21  or  claim 22  wherein the monitoring or detecting of the fluorescence of the labelled-organic compound is by transillumination, spectroscopy, microscopy or cytometry.