Novel tricarbocyanine-cyclodextrin(s) conjugates and use thereof

Abstract

The invention relates to novel tricarbocyanine-cyclodextrin(s) conjugates useful as markers in the diagnosis of kidney diseases, diagnostic compositions comprising the conjugates, their use, and their production.

1 . A fluorescent compound of formula (I)
   F-L n -CD n   (I)
   wherein   F is a tricarbocyanine residue of formula (II)   
       
         
           
           
               
               
           
         
         wherein 
         R 1  and R 2  are independently selected from H, SO 3 H, CO 2 H, SO 2 NH 2 , CH 2 COOH, NH 2 , NHCOCH 2 I, NO 2 , Br, Cl, CH 3 ; 
         R 3  and R 4  are independently selected from C 1-4  alkyl, (CH 2 ) 3 C≡CH, (CH 2 ) 4 C≡CH (CH 2 ) 5 COOH, (CH 2 ) 3 SO 3 H, (CH 2 ) 4 SO 3 H, (CH 2 ) 3 NH 2 , (CH 2 ) 4 NH 2 , (CH 2 ) 3 N + (CH 3 ) 3 , (CH 2 ) 5 N + (CH 3 ) 3 , (CH 2 ) 3 N 3 , (CH 2 ) 4 N 3 , (CH 2 ) 3 NHCOCH 2 I, (CH 2 ) 4 NHCOCH 2 I, (CH 2 CH 2 O) 2 CH 3 , (CH 2 CH 2 O) 3 CH 3 , (CH 2 CH 2 O) 4 CH 3 ; 
         R 5 , is H, Cl, or 
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein 
         X is selected from NH, O, S; 
         j is an integer from 1 to 4; 
         k is an integer from 1 to 4; 
         CD is a cyclodextrin residue of formula (III) 
       
       
         
           
           
               
               
           
         
         wherein 
         m is and integer equal to 6, 7 or 8, 
         R′, R″, R′″ are independently selected from OH, OCH 3 , OCH 2 CH 3 , OCH 2 CHOHCH 3 , OCHOHCH 3 , OCH 2 COOH, O(CH 2 ) 4 SO 3 H, N 3 , NH 2 , NHCOCH 3 , OCH 2 C≡CH, SH; 
         L is a linker group resulting from the coupling of the tricarbocyanine of formula (II) to the cyclodextrin(s) of formula (III) according to the following Table: 
       
       
         
           
                 
                 
                 
               
                     
                 
                   Functional group of the 
                   Functional group of the 
                     
                 
                   tricarbocyanine (F) in any 
                   cyclodextrin (CD) in any 
                     
                 
                   of the groups 
                   of the groups 
                     
                 
                   R 3 , R 4  or R 5   
                   R′, R″ or R′′′ 
                   Linker group (L) 
                 
                     
                 
                   COOH 
                   OH 
                   —C(O)O— 
                 
                   COOH 
                   NH 2   
                   —C(O)NH— 
                 
                   NCS 
                   OH 
                   —NC(S)O— 
                 
                   NCS 
                   NH 2   
                   —NC(S)NH— 
                 
                   NH 2   
                   COOH 
                   —NHC(O)— 
                 
                   NHCOCH 2 I 
                   SH 
                   NHC(O)CH 2 S— 
                 
                     
                 
                   C≡CH 
                   N 3   
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
                   N 3   
                   C≡CH 
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
                   dichlorotriazine 
                   OH 
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
                   dichlorotriazine 
                   OH, OH 
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
                   dichlorotriazine 
                   OH, NH 2   
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
             
                
                
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         n is an integer from 1 to 4, 
         and salts thereof. 
       
     
     
         2 . The fluorescent compound according to  claim 1 , wherein m is equal to 7 or 8. 
     
     
         3 . The fluorescent compound according to  claim 1 , wherein at least one of the R′, R″, R′″ groups is selected from OCH 3 , OCH 2 CH 3 , OCH 2 CHOHCH 3 , whit the provision that at least one group from R′, R″, R′″ is OH. 
     
     
         4 . The fluorescent compound according to  claim 1 , wherein R′ and R″ are OH, and R′″ is selected from OCH 3 , OCH 2 CH 3 , OCH 2 CHOHCH 3 , wherein the substitution degree of R′″ is comprised between 0.5 and 1.5 for each unit of Formula (III). 
     
     
         5 . The fluorescent compound according to  claim 4 , wherein R′″ is OCH 2 CHOHCH 3.    
     
     
         6 . The fluorescent compound according to  claim 1 , wherein R 1  and R 2  are independently selected from H, SO 3 H and COOH. 
     
     
         7 . The fluorescent compound according to  claim 1 , wherein R 3  and R 4  are independently selected from methyl, ethyl, (CH 2 ) 5 COOH, (CH 2 ) 4 SO 3 H, (CH 2 ) 3 N + (CH 3 ) 3 . 
     
     
         8 . The fluorescent compound according to  claim 1 , wherein R 5  is selected from H, Cl, or 
       
         
           
           
               
               
           
         
       
     
     
         9 . The fluorescent compound according to  claim 1 , wherein L is selected from an ester, an ether, an amide or 
       
         
           
           
               
               
           
         
       
     
     
         10 . The fluorescent compound according to  claim 1 , selected from:
 a compound of formula (IV):   
       
         
           
           
               
               
           
         
       
       wherein CD is 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linker L of formula (I) is an ester bond formed in the coupling reaction of the carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD;
 a compound of formula (V): 
 
       
         
           
           
               
               
           
         
       
       wherein CD is 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linker L of Formula is an ester bond formed in the coupling reaction of the carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD;
 a compound of formula (VI): 
 
       
         
           
           
               
               
           
         
       
       wherein CD is 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linker L of Formula is an ester bond formed in the coupling reaction of the carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD;
 a compound of formula (VII): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1  and CD 2  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ether bonds formed in the coupling reaction of a dichlorotriazine group of the tricarbocyanine with a residue group R′″═OH of CD 1  and CD 2 , respectively;
 a compound of formula (VIII): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1  and CD 2  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD 1  and CD 2 , respectively;
 a compound of formula (IX): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1  and CD 2  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD 1  and CD 2 , respectively;
 a compound of formula (X): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1 , CD 2  and CD 3  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD 1 , CD 2 , and CD 3 , respectively;
 a compound of formula (XI): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1 , CD 2  and CD 3  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD 1 , CD 2  and CD 3 , respectively.
 a compound of formula (XII): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1  and CD 2  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD 1  and CD 2 , respectively;
 a compound of formula (XIII): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1  and CD 2  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R″═OH of CD 1  and CD 2 , respectively;
 a compound of formula (XIV): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1 , CD 2 , CD 3  and CD 4  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD 1 , CD 2 , CD 3  and CD 4 , respectively;
 a compound of formula (XV): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1  and CD 2  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R″═OH of CD 1  and CD 2 , respectively;
 a compound of formula (XVI): 
 
       
         
           
           
               
               
           
         
       
       wherein CD 1  and CD 2  are, independently, 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linkers L of Formula (I) are ester bonds formed in the coupling reaction of a carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD 1  and CD 2 , respectively;
 a compound of formula (XVII): 
 
       
         
           
           
               
               
           
         
       
       wherein CD is a 2-hydroxypropyl-β-cyclodextrin (HβCD) or 2-hydroxypropyl-γ-cyclodextrin (HγCD) and the linker L of Formula (I) is an ester bond formed in the coupling reaction of the carboxyl group of the tricarbocyanine with a residue group R′″═OH of CD. 
     
     
         11 . A fluorescent compound according to  claim 1  for use in the diagnosis of kidney function in a mammal. 
     
     
         12 . The fluorescent compound according to  claim 9  for use in determining the glomerular filtration rate. 
     
     
         13 . A diagnostic formulation for use in the diagnosis of kidney function, comprising at least one fluorescent compound according to  claim 1 . 
     
     
         14 . A method for diagnosing kidney function of a mammal, comprising:
 (a) administering to the mammal at least one fluorescent compound according to  claim 1 , and   (b) detecting and measuring the fluorescence emission of the at least one fluorescent compound.   
     
     
         15 . The method according to  claim 14 , wherein the administration is intravenous. 
     
     
         16 . The method according to  claim 14 , wherein the fluorescence detection and measurement comprise the detection and measurement of fluorescence emerging from the mammalian skin in response to excitation with a red light or near infrared light source. 
     
     
         17 . The method according to  claim 14 , wherein fluorescence is detected and measured by placing a sensor device onto the mammalian skin. 
     
     
         18 . The method according to  claim 14 , wherein the method is carried out over a clinically relevant measuring time. 
     
     
         19 . A screening method to identify a test agent suitable for the treatment of a chronic kidney disease in a mammal comprising:
 i) administering to an animal model of chronic kidney disease the test agent and at least one fluorescent compound according to  claim 1 , wherein the at least one fluorescent compound is administered subsequently to the test agent;   ii) measuring the glomerular filtration rate (GFR) of the animal model by detecting and measuring the fluorescence emission of the at least one fluorescent compound, wherein the detection and measurement of the fluorescence comprises the detection and measurement of the fluorescent emission emerging from the skin of the animal model in response to excitation with a red light or near infrared light source;   iii) selecting the test agent that increases the glomerular filtration rate.