Composition and device for detecting leukocytes in urine

Abstract

Thiazole esters are suitable for detecting the presence of leukocytes in urine. Such thiazole esters are suitable for use in compositions, diagnostic devices, and methods for detecting the presence of leukocytes. A thiazole ester of the invention is of the formula: or a salt or solvated salt thereof, in which A is an N-blocked amino acid residue or N-blocked peptide chain, preferably an alanine residue or polyalanine chain; and R 1 and R 2 are each independently hydrogen, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, unsubstituted or substituted alkyl, unsubstituted or substituted alkenyl, unsubstituted or substituted alkoxy, amino, unsubstituted or substituted acyl, halo, nitro, cyano, —SO 3 H, or hydroxy, with the proviso that R 1 and R 2 are not both hydrogen. In one embodiment, at least one of R 1 and R 2 is methoxy, ethoxy, propoxy, or butoxy. In still another embodiment, R 1 is hydrogen and R 2 is methoxy, ethoxy, propoxy, or butoxy.

What is claimed is:  
     
         1 . A compound of the formula:  
       
         
           
           
               
               
           
         
       
       or a salt or solvated salt thereof, wherein 
 A is an N-blocked amino acid residue or N-blocked peptide chain; and  
 R 1  and R 2  are each independently hydrogen, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, unsubstituted or substituted alkyl, unsubstituted or substituted alkenyl, unsubstituted or substituted alkoxy, amino, unsubstituted or substituted acyl, halo, nitro, cyano, —SO 3 H, or hydroxy, wherein R 1  and R 2  are not both hydrogen.  
 
     
     
         2 . The compound of  claim 1 , wherein at least one of R 1  and R 2  is selected from the group consisting of methoxy, ethoxy, propoxy, and butoxy.  
     
     
         3 . The compound of  claim 1 , wherein R 1  is hydrogen and R 2  is methoxy, ethoxy, propoxy, or butoxy.  
     
     
         4 . The compound of  claim 3 , wherein the position of R 2  is meta or ortho relative to thiazolyl.  
     
     
         5 . The compound of  claim 1 , wherein the compound is of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         6 . The compound of  claim 1 , wherein the compound is of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         7 . The compound of  claim 1 , wherein A is N-blocked alanine or N-blocked polyalanine.  
     
     
         8 . A compound of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         9 . A compound of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         10 . A composition comprising: 
 (a) a compound of the formula                          or a salt or solvated salt thereof, wherein    A is an N-blocked amino acid residue or N-blocked peptide chain; and    R 1  and R 2  are each independently hydrogen, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, unsubstituted or substituted alkyl, unsubstituted or substituted alkenyl, unsubstituted or substituted alkoxy, amino, unsubstituted or substituted acyl, halo, nitro, cyano, —SO 3 H, or hydroxy, wherein R 1  and R 2  are not both hydrogen; and    (b) a diazonium salt.    
     
     
         11 . The composition of  claim 10 , wherein the compound has at least one of R 1  and R 2  that is selected from the group consisting of methoxy, ethoxy, propoxy, and butoxy.  
     
     
         12 . The composition of  claim 10 , wherein the compound has R 1  that is hydrogen and R 2  that is methoxy, ethoxy, propoxy, or butoxy.  
     
     
         13 . The composition of  claim 12 , wherein the compound has R 2  positioned meta or ortho relative to thiazolyl.  
     
     
         14 . The composition of  claim 10 , wherein the compound is of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         15 . The composition of  claim 10 , wherein the compound is of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         16 . The composition of  claim 10 , wherein the compound has A that is N-blocked alanine or N-blocked polyalanine.  
     
     
         17 . The composition of  claim 10 , wherein the diazonium salt is selected from the group consisting of 1-diazo-8-naphtol-3,6-disulfonic acid, chloride, zinc chloride double salt; 6-diazo-1-naphtol-3-sulfonic acid, chloride double salt; and 2-methoxy-4-morpholinobenzene diazonium chloride, zinc chloride double salt.  
     
     
         18 . The composition of  claim 10 , wherein the composition is free of accelerating salt.  
     
     
         19 . A diagnostic device for detecting leukocytes and proteinase in urine, comprising: 
 an inert carrier having deposited thereon a compound of the formula:                          or a salt or solvated salt thereof, wherein    A is an N-blocked amino acid residue or N-blocked peptide chain; and    R 1  and R 2  are each independently hydrogen, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, unsubstituted or substituted alkyl, unsubstituted or substituted alkenyl, unsubstituted or substituted alkoxy, amino, unsubstituted or substituted acyl, halo, nitro, cyano, —SO 3 H, or hydroxy, wherein R 1  and R 2  are not both hydrogen.    
     
     
         20 . The device of  claim 19 , wherein the compound has at least one of R 1  and R 2  that is selected from the group consisting of methoxy, ethoxy, propoxy, and butoxy.  
     
     
         21 . The device of  claim 19 , wherein the compound has R 1  that is hydrogen and R 2  that is methoxy, ethoxy, propoxy, or butoxy.  
     
     
         22 . The device of  claim 21 , wherein the compound has R 2  that is at a position meta or ortho relative to thiazolyl.  
     
     
         23 . The device of  claim 19 , wherein the compound is of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         24 . The device of  claim 19 , wherein the compound is of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         25 . The device of  claim 19 , wherein the compound has A that is N-blocked alanine or N-blocked polyalanine.  
     
     
         26 . The device of  claim 19 , wherein the inert carrier has a diazonium salt deposited thereon.  
     
     
         27 . The device of  claim 26 , wherein the diazonium salt is selected from the group consisting of 1-diazo-8-naphtol-3,6-disulfonic acid, chloride, zinc chloride double salt; diazonium salt is 6-diazo-1-naphtol-3-sulfonic acid, chloride double salt; and 2-methoxy-4-morpholinobenzene diazonium chloride, zinc chloride double salt.  
     
     
         28 . The device of  claim 19 , wherein the inert carrier is filter paper.  
     
     
         29 . The device of  claim 19 , wherein the device is free of accelerating salt.  
     
     
         30 . A method for detecting the presence of leukocytes in urine, comprising contacting a urine sample with a diazonium salt and a compound of the formula:  
       
         
           
           
               
               
           
         
       
       or a salt or solvated salt thereof, wherein 
 A is an N-blocked amino acid residue or N-blocked peptide chain; and  
 R 1  and R 2  are each independently hydrogen, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, unsubstituted or substituted alkyl, unsubstituted or substituted alkenyl, unsubstituted or substituted alkoxy, amino, unsubstituted or substituted acyl, halo, nitro, cyano, —SO 3 H, or hydroxy, wherein R 1  and R 2  are not both hydrogen.  
 
     
     
         31 . The method of  claim 30 , wherein the diazonium salt is selected from the group consisting of 1-diazo-8-naphtol-3,6-disulfonic acid, chloride, zinc chloride double salt; 6-diazo-1-naphtol-3-sulfonic acid, chloride double salt; and 2-methoxy-4-morpholinobenzene diazonium chloride, zinc chloride double salt.  
     
     
         32 . The method of  claim 30 , wherein the compound has at least one of R 1  and R 2  that is selected from the group consisting of methoxy, ethoxy, propoxy, and butoxy.  
     
     
         33 . The compound of  claim 30 , wherein the compound has R 1  that is hydrogen and R 2  that is methoxy, ethoxy, propoxy, or butoxy.  
     
     
         34 . The compound of  claim 33 , wherein the compound has R 2  at a position that is meta or ortho relative to thiazolyl.  
     
     
         35 . The method of  claim 30 , wherein the compound is of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         36 . The method of  claim 30 , wherein the compound is of the formula:  
       
         
           
           
               
               
           
         
       
     
     
         37 . The method of  claim 30 , wherein the compound has A that is N-blocked alanine or N-blocked polyalanine.  
     
     
         38 . The method of  claim 30 , wherein the steps of the method are free of use of accelerating salt.