US2014162309A1PendingUtilityA1

Arsenical Fluorescent Agents and Assays

Assignee: STANFORD RES INST INTPriority: Mar 19, 2010Filed: Feb 14, 2014Published: Jun 12, 2014
Est. expiryMar 19, 2030(~3.6 yrs left)· nominal 20-yr term from priority
G01N 33/582C09B 21/00G01N 2333/44C12Q 1/04C09B 19/00C07F 9/88C09B 17/00C07F 9/80C09B 11/24
49
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Claims

Abstract

The invention provides methods and compositions for labeling dithiol-containing analytes, including a substituted, optionally hydro-, optionally hetero-, monoarsenical anthracene compound comprising a 4′ rotation blocking group and a 5′ arsenic, and that exhibits a detectable increase or shift in fluorescence when the arsenic reacts with two thiols of a rotation-blocking binding target molecule.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A substituted, optionally hydro-, optionally hetero-, monoarsenical anthracene compound comprising a 4′ rotation blocking group and a 5′ arsenic, and that exhibits a detectable increase or shift in fluorescence when the arsenic reacts with two thiols of a rotation-blocking binding target molecule, the compound having the structure I: 
       
         
           
           
               
               
           
         
         wherein: 
         R1 is O, S, NRα, or CRα 2 ; 
         R2 is C or N; 
         R3 is H, optionally substituted-, optionally hetero-alkyl, optionally substituted-, optionally hetero-alkenyl, optionally substituted-, optionally hetero-alkynyl, optionally substituted-, optionally hetero-aryl, or optionally substituted-, optionally hetero-alkoxy; 
         R4 and R5 are independently carbonyl, carbonothioyl, NRα 2 , ORα, or SRα; 
         R6 and R7 are Rα, halide, ORα, NRα 2 , ORα, SRα, or nitro (—NO 2 ); 
         R8 and R9 are Rα, halide, ORα, NRα 2 , ORα, SRα, or nitro (—NO 2 ); 
         R10 is a rotational blocking group; 
         R11 is an arsenic protecting group displaced by reaction of the arsenic with the thiols of the target molecule; 
         Rα groups are independently H, optionally substituted-, optionally hetero-alkyl, optionally substituted-, optionally hetero-alkenyl, optionally substituted-, optionally hetero-alkynyl, optionally substituted-, optionally hetero-aryl, or optionally substituted-, optionally hetero-alkoxy; 
         wherein one or more of the pairs R4-R6, R6-R8, R5-R7, and R7-R9 may be covalently joined in one or more rings; 
         and tautomers, anhydrides and salts of the compound. 
       
     
     
         2 . The compound of  claim 1  wherein R11 is carbonyl, 1,2 ethanedithiol, or dihydroxyl. 
     
     
         3 . The compound of  claim 1  wherein the rotation blocking group is an optionally substituted-, optionally hetero-alkyl, optionally substituted-, optionally hetero-alkenyl, optionally substituted-, optionally hetero-alkynyl, optionally substituted-, optionally hetero-aryl, or optionally substituted-, optionally hetero-alkoxy. 
     
     
         4 . The compound of  claim 1  wherein:
 R11 is carbonyl, 1,2 ethanedithiol, or dihydroxyl; and 
 the rotation blocking group is an optionally substituted-, optionally hetero-alkyl, optionally substituted-, optionally hetero-alkenyl, optionally substituted-, optionally hetero-alkynyl, optionally substituted-, optionally hetero-aryl, or optionally substituted-, optionally hetero-alkoxy. 
 
     
     
         5 . The compound of  claim 1  wherein:
 R1 is O, S, N, or C; 
 
     
     
         6 . The compound of  claim 1  wherein:
 R1 is O, S, N, or C; 
 R11 is carbonyl, 1,2 ethanedithiol, or dihydroxyl; and 
 the rotation blocking group is an optionally substituted-, optionally hetero-alkyl, optionally substituted-, optionally hetero-alkenyl, optionally substituted-, optionally hetero-alkynyl, optionally substituted-, optionally hetero-aryl, or optionally substituted-, optionally hetero-alkoxy. 
 
     
     
         7 . The compound of  claim 1  wherein: 
       
         
           
                 
                 
                 
               
                     
                 
                   R1 is O; 
                   R1 is O; 
                   R1 is O; 
                 
                   R2 is C; 
                   R2 is C; 
                   R2 is C; 
                 
                   R3 is 2-carboxyphenyl; 
                   R3 is 2-carboxyphenyl; 
                   R3 is 2-carboxyphenyl; 
                 
                   R4 is hydroxyl; 
                   R4 is hydroxyl; 
                   R4 is amine; 
                 
                   R5 is carbonyl; 
                   R5 is carbonyl; 
                   R5 is amine; 
                 
                   R6 and R7 are H; and 
                   R6 and R7 are H or Cl; 
                   R6 and R7 are H; and 
                 
                   R8 and R9 are H; 
                   R8 and R9 are H; and 
                   R8 and R9 are H; 
                 
                     
                   R10 is methyl or benzyl; 
                 
                   R1 is O; 
                   R1 and R2 are N; 
                   R1 is O; 
                 
                   R2 is C; 
                   R3 is H or methyl; 
                   R2 is N; 
                 
                   R3 is 2-carboxyphenyl; 
                   R4 and R5 are O, S, or N; 
                   R3 is H or methyl; 
                 
                   R4 is hydroxyl; 
                   R6 and R7 are H; and 
                   R4 is hydroxyl; 
                 
                   R5 is carbonyl; 
                   R8 and R9 are H; 
                   R5 is carbonyl; 
                 
                   R6 and R7 are Br or nitro (—NO 2 ); 
                     
                   R6 and R7 are H; and 
                 
                   and R8 and R9 are H; 
                     
                   R8 and R9 are H; 
                 
                   R1 is S; 
                   R1 is S; 
                   R1 and R2 are C; 
                 
                   R2 is N; 
                   R2 is C; 
                   R3 is H or C; 
                 
                   R3 is H or C; 
                   R3 is 2-carboxyphenyl; 
                   R4 and R5 are O, S, N; 
                 
                   R4 is hydroxyl; 
                   R4 is hydroxyl; 
                   R6 and R7 are H; and 
                 
                   R5 is carbonyl; 
                   R5 is carbonyl; 
                   R8 and R9 are H; 
                 
                   R6 and R7 are H; and 
                   R6 and R7 are H; and 
                 
                   R8 and R9 are H; 
                   R8 and R9 are H; 
                 
                   R1 is N; 
                   R1 is N; 
                   R1 is N; 
                 
                   R2 is C; 
                   R2 is C; 
                   R2 is C; 
                 
                   R3 is H or C; 
                   R3 is H; 
                   R3 is H; 
                 
                   R4 and R5 are O, S, or N; 
                   R4 and R5 are NH 2 ; 
                   R4 and R5 are N(CH 3 ) 2 ; 
                 
                   R6 and R7 are H; and 
                   R6 and R7 are CH 3 ; and 
                   R6 and R7 are H; and 
                 
                   R8 and R9 are H; 
                   R8 and R9 are H; or 
                   R8 and R9 are H. 
                 
                     
                 
             
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         8 . The compound of  claim 7  wherein:
 R11 is carbonyl, 1,2 ethanedithiol, or dihydroxyl; and 
 the rotation blocking group is an optionally substituted-, optionally hetero-alkyl, optionally substituted-, optionally hetero-alkenyl, optionally substituted-, optionally hetero-alkynyl, optionally substituted-, optionally hetero-aryl, or optionally substituted-, optionally hetero-alkoxy. 
 
     
     
         9 . A compound of  claim 1  selected from: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         10 . The compound of  claim 1  wherein the arsenic is reacted with two thiols of a rotation-blocking binding target molecule forming a conjugate having the general structure II: 
       
         
           
           
               
               
           
         
         wherein AC is the anthracene core, RBG is the rotation blocking group and TM is the target molecule, and tautomers, anhydrides and salts of the compound. 
       
     
     
         11 . A method of using a compound of  claim 1  comprising the step of:
 contacting the compound with the target molecule wherein the arsenic reacts with the thiols.

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