Luciferin derivatives from bicyclic reactants and aminothiol derivatives and methods of use thereof

Abstract

The present disclosure features a condensation reaction and a luciferin-unmasking reaction that can be carried out under physiological conditions. In general, the condensation reaction involves reacting a bicyclic reactant with an aminothiol derivative, generating a luciferin or luciferin derivative. A luciferin can provide detectable luminescence. A luciferin derivative can be unmasked to provide detectable luminescence in a luciferin-unmasking reaction. The present disclosure provides bicyclic reactants and aminothiol derivatives suitable for use in the condensation reaction. The condensation and luciferin-unmasking reactions find use in a variety of applications, which are also provided.

What is claimed is: 
     
         1 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q 1  is a heteroatom functional group selected from —O— and —NR Q1 —; 
 R Q1  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 1  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; and 
 PG I  is an optional protecting group for cyano group. 
 
     
     
         2 . The compound of  claim 1  having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         3 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q 2  is a heteroatom functional group selected from —O— and —NR Q2 —; 
 R Q2  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 2  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; 
 Q 3  is —O— or —S—; and 
 PG I  is an optional protecting group for cyano group. 
 
     
     
         4 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q 4  is a heteroatom functional group selected from —O— and —NR Q4 —; 
 R Q4  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 3  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; 
 Q 5  is —O— or —S—; and 
 PG IV  is an optional protecting group for cyano group. 
 
     
     
         5 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q 6  is a heteroatom functional group selected from —O— and —NR Q6 —; 
 R Q6  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 4  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; 
 Q 7  is —O— or —S—; and 
 PG I  is an optional protecting group for cyano group. 
 
     
     
         6 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q is a heteroatom functional group selected from —O— and —NR Q8 —; 
 R Q8  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 5  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; 
 Q 9  is —O— or —S—; and 
 PG I  is an optional protecting group for cyano group. 
 
     
     
         7 . The compound of any of  claims 1  and  3 - 6 , wherein at least one of PG I  and PG IV  is a protecting group. 
     
     
         8 . The compound of any of  claims 1  and  3 - 6 , wherein—PG IV  is a protecting group. 
     
     
         9 . The compound of any of  claims 1  and  3 - 6 , wherein—PG IV  is a protecting group. 
     
     
         10 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 PG II  is hydrogen or a protecting group for an amino group; 
 PG III  is hydrogen or a protecting group for a thiol group; 
 PG V  is hydrogen or a protecting group for a carboxyl group; and 
 L is C 1-2  alkylene, C 1-2  alkenylene, or C 1-2  alkynylene. 
 
     
     
         11 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 PG II  is hydrogen or a protecting group for an amino group; 
 PG III  is hydrogen or a protecting group for a thiol group; 
 PG V  is hydrogen or a protecting group for a carboxyl group. 
 
     
     
         12 . The compound of any of  claims 10 - 11 , wherein PG V  is hydrogen. 
     
     
         13 . The compound of any of  claims 10 - 11 , wherein PG V  is a protecting group. 
     
     
         14 . The compound of any of  claims 10 - 11 , wherein PG II  is hydrogen. 
     
     
         15 . The compound of any of  claims 10 - 11 , wherein PG II  is a protecting group. 
     
     
         16 . The compound of  claim 15 , wherein PG II  forms a masked amino group in which the masked amino group is a carbamate or an amide. 
     
     
         17 . The compound of  claim 15 , wherein PG II  forms a masked amino group in which the masked amino group is an amino group that is connected to an amino acid, peptide, or protein. 
     
     
         18 . The compound of  claim 15  having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         19 . The compound of any of  claims 10 - 11 , wherein PG III  is hydrogen. 
     
     
         20 . The compound of any of  claims 10 - 11 , wherein PG III  is a protecting group. 
     
     
         21 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q 1  is a heteroatom functional group selected from —O— and —NR Q1 —; 
 R Q1  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 1  is selected from hydrogen, halogen, hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; 
 PG V  is hydrogen or a protecting group for a carboxyl group; and 
 L is C 1-2  alkylene, C 1-2  alkenylene, or C 1-2  alkynylene. 
 
     
     
         22 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q 2  is a heteroatom functional group selected from —O— and —NR Q2 —; 
 R Q2  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 2  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; 
 Q 3  is —O— or —S—; 
 PG V  is hydrogen or a protecting group for a carboxyl group; and 
 L is C 1-2  alkylene, C 1-2  alkenylene, or C 1-2  alkynylene. 
 
     
     
         23 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q 4  is a heteroatom functional group selected from —O— and —NR Q4 —; 
 R Q4  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 3  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, substituted amino, a moiety that comprises a reactive group that facilitates covalent attachment of a molecule of interest; and a molecule of interest; 
 Q 5  is —N— or —CH—; 
 PG V  is hydrogen or a protecting group for a carboxyl group; and 
 L is C 1-2  alkylene, C 1-2  alkenylene, or C 1-2  alkynylene. 
 
     
     
         24 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q 6  is a heteroatom functional group selected from —O— and —NR Q6 —; 
 R Q6  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 4  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; 
 Q 7  is —N— or —CH—; 
 PG V  is hydrogen or a protecting group for a carboxyl group; and 
 L is C 1-2  alkylene, C 1-2  alkenylene, or C 1-2  alkynylene. 
 
     
     
         25 . A compound of formula: 
       
         
           
           
               
               
           
         
       
       wherein
 Q is a heteroatom functional group selected from —O— and —NR Q8 —; 
 R Q8  is selected from hydrogen, alkyl, and substituted alkyl; 
 PG IV  is hydrogen or a protecting group for the heteroatom functional group; 
 R 5  is selected from hydrogen, halogen hydroxyl, alkyl, substituted alkyl, alkoxy, amino, and substituted amino; 
 Q 9  is —N— or —CH—; 
 PG V  is hydrogen or a protecting group for a carboxyl group; and 
 L is C 1-2  alkylene, C 1-2  alkenylene, or C 1-2  alkynylene. 
 
     
     
         26 . A method for detection of a biological process or biomolecule in a test subject, the method comprising contacting test subject with a compound of any of Formula I-V and a compound of Formula VI. 
     
     
         27 . The method of  claim 26 , wherein the test subject is contacted with the compound of  claim 2  and  claim 18 . 
     
     
         28 . The method of  claim 26 , further comprising monitoring the test subject for luminescence, wherein luminescence indicates the presence of the biological process or biomolecule. 
     
     
         29 . The method of  claim 28 , wherein the biomolecule is caspase-8 and/or H 2 O 2 . 
     
     
         30 . The method of  claim 28 , wherein the biological process is cancer, a cardiovascular disorder, diabetes, a neurodegenerative disease, or an inflammatory response.