Substituted imidazo[1,2-a]pyrazines as luciferase substrates and preparation method thereof
Described are substituted imidazo[1,2-a]pyrazine compounds, which are coelenterazine analogs, kits comprising the compounds, and methods of using the compounds for the detection of luminescence in luciferase-based assays. Also described are methods for making the compounds, such as a method using aminopyrazine acetophosphonates as synthesis intermediates.
1. A method for preparing a compound of formula (I):
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl; and
q is 1;
the method comprising:
(i) reacting a compound of formula A:
wherein:
R 5 is methyl;
with a compound of formula v:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl;
in the presence of 1,1,3,3-tetramethylguanidine and a solvent, to provide a compound of formula vi:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl; and
(ii) reducing the compound of formula vi above with sodium borohydride, to provide a compound of formula (I); or
(i) reacting a compound of formula B:
wherein:
R 5 is benzyl;
with a compound of formula v:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl;
in the presence of 1,1,3,3-tetramethylguanidine and a solvent, to provide a compound of formula vi:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl; and
(ii) reducing the compound of formula vi above with sodium borohydride, to provide a compound of formula (I); or
(i) reacting a compound of formula C:
wherein:
R 5 is tert-butyl;
with a compound of formula v:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl;
in the presence of 1,1,3,3-tetramethylguanidine and a solvent, to provide a compound of formula vii:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl;
(ii) reacting the compound of formula vii above with trifluoroacetic acid, to provide a compound of formula ix:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl; and
(iii) reacting the compound of formula ix above with acetic anhydride followed by sodium borohydride, to provide a compound of formula (I); or
(i) reacting a compound of formula C:
wherein:
R 5 is tert-butyl;
with a compound of formula v:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl;
in the presence of 1,1,3,3-tetramethylguanidine and a solvent, to provide a compound of formula vii:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl;
(ii) hydrogenating the compound of formula vii above in the presence of Rh(PPh 3 ) 3 Cl, to provide a compound of formula viii:
wherein:
R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl or phosphonate, wherein, at each occurrence, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, heteroarylcarbonyl and phosphonate are independently and optionally substituted with 1, 2, 3, 4, 5, 6 or 7 functional groups independently selected from the group consisting of halogen, oxo, thioxo, cyano, carbamate, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, allyloxy, alkoxyalkyl, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, alkylsulfonyl, arvlsulfonyl, aminosulfonyl, alkylsulfinyl, —COOH, amido, silyl, silyloxy, alkylsulfanyl, arylsulfanyl, thiotriazolyl and acyl; and
(iii) reacting the compound of formula viii above with trifluoroacetic acid followed by carbonyldiimidazole, to provide a compound of formula (I).
2. The method of claim 1 , wherein R 1 is phenyl, bicyclic aryl, tricyclic aryl, monocyclic heteroaryl, bicyclic heteroaryl or tricyclic heteroaryl, wherein, at each occurrence, the phenyl, bicyclic aryl, tricyclic aryl, monocyclic heteroaryl, bicyclic heteroaryl and tricyclic heteroaryl are independently and optionally substituted.
3. The method of claim 1 , wherein R 1 is phenyl or monocyclic heteroaryl, wherein, at each occurrence, the phenyl and monocyclic heteroaryl are independently and optionally substituted.
4. The method of claim 1 , wherein R 1 is 2-furanyl.
5. The method of claim 1 , wherein R 5 is methyl.
6. The method of claim 5 , wherein the solvent is methanol.
7. The method of claim 1 , wherein R 5 is benzyl.
8. The method of claim 7 , wherein the solvent is methanol.
9. The method of claim 1 , wherein R 5 is tert-butyl.
10. The method of claim 9 , wherein the solvent is methanol.