Optical molecular sensors for cytochrome P450 activity
The invention provides a compound, useful as an optical probe or sensor of the activity of at least one cytochrome P450 enzyme, and methods of using the compound to screen candidate drugs, and candidate drugs identified by these methods. The optical probe of the invention is a compound having the generic structure Y-L-Q, wherein Q is a chemical moiety that gives rise to optical properties in its hydroxy or hyrdoxylate, phenol or phenoxide form that are different from the optical properties that arise from its ether form.
1. A process for preparing a compound useful as a sensor for cytochrome P450 activity having the structure
R 3 -L-Q
said process comprising the step of reacting a compound of the formula
H-Q
in the presence of DMF/K 2 CO 3 or diisopropylethylamine/DMF at temperatures at about 0° C. to 5° C., with a compound of the formula
R 3 -L-X
wherein:
X is a suitable leaving group selected from the group consisting of a halogen atom, a tosyl group, a mesyl group, and a triflate group;
R 3 is an optionally substituted aryl;
L has the chemical structure (−OCR 2 H) p —, wherein each R 2 is separately selected from the group consisting of a hydrogen atom, saturated C 1 -C 20 alkyl, unsaturated C 2 -C 20 alkenyl, unsaturated C 2 -C 20 alkynyl, substituted saturated C 1 -C 20 alkyl, substituted unsaturated C 2 -C 20 alkenyl, substituted unsaturated C 2 -C 20 alkynyl, C 3 -C 20 cycloalkyl, C 3 -C 20 cycloalkenyl, substituted saturated C 3 -C 20 cycloalkyl, substituted unsaturated C 3 -C 20 cycloalkenyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl, and p is a positive integer selected from one to twelve;
Q is attached to L through an ether linkage via the oxygen atom indicated by the following arrow, and has a structure selected from the group consisting of the following structures:
wherein:
R a , R b , R c , R d , and R e , are each separately selected from the group consisting of a hydrogen atom, a halogen atom, C 1 -C 20 alkyl, substituted C 1 -C 20 alkyl, perhalogenated alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, benzyl, heteroaryl, substituted heteroaryl, cyano, nitro, azido, —SR S , —OR O , —NR n1 R n2 , —N + R n1 R n2 R n3 , —P + R n1 R n2 R n3 , —COR C , —C(═NOR O )R C , —CSR C , —OCOR C , —OCONR n1 R n2 , —OCO 2 R C , —CONR n1 R n2 , —C(═N)NR n1 R n2 , —CO 2 R O , —SO 2 NR n1 R n2 , —SO 3 R O , —SO 2 R O , —PO(OR O ) 2 , —NR n1 CSNR n2 R n3 , —NR n1 C(═N)NR n2 R n3 , —NR n1 CONR n2 R n3 , —NR n1 COR C and —NR n1 S(═O) 2 R S ;
R n1 , R n2 , R n3 , R O and R S are each separately selected from the group consisting of a hydrogen atom, C 1 -C 20 alkyl, substituted C 1 -C 20 alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, benzyl, heteroaryl, and substituted heteroaryl;
R C is selected from the group consisting of a hydrogen atom, C 1 -C 20 alkyl, substituted C 1 -C 20 alkyl, perhalogenated alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, benzyl, heteroaryl, substituted heteroaryl, and cyano; and
T is an oxygen atom.
2. The process of claim 1 , wherein Q is a 7-hydroxycoumarin.
3. The process of claim 1 , wherein Q is a cyanocoumarin.
4. The process of claim 1 , wherein Q is a trifluorocoumarin.
5. The process of claim 1 , wherein R 2 is a hydrogen atom for all values of p.
6. The process of claim 1 , wherein p is equal to one.