Probe for selectively characterizing enzymes involved in xenobiotic metabolism and method of making and using the same

1. A probe having a structure satisfying Formula II

wherein

ERG, if present, is

S(O) 2 OH, or an anionic form thereof; P(O(OH) 2 , or an anionic form thereof; a halogen; or —OPh-CH 2 —ONO 2 ;

EBG has a structure satisfying a formula selected from Formulas IIA EBG -IIJ EBG

wherein Y is CH 3 or CF 3 ; Y′ is O, NO 2 , or —N═NR″, wherein R″ is a dye or other reporting moiety;

and m′ is an integer selected from 0 to 5; and R′ is selected from aldehyde, ketone, ester, carboxylic acid, acyl, acyl halide, cyano, sulfonate, nitro, nitroso, quaternary amine, CF 3 , alkyl halide, or combinations thereof;

Linker a comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;

each of Linker b , and Linker c , if present, independently comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;

R, if present, is a group comprising a carbamate or a carbonate; and

each of n, m, p, and q independently is 0 or 1;

provided that one of the following applies:

(a) one or more Tags are present, and each Tag is independently a fluorophore, a binding partner of an affinity-based binding pair, a quantum dot, or a dye; or

(b) one or more p Tags are present and each p Tag is independently an azide or an alkyne.

2. The probe of claim 1 , wherein Linker a is a linker group having a structure satisfying a formula selected from Formulas IIA Linkera -IIH Linkera

wherein each n′ independently is an integer ranging from 1 to 50; Z is oxygen or NR′″, wherein R′″ is hydrogen, aliphatic, or aromatic; and Q is carbon, oxygen or NR′″, wherein R′″ is hydrogen, aliphatic, or aromatic.

3. The probe of claim 1 , wherein Linker b is present and comprises a —(CH 2 ) n′ — group, wherein n′ is an integer ranging from 1 to 50; an amide group; or a combination thereof.

4. The probe of claim 1 , wherein Linker c is present and has a structure satisfying Formula IIA Linkerc or Formula IIB Linkerc

wherein n′ is an integer ranging from 0 to 50; and Z is oxygen or NR′″, wherein R′″ is hydrogen, aliphatic, or aromatic.

5. The probe of claim 1 , wherein R is present and has a structure satisfying any one or more Formulas IIA R -IID R

wherein Z and Z′ independently are oxygen or NR′″, wherein R′″ is hydrogen, aliphatic, or aromatic; and n′ is an integer ranging from 0 to 50.

6. The probe of claim 1 , wherein the one or more Tags independently are rhodamine, fluorescein, or biotin.

7. The probe of claim 1 , wherein the compound has a structure satisfying Formula IIIA

8. The probe of claim 7 , wherein the ERG is present and is iodo; —OPh-CH 2 —ONO 2 ;

9. The probe of claim 7 , wherein Linker a is an ester group, an —O(CH 2 ) n′ NR′″C(O)(CH 2 ) n′ — group, or a —(CH 2 ) n′ — group, wherein each n′ independently is an integer ranging from 1 to 20 and wherein R′″ is hydrogen, aliphatic, or aromatic.

10. The probe of claim 7 , wherein m is 1 and Linker c is an —NR′″C(O)(CH 2 ) n′ — group or an —NR′″C(O)CH 2 [O(CH 2 ) 2 ] n′ OCH 2 — group, wherein each n′ independently is an integer ranging from 1 to 20 and wherein R′″ is hydrogen, aliphatic, or aromatic.

11. The probe of claim 7 , wherein m is 1.

12. A probe selected from:

13. A probe having a structure satisfying Formula II

wherein

ERG is

EBG has a structure satisfying a formula selected from

wherein Y′ is O; m′ is an integer selected from 0 to 5; and each occurrence of R′ is independently selected from aldehyde, ketone, ester, carboxylic acid, acyl, acyl halide, cyano, sulfonate, nitro, nitroso, quaternary amine, CF 3 , alkyl halide, or combinations thereof;

Linker a comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;

each of Linker b , and Linker c , if present, independently comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;

R, if present, is a group comprising a carbamate or a carbonate;

each of n, m, p, and q independently is 0 or 1; and

provided that one of the following applies:

(a) one or more Tags are present, and each Tag is independently a fluorophore, a binding partner of an affinity-based binding pair, a quantum dot, or a dye; or

(b) one or more p Tags are present, and each p Tag is independently an azide or an alkyne.

14. The probe of claim 13 , wherein the ERG is

15. The probe of claim 13 , wherein the EBG has a structure satisfying a formula selected form

16. The probe of claim 13 , wherein the EBG has a structure satisfying a formula of

17. The probe of claim 13 , wherein Linker b and R are absent and wherein q and m are 0.

18. The probe of claim 13 , wherein Linker a is an —O(CH 2 ) n′ NR′″C(O)(CH 2 ) n′ — group, or a —(CH 2 ) n′ — group, wherein each n′ independently is an integer ranging from 1 to 20 and wherein R′″ is hydrogen, aliphatic, or aromatic.

19. The probe of claim 13 , wherein the one or more Tags are present.

20. The probe of claim 13 , wherein the one or more p Tags are present.

21. The probe of claim 13 , wherein n is 1.

22. A probe selected from: