Claims
- 1. A compound having the formula
- 2. A compound in accordance with claim 1 in which R2 is H.
- 3. A compound in accordance with claim 1 in which R1 is a residue of a fluorescent aldehyde.
- 4. A compound in accordance with claim 1 in which R1 is a residue of a UV light-absorbing aldehyde.
- 5. A compound in accordance with claim 1 in which R1 is a residue of a colorimetric aldehyde.
- 6. A compound in accordance with claim 1 in which R3 is a member selected from the group consisting of alkyl, substituted alkyl, aralkyl, and substituted aralkyl.
- 7. A compound in accordance with claim 6 in which R3 is a member selected from the group consisting of C1-C6 alkyl, substituted C1-C6 alkyl, phenyl-(C1-C6 alkyl), and substituted phenyl-(C1-C6 alkyl), and R2 is H.
- 8. A compound in accordance with claim 6 in which R3 is a member selected from the group consisting of C1-C6 alkyl, hydroxy-substituted C1-C6 alkyl, (C1-C3 alkoxy)-substituted C1-C6 alkyl, halo-substituted C1-C6 alkyl, phenyl-(C1-C6 alkyl), and hydroxy-substituted phenyl (C1-C6 alkyl), (C1-C3 alkoxy)-substituted phenyl-(C1-C6 alkyl), and halo-substituted phenyl-(C1-C6 alkyl), and R2 is H.
- 9. A compound in accordance with claim 1 in which R3 is
- 10. A compound in accordance with claim 9 in which R4 is a member selected from the group consisting of alkyl and substituted alkyl.
- 11. A compound in accordance with claim 10 in which R4 is a member selected from the group consisting of C1-C12 alkyl, hydroxy-substituted C1-C12 alkyl, (C1-C3 alkoxy)-substituted C1-C12 alkyl, and halo-substituted C1-C12 alkyl, and R2 is H.
- 12. A compound in accordance with claim 10 in which R4 is a member selected from the group consisting of C1-C6 alkyl, hydroxy-substituted C1-C6 alkyl, (C1-C3 alkoxy)-substituted C1-C6 alkyl, and halo-substituted C1-C6 alkyl, and R2 is H.
- 13. A compound in accordance with claim 9 in which R4 is a member selected from the group consisting of aralkyl.and substituted aralkyl.
- 14. A compound in accordance with claim 13 in which R4 is a member selected from the group consisting of phenyl-(C1-C6 alkyl) and substituted phenyl-(C1-C6alkyl).
- 15. A compound in accordance with claim 13 in which R4 is a member selected from the group consisting of phenyl-(C1-C6 alkyl), hydroxy-substituted phenyl-(C1-C6 alkyl), (C1-C3 alkoxy)-substituted phenyl-(C1-C6 alkyl), and halo-substituted phenyl-(C1-C6 alkyl), and R2 is H.
- 16. A compound in accordance with claim 13 in which R4 is a member selected from the group consisting of benzyl and phenylethyl.
- 17. A compound in accordance with claim 9 in which R4 is a member selected from the group consisting of oxiranylalkyl, substituted oxiranylalkyl, oxiranylalkoxy, substituted oxiranylalkoxy, oxiranylamino, and substituted oxiranylamino.
- 18. A compound in accordance with claim 17 in which R4 is a member selected from the group consisting of oxiranyl-(C1-C3 alkyl), C1-C3 alkyl-substituted oxiranyl-(C1-C3 alkyl), oxiranyl-(C1-C3 alkoxy), C1-C3 alkyl-substituted oxiranyl-(C1-C3 alkoxy), oxiranylamino, and C1-C3 alkyl-substituted oxiranylamino, and R2 is H.
- 19. A compound in accordance with claim 1 in which R1 is a member selected from the group consisting of naphthyl, substituted naphthyl, anthryl, substituted anthryl, phenanthryl, substituted phenanthryl, phenalenyl, substituted phenalenyl, quinolinyl, substituted quinolinyl, N-alkyl quinolinium, substituted N-alkyl quinolinium, acridinyl, substituted acridinyl, N-alkyl acridinium, substituted N-alkyl acridinium, coumarinyl, substituted coumarinyl, furanocoumarinyl, substituted furanocoumarinyl, fluorenyl, substituted fluorenyl, fluorenonyl, substituted fluorenonyl, acetaminophenyl, indolyl, substituted indolyl, trans-retinyl, resorufinyl, substituted resorufinyl, polymethenyl, and substituted polymethenyl.
- 20. A compound in accordance with claim 19 in which R1 is a member selected from the group consisting of naphthyl, anthryl, phenanthryl, quinolinyl, coumarinyl, furanocoumarinyl, fluorenyl, fluorenonyl, acetaminophenyl, indolyl, trans-retinyl, resorufinyl, polymethenyl, and substituted analogs of all of the above in which the substituents are members selected from the group consisting of hydroxy, halo, nitro, amino. C1-C3 alkylamino, di-(C1-C3-alkyl)-amino, and C1-C3 alkoxy.
- 21. A method of detecting the presence of enzyme activity capable of converting an —O—R group in a substrate, where R is other than H, to —OH, said method comprising:
(a) contacting a sample suspected of containing such activity with a reporter having the formula 44 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; R2 is a member selected from the group consisting of H, C1-C2 alkyl, benzyl, and substituted benzyl; and R3 is R or a mimic of R that provides said reporter with a molecular structure that is susceptible to said enzyme activity whereby the —O—R3 of said reporter formula converts to —OH, R3 further being a member selected from the group consisting of alkyl, substituted alkyl, aralkyl, substituted aralkyl, phosphate, sulfate, carbohydrate residues, substituted carbohydrate residues, and 45in which R4 is a member selected from the group consisting of alkyl, substituted alkyl, aralkyl, substituted aralkyl, oxiranylalkyl, substituted oxiranylalkyl, oxiranylalkoxy, substituted oxiranylalkoxy, alkylamino, substituted alkylamino, peptidylamino, cycloalkylamino, substituted cycloalkylamino, oxiranylamino, substituted oxiranylamino, alkoxy, substituted alkoxy, and a residue of the acid moiety of a pyrethroid, under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 46 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 47 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter.
- 22. A method in accordance with claim 21 in which R2 is H.
- 23. A method in accordance with claim 21 in which R3 is a member selected from the group consisting of alkyl, substituted alkyl, aralkyl, and substituted aralkyl.
- 24. A method in accordance with claim 23 in which R3 is a member selected from the group consisting of C1-C6 alkyl, substituted C1-C6 alkyl, phenyl-(C1-C6 alkyl), and substituted phenyl-(C1-C6 alkyl), and R2 is H.
- 25. A method in accordance with claim 21 in which R3 is
- 26. A method in accordance with claim 25 in which R4 is a member selected from the group consisting of alkyl and substituted alkyl.
- 27. A method in accordance with claim 25 in which R4 is a member selected from the group consisting of C1-C6 alkyl, hydroxy-substituted C1-C6 alkyl, (C1-C3 alkoxy)-substituted C1-C6 alkyl, and halo-substituted C1-C6 alkyl, and R2 is H.
- 28. A method in accordance with claim 25 in which R4 is a member selected from the group consisting of aralkyl and substituted aralkyl.
- 29. A method in accordance with claim 25 in which R4 is a member selected from the group consisting of phenyl-(C1-C6 alkyl), hydroxy-substituted phenyl-(C1-C6 alkyl), (C1-C3 alkoxy)-substituted phenyl-(C1-C6 alkyl), and halo-substituted phenyl-(C1-C6 alkyl), and R2 is H.
- 30. A method in accordance with claim 25 in which R4 is a member selected from the group consisting of oxiranylalkyl, substituted oxiranylalkyl, oxiranylalkoxy, substituted oxiranylalkoxy, oxiranylamino, and substituted oxiranylamino.
- 31. A method in accordance with claim 25 in which R4 is a member selected from the group consisting of oxiranyl-(C1-C3 alkyl), C1-C3 alkyl-substituted oxiranyl-(C1-C3 alkyl), oxiranyl-(C1-C3 alkoxy), C1-C3 alkyl-substituted oxiranyl-(C1-C3 alkoxy), oxiranylamino, and C1-C3 alkyl-substituted oxiranylamino, and R2 is H.
- 32. A method in accordance with claim 25 in which R4 is a residue of the acid moiety of a pyrethroid.
- 33. A method in accordance with claim 32 in which said residue is a member selected from the group consisting of (2′,2′-dimethylvinyl)-2,2-dimethylcyclopropyl, (2′,2′-dichlorovinyl)-2,2-dimethylcyclopropyl, and (2′,2′-dibromovinyl)-2,2-dimethyl-cyclopropyl, and R2 is H.
- 34. A method in accordance with claim 25 in which R1 is a member selected from the group consisting of naphthyl, substituted naphthyl, anthryl, substituted anthryl, phenanthryl, substituted phenanthryl, phenalenyl, substituted phenalenyl, quinolinyl, substituted quinolinyl, N-alkyl quinolinium, substituted N-alkylquinolinium, N-alkyl acridinium, substituted N-alkyl acridinium, coumarinyl, substituted coumarinyl, furanocoumarinyl, substituted furanocoumarinyl, fluorenyl, substituted fluorenyl, fluorenonyl, substituted fluorenonyl, acetaminophenyl, indolyl, substituted indolyl, trans-retinyl, resorufinyl, substituted resorufinyl, polymethenyl, and substituted polymethenyl.
- 35. A method in accordance with claim 25 in which R1 is a member selected from the group consisting of naphthyl, anthryl, phenanthryl, quinolinyl, coumarinyl, furanocoumarinyl, fluorenyl, fluorenonyl, acetaminophenyl, indolyl, trans-retinyl, resorufinyl, polymethenyl, and substituted analogs of all of the above in which the substituents are members selected from the group consisting of hydroxy, halo, nitro, amino C1-C3 alkylamino, di-(C1-C3-alkyl)-amino, and C1-C3 alkoxy.
- 36. A method for the detection of esterase activity in a sample, said method comprising
(a) contacting said sample with a reporter having the formula 49 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; R2 is a member selected from the group consisting of H, C1-C2 alkyl, benzyl. and substituted benzyl; and R4 is a member selected from the group consisting of alkyl, substituted alkyl, aralkyl, substituted aralkyl, oxiranylalkyl, substituted oxiranylalkyl, oxiranylalkoxy, substituted oxiranylalkoxy, alkylamino, substituted alkylamino, peptidylamino, cycloalkylamino, substituted cycloalkylamino, oxiranylamino, substituted oxiranylamino, alkoxy, substituted alkoxy, and a residue of the acid moiety of a pyrethroid, R1, R2, and R4 further being selected to render said reporter a substrate for said esterase activity, under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 50 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 51 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter, and correlating said detection of said carbonyl compound with said esterase activity.
- 37. A method for the detection of cytochrome P450 activity in a sample, said method comprising
(a) contacting said sample with a reporter having the formula 52 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; R2 is a member selected from the group consisting of H, C1-C2 alkyl, benzyl, and substituted benzyl; and R3 is a member selected from the group consisting of alkyl, substituted alkyl, aralkyl, and substituted aralkyl, R1, R2, and R4 further being selected to render said reporter a substrate for said cytochrome P450 activity, under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 53 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 54 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter, and correlating said detection of said carbonyl compound with said esterase activity.
- 38. A method for the detection of epoxide hydrolase activity in a sample, said method comprising
(a) contacting said sample with a reporter having the formula 55 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; R2 is a member selected from the group consisting of H, C1-C2 alkyl, benzyl, and substituted benzyl; R5 is a member selected from the group consisting of CH2, O, and NH; and R6 is a member selected from the group consisting of H, alkyl, substituted alkyl, aralkyl, and substituted aralkyl, under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 56 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 57 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter, and correlating said detection of said carbonyl compound with said epoxide hydrolase activity.
- 39. A method for the detection of glutathione S-transferase activity in a sample, said method comprising
(a) contacting said sample with a reporter having the formula 58 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; and R2 is a member selected from the group consisting of H, C1-C2 alkyl, benzyl, and substituted benzyl; under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 59 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 60 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter, and correlating said detection of said carbonyl compound with said glutathione S-transferase activity.
- 40. A method for the detection of lipase activity in a sample, said method comprising
(a) contacting said sample with a reporter having the formula 61 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; R2 is a member selected from the group consisting of H, C1-C2 alkyl, benzyl, and substituted benzyl; and R7 is a fatty acid residue; under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 62 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 63 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter, and correlating said detection of said carbonyl compound with said lipase activity.
- 41. A method for the detection of protease activity in a sample, said method comprising
(a) contacting said sample with a reporter having the formula 64 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; R2is a member selected from the group consisting of H, C1-C2 alkyl, benzyl. and substituted benzyl; and R7 is a polypeptide; under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 65 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 66 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter, and correlating said detection of said carbonyl compound with said protease activity.
- 42. A method for the detection of β-glucuronidase activity in a sample, said method comprising
(a) contacting said sample with a reporter having the formula 67 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; and R2 is a member selected from the group consisting of H, C1-C2 alkyl, benzyl, and substituted benzyl; under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 68 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 69 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter, and correlating said detection of said carbonyl compound with said β-glucuronidase activity.
- 43. A method for the detection of alkaline phosphatase activity in a sample, said method comprising
(a) contacting said sample with a reporter having the formula 70 in which:
R1 is a residue of a member selected from the group consisting of aldehydes and ketones whose optical detectability by fluorescence emission or light absorptivity is measurably greater than the optical detectability of the cyanohydrin precursors of said aldehydes and ketones; and R2 is a member selected from the group consisting of H, C1-C2 alkyl, benzyl, and substituted benzyl; under conditions promoting the conversion of said reporter to a cyanohydrin of the formula 71 and the rearrangement of said cyanohydrin to a carbonyl compound of the formula 72 and (b) detecting said carbonyl compound by optical detection in a manner differentiating said carbonyl compound from said cyanohydrin and said reporter, and correlating said detection of said carbonyl compound with said alkaline phosphatase activity.
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0001] This invention was made at least in part with Government funding under Contract No. ES02710 awarded by the National Institute of Environmental Health Services, and Contract No. ESO4699 awarded by the National Institute of Environmental Health Services Superfund Program. The Government has certain rights to this invention.
Divisions (1)
|
Number |
Date |
Country |
Parent |
09971105 |
Oct 2001 |
US |
Child |
10338486 |
Jan 2003 |
US |