Claims
- 1. A compound having the general formula:
- 2. The compound of claim 1, wherein the donor fluorescent moiety is selected from the group consisting of:
- 3. The compound of claim 2, wherein the linker is selected from the group consisting of a direct bond to a heteroatom in the fluorescent moiety, —O(CH2)n—, —S(CH2)n—, —NR2(CH2)n—, —N+R2(CH2)n, —OCONR2(CH2)—,—O2C(CH2)n—, —SCSNR2(CH2)n—, —SCSO(CH2)n—, —S(CH2)nCONR2(CH2)m, —S(CH2)nNR2CO(CH2)m, and
- 4. The compound of claim 1, wherein the compound has the structure:
- 5. A method for detecting the presence of β-lactamase activity in a sample, comprising:
contacting the sample with at least one compound of general formula I: 15in which R is a benzyl, 2-thienylmethyl, or cyanomethyl group, or a quencher; R′ is selected from the group consisting of H, physiologically acceptable salts or metal, ester groups, ammonium cations, —CHR2OCO(CH2)nCH3, —CHR2OCOC(CH3)3, acylthiomethyl, acyloxy-alpha-benzyl, deltabutyrolactonyl, methoxycarbonyloxymethyl, phenyl, methylsulphinylmethyl, β-morpholinoethyl, dialkylaminoethyl, and dialkylaminocarbonyloxymethyl, in which R2 is selected from the group consisting of H and lower alkyl; A is selected from the group consisting of S, O, SO, SO2 and CH2 ; and Z is a donor fluorescent moiety.
- 6. The method of claim 5, wherein said sample has a β-lactamase reporter gene.
- 7. The method of claim 6, wherein said β-lactamase reporter gene is in a mammalian cell.
- 8. The method of claim 5, wherein samples having β-lactamase activity are separated from samples having no β-lactamase activity by fluorescent-activated cell sorting.
- 9. The method of claim 5, wherein the β-lactamase activity results from a β-lactamase enzyme that was prepared by mutagenesis of another β-lactamase enzyme.
- 10. The method of claim 5, wherein said compound is a membrane permeant derivative.
- 11. The method of claim 5, wherein the donor fluorescent moiety is selected from the group consisting of:
- 12. The method of claim 11, wherein the linker is selected from the group consisting of a direct bond to a heteroatom in the fluorescent moiety, —O(CH2)n—, —S(CH2)n—, —NR2(CH2) n—, —N+R2(CH2)n, —OCONR2(CH2)n—, —O2C(CH2)n—, —SCSNR2(CH2)n,—SCSO(CH2)n—, —S(CH2)nCONR2(CH2)m, —S(CH2)nNR2CO(CH2)m, and
- 13. The method of claim 5, wherein the compound has the structure:
- 14. A method for determining whether a compound of claim 1 is a substrate for a β-lactamase enzyme, comprising: contacting said compound with a sample containing said β-lactamase enzyme; exciting at the wavelength for the said compound when cleaved; and measuring fluorescence.
- 15. The method of claim 14, wherein said compound is a membrane permeant derivative.
- 16. The method of claim 14, wherein said β-lactamase enzyme has been prepared by mutagenesis of another β-lactamase enzyme.
RELATED APPLICATION DATA
[0001] This application claims priority under 35 U.S.C. § 119 to U.S. Provisional Application Serial No. 60/261,313, filed Jan. 12, 2001 the contents of which is incorporated by reference in its entirety herein.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH
[0002] This invention was made in part with government support under Grant No. NS27177 awarded by the National Institutes of Health. The government may have certain rights in the invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60261313 |
Jan 2001 |
US |