US5262545A
Expired / Lapsed
US
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Fluorescent chloramphenicol derivatives for determination of chloramphenicol acetyltransferase activity
Assignee: Medical Research Foundation of Oregon
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Filing: 1991-06-18
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Granted: 1993-11-16
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Expires: 2011-06-18
Abstract
Abstract Fluorescent compounds useful in the determination of chloramphenicol acetyltransferase (CAT) enzyme activity are described. The compounds are fluorescent derivatives related in structure to chloramphenicol comprising a base, <IMAGE> substituted at one to five aromatic ring positions by substituents, which may be the same or different, that are alkyl-, hydroxy-, alkoxy-, aryl-, halo-, nitro-, amino-, alkylamido-, or arylamido-; and a fluorescent moiety linked to the base at -NH- by an aliphatic chain of 1-12 atoms other than hydrogen, where such atoms are carbon, nitrogen, oxygen, or sulfur, or combinations thereof. The substrate compounds are acylated in the presence of CAT to produce fluorescent mono- and diacylated products, which are then physically separated from the reaction mixture and quantitated by means of their fluorescence and/or absorbance. Fluorescent molecules conjugated to chloramphenicol include derivatives of fluorescein, rhodamine, coumarin, dimethylaminonaphthalene sulfonic acid (dansyl), pyrene, anthracene, nitrobenzoxadiazole (NBD), acridine and dipyrrometheneboron difluoride.
Inventors
- 👤 Richard P. Haughland
- 👤 Hee C. Kang
- 👤 Steven L. Young
- 👤 Michael H. Melner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 13:19
CPC Classifications
C; C07; C07D; C07D493/00; C07D493/02; C07D493/08; C07C; C07C233/00; C07C233/01; C07C233/34; C07C233/42; C07C233/43; C07C311/00; C07C311/30; C07C311/37; C07C311/38; C07C311/39; C07C311/42; C07D219/00; C07D219/04; C07D219/06; C07D271/00; C07D271/12; C07D311/00; C07D311/02; C07D311/04; C07D311/06; C07D311/08; C07D311/16; C07D311/78; C07D311/80; C07D311/82; C07F; C07F5/00; C07F5/02; C07F5/022; C12; C12Q; C12Q1/00; C12Q1/48; C07C2603/00; C07C2603/02; C07C2603/04; C07C2603/22; C07C2603/24; C07C2603/40; C07C2603/42; C07C2603/50; Y; Y10; Y10S; Y10S435/00; Y10S435/968
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
134 words
A compound of the formula: BASE--N.sub.s --*X a) where BASE is: ##STR4## substituted at one to five aromatic ring positions by substituents, which may be the same or different, that are alkyl-, hydroxy-, alkoxy-, aryl-, halo-, nitro-, amino-, alkylamido- (where the alkyl portion contains 1-3 carbons, or arylamido- (where the aryl portion is a phenyl or alkyl-substituted phenyl) and n is greater than 0 and less than 6; and b) *X is a non-reduced tricyclic difluoroboradiazaindacene fluorophore covalently bound to the terminal CH 2 of BASE through a linker N s ; such that c) N s --*X is amino-linked (--NH--*X), acetamido-linked (--NHCOCH 2 --*X), propionamido-linked (--NHCOCH 2 CH 2 --*X), sulfonamido-linked (--NHSO 2 --*X), carboxamido-linked (--NHCO--*X), ureidyl-linked (--NHCONH--*X), thioureidyl-linked (--NHCSNH--*X), or linked by a covalent bond, to said terminal CH 2 of BASE.
Claim 2
Dependent Claim
134 words
A compound of the formula: BASE--N.sub.s --*X a) where BASE is: ##STR4## substituted at one to five aromatic ring positions by substituents, which may be the same or different, that are alkyl-, hydroxy-, alkoxy-, aryl-, halo-, nitro-, amino-, alkylamido- (where the alkyl portion contains 1-3 carbons, or arylamido- (where the aryl portion is a phenyl or alkyl-substituted phenyl) and n is greater than 0 and less than 6; and b) *X is a non-reduced tricyclic difluoroboradiazaindacene fluorophore covalently bound to the terminal CH 2 of BASE through a linker N s ; such that c) N s --*X is amino-linked (--NH--*X), acetamido-linked (--NHCOCH 2 --*X), propionamido-linked (--NHCOCH 2 CH 2 --*X), sulfonamido-linked (--NHSO 2 --*X), carboxamido-linked (--NHCO--*X), ureidyl-linked (--NHCONH--*X), thioureidyl-linked (--NHCSNH--*X), or linked by a covalent bond, to said terminal CH 2 of BASE.
Claim 3
Dependent Claim
117 words
A compound of the formula: BASE--N.sub.s --*X a) where BASE is: ##STR5## where R is alkyl-, hydroxy-, alkoxy-, aryl-, halo-, nitro-, amino-, alkylamido- (where the alkyl portion contains 1-3 carbons), or arylamido- (where the aryl portion is a phenyl or alkyl-substituted phenyl), and n is greater than 0 and less than 6; b) *X is a non-reduced tricyclic difluoroboradiazaindacene fluorophore covalently bound to the terminal CH 2 of BASE through a linker N s ; such that c) N s --*X is amino-linked (--NH--*X), acetamido-linked (--NHCOCH 2 --*X), propionamido-linked (--NHCOCH 2 CH 2 --*X), sulfonamido-linked (--NHSO 2 --*X), carboxamido-linked (--NHCO--*X), ureidyl-linked (--NHCONH--*X), thioureidyl-linked (--NHCSNH--*X), or linked by a covalent bond, to said terminal CH 2 of BASE.
Claim 4
Dependent Claim
117 words
A compound of the formula: BASE--N.sub.s --*X a) where BASE is: ##STR5## where R is alkyl-, hydroxy-, alkoxy-, aryl-, halo-, nitro-, amino-, alkylamido- (where the alkyl portion contains 1-3 carbons), or arylamido- (where the aryl portion is a phenyl or alkyl-substituted phenyl), and n is greater than 0 and less than 6; b) *X is a non-reduced tricyclic difluoroboradiazaindacene fluorophore covalently bound to the terminal CH 2 of BASE through a linker N s ; such that c) N s --*X is amino-linked (--NH--*X), acetamido-linked (--NHCOCH 2 --*X), propionamido-linked (--NHCOCH 2 CH 2 --*X), sulfonamido-linked (--NHSO 2 --*X), carboxamido-linked (--NHCO--*X), ureidyl-linked (--NHCONH--*X), thioureidyl-linked (--NHCSNH--*X), or linked by a covalent bond, to said terminal CH 2 of BASE.
Claim 5
Dependent Claim
52 words
A compound, as claimed in claim 1, wherein the fluorophore *X has the formula: ##STR6## where BASE-N s -- is attached to said fluorophore at one of S 1 -S 7 , and the remainder of S 1 -S 7 , which may be the same or different, are hydrogen or alkyl-.
Claim 6
Dependent Claim
21 words
A compound, as claimed in claim 3, where BASE is substituted at one aromatic position by a substituent that is alkylamido-.
Claim 7
Dependent Claim
45 words
A compound, as claimed in claim 4, where n=5 and N s --*X is acetamido-linked (--NHCOCH 2 --*X) or propionamido-linked (--NHCOCH 2 CH 2 --*X) to said terminal CH 2 of BASE; or n is 1 or 2 and N s is a covalent bond.
Claim 8
Dependent Claim
52 words
A compound, as claimed in claim 2, wherein the fluorophore *X has the formula: ##STR7## where BASE--N s -- is attached to said fluorophore at one of S 1 -S 7 , and the remainder of S 1 -S 7 , which may be the same or different, are hydrogen or alkyl-.
Claim 9
Dependent Claim
11 words
A compound, as claimed in claim 6, wherein R is acetamido-.
Claim 10
Dependent Claim
38 words
A compound, as claimed in claim 7, wherein n is 5 and N s is acetamido (--CH 2 CONH--), or propionamido (--CH 2 CH 2 CONH--), or n is less than 3 N x is a covalent bond.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1996-11-15 | 1997-05-17 | 1997-11-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-11-15 | 2001-05-17 | 2001-11-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-11-15 | 2005-05-17 | 2005-11-16 | $7700.00 | Window Closed / Lapsed |