US4409140A
Expired / Lapsed
US
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Substrates and method for determining enzymes
Assignee: Individual
•
Filing: 1981-06-15
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Granted: 1983-10-11
•
Expires: 2001-06-15
Abstract
Abstract A method is disclosed for determining the presence of an enzyme in a biological fluid, which includes the steps of contacting the fluid with a synthetic chromogenic substrate, which is an amino acid derivative of 7-amino-4-trifluoromethylcoumarin; incubating the substrate-containing fluid to effect enzymatic hydrolysis; and fluorometrically determining the presence of the free 7-amino-4-trifluoromethylcoumarin chromophore in the hydrolyzate.
Inventors
- 👤 Robert E. Smith
- 👤 Eugene R. Bissell
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 22:31
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/37; C07; C07D; C07D311/00; C07D311/02; C07D311/04; C07D311/06; C07D311/08; C07D311/16; C07K; C07K5/00; C07K5/04; C07K5/06; C07K5/06191; C07K5/08; C07K5/0827; C07K5/10; C07K5/1027; C12Q2337/00; C12Q2337/20; C12Q2337/24
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
58 words
A proteolytic enzyme substrate of the formula ##STR6## wherein R is an amino acid, a peptide, an amino acid which has been reacted with a blocking group, or a peptide, the terminal amino acid of which has been reacted with a blocking group; and R is releaseable therefrom by enzymatic hydrolysis to form a measureable amount of 7-amino-4-trifluoromethylcoumarin.
Claim 2
Dependent Claim
58 words
A proteolytic enzyme substrate of the formula ##STR6## wherein R is an amino acid, a peptide, an amino acid which has been reacted with a blocking group, or a peptide, the terminal amino acid of which has been reacted with a blocking group; and R is releaseable therefrom by enzymatic hydrolysis to form a measureable amount of 7-amino-4-trifluoromethylcoumarin.
Claim 3
Dependent Claim
20 words
The substrate of claim 1, wherein R is an amino acid chain comprising from 1 to about 12 amino acids.
Claim 4
Dependent Claim
20 words
The substrate of claim 1, wherein R is an amino acid chain comprising from 1 to about 6 amino acids.
Claim 5
Dependent Claim
9 words
The substrate of claim 1 wherein R is gly-gly-arg-.
Claim 6
Dependent Claim
9 words
The substrate of claim 1, wherein R is d-ala-leu-lys-.
Claim 7
Dependent Claim
9 words
The substrate of claim 1, wherein R is leu-.
Claim 8
Dependent Claim
9 words
The substrate of claim 1, wherein R is Cbz-Val-Lys-Lys-Arg-.
Claim 9
Dependent Claim
24 words
The substrate of any one of claims 2, 3, 4, 5, or 6, wherein a blocking group is bonded to the terminal amino acid.
Claim 10
Dependent Claim
18 words
The substrate of claim 8, wherein said blocking group is carbobenzoxy, benzoyl, glutaryl, t-butylcarbonyl, d-proline, d-valine, or d-alanine.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-10-10 | 1987-04-11 | 1987-10-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-10-10 | 1991-04-11 | 1991-10-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-10-10 | 1995-04-11 | 1995-10-11 | $7700.00 | Window Closed / Lapsed |