US4275153A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Analytical fluorogenic substrates for proteolytic enzymes
Assignee: Siemens Healthcare Diagnostics GmbH Switzerland
•
Filing: 1978-08-03
•
Granted: 1981-06-23
•
Expires: 1998-08-03
Abstract
Abstract Fluorogenic substrates for proteolytic enzymes having the formula: <IMAGE> or acid salts thereof wherein: R1 is hydrogen-L, hydrogen-D, benzoyl, benzenesulfonyl, glutaryl, pyroglutamyl, carbobenzoxy, D-serine, or carbobenzoxy-serine; R2 is hydrogen, phenyl, a straight, branched or cyclic alkyl having 1 to 4 carbons, or propionic acid; R3 is hydrogen, straight or branched or cyclic alkyl having 1 to 4 carbons, 4-aminobutane, or 3-guanidylpropane; R4 is methyl, 4-aminobutane, or 3-guanidylpropane; R5 is a fluorogenic moiety, severable from said compound by a proteolytic enzyme and having different fluorescent property when severed from said compound than when forming part of said compound.
Inventors
- 👤 Robert J. Gargiulo
- 👤 Gary A. Mitchell
- 👤 Patricia M. Hudson
- 👤 Sharon P. Pochron
- 👤 Rolf M. Huseby
- 👤 Robert E. Smith
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 00:32
CPC Classifications
C; C07; C07K; C07K5/00; C07K5/04; C07K5/08; C07K5/0821; C07K5/0823; C07K5/0802; C07K5/0804; C07K5/0812; C07K5/0819; C07K5/10; C07K5/1002; C07K5/1005; C07K5/1013; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/37; C12Q1/56; C12Q2337/00; C12Q2337/10; C12Q2337/30; G; G01; G01N; G01N2333/00; G01N2333/90; G01N2333/914; G01N2333/948; G01N2333/968; G01N2333/974; G01N2400/00; G01N2400/10; G01N2400/38; G01N2400/40; Y; Y10; Y10S; Y10S530/00; Y10S530/802; Y10S930/00; Y10S930/01; Y10S930/28
Claims Analysis
Total Claims: 8
Claim 1
Independent Claim
103 words
A method for determining plasmin or plasmin-like enzymes in a fluid comprising: A. mixing said fluid with a fluorogenic substrate of the formula ##STR3## or acid salts thereof, wherein R is hydrogen-L, hydrogen-D, benzoyl, benzene sulfonyl, glutaryl, pyroglutamyl, carbobenzoxy, D-serine or carbobenzoxy-serine, thereby forming a substrate-containing fluid, whereby said substrate is enzymatically hydrolyzable by said plasmin, or plasmin-like enzymes to yield 5-aminoisophthalic acid dimethyl ester; B. exposing said substrate-containing fluid to light of a wavelength suitable for causing fluorescent emission by said 5-aminoisophthalic acid dimethyl ester; and C. while exposing said substrate-containing fluid to said light, measuring the fluorescent emission of said fluid.
Claim 2
Independent Claim
103 words
A method for determining plasmin or plasmin-like enzymes in a fluid comprising: A. mixing said fluid with a fluorogenic substrate of the formula ##STR3## or acid salts thereof, wherein R is hydrogen-L, hydrogen-D, benzoyl, benzene sulfonyl, glutaryl, pyroglutamyl, carbobenzoxy, D-serine or carbobenzoxy-serine, thereby forming a substrate-containing fluid, whereby said substrate is enzymatically hydrolyzable by said plasmin, or plasmin-like enzymes to yield 5-aminoisophthalic acid dimethyl ester; B. exposing said substrate-containing fluid to light of a wavelength suitable for causing fluorescent emission by said 5-aminoisophthalic acid dimethyl ester; and C. while exposing said substrate-containing fluid to said light, measuring the fluorescent emission of said fluid.
Claim 3
Dependent Claim
9 words
The method of claim 1 wherein R is hydrogen-D.
Claim 4
Dependent Claim
35 words
The method of claim 1 or 2 wherein said substrate-containing fluid is exposed to light of a wavelength of about 335 nanometers, and said fluorescent emission is measured at a wavelength of about 430 nanometers.
Claim 5
Independent Claim
109 words
A method for determining thrombin, trypsin or thrombin or trypsin-like enzymes in a fluid comprising: A. mixing said fluid with a fluorogenic substrate of the formula ##STR4## or acid salts thereof, wherein R is hydrogen-L, hydrogen-D benzoyl, benzene sulfonyl, glutaryl, pyroglutamyl, carbobenzoxy, D-serine or carbobenzoxy-serine, thereby forming a substrate-containing fluid, whereby said substrate is enzymatically hydrolyzable by said thrombin, trypsin, or thrombin or trypsin-like enzymes to yield 5-aminoisophthalic acid dimethyl ester; B. exposing said substrate-containing fluid to light of a wavelength suitable for causing fluorescent emission by said 5-aminoisophthalic acid dimethyl ester; and C. while exposing said substrate-containing fluid to said light, measuring the fluorescent emission of said fluid.
Claim 6
Independent Claim
109 words
A method for determining thrombin, trypsin or thrombin or trypsin-like enzymes in a fluid comprising: A. mixing said fluid with a fluorogenic substrate of the formula ##STR4## or acid salts thereof, wherein R is hydrogen-L, hydrogen-D benzoyl, benzene sulfonyl, glutaryl, pyroglutamyl, carbobenzoxy, D-serine or carbobenzoxy-serine, thereby forming a substrate-containing fluid, whereby said substrate is enzymatically hydrolyzable by said thrombin, trypsin, or thrombin or trypsin-like enzymes to yield 5-aminoisophthalic acid dimethyl ester; B. exposing said substrate-containing fluid to light of a wavelength suitable for causing fluorescent emission by said 5-aminoisophthalic acid dimethyl ester; and C. while exposing said substrate-containing fluid to said light, measuring the fluorescent emission of said fluid.
Claim 7
Dependent Claim
9 words
The method of claim 4 wherein R is hydrogen-D.
Claim 8
Dependent Claim
35 words
The method of claim 4 or 5 wherein said substrate-containing fluid is exposed to light of a wavelength of about 335 nanometers and said fluorescent emission is measured at a wavelength of about 430 nanometers.
References Cited (Backward Citations)
US3884896A
Cited
US3886136A
Cited
US4016042A
Cited
US4028318A
Cited
US4070245A
Cited
US4137225A
Cited
US4169015A
Cited
US4167449A
Cited
US4155916A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-06-22 | 1984-12-22 | 1985-06-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-06-22 | 1988-12-22 | 1989-06-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-06-22 | 1992-12-22 | 1993-06-23 | $7700.00 | Window Closed / Lapsed |