US4276375A
Expired / Lapsed
US
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Determination of serine proteases
Assignee: Phadia AB
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Filing: 1979-10-22
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Granted: 1981-06-30
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Expires: 1999-10-22
Abstract
Abstract Serine proteases in a substance such as blood are determined by contacting the substance with a novel chromogenic or fluorgenic substrate for serine proteinases and spectrophotometrically measuring the quantity of a chromophore or fluorescent compound released from the substrate by serine proteases in the substance. The novel substrate has the amino acid sequence-ILe-A-Gly-Arg-wherein A is Asp or Glu substituted in the carboxylic group by esterification or amidation. The novel substrate increases sensitivity and accuracy of the determination.
Inventors
- 👤 Karl G. Claeson
- 👤 Leif E. Aurell
- 👤 Leif R. Simonsson
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; C07K5/10; C07K5/1002; C07K5/1005; C07K5/101; C12; C12Q; C12Q1/00; C12Q1/56; C12Q2337/00; C12Q2337/10; C12Q2337/12; Y; Y10; Y10S; Y10S530/00; Y10S530/802; Y10S930/00; Y10S930/01; Y10S930/28
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
158 words
A method for diagnostic determination of serine proteases which comprises contacting a substance in which said determination of serine poteases is desired with a chromogenic or fluorogenic enzyme substrate containing the sequence -Ile-A-Gly-Arg-R 2 where A is Asp or Glu, substituted in the carboxylic group by esterification or amidation, wherein the esters contain a short chain alkyl or hydroxyalkyl or alkyl-substituted aminoalkyl or cycloalkyl group containing a total of 2-6 carbon atoms, and the amides contain mono or di-substituted short chain alkyl or hydroxyalkyl or alkyl-substituted aminoalkyl group containing a total of 1-6 carbon atoms or a heterocyclic group in which the amido nitrogen forms part of a piperidine, a morpholine or a piperazine ring, and R 2 is a group which gives a chromophore or fluorescent compound upon enzymatic hydrolysis, and then spectrophotometrically measuring the quantity of chromophore or fluorescent compound released from said substrate by enzymatic hydrolysis of the substrate by serine proteases in said substance.
Claim 2
Independent Claim
158 words
A method for diagnostic determination of serine proteases which comprises contacting a substance in which said determination of serine poteases is desired with a chromogenic or fluorogenic enzyme substrate containing the sequence -Ile-A-Gly-Arg-R 2 where A is Asp or Glu, substituted in the carboxylic group by esterification or amidation, wherein the esters contain a short chain alkyl or hydroxyalkyl or alkyl-substituted aminoalkyl or cycloalkyl group containing a total of 2-6 carbon atoms, and the amides contain mono or di-substituted short chain alkyl or hydroxyalkyl or alkyl-substituted aminoalkyl group containing a total of 1-6 carbon atoms or a heterocyclic group in which the amido nitrogen forms part of a piperidine, a morpholine or a piperazine ring, and R 2 is a group which gives a chromophore or fluorescent compound upon enzymatic hydrolysis, and then spectrophotometrically measuring the quantity of chromophore or fluorescent compound released from said substrate by enzymatic hydrolysis of the substrate by serine proteases in said substance.
Claim 3
Dependent Claim
12 words
The method of claim 1 wherein said serine proteases is factor Xa.
Claim 4
Dependent Claim
20 words
The method of claim 1 wherein said substrate has the formula R 1 -Ile-A-Gly-Arg-NH-R 2 wherein R 1 is acyl.
Claim 5
Dependent Claim
10 words
The method of claim 3 wherein R 1 is acetyl.
Claim 6
Dependent Claim
10 words
The method of claim 3 wherein R 1 is benzoyl.
Claim 7
Dependent Claim
18 words
The method of claim 3 wherein R 2 is p-nitrophenyl or β-naphthyl or 4 methoxy-β-naphthyl or salt thereof.
Claim 8
Dependent Claim
16 words
The method of claim 1 wherein A is Asp or Glu substituted with a methoxy group.
Claim 9
Dependent Claim
16 words
The method of claim 1 wherein A is Asp or Glu substituted with an ethoxy group.
Claim 10
Dependent Claim
16 words
The method of claim 1 wherein A is Asp or Glu substituted with an isopropoxy group.
Claim 11
Dependent Claim
16 words
The method of claim 1 wherein A is Asp or Glu substituted with a cyclohexoxy group.
Claim 12
Dependent Claim
16 words
The method of claim 1 wherein A is Asp or Glu substituted with a dimethylaminoethoxy group.
Claim 13
Dependent Claim
18 words
The method of claim 1 wherein A is Asp or Glu substituted with CONHCH(CH 3 ) 2 .
Claim 14
Dependent Claim
14 words
The method of claim 1 wherein A is Asp or Glu substituted with ##STR7##
Claim 15
Dependent Claim
20 words
The method of claim 1 wherein A is Asp or Glu substituted with CON(CH 2 CH 2 OH) 2 .
Claim 16
Independent Claim
27 words
The method of any of claims 7-14 wherein said substrate has the formula R 1 -Ile-A-Gly-Arg-NH-R 2 wherein R 1 is benzoyl and R 2 is p-nitrophenyl.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-06-29 | 1984-12-29 | 1985-06-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-06-29 | 1988-12-29 | 1989-06-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-06-29 | 1992-12-29 | 1993-06-30 | $7700.00 | Window Closed / Lapsed |