US4162941A
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Method for determining a proteolytic enzyme
Assignee: Chromogenix AB
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Filing: 1977-03-04
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Granted: 1979-07-31
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Expires: 1997-03-04
Abstract
Abstract The proteolytic enzyme designated as factor Xa is diagnostically determined by contacting the enzyme with a chromogenic substrate represented by the formula R1-A1-A2-Gly-Arg-NH-R2 or its salts, where R1 is hydrogen or alkanoyl having from 1 to 12 carbon atoms or cyclohexylcarbonyl or benzoyl or benzoyl substituted with one or two halogen atoms, methylamine or phenyl groups or benzene sulphonyl or toluenesulphonyl, R2 is nitrophenyl or naphthyl or nitronaphthyl or methoxynaphthyl or quinolyl or nitroquinolyl, A1 is a single bond or one of the amino acids Gly, Ala, Val, Leu, Ileu, Pro, Met, Phe or Tyr, and A2 is one of the amino acids Glu, Gln, Asp, or Asn and spectrophotometrically measuring the degree of enzymatic hydrolysis.
Inventors
- 👤 Leif E. Aurell
- 👤 Karl G. Claeson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 23:31
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/56; C12Q2337/00; C12Q2337/10; C12Q2337/12; C12Q2337/30; G; G01; G01N; G01N2333/00; G01N2333/90; G01N2333/914; G01N2333/948; G01N2333/95; G01N2333/964; G01N2333/96425; G01N2333/96427; G01N2333/9643; G01N2333/96433; G01N2333/96441; G01N2333/96444; G01N2333/968; G01N2333/974; G01N2333/976; Y; Y10; Y10S; Y10S530/00; Y10S530/802; Y10S930/00; Y10S930/01; Y10S930/28
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
145 words
A method for diagnostic determination of a proteolytic enzyme designated as Xa factor which comprises contacting said proteolytic enzyme with a chromogenic substrate represented by the following formula: R.sub.1 -A.sub.1 -A.sub.2 -Gly-Arg-NH-R.sub.2 or its salts, where R 1 is selected from the group consisting of hydrogen, alkanoly having from 1 to 12 carbon atoms, cyclohexylcarbonyl, benzoyl, benzoyl substituted with one or two halogen atoms, methylamine or phenyl groups, benzene sulphonyl and toluenesulphonyl; R 2 is a chromophoric group; Al is selected from the group consisting of a single bond, and the amino acids selected from the group consisting of Gly, Ala, Val, Leu, Ileu, Pro, Met, Phe and Tyr; and A 2 is selected from the group consisting of the amino acids Glu, Gln, Asp, and Asn, and then spectrophotometrically measuring the degree of enzymatic hydrolysis caused by said proteolytic enzyme acting on said substrate.
Claim 2
Independent Claim
145 words
A method for diagnostic determination of a proteolytic enzyme designated as Xa factor which comprises contacting said proteolytic enzyme with a chromogenic substrate represented by the following formula: R.sub.1 -A.sub.1 -A.sub.2 -Gly-Arg-NH-R.sub.2 or its salts, where R 1 is selected from the group consisting of hydrogen, alkanoly having from 1 to 12 carbon atoms, cyclohexylcarbonyl, benzoyl, benzoyl substituted with one or two halogen atoms, methylamine or phenyl groups, benzene sulphonyl and toluenesulphonyl; R 2 is a chromophoric group; Al is selected from the group consisting of a single bond, and the amino acids selected from the group consisting of Gly, Ala, Val, Leu, Ileu, Pro, Met, Phe and Tyr; and A 2 is selected from the group consisting of the amino acids Glu, Gln, Asp, and Asn, and then spectrophotometrically measuring the degree of enzymatic hydrolysis caused by said proteolytic enzyme acting on said substrate.
Claim 3
Dependent Claim
14 words
The method of claim 1 wherein the spectrophotometrically measuring is conducted at 405 nm.
Claim 4
Dependent Claim
10 words
The method of claim 1 wherein the substrate is benzoyl-Ileu-Glu-Gly-Arg-p-nitroanilide.
Claim 5
Dependent Claim
10 words
The method of claim 1 wherein the substrate is H-Phe-Glu-Gly-Arg-p-nitroanilide.
Claim 6
Dependent Claim
10 words
The method of claim 1 wherein the substrate is benzoyl-Leu-Glu-Gly-Arg-p-nitroanilide.
Claim 7
Dependent Claim
10 words
The method of claim 1 wherein the substrate is benzoyl-Val-Glu-Gly-Arg-p-nitroanilide.
Claim 8
Dependent Claim
10 words
The method of claim 1 wherein the substrate is benzoyl-Leu-Asp-Gly-Arg-p-nitroanilide.
Claim 9
Dependent Claim
10 words
The method of claim 1 wherein the substrate is benzoyl-Ileu-Glu-Gly-Arg-2-naphthylamide.
Claim 10
Dependent Claim
10 words
The method of claim 1 wherein the substrate is benzoyl-Ileu-Glu-Gly-Arg-4-methoxy-2-naphthylamide.
Claim 11
Dependent Claim
10 words
The method of claim 1 wherein the substrate is benzoyl-Ileu-Gln-Gly-Arg-p-nitroanilide.
Claim 12
Dependent Claim
22 words
The method of claim 1 wherein R 2 is selected from the group consisting of nitrophenyl, naphthyl, nitronaphthyl, methoxynaphthyl, quinolyl and nitroquinolyl.
Claim 13
Dependent Claim
12 words
The method of claim 1 wherein R 2 is nitrophenyl or naphthyl.
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) | 1982-07-30 | 1983-01-29 | 1983-07-31 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1986-07-30 | 1987-01-29 | 1987-07-31 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1990-07-30 | 1991-01-29 | 1991-07-31 | $7700.00 | Window Closed / Lapsed |