Claims
- 1. 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.sub.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.sub.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.
- 2. The method of claim 1 wherein said serine proteases is factor Xa.
- 3. The method of claim 1 wherein said substrate has the formula R.sub.1 -Ile-A-Gly-Arg-NH-R.sub.2 wherein R.sub.1 is acyl.
- 4. The method of claim 3 wherein R.sub.1 is acetyl.
- 5. The method of claim 3 wherein R.sub.1 is benzoyl.
- 6. The method of claim 3 wherein R.sub.2 is p-nitrophenyl or .beta.-naphthyl or 4 methoxy-.beta.-naphthyl or salt thereof.
- 7. The method of claim 1 wherein A is Asp or Glu substituted with a methoxy group.
- 8. The method of claim 1 wherein A is Asp or Glu substituted with an ethoxy group.
- 9. The method of claim 1 wherein A is Asp or Glu substituted with an isopropoxy group.
- 10. The method of claim 1 wherein A is Asp or Glu substituted with a cyclohexoxy group.
- 11. The method of claim 1 wherein A is Asp or Glu substituted with a dimethylaminoethoxy group.
- 12. The method of claim 1 wherein A is Asp or Glu substituted with CONHCH(CH.sub.3).sub.2.
- 13. The method of claim 1 wherein A is Asp or Glu substituted with ##STR7##
- 14. The method of claim 1 wherein A is Asp or Glu substituted with CON(CH.sub.2 CH.sub.2 OH).sub.2.
- 15. The method of any of claims 7-14 wherein said substrate has the formula R.sub.1 -Ile-A-Gly-Arg-NH-R.sub.2 wherein R.sub.1 is benzoyl and R.sub.2 is p-nitrophenyl.
Priority Claims (1)
Number |
Date |
Country |
Kind |
7613463 |
Dec 1976 |
SEX |
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Parent Case Info
This is a division of application Ser. No. 852,006, filed Nov. 16, 1977, now U.S. Pat. No. 4,207,232.
US Referenced Citations (5)
Divisions (1)
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Number |
Date |
Country |
Parent |
852006 |
Nov 1977 |
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