Trypsin substrate and diagnostic device, and method of using same
View Patent ↗A compound of the formula (I) wherein R 1 is a protecting group for Nα, R 2 is a protecting group for N G , and R 3 is aryl, and wherein the compound of formula (I) is a trypsin substrate such that trypsin cleaves the O—C single bond, which liberates R 3 —OH; a diagnostic device comprising same; a method for preparing the diagnostic device; and a method of using the diagnostic device to detect levels of urinary trypsin inhibitor in a biological sample; and a diagnostic kit for detecting levels of urinary trypsin inhibitor in a biological sample.
1. A diagnostic device comprising:
a carrier matrix; and
a compound of the formula (I)
wherein R 1 is a protecting group for Nα;
R 2 is a protecting group for N G ; and
R 3 is aryl;
wherein the compound of formula (I) is a trypsin substrate such that trypsin cleaves the O—C single bond, which liberates R 3 —OH.
2. The diagnostic device of claim 1 wherein R 1 is selected from the group consisting of acyl, arene sulfonyl, and carbamoyl derivatives.
3. The diagnostic device of claim 1 wherein R 1 is selected from the group consisting of t-butyloxycarbonyl and derivatives, benzyloxycarbonyl and derivatives, benzoyl and derivatives, and benzene sulfonyl and derivatives.
4. The diagnostic device of claim 1 wherein R 2 is selected from the group consisting of nitro, arene sulfonyl, carbamoyl, and acyl.
5. The diagnostic device of claim 1 wherein R 2 is selected from the group consisting of nitro, benzene sulfonyl and derivatives, tosyl, carbobenzyloxy and derivatives, and benzoyl and derivatives.
6. The diagnostic device of claim 1 wherein R 3 comprises a heterocyclic aromatic moiety.
7. The diagnostic device of claim 6 wherein R 3 is a fused ring system.
8. The diagnostic device of claim 1 wherein R 3 is carbocyclic.
9. The diagnostic device of claim 8 wherein R 3 is 1-naphthol and derivatives thereof.
10. The diagnostic device of claim 1 wherein R 3 is selected from the group consisting of phenylpyrrole and derivatives thereof, coumarin and derivatives thereof, phenylthiophene and derivatives thereof, indole and derivatives thereof, and 2-phenyl-5H-thiazol and derivatives thereof.
11. The diagnostic device of claim 1 wherein the carrier matrix is filter paper.
12. The diagnostic device of claim 1 wherein the carrier matrix contains a diazonium salt.
13. The diagnostic device of claim 12 wherein R 3 —OH reacts with a diazonium salt to form a visible color.
14. The diagnostic device of claim 12 wherein the diazonium salt has the structure:
R 4 —N 2 + An −
wherein R 4 is aryl; and
wherein An − is an anion.
15. The diagnostic device of claim 14 wherein R 4 is morpholinobenzene and derivatives thereof.
16. The diagnostic device of claim 12 wherein the diazonium salt is a zwitter ion having the structure
wherein D − is an anion;
wherein G is independently H, C 1-6 alkyl, or in which the two G moieties together form a fused ring system; and
wherein B is H or OH.
17. The diagnostic device of claim 1 wherein R 3 —OH is optically distinct from the compound of formula (I).