Method of detecting cysteine proteases
AssigneePrototek, Inc.
InventorsKyujin Cho, David W. Rasnick
Life of this patent
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Abstract
Peptide thioneamides are provided as synthetic substrates for cysteine proteases. The compounds of the invention are peptides consisting of one or more blocked or unblocked amino acids wherein the terminal carboxy of the peptides are thionated and forms a thioneamide linkage with a fluorogenic or chromogenic leaving group. The alpha carbonyls of the remaining amino acids are thionated or unthionated. The invention includes methods and kits for using the peptide thioneamides.
Claims9
A method of detecting cysteine proteases in a sample fluid, the method comprising the steps of:
- forming a mixture comprising a buffering agent, the sample fluid, and an amount effective as a substrate for cysteine proteases of a compound, or salt thereof, defined by the formula:
- (AA).sub.n --AA.psi.[CS]-W
- wherein:
- (AA).sub.n is a blocked or unblocked, thionated or unthionated amino acid whenever n=1, and (AA).sub.n is a peptide consisting of blocked or unblocked, thionated or unthionated amino acids whenever n is greater than one,
- n is in the range between 0 and 12 inclusive,
- AA.psi.[CS] is a blocked or unblocked amino acid whose alpha carbonyl oxygen has been replaced with a sulfur atom, and whose alpha amine forms a peptide bond with the alpha carbonyl of the adjacent amino acid of the (AA).sub.n moiety whenever n is greater than or equal to one, and
- W is a chromogenic or fluorogenic leaving group covalently attached to the carbon atom of the alpha carbonyl of AA.psi.[CS];
- and
- relating the change in color or fluorescence of the mixture to the activity of the cysteine protease as said compound is hydrolysed thereby.
A kit for detecting the presence of a cysteine protease in a sample fluid, the kit comprising:
- a buffering agent;
- and
- an amount effective as a substrate for cysteine proteases of a compound, or salt thereof, defined by the formula:
- (AA).sub.n --AA.psi.[CS]-W
- wherein:
- (AA).sub.n is a blocked or unblocked, thionated or unthionated amino acid whenever n=1, and (AA).sub.n is a peptide consisting of blocked or unblocked, thionated or unthionated amino acids whenever n is greater than one;
- n is in the range between 0 and 12 inclusive;
- AA.psi.[CS] is a blocked or unblocked amino acid whose alpha carbonyl oxygen has been replaced by a sulfur atom, and whose alpha amine forms a peptide bond with the alpha carbonyl of the adjacent amino acid of the (AA).sub.n moiety whenever n is greater than or equal to one;
- and
- W is a chromogenic or fluorogenic leaving group covalently attached to the carbon atom of the alpha carbonyl of AA.psi.[CS].
Dependencies are inferred from the claim text.