Methods for detecting LP-PLA2 activity and inhibition of LP-PLA2 activity
AssigneeGlaxo Group Limited
InventorsYaping Shou, Yin Fai Amy Siu, George T. Walker
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Abstract
This invention relates to methods for determining the activity of Lp-PLA2 in at least one sample from an animal. The invention also relates to methods for determining the inhibition of Lp-PLA2 activity in samples from animals that are administered an Lp-PLA2 inhibitor.
Claims23
A colorimetric method of determining the percentage of inhibition of Lp-PLA2 activity, the method comprising:
- preparing a solution comprising a substrate for Lp-PLA2 having a colorimetric detectable moiety, which substrate is 1-myristoyl-2-(4-nitrophenylsuccinyl)phosphatidylcholine;
- contacting a first blood sample that is taken from a human subject at a first time with the solution;
- colorimetrically detecting Lp-PLA2 activity from the first blood sample wherein the first blood sample is diluted less than 33 times;
- contacting a second blood sample that is taken from the subject at a second time one hour or more after the first time with the solution, wherein the second blood sample is taken after the subject has been administered a drug that inhibits Lp-PLA2;
- colorimetrically detecting Lp-PLA2 activity from the second blood sample wherein the second blood sample is diluted less than 33 times;
- and
- detecting greater than 30% inhibition of Lp-PLA2 activity between the first and second blood samples by comparing the activity of the Lp-PLA2 between the first and second blood samples, indicating that the subject has taken a drug that inhibits Lp-PLA2.
A method of clinically monitoring a patient to assess the efficacy of an inhibitor of lipoprotein-associated phospholipase A2 (Lp-PLA2), the method comprising:
- receiving a sample from a human patient that has been administered an Lp-PLA2 inhibitor;
- and
- colorimetrically detecting greater than 30 percent inhibition of Lp-PLA2 from the sample by contacting the sample with a solution comprising a substrate for Lp-PLA2 including a colorimetric detectable moiety, which substrate is 1-myristoyl-2-(4-nitrophenylsuccinyl)phosphatidylcholine and wherein the sample is diluted less than 33 fold.
A method of determining inhibition of lipoprotein-associated phospholipase A2 (Lp-PLA2) enzyme activity from a sample, the method comprising:
- preparing a solution comprising a substrate for Lp-PLA2 having a colorimetric detectable moiety, which substrate is 1-myristoyl-2-(4-nitrophenylsuccinyl)phosphatidylcholine;
- contacting a sample from a human that has been administered an Lp-PLA2 inhibitor, wherein the sample is diluted less than 33 fold by contact with the solution;
- and
- colorimetrically detecting Lp-PLA2 activity.
Dependencies are inferred from the claim text.