HPLC assay with UV spectrometric detection for the evaluation of inhibitors of cytosolic phospholipase A2

1‐(3‐Biaryloxy‐2‐oxopropyl)indole‐5‐carboxylic Acids and Related Compounds as Dual Inhibitors of Human Cytosolic Phospholipase A2α and Fatty Acid Amide Hydrolase

“…The ability of the test compounds to inhibit cPLA 2 a isolated from human platelets was performed as described earlier. [32] Briefly, 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (200 mm) sonicated with 1,2-dioleoyl-sn-glycerol (100 mm) in a bath sonicator at 30-35 8C was used as substrate. Enzyme reaction was terminated after 60 min by addition of a mixture of acetonitrile, methanol, and 0.1 m aqueous Na 2 EDTA solution, which contained 4-undecyloxybenzoic acid as internal standard and nordihydroguaiaretic acid (NDGA) as oxygen scavenger.…”

“…[29,30] cPLA 2 a inhibition was evaluated by applying cPLA 2 a isolated from human platelets. [32] Enzyme activity was measured in a solution containing co-vesicles of 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (SAPC) and 1,2-dioleoyl-sn-glycerol (DOG). Inhibitory potency of the test compounds was assessed by comparing the amount of arachidonic acid released from SAPC in their absence and presence after an incubation time of 60 min with reversed-phase HPLC and UV detection.…”

Design and synthesis of 3-pyrrol-3-yl-3H-isobenzofuran-1-ones as inhibitors of human cytosolic phospholipase A2α

“…The ability of test compounds to inhibit cPLA 2 a isolated from human platelets was performed as previously described [10]. Briefly, sonicated covesicles consisting of 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (0.2 mM) and 1,2-dioleoyl-sn-glycerol (0.1 mM) were used as substrate.…”

“…All newly synthesized isobenzofuran-1-one derivatives were evaluated in an assay applying cPLA 2 a isolated from human platelets [10]. Sonicated covesicles consisting of 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycerol were used as enzyme substrate.…”

Determination of arachidonic acid by on-line solid-phase extraction HPLC with UV detection for screening of cytosolic phospholipase A2α inhibitors

“…We have published a screening method for cPLA 2 ␣ inhibitors detecting the enzyme product arachidonic acid released from the natural phospholipid 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (SAPC) by HPLC and UV detection at 200 nm [30]. To increase enzyme activity, 1,2-dioleoyl-sn-glycerol was added to the substrate in this assay.…”