Sequence specific inhibition of human type II phospholipase A2 enzyme activity by phosphorothioate oligonucleotides.
Phosphorothioate oligonucleotides were identified which directly inhibited human type II phospholipase A2 (PLA2) enzyme activity in a sequence specific manner. The minimum pharmacophore common to all oligonucleotides which inhibited PLA2 enzyme activity consisted of two sets of three or more consecutive guanosine residues in a row. These oligonucleotides appear to form G quartets resulting in the formation of oligonucleotide aggregates. Additionally, a phosphorothioate backbone was required to be effective inhibitors of type II PLA2. The activity of one oligodeoxynucleotide, IP 3196 (5'-GGGTGGGTATAGAAGGGCTCC-3') has been characterized in more detail. IP 3196 inhibited PLA2 enzyme activity when the substrate was presented in the form of a phospholipid bilayer but not when presented in the form of a mixed micelle with anionic detergents. Human type II PLA2 was 50-fold more sensitive to inhibition by IP 3196 than venom and pancreatic type I enzymes. These data demonstrate that phosphorothioate oligonucleotides can specifically inhibit human type II PLA2 enzyme activity in a sequence specific manner.
Bennett
C F
CF
Department of Molecular Pharmacology, ISIS Pharmaceuticals, Carlsbad, CA 92008.
Chiang
M Y
MY
Wilson-Lingardo
L
L
Wyatt
J R
JR
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000494:pharmacology*; D010179:Pancreas / Q000201:enzymology; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D011189:Potassium Chloride / Q000494:pharmacology; D012910:Snake Venoms / Q000737:chemistry; D012965:Sodium Chloride / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013873:Thionucleotides
D009838:Oligodeoxyribonucleotides; D012910:Snake Venoms; D013873:Thionucleotides; C088940:IP 3196; D012965:Sodium Chloride; D011189:Potassium Chloride; D010741:Phospholipases A; D054467:Phospholipases A2