The effect of pravastatin on hepatic 3-hydroxy-3-methylglutaryl CoA reductase obtained from poloxamer 407-induced hyperlipidemic rats.
A single 300-mg intraperitoneal injection of poloxamer 407 (P-407) to rats produces a marked hypercholesterolemia for a minimum of 96 hours and increases the activity of hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase compared with the enzyme activity found in microsomal homogenates of control animals. We attempted to determine whether inhibition of microsomal HMG-CoA reductase by pravastatin sodium would yield similar values for the maximum reaction in velocity (Vmax) and the HMG-CoA reductase-pravastatin dissociation constant (Ki) when the enzyme was in the activated state compared with the control state. Knowledge of the respective values for Vmax and Ki would allow us to determine whether P-407-induced hypercholesterolemia in the rat was refractory to pravastatin treatment. Over a pravastatin concentration range of 0.5-50 nM, enzyme activity in vitro decreased as the drug's concentration increased. A standard Dixon plot of mean values of reciprocal reaction velocity versus pravastatin concentration yielded Ki of 3.7 and 4.1 nM for the control and activated states, respectively. The Vmax for conversion of HMG-CoA to mevalonate in vitro in the presence of pravastatin was 3.5-fold greater when assayed in microsomal homogenates obtained from P-407-injected rats compared with control animals. Dixon plot analysis of the data resulted in Vmax of 58.1 and 202 pmol.min-1.mg-1 for the control and activated states, respectively. These data suggest that whereas the Vmax is affected, injection of P-407 to rats does not alter the binding affinity of pravastatin for receptor(s) contained in HMG-CoA reductase as reflected by similar Ki values. This experimental animal model may be an additional screen with which to rank order the relative potency of HMG-CoA reductase inhibitors by determining the drug's effectiveness when HMG-CoA reductase is in an activated state.
Johnston
T P
TP
Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City 64110-2499, USA. tjohnston/cctr.umkc.edu
Palmer
W K
WK
D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D006937:Hypercholesterolemia / Q000139:chemically induced / Q000503:physiopathology*; D066298:In Vitro Techniques; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology; D011060:Poloxalene / Q000633:toxicity; D017035:Pravastatin / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013501:Surface-Active Agents / Q000633:toxicity
D000924:Anticholesteremic Agents; D004791:Enzyme Inhibitors; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D013501:Surface-Active Agents; D011060:Poloxalene; D017035:Pravastatin