By 1023/SK&F 96022: biochemistry of a novel (H+ + K+)-ATPase inhibitor.
The mechanism by which the substituted benzimidazole sulphoxide BY 1023/SK&F 96022 inhibited the (H+ + K+)-ATPase, the enzyme responsible for hydrogen ion secretion in the stomach, was studied in a variety of in vitro preparations. In gastric preparations that were capable of active hydrogen ion transport with consequent lumenal acidification, BY 1023/SK&F 96022 inhibited with high potency and in a time-dependent manner consistent with the acid-induced conversion of the parent benzimidazole sulphoxide to a covalent inhibitor (cyclic sulphenamide). The following IC50 values were obtained for the inhibition of aminopyrine accumulation: intact gastric glands stimulated with 1 mM dibutyryl cAMP, 1.0 microM; permeabilized gastric glands stimulated with 5 mM ATP, 0.42 microM; intact gastric vesicles stimulated with 150 mM KCl, 9 microM valinomycin and 2 mM MgATP, 3.5 microM. In a preparation that could not generate pH gradients, lyophilized gastric vesicles at pH 7.4, BY 1023/SK&F 96022 inhibited K(+)-stimulated ATPase activity with relatively low potency, 70 microM, indicating its good chemical stability at neutral pH. As assessed by ATPase inhibition, this stability was three times greater than that of omeprazole. Inhibition by BY 1023/SK&F 96022 was not reversed by dilution in either permeabilized gastric glands or intact gastric vesicles. Inhibition could, however, be completely reversed by subsequent incubation with 20 mM beta-mercaptoethanol (intact gastric glands) or 100 mM dithiothreitol (intact gastric vesicles) suggesting a disulphide link between inhibitor and enzyme. The concentration of glutathione needed to protect against inhibition by BY 1023/SK&F 96022 was 10,000 times higher in intact, compared with lyophilized, gastric vesicles indicating an interaction with the lumenal (extra-cellular) face of the (H+ + K+)-ATPase. BY 1023/SK&F 96022 and omeprazole were also found to inhibit acidification in purified kidney lysosomes with IC50 values of 194 and 75 microM, respectively. Protection by 10 microM glutathione suggested that this did not result from intralysosomal activation of these inhibitors. Thus, BY 1023/SK&F 96022 has the combined properties of good chemical stability at neutral pH and effective conversion to the cyclic sulphenamide at acidic pH. In this way the activation to the cyclic sulphenamide may be optimally restricted to the parietal cell canaliculus.
Simon
W A
WA
Smith Kline & French Research Ltd, Welwyn, Herts, U.K.
Keeling
D J
DJ
Laing
S M
SM
Fallowfield
C
C
Taylor
A G
AG
D053799:2-Pyridinylmethylsulfinylbenzimidazoles; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000632:Aminopyrine / Q000378:metabolism; D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology*; D005753:Gastric Mucosa / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D005978:Glutathione / Q000494:pharmacology; D017506:H(+)-K(+)-Exchanging ATPase; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D009853:Omeprazole / Q000494:pharmacology; D000077402:Pantoprazole; D011725:Pyridines / Q000494:pharmacology*; D011817:Rabbits; D013454:Sulfoxides / Q000494:pharmacology*; D005466:Fluorocarbons
D053799:2-Pyridinylmethylsulfinylbenzimidazoles; D000251:Adenosine Triphosphatases; D000632:Aminopyrine; D001562:Benzimidazoles; D005978:Glutathione; D017506:H(+)-K(+)-Exchanging ATPase; D008274:Magnesium; D009853:Omeprazole; D000077402:Pantoprazole; D011725:Pyridines; D013454:Sulfoxides; C061512:SK&F 95448; D005466:Fluorocarbons