Inhibition of gastric H⁺,K⁺-ATPase by new dihydropyrazole derivative KYY-008

The gastric proton pump (H⁺,K⁺-ATPase) responsible for the H⁺ secretion of gastric acid is an essential therapeutic target for acid-related diseases. H⁺,K⁺-ATPase belongs to a P₂-type ATPase family. Here, we examined the effects of a newly synthesized dihydropyrazole derivative KYY-008 on the H⁺,K⁺-ATPase. KYY-008 concentration-dependently inhibited the enzyme activity of the ATPase in the membrane fractions prepared from isolated hog gastric mucosa and from human kidney HEK293 cells in which gastric H⁺,K⁺-ATPase is exogenously expressed. The IC₅₀ values in these samples were 3.4 μM and 3.7 μM, respectively. In addition, KYY-008 significantly inhibited the H⁺,K⁺-ATPase-derived H⁺ uptake into the tightly sealed vesicles prepared from the hog gastric mucosa. In contrast, KYY-008 has no effect on the activities of other P₂-type ATPases such as Na⁺,K⁺-ATPase and Ca²⁺-ATPase. KYY-008 did not change the ionic currents of voltage-dependent Ca²⁺ channels, that were potential targets for some dihydropyrazole derivatives. Together, we discovered a new dihydropyrazole derivative which acts as a selective inhibitor of gastric H⁺,K⁺-ATPase.