Differential effects of tamoxifen-like compounds on osteoclastic bone degradation, H(+)-ATPase activity, calmodulin-dependent cyclic nucleotide phosphodiesterase activity, and calmodulin binding.
We studied effects of calmodulin antagonists on osteoclastic activity and calmodulin-dependent HCl transport. The results were compared to effects on the calmodulin-dependent phosphodiesterase and antagonist-calmodulin binding affinity. Avian osteoclast degradation of labeled bone was inhibited approximately 40% by trifluoperazine or tamoxifen with half-maximal effects at 1-3 microM. Four benzopyrans structurally resembling tamoxifen were compared: d-centchroman inhibited resorption 30%, with half-maximal effect at approximately 100 nM, cischroman and CDRI 85/287 gave 15-20% inhibition, and l-centchroman was ineffective. No benzopyran inhibited cell attachment or protein synthesis below 10 microM. However, ATP-dependent membrane vesicle acridine transport showed that H(+)-ATPase activity was abolished by all compounds with 50% effects at 0.25-1 microM. All compounds also inhibited calmodulin-dependent cyclic nucleotide phosphodiesterase at micromolar calcium. Relative potency varied with assay type, but d- and l-centchroman, surprisingly, inhibited both H(+)-ATPase and phosphodiesterase activity at similar concentrations. However, d- and l-centchroman effects in either assay diverged at nanomolar calcium. Of benzopyrans tested, only the d-centchroman effects were calcium-dependent. Interaction of compounds with calmodulin at similar concentrations were confirmed by displacement of labeled calmodulin from immobilized trifluoperazine. Thus, the compounds tested all interact with calmodulin directly to varying degrees, and the observed osteoclast inhibition is consistent with calmodulin-mediated effects. However, calmodulin antagonist activity varies between specific reactions, and free calcium regulates specificity of some interactions. Effects on whole cells probably also reflect other properties, including transport into cells.
Williams
J P
JP
Department of Pathology, University of Alabama at Birmingham 35294, USA.
McDonald
J M
JM
McKenna
M A
MA
Jordan
S E
SE
Radding
W
W
Blair
H C
HC
D000818:Animals; D016588:Anticarcinogenic Agents / Q000494:pharmacology; D001578:Benzopyrans / Q000494:pharmacology; D001862:Bone Resorption; D002147:Calmodulin / Q000187:drug effects / Q000378:metabolism*; D002486:Centchroman / Q000494:pharmacology; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D004305:Dose-Response Relationship, Drug; D005260:Female; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism*; D010727:Phosphoric Diester Hydrolases / Q000187:drug effects / Q000378:metabolism*; D010880:Piperidines / Q000494:pharmacology; D006180:Proton-Translocating ATPases / Q000187:drug effects / Q000378:metabolism*; D013629:Tamoxifen / Q000494:pharmacology*; D014268:Trifluoperazine / Q000494:pharmacology
D016588:Anticarcinogenic Agents; D001578:Benzopyrans; D002147:Calmodulin; D010880:Piperidines; D013629:Tamoxifen; C066624:CDRI 85-287; D014268:Trifluoperazine; D002486:Centchroman; D010727:Phosphoric Diester Hydrolases; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D006180:Proton-Translocating ATPases