Naphthalenesulfonamides as calmodulin antagonists and protein kinase inhibitors.
N-(6-Aminoethyl)-5-chloro-1-naphthalenesulfonamide (A-3), which is a shorter alkyl chain derivative of the calmodulin (CaM) antagonist, W-7, was found to inhibit smooth muscle myosin light chain kinase (MLC-kinase) through a mechanism different from that related to W-7. Both the holoenzyme and the catalytic fragment, which is active without CaM, were susceptible to A-3 with a similar concentration dependency, thereby indicating that the inhibitory effect is due to the direct interaction of the compound with the enzyme molecule and not with the enzyme activator. Naphthalenesulfonamides are both CaM antagonists and direct inhibitors of MLC-kinase, and these actions depend on the length of the alkyl chain (C2-C6). Although the potencies in inhibiting CaM functions increased, the direct effects on MLC-kinase decreased with extension of the carbon chain of the derivatives. Kinetic studies indicated that A-3 inhibited MLC-kinase competitively with respect to ATP and that the Ki value was 7.4 microM. A-3 was also a competitive inhibitor of cAMP-dependent protein kinase, cGMP-dependent protein kinase, protein kinase C, casein kinase I, and casein kinase II, with respect to ATP. The Ki values of naphthalenesulfonamides for these enzymes also increased with extension of the carbon chain of the derivatives. These results suggest that naphthalenesulfonamides inhibit protein phosphorylation not only by inhibition of the enzyme-activating process but also by inhibition of the catalytic process. The mode of interaction between the derivatives and protein kinases differs from the interaction between the derivatives and CaM.
Inagaki
M
M
Kawamoto
S
S
Itoh
H
H
Saitoh
M
M
Hagiwara
M
M
Takahashi
J
J
Hidaka
H
H
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002147:Calmodulin / Q000037:antagonists & inhibitors*; D002645:Chickens; D006162:Guanylate Cyclase / Q000378:metabolism; D066298:In Vitro Techniques; D009219:Myosin-Light-Chain Kinase; D010726:Phosphodiesterase Inhibitors; D047428:Protein Kinase Inhibitors; D013329:Structure-Activity Relationship; D013449:Sulfonamides / Q000494:pharmacology*
D002147:Calmodulin; D010726:Phosphodiesterase Inhibitors; D047428:Protein Kinase Inhibitors; D013449:Sulfonamides; C017967:W 7; D009219:Myosin-Light-Chain Kinase; D000251:Adenosine Triphosphatases; D000262:Adenylyl Cyclases; D006162:Guanylate Cyclase