Morpholino derivatives of benzyl-benzodioxole, a study of structural requirements for drug interactions at the colchicine/podophyllotoxin binding site of tubulin.

We performed a structure-activity evaluation of the effects of methoxy substituents in the benzyl moiety of a series of morpholinyl Mannich base derivatives of 6-benzyl-1,3-benzodioxol-5-ol ("morpholino compounds") on the ability of these compounds to inhibit tubulin polymerization in vitro. Structurally these agents are most similar to the natural product podophyllotoxin and, like podophyllotoxin, they inhibited in vitro tubulin polymerization, tubulin-dependent GTP hydrolysis, and the binding of colchicine to tubulin. The benzyl ring (C ring) of these compounds appeared to be analogous to the trimethoxybenzene ring (E ring) of podophyllotoxin (with its methoxy substituents at the 3', 4' and 5' positions), but the morpholino compound superficially most similar to podophyllotoxin (with 3', 4' and 5' methoxy substituents) was the least active in the series. The most potent methoxy-substituted morpholino compounds bear these substituents either at the 2' and 4' positions (NSC 370277) or at the 2', 4' and 6' positions (NSC 381577). NSC 370277 and NSC 381577 were essentially identical in their inhibitory effects on tubulin polymerization, but the latter compound was considerably more effective as an inhibitor of the binding of colchicine to tubulin. The most active of the monomethoxy substituted compounds bore this group at position 4'. A number of compounds with alternative substituents at this position (in particular, alkyl-substituted amines) also had significant in vitro inhibitory effects on tubulin polymerization. Although the morpholino compounds appear to possess only limited cytotoxicity, these findings suggest possible modifications of the antimitotic benzyl-benzodioxole compounds described previously [Batra et al., Molec. Pharmac. 27, 94 (1985)] to enhance their antineoplastic activity.