Minor groove binders are one of the most widely studied class of agents characterized by a high level of sequence specificity and they are still an interesting class of DNA ligands which demonstrate to possess several biological activities. It is important to underlie that in order to be effective as an anticancer agent, a minor groove binder should covalently and irreversibly bind the DNA with a long permanence of DNA lesion. This presentation will focus on cytotoxics structurally derivates from main minor groove binders such as distamycin A (1) , anthramycin (2) and CC-1065 (3). In particular will be underline the synthesis and biological activity of molecular hybrids deriving among the combination of the distamycin A and naturally occurring alkylating agents (1). A series of 2-(3’,4’,5’-trimethoxybenzoyl)-3-amino-5-aryl/heteroaryl thiophene derivatives and its corresponding isomers, with a 2-amino-3-(3’,4’,5’-trimethoxybenzoyl)-5-phenyl thiophene core, were synthesized and evaluated for antiproliferative activity, inhibition of tubulin polymerization and their effects on the cell cycle. The SAR was elucidated with various substitutions on the aryl moiety at the 5-position of the thienyl ring (2).