It is estimated that over 80% of microbial infections in the human body are biofilms related infections. Since bacteria in biofilms are less susceptible to antibiotics than are bacteria in the planktonic state, biofilm associated infections pose a major health threat, and there is a pressing need for anti-biofilm agents. Recent studies have shown that cationic amphiphiles inhibit the biofilm formation. In this context, a series of water soluble cationic amphiphiles have been synthesized and their biofilm inhibition properties towards six clinically important Gram positive pathogens were studied. These conjugates showed excellent anti-biofilm activity without exhibiting any bacteriostatic or bactericidal effect and mammalian cell membrane damage. The synthesis and the biofilm inhibition properties of the cationic amphiphiles will be presented.