Spider venoms are complex mixtures composed of a variety of compounds, including salts, small organic molecules, peptides, and proteins. But, the venom of a few species is dangerous to humans. High levels of chemical diversity make spider venoms attractive subjects for chemical prospecting. Many spider venom components show potential activity against a wide range of human diseases. However, the development of novel venom-derived therapeutics requires an understanding of their mechanisms of action. This review will highlight the structures, activities and the possible mechanisms of action of spider venoms and their components against cancer, microbial infections, and pain.