By screening a library of sulphated compounds of low molecular weight, we have found that several cyclitol derivatives, each modified with two sulphate groups in addition to pyrrole and various aromatic moieties, inhibited infectivity of herpes simplex virus (HSV) at concentrations approximately 100 times lower than those toxic for cultured cells. These disulphated cyclitols interfered with HSV-1 attachment to cells, and efficiently reduced the cell-to-cell spread of the virus. This effect is most likely due to their low molecular weight and associated with the compounds' capability to access the narrow intercellular spaces. Furthermore, these disulphated cyclitols also inactivated infectivity of HSV. However, the virus-inactivating activities of these compounds were to some extent diminished in the presence of human cervical secretions or other protein-rich solutions suggesting that disulphated cyclitols may have some features of surfactant-type virucides. In conclusion, this new class of anti-HSV compounds offers potential for further development.