The invention discloses crowe saponins as well as a preparation method and application thereof in resisting highly pathogenic H5N1 influenza virus. In the invention, natural product yam aglycon is taken as a raw material, hydroxyl is introduced on C-6 through structural modification to obtain crowe sapogenin, glycosyl is introduced on C-3 to obtain series of crowe saponins, and finally different acyl groups are introduced on C-6 hydroxy to obtain series of the compounds. Pharmacological experiments show that the crowe saponins have obvious inhibition on the process of invading host cells by H5N1 highly pathogenic influenza virus, and the crowe saponins can serve as a medicine for preventing or treating influenza virus. The invention also relates to a combination of the compounds and other antiviral agents as well as medicines containing the same.