The invention discloses an application of diphyllin in preparation of drugs for inhibiting poxvirus and preventing or treating poxvirus infection, the poxvirus is common orthopoxvirus poxvirus, the poxvirus comprises
vaccinia virus (VACV),
vaccinia virus (CPXV) and rabbit poxvirus mucinoma
virus (MYXV), the content of diphyllin is in a range of 1.25-2.5 [mu] M / mL, and the content of diphyllin is in a range of 1.25-2.5 [mu] M / mL. The compound has a remarkable inhibiting effect on infection of poxvirus family members such as CPXV, is expected to be used as a
medicine for preventing and treating infection of CPXV and other poxviruses, and has important significance on prevention and control of diseases related to poxvirus infection.