The resurgence of mpox virus (MPXV) poses a significant challenge to global public health. Currently, there is a limited understanding of the evolutionary details of MPXV during its epidemics, and no specific drugs have been developed for it. Herein, analysis of mutations and positive selection sites (PSSs) within the MPXV genomes revealed 799 mutations and 40 PSSs. Visualization analysis indicated that these mutations and PSSs may affect protein structure. Additionally, a protein–protein interaction (PPI) network between human and MPXV was established, identifying 346 MPXV-interacting human proteins (MIHPs). An interaction network involving MIHPs and other viruses confirmed that these proteins can interact with various viruses that infect humans. Functional analysis of MIHPs suggested their enrichment in host immunity pathways. Lastly, two drugs targeting MIHPs and four compounds targeting MPXV proteins were screened as candidate antivirals against MPXV. These findings not only deepen our understanding of MPXV evolution but also aid in the development of anti-MPXV drugs.