Provided are a streptovaricin derivative, a preparation method therefor, and an application thereof, pertaining to the field of bio-pharmaceuticals. The novel streptovaricin derivative 1-4 is obtained by using Streptomyces spectabilis XW-1 (accession number: CCTCC NO: M2017417) as a starting strain, and carrying out single inactivation or deletion on P450 genes (svP4 and stvP5) of a post-modification system in a streptovaricins biosynthesis gene cluster of the strain, followed by carrying out separation and purification on a crude extract of fermentation products of the two mutant strains, to give the novel streptovaricins derivative 1-4. The novel streptovaricins derivative of the present invention has good antibacterial effects against methicillin-resistant Staphylococcus aureus, Escherichia coli, and Acinetobacter baumannii, provides a new solution to the issue of antibiotic resistance, and provides a basis for structural modification of streptovaricins antibiotics.