The invention provides a glycoside molecular derivative as well as a preparation method and application thereof, and belongs to the technical field of aminoglycoside antibiotics. The invention provides an etimicin derivative, and the etimicin derivative is a compound represented by the following formula and a pharmaceutically acceptable salt or prodrug thereof, wherein R1 represents a hydrogen atom, an ethyl group or an acetyl group, R2 represents a hydrogen atom, an ethyl group or an acetyl group, R3 represents a hydrogen atom, an ethyl group or an acetyl group, R4 represents a hydrogen atom,an ethyl group or an acetyl group, and R5 represents a hydrogen atom, an ethyl group or an acetyl group; wherein at least one of R1, R2, R3 is an ethyl group or an acetyl group; when R1 is an ethyl group, at least one of R2 and R3 is an ethyl group or an acetyl group; when R1 is an acetyl group, R2 and R3 can be hydrogen at the same time. The derivative can be used as a new aminoglycoside antibiotics product with low toxicity and resistance to drug-resistant bacteria, has good resistance to drug-resistant bacteria, and has a significant effect on resisting drug-resistant bacteria. The formulais shown in the description.