Provided are a PEGylated analogue protein of canine urate oxidase, preparation method and use thereof. The analogue protein of canine urate oxidase is a canine urate oxidase, or a chimeric protein comprising part of the amino acid sequence of a canine urate oxidase and part of the amino acid sequence of a human urate oxidase, or a mutant protein thereof. The PEGylated analogue protein of canine urate oxidase and pharmaceutical compositions thereof according to the present invention can be used for the prevention and/or treatment of hyperuricemia and chronic gout.