Disclosed is a 3-substituted nalidixic acid analog compound and its preparation method and uses in pharmacy, wherein the compound has antineoplastic activity and a general formula (I) with the 3-position substituted by benzimidazole and benzothiazole, the preparation process of the compound comprises condensing various substituted naphthyridine ketone-3-formylic acid, o-phenylenediamine, o-aminophenol, o-aminobenzenethiol and into polyphosphoric acids. The compound can be used in preparing anti-cancer medicaments.