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The application belongs to the field of biological medicine, and discloses a pharmaceutical composition of water channel protein inhibitor combined with temozolomide for treating brain glioma, wherein the pharmaceutical composition comprises AQP4 inhibitor AER270 and TMZ. The application further discloses a pharmaceutical preparation comprising the above pharmaceutical composition and application of the pharmaceutical preparation in preparation of a medicine for treating brain glioma. The composition provided by the application provides an effective drug combination strategy for treating brain glioma, and it is found for the first time that AER270 and TMZ have a synergistic effect on the treatment of brain glioma.