The invention relates to an ofloxacin molecular inclusion nanometer preparation and a preparation method thereof, belonging to the technical field of medicines. The main raw materials of the molecular inclusion nanometer preparation are cyclodextrin, solubilizer, polymer materials, ofloxacin and the like. The preparation method comprises the steps of: adding the cyclodextrin after dissolving the ofloxacin in a solution containing the solubilizer and preparing a molecular inclusion compound through a gradient ultrasonic grinding method; then adding the polymer materials for grinding; and drying and smashing to obtain the ofloxacin molecular inclusion nanometer preparation. The average particle size of the molecular inclusion nanometer preparation is 252.3 nm; and the drug content is 1%-25%. The preparation method, disclosed by the invention, has the advantages of being simple in a preparation process, high in drug carrying amount and stability, controllable in quality, suitable for industrial production and the like. The ofloxacin molecular inclusion nanometer preparation prepared by the method has the advantages of efficiently improving stability of the drug and dissolution degree as well as dissolution speed of the drug in a gastrointestinal tract, improving a drug absorption rate, realizing rapid and efficient effects, masking bitter taste of the drug, reducing drug irradiation and improving drug palatability and drug availability.