OBJECTIVE: To prepare nanostructured lipid carrier of adefovir dipivoxil (ADV-NLC), and optimize the formulation. METHODS: Using stearic acid and glycerin monostearate as solid lipid, oleic acid as liquid lipid, Gemini surfactant and polysorbate 80 as emulsifier, sodium dodecyl sulfate (SDS) as stabilizer, solvent dispersion ultrasonic method was used to prepare ADV-NLC. And using particle size, polydispersity index, Zeta potential, encapsulation efficiency as indexes, single factor test was conducted to screen Gemini surfactant-polysorbate 80 ratio, emulsifier dosage (ratio of emulsifier to water phase), drug-lipid ratio, solid-liquid lipid ratio. RESULTS: The formula was as follow as 3% emulsifier (Gemini surfactant-polysorbate 80 ratio of 1 ∶ 2), 4.5% drug-lipid ratio, solid-liquid lipid ratio of 6 ∶ 5. The average particle size of the prepared ADV-NLC was (48.83±2.65) nm, polydispersity index<0.3, Zeta potential was (-28.7±1.8) mV, encapsulation efficiency was (77.65±0.03)% (n=3). CONCLUSIONS: ADV-NLC is successfully prepared, and the formulation is reasonable and feasible.