Background <p>This study aimed to evaluate the safety and immunogenicity of 5-dose and 4-dose schedules of a freeze-dried human rabies vaccine (MRC-5 cells) in participants aged 10–60</p> Methods <p>This study adopts a randomized, double-masked, single-center, non-inferiority trial design with similar vaccines. This phase III clinical trial consisted of two stages, the first stage recruited 80 participants, and the second stage recruited 1,800 participants. In stage 1, 40 participants aged 18–60 years were enrolled to vaccinate with the investigational vaccine firstly, and then 40 people aged 10–17 years were enrolled for safety analysis. The number of participants who received either a 5-dose or a 4-dose immunization regimen was 40. During stage 2, subjects were assigned to 5-dose Essen group, 4-dose Zagreb group, 5-dose control group to conduct rabies virus neutralization antibody detection and adverse events(AEs) collection.</p> Results <p>In stage 2, 14 days after the initial dose of immunization and 14 days after completing the immunization program, the differences in the seroconversion rates (5-dose Essen group vs. 5-dose control group, 4-dose Zagreb group vs. 5-dose control group) were both 0.00, with the lower limits of the 95% confidence intervals exceeding the non-inferiority margin of −5%. AEs occurred at a total rate of 35.17% in the 5-dose Essen group, 36.50% in the 5-dose control group, and 30.00% in the 4-dose Zagreb group.</p> Conclusions <p>The immunogenicity of 5-dose and 4-dose schedules of the freeze-dried human rabies vaccine (MRC-5 cells) was non-inferior to that of the human diploid cell vaccine, and the safety was good after vaccination.</p> Trials registration <p>NCT06078423, A Clinical Trial of Freeze-dried Human Rabies Vaccine (MRC-5 Cells), “retrospectively registered” on 2023–10-11.</p>