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Background: The present study aimed to compare the clinical efficacy and safety profile of Ziferon and Betaferon. Objectives: In total, 41 consecutive patients with relapsing forms of Multiple Sclerosis (MS) were selected from the MS outpatient clinic affiliated to Tehran University of Medical Sciences. The patients were randomly assigned into two groups. Methods: Each group either received Ziferon 250 mcg Subcutaneously (SC) in alternate days or Betaferon 250 mcg SC on alternate days. Clinical and para-clinical outcomes, such as mean relapse rate/year score, mean Expanded Disability Status Scale (EDSS)/year score, the cumulative number, and the volume of gadolinium-enhancing lesions, in addition to the cumulative number of new T2 lesions and safety profile were evaluated for each group during the years of treatment. Results: There were no significant differences in Magnetic Resonance Imaging (MRI) outcomes (change in total lesion volume, new lesion per T2-weighted scan, and gadoliniumenhancing lesions per T1-weighted scan from baseline; P=0.236, P=0.56, & P=0.496, respectively was observed). There was no significant difference in the relapse rate between Ziferon and Betaferon treated groups (P=0.56). There were no unexpected safety events. The number of patients who discontinued the study due to adverse events occurrence was similar between the two groups. Conclusion: Evidence demonstrates the non-inferiority and bio-similarity of Ziferon (interferon beta-1b) to Betaferon in terms of efficacy and safety profile.