Background: Vulvovaginal candidiasis is a communal problem in virtually all the women which is caused by Candida albicans. Objective: Aim of the present study was to prepare and characterize vaginal suppositories of fluconazole for the treatment of vulvovaginal candidiasis. Methods: Fluconazole suppositories were prepared using water soluble and fatty bases. Bases and their proportions used were selected in such a way that they have flexibility in storage conditions unlike conventional suppositories. Suppositories were prepared and examined for physical characteristics and in vitro release studies. Results: Present study showed ultimate results with respect to the physical characteristics of suppositories and in vitro drug release studies. In vitro drug release from the prepared suppositories was in the following order FVS 3 (100.00 ± 3.7% in 1.0 h) > FVS 1 (86.29 ± 4.9% in 12.0 h) > FVS 2 (80.47 ± 2.4% in 12.0 h) > FVS 5 (22.51 ± 0.42% in 24.0 h) > FVS 4 (18.09 ± 1.31% in 24.0 h). These drug release results are supported by the disintegration time of suppositories. Lesser the disintegration time faster the drug release. Conclusion: Study concludes that it may be fruitful to explore the in vivo activities of suppositories prepared with the combination of agar and HPMC as it showed around 80% drug release over 12.0 h. Represented combination may also be more effective for the treatment of vulvovaginal candidiasis.