Extracts of . Cyclopia species, indigenous South African fynbos plants used for the production of honeybush tea, have potential as anti-obesity nutraceutical ingredients. Previously, we demonstrated that aqueous extracts of . C. maculata and . C. subternata exhibited anti-obesity effects in 3T3-L1 adipocytes. In this study, we further explored these anti-obesity effects of . C. maculata and . C. subternata as well as . C. intermedia for the first time. Extracts were prepared using a 40% methanol-water mixture (40% MeOH) in order to enhance the polyphenolic content of the extracts. Moreover, these extracts were separated into aqueous and organic fractions using liquid-liquid partitioning with . n-butanol and water to further enrich the polyphenol content of the organic fractions. Extracts of all three . Cyclopia species decreased the lipid content in 3T3-L1 adipocytes, although differences in bioactivity of their aqueous and organic fractions were observed. The organic fraction of . C. intermedia was further investigated. This fraction dose-dependently decreased the lipid content in 3T3-L1 adipocytes without affecting cell viability, while increasing mRNA expression of . HSL (1.57-fold, P. <. 0.05), . SIRT1 (1.5-fold, P. =0.07), . UCP3 (1.5-fold, P. <. 0.05) and . PPARγ (1.29-fold, P. <. 0.05). Daily treatment of obese db/db mice with 351.5. mg/kg bodyweight of the organic . C. intermedia fraction for 28. days decreased bodyweight gain by 21% (P. <. 0.05) without any effect on food or water consumption. The organic fraction was enriched in phenolic content relative to the extract with neoponcirin, a flavanone not previously identified in . Cyclopia species, mangiferin, isomangiferin and hesperidin comprising 17.37% of the organic fraction of . C. intermedia compared to 4.96% of its large scale prepared 40% MeOH extract. Their specific roles as anti-obesity agents in these models needs to be studied to guide product development.