Reduced brown adipose tissue (BAT) activity is considered to contribute to obesity development. BAT activity is decreased in both women with PCOS and PCOS animal models, possibly due to androgen excess. In addition to ovarian androgens, women with PCOS may also have elevated adrenal 11-oxygenated androgens. Here we aim to unravel whether 11-oxygenated androgens, also known as 11-keto-androgens, affect BAT metabolism. The mouse brown adipocyte cell line T37i was treated with increasing concentrations (10^-7–10^-5M) of testosterone (T), dihydrotestosterone (DHT), 11-keto-testosterone (KT) or 11-keto-dihydrotestosterone (KDHT) during or after differentiation. Adipocyte differentiation was assessed by lipid accumulation and gene expression. In addition, female mice received a single injection of vehicle, DHT, KT or KDHT (0.1mg). BAT was collected 24 hours later for RNAseq analysis to identify differentially expressed genes (DEGs) and enriched pathways by Gene Set Enrichment Analysis (GSEA). T, DHT, KT and KDHT treatment during differentiation dose-dependently reduced lipid droplet accumulation and inhibited mRNA expression of Ucp1, Prdm16 and Pgc1a by almost 2-fold (all P<0.05). Similar results were obtained in mature T37i cells. RNAseq analysis of DHT-exposed BAT identified 374 DEGs, of which 46 were downregulated and 328 were upregulated. KT and KDHT treatment resulted in only 100 and 166 DEGs respectively, with a 50% distribution in up- and downregulated genes. Intriguingly, of these DEGs, only 4 genes related to adipogenesis were shared between DHT, KT and KDHT treatment. DHT and KDHT treatment resulted in 49 shared DEGs, which were involved in inflammation. GSEA revealed downregulation of oxidative phosphorylation and fatty acid metabolism pathways upon DHT or KT treatment, while KDHT-induced DEGs were enriched in proliferation-related pathways. Our results demonstrate that 11-oxygenated androgens inhibit differentiation and function of brown adipocytes in vitro. The in vivo effect of 11-oxygenated androgens on BAT transcriptomics appeared to differ from DHT.