Enhancing economic security of regions is crucial for the development of the whole country, which is what makes research in this sphere particularly important. This study aims to analyze and compare the economic security data on the regions constituting the Ural Federal District (Russia). In contrast with current studies in the field, we are conducting detailed analysis of the factors that affect the development of small and medium-sized enterprises (SMEs) and business climate in the regions. The conceptual framework of this research relies on entrepreneurship theories and theoretical approaches to analysis and evaluation of regional economic security. We develop methodology based on sets of quantitative and qualitative indicators and apply analytical, comparative and statistical methods as well as the method of expert evaluation. The data are provided by the regional statistic services and business support foundations. We also analyze regional support programs for small and medium-sized businesses. We found that all regions of the Ural Federal District are characterized by the medium (acceptable) level of economic security and moderate risk. In the economic security ranking, Tyumen region is at the top while the second place is occupied by Sverdlovsk region; Chelyabinsk and Kurgan regions are at the bottom.