The contribution is focused on application of cluster analysis to assessment of business environment of selected regions in Czech Republic. Four indicators were created from the set of economic variables. They are: Indicator of economic performance, Indicator of human resources quality, Indicator of infrastructure and information technology, Indicator of environment and attractiveness. Economic performance covers the growth performance, labour and wages. Human resources quality includes the qualification and skills of population, participation in education. Other two indicators include science and technology performance, information technology and quality of environment in regions. Quality of business environment is significant attribute in the decision making of entrepreneur about process of establishment business organization. The aim of the contribution is to make analysis of business environment which offers relevant informations to subjects of executive and possible investors.