This study aims to contribute to advancing the understanding of entrepreneurial ecosystems, Latin American development, and the evolution and future perspectives of the Middle East. It used a cross-sectional research design and quantitative data with 750 observations from the Global Entrepreneurship Monitor, specifically the National Experts Survey and the Adult Population Survey. The results were analyzed by the Partial Least Squares Technique (PLS-SEM) by grouping countries into two balanced samples of underexplored regions: the Middle East and Latin America. The two regions, Latin America and the Middle East, have diverse entrepreneurial ecosystems; each condition impacts entrepreneurial activities in different ways. In Latin America, the most significant variable is “Physical Infrastructure”, while in the Middle East, the most significant determinants are “Commercial and Professional Infrastructure” and “Entrepreneurship Culture”. This study shows that to support entrepreneurial activities, each region requires different settings for their entrepreneurial ecosystems. It contributes to the literature on regional entrepreneurial ecosystems and to less explored regions to advance our understanding, which will drive better policies and actions.