Access to finance is a critical factor for the growth and sustainability of women-owned enterprises, yet women entrepreneurs in developing countries, including Uganda, often face significant barriers in securing business finance. This study investigated the challenges faced by women entrepreneurs in accessing business finance in Nakaseke District, with particular focus on the influence of collateral requirements, interest rates, financial literacy, entrepreneurial networks, and socio-cultural and institutional factors. The study employed a descriptive cross-sectional research design using a mixed-methods approach. Quantitative data were collected from 145 women entrepreneurs through structured questionnaires, while qualitative insights were obtained via semistructured interviews with key informants, including microfinance officers, local government officials, and leaders of women entrepreneurs’ associations. Data were analyzed using descriptive and inferential statistics, supplemented with thematic analysis of qualitative responses. The findings revealed that high collateral requirements and elevated interest rates constitute the most significant barriers to formal financial access for women entrepreneurs. Financial literacy deficits and inadequate business record-keeping moderately limited access, while socio-cultural norms and discriminatory practices further constrained women’s financial autonomy. In response, women entrepreneurs relied heavily on informal financing mechanisms such as family support, rotating savings groups, and participation in women’s networks. Participation in financial literacy programs and entrepreneurial networks was found to enhance financial access, although the reach of these interventions remains limited. The study concludes that access to finance for women entrepreneurs in Nakaseke District is influenced by a combination of financial, institutional, and socio-cultural factors. Policy interventions that reduce collateral requirements, lower interest rates, promote financial literacy, strengthen entrepreneurial networks, and address socio-cultural constraints are critical to improving women’s financial inclusion. The findings provide practical guidance for policymakers, financial institutions, and development partners seeking to empower women entrepreneurs and enhance economic development at the district and national levels.