Apex predators ideally require vast intact spaces that support sufficient prey abundances to sustain them. In a developing world, however, it is becoming extremely difficult to maintain large enough areas to facilitate apex predators outside of protected regions. Free-roaming leopards (Panthera pardus) are the last remaining apex predator in the Greater Cape Floristic Region, South Africa, and face a multitude of threats attributable to competition for space and resources with humans. Using camera-trap data, we investigated the influence of anthropogenic land modification on leopards and the availability of their natural prey species in two contrasting communities—primarily protected (Cederberg) and agriculturally transformed (Piketberg). Potential prey species composition and diversity were determined, to indicate prey availability in each region. Factors influencing spatial utilisation by leopards and their main prey species were also assessed. Estimated potential prey species richness (Cederberg = 27, Piketberg = 26) and diversity indices (Cederberg—H′ = 2.64, Ds = 0.90; Piketberg—H′ = 2.46, Ds = 0.89), supported by both the Jaccard’s Index (J = 0.73) and Sørensen’s Coefficient (CC = 0.85), suggested high levels of similarity across the two regions. Main leopard prey species were present in both regions, but their relative abundances differed. Grey rhebok, klipspringer, and rock hyrax were more abundant in the Cederberg, while Cape grysbok, Cape porcupine, chacma baboon, and common duiker were more abundant in Piketberg. Leopards persisted across the agriculturally transformed landscape despite these differences. Occupancy modelling revealed that the spatial dynamics of leopards differed between the two regions, except for both populations preferring areas further away from human habitation. Overall, anthropogenic factors played a greater role in affecting spatial utilisation by leopards and their main prey species in the transformed region, whereas environmental factors had a stronger influence in the protected region. We argue that greater utilisation of alternative main prey species to those preferred in the protected region, including livestock, likely facilitates the persistence of leopards in the transformed region, and believe that this has further implications for human-wildlife conflict. Our study provides a baseline understanding of the potential direct and indirect impacts of agricultural landscape transformation on the behaviour of leopards and shows that heavily modified lands have the potential to facilitate mammalian diversity, including apex predators. We iterate that conservation measures for apex predators should be prioritised where they are present on working lands, and encourage the collaborative development of customised, cost-effective, multi-species conflict management approaches that facilitate coexistence.