<p>Urbanization affects wildlife species, particularly in wildland-urban interfaces. In the rapidly urbanizing landscape of Assam in India, we explored the relationship between urban expansion and wildlife conservation, focusing on the Indian leopard (<i>Panthera pardus fusca</i>). We assessed the land use and land cover changes, human-leopard interactions, and habitat suitability analysis of leopard. We also estimated the relative leopard abundance based on suitable patches within the free space in Guwahati using human building dataset. Our analysis shows that built-up areas have expanded from 6.23% in 1989 to 16.99% in 2019, while natural habitats vital for leopard have substantially reduced from 11.39 to 0.33%, respectively. Our survey in buffer zones around key forest areas indicates a growing trend in interactions, leading to a decrease in free space vital for leopard abundance. We estimated that Guwahati city can currently support 14 leopards (38–8 SD) within the existing landscape, but increased urban development could reduce this number to 4 (10–2 SD). Our study highlights the challenges of wildlife conservation in urban landscapes and the need for strategies that balance urban development with biodiversity protection. Our findings align with Target 4 of the Kunming-Montreal Global Biodiversity Framework, which emphasizes managing human-wildlife interactions to reduce conflicts and foster coexistence by 2030.</p>