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Rapid urbanization has a comprehensive impact on the patterns of regional ecological security (ESPs) in the Sino-Singapore Guangzhou Knowledge City, China (SSGKC). This study aims to propose recommendations for ensuring regional ecological security by conducting a spatiotemporal dynamic assessment of ESPs. The findings reveal that SSGKC has experienced rapid urbanization in the past decade, resulting in more than doubling of construction land. The results indicate that the area of the largest 10 patches of ESs decreased by 13.5%. Two largest patches of ESs in the southwest became smaller and were out of the top 10 patches in 2020. The expansion of construction land has encroached upon key ecological corridors (ECs). Two ECs connecting the eastern and central regions have shifted southward. Landscape pattern indexes demonstrate that not only has the area of ESs decreased, but they have also become more fragmented. This study also utilized the Urban Development Boundary (UDB) and FLUS model to simulate and predict the potential impact of future land use change on ESPs. Policymakers are advised to optimize the UDB by excluding areas that overlap with ESs and key ECs, in order to minimize the potential occupation of ESs and ECs in the future. The circular ECs connecting the eastern and central regions should be given priority planned and protection.