Cities incorporating navigable canals have played a crucial role in global trade and provided a platform for a range of activities for people from various locations. This research aims to comprehend the role of inner-city canals, formed as branches of shipping canals, in the spatial accessibility and functional structure of two contemporary urban systems: London and Amsterdam. Both cities are major post-industrial hubs in Europe and their spatial development and socioeconomic conditions have been greatly influenced by waterways. While the canal network in Amsterdam was planned alongside street layout planning in the early 17th century, serving commercial purposes, canals were integrated into London’s pre-existing urban form mainly for transportation in the 19th century. The current situation in these cities is impacted by this disparity in three ways: (a) the potential use of canals in the urban transportation system; (b) the spatial accessibility of street networks; and (c) the correlations between street accessibility and land use patterns in canal neighbourhoods. The research employs analytical methods of space syntax, geographic information systems, and statistical techniques to create and apply integrated urban models, incorporating spatial network measures, retail density, and functional diversity for street segments, to compare various urban conditions. The research reveals the crucial finding that the incorporation of canals into the street system leads to a substantial increase in the mean values of street network accessibility in Amsterdam. Additionally, the study highlights the vital contribution of diagonal streets linked with canal networks towards retail density in this city. In contrast, the accessibility measures and spatial patterns of urban functions in London are predominantly influenced by proximity to canals.