Several research and innovation initiatives have been pursued worldwide for the development of autonomous and unmanned ships. However, these ships’ wider adoption is limited by the existing regulatory framework, which presently does not provide clear guidelines and requirements for the design and operation of autonomous ships. The aim of this study is to comprehensively analyse the existing maritime safety and security regulatory framework including the national and international regulations for designing, building, testing and operating the unmanned next-generation inland waterways (IWW) vessel considered in the AUTOSHIP project operating in the Flemish inland waters. This study initiates with the identification of the regulatory bodies controlling the operation and testing of the investigated vessel and then identifies barriers in regulations where amendments or new developments are required. Subsequently, a strategy for overcoming these barriers is proposed. The main regulatory gaps that are identified include the requirements for navigation, emergency and environment protection functions, where new definitions are required for unmanned ship operations (e.g. master, crew, remote control centre). Moreover, some of the regulations explicitly specify the existence of crew on the ships for navigation, emergency and environment protection functions. A three-phase strategy is proposed to overcome the current regulatory barriers.