The importance of the Acacia genus is due to its usage in nutrition, and in folk and modern medicine. They are considered as promising sources of bioactive metabolites displaying beneficial to human health. The chemical and biological investigation of some Acacia species helped in the design of food supplements and drugs. A. cyanophylla has been introduced in Tunisia mainly to rehabilitate rangelands, especially in semi-arid areas, and also as a potential forage resource during periods of drought. The aim of this study was to investigate, in vivo, the analgesic, anti-inflammatory and gastroprotective potentials of the EtOAc extract and one of its major glycoside constituents, isosalipurposide (1), from the flowers of A. cyanophylla. The gastroprotective activity was evaluated using HCl/EtOH induced gastric ulcers in rats. The EtOAc extract exhibited interesting gastroprotective, analgesic and anti-inflammatory activities in a dose-dependent manner. Similarly, isosalipurposide (1) displayed promising anti-inflammatory and analgesic potentials and showed a protection of stomach against ulcer induced by HCl/EtOH. These data suggest that the pharmacological potential of the EtOAc extract could be mainly due to its main component isosalipurposide (1), which leads to conclude that the flowers of A. cyanophylla may be a possible source for the quantitative isolation of a pharmacoactive compound of interest.