Maytenus dhofarensis is a Celastraceous herb naturally growing in the southern region of Oman, locally known as "Serbit." Ethnobotanical information indicates that when goats consume the plant's fruits, they experience illness marked by shivering attacks. Despite not being recognized as a medicinal plant in Omani herbal culture, other Maytenus species worldwide possess medicinal values. The absence of phytochemical studies on M. dhofarensis prompted our interest in exploring this plant for structurally novel compounds and their bioactivities against kinases related to cell survival and proliferation signaling pathways. In this study, hexane and chloroform extracts from the fruits of M. dhofarensis were chemically investigated, leading to the isolation of seven compounds, monoacetyldiglyceride (1), triacylglyceride (2), palmitic acid (3), hydroxylated monoacetyldiglyceride (4), two dihydro-β-agarofuran-type sesquiterpene pyridine alkaloids (5) and (6), and alkenyl isothiocyanate (7). Compounds (1–7) represent new isolates from M. dhofarensis. The structures of these compounds were determined through comprehensive IR, 1D and 2D NMR, UV, and ESI-MS analyses. The relative configurations of compound 5 were deduced from density function theory (DFT) calculations and NMR experiments. Additionally, hydroxylated fatty acid residues, palmitic acid, stearic acid, and oleic acid (18:9) were identified in the seed oil of the plant using GC-MS DMOX technique. Compound 5 exhibits potential anti-cancer effects, inducing morphological changes and a slight inhibition of colonies formation in MCF-7 cells. Molecular pathway analysis reveals a significant impact on p-Akt expression in MCF-7 cells after treatment with compound 5, suggesting a targeted mechanism.