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Archaeological and palaeoenvironmental evidence from the Armenian Highlands and wider southern Caucasus region emphasises the significance of Marine Oxygen Isotope Stage 3 (c. 57–29 ka) as a crucial period for understanding hominin behaviours amidst environmental fluctuations. Ararat-1 cave, situated in the Ararat Depression, Republic of Armenia, presents potential for resolving emerging key debates regarding hominin land use adaptations during this interval, due to its well-preserved lithic artefacts and faunal assemblages. We present the first results of combined sedimentological, geochronological (luminescence and radiocarbon), archaeological and palaeoecological (macrofauna, microfauna and microcharcoal) study of the Ararat-1 sequence. We demonstrate sediment accumulation occurred between 52 and 35 ka and was caused by a combination of aeolian activity, cave rockfall and water action. Whilst the upper strata of the Ararat-1 sequence experienced post-depositional disturbance due to faunal and anthropogenic processes, the lower strata remain relatively undisturbed. We suggest that during a stable period within MIS 3, Ararat-1 was inhabited by Middle Palaeolithic hominins amidst a mosaic of semi-arid shrub, grassland, and temperate woodland ecosystems. These hominins utilised local and distant toolstone raw materials, indicating their ability to adapt to diverse ecological and elevation gradients. Through comparison of Ararat-1 with other sequences in the region, we highlight the spatial variability of MIS 3 environments and its on hominin land use adaptations. This demonstrates the importance of the Armenian Highlands for understanding regional MP settlement dynamics during a critical period of hominin dispersals and evolution.