After eight decades since its discovery in 1939, new investigations have been undertaken at Grotta Guattari (Latium, central Italy), a coastal cave by the Tyrrhenian Sea coast and one of the iconic sites of the Italian prehistory, as it yielded an almost complete skull and other remains of Neanderthals. The new excavations of the innermost and untouched cave deposits resulted in an outstanding amount of mammal bones, 40 out of which attributable to Neanderthal, including new large portions of cranial remains. Preliminary taphonomic hints and the collected stratigraphic evidence strongly indicate that the impressive accumulation of the large mammal bones was the work of spotted hyena, in a period in which human frequentation was really sporadic or even completely absent. The new acquired U/Th chronology of Grotta Guattari speleothems provided new constraints for reconstructing the sedimentary and paleoenvironmental history of the archaeological successions and human remains. The accumulation of terrestrial sediments started at ∼112 ka, immediately after the end of the Last Interglacial sea-level highstand (∼116 ka). However, the hyena frequentation, and thus the bone accumulations, occurred several thousands of years after and lasted for a very short time interval, precisely between slightly before 66 ka and 65 ka. The cave became abandoned by hyena after ∼65 ka and before ∼59 ka, because of the cave obstruction and/or the altered environmental conditions related to the Heinrich Event 6 (∼64–60 ka). The regional paleoclimatic records indicate severe conditions during the short interval of the hyena frequentation. In contrast, though providing evidence of open and arid environments, the faunal assemblage and the pollen from Grotta Guattari reveal that the local conditions were less severe, likely because of the mitigating effect of the Tyrrhenian Sea. In the framework of the Italian findings, Grotta Guattari results the richest and better chronologically constrained site of Neanderthal remains, posing it as one of the rare sites of the European prehistory that allows putting into the context an extraordinary large sample of the population of Neanderthals, and of other large mammals, in a very narrow and precisely dated temporal interval of the early Pleniglacial.