We describe three new species of Bryophryne from the Region of Cusco in southern Peru, increasing the number of currently known Bryophryne to six. One of the new species differs from all species of Bryophryne in having a tympanic annulus and tympanic membrane. Males of this species have vocal slits, a vocal sac, and produce a call, which we analyze herein. This species is found at San Luis, a montane cloud forest along the road from Abra Malaga to Quillabamba at elevations between 3272 and 3354 m. The second new species has an orange throat and groin and is found along the Ericsson trail that connects Acjanaco to Pillahuata in Manu National Park and near Abra Acjanaco along the Paucartambo-Pilcopata road at elevations between 3266 and 3430 m. The third new species has the throat and chest mottled pale gray and tan, whereas the belly is black with white flecks. This species is only known from the upper Marcapata valley along the road from Abra Huallahualla to Quincemil at elevations between 3129 and 3285 m. Ecological observations for all new species are described and a map showing the type localities of all currently known species of Bryophryne is presented.