A new frog of the genus Psychrophrynella is described based on specimens from the Cordillera de Vilcabamba, in the department of Cusco in southeastern Peru. The new species inhabits the humid puna and is only known from its type locality in Challcha, near the road between Vilcabamba and Pampaconas, at 3,707 m asl. This new taxon is assigned to the genus Psychrophrynella based on a narrowest genetic distance of 16S rRNA with P. glauca (8.3&) and the presence of a fold-like tubercle on the inner edges of the tarsus. The description of Psychrophrynella vilcabambensis sp. nov. is based on three individuals. This new species can be differentiated from other members of the genera Psychrophrynella and Noblella by the combination of the following characters: light reddish-brown to tan coloration on the dorsum with dark brown markings, the presence of a thoracic fold, ulnar tubercles, a tubercle on the heel, three tubercles on outer edge of tarsus, and toes with lateral fringes. The SVL of male and female specimens are 16.5 and 16.6 mm, respectively.