The Andean Cordilleras of Colombia, especially the Cordillera Occidental, are among the areas with the highest diversity of Pristimantis frogs in the world. Within the Cordillera Occidental, the Serranía de los Paraguas is famous for its high diversity of sympatric species of Pristimantis frogs. In this study, we investigated acoustic frequency segregation in five sympatric and syntopic species of the genus Pristimantis inhabiting this Serranía. It is important to point out that three of the five species' calls are described for the first time: Pristimantis brevifrons, P. ptochus, and P. silverstonei. Our results showed that acoustic frequency segregation occurs among these five Pristimantis species. Another notable result was the inverse relationship between dominant frequency and male body size, which was corroborated in all species except in P. alius. Finally, we suggested that for improving our knowledge on species coexistence in Pristimantis, future research should also focus on species history and character evolution, given that the evolutionary history is also fundamental for explaining species coexistence.