The "Paddington" movies are a well-established cinematic entity at this point. As such, most of the principal human, non-CGI cast will play the same roles they have twice before in "Paddington in Peru." Hugh Bonneville will once again play London dad Henry Brown, alongside returning actors Samuel Joslin as Jonathan Brown, Madeline Harris as Judy Brown, Julie Walters as Mrs. Bird, and Jim Broadbent as Mr. Gruber. On the voice side, Ben Whishaw will reprise his audio-only part of Paddington Bear, and Imelda Staunton is set to portray Paddington's Aunt Lucy once more.

As far as new cast members, Emily Mortimer will step into the role of Mary Brown, vacated by Sally Hawkins, who is moving on after playing that part in the first two "Paddington" movies. Oscar-winner Olivia Colman joins the cast as the bubbly, guitar-playing nun The Reverend Mother, who operates Aunt Lucy's Home for Retired Bears. Antonio Banderas is also making his "Paddington" debut as riverboat captain and adventurer Hunter Cabot. Rachel Zegler of "West Side Story" was initially cast as Banderas' daughter, Gina Cabot, but had to drop out due to the SAG-AFTRA strike. Spain-based actor Carla Tous has since landed the role.