Holland, 23, and Smith, 50, play scientist Walter Beckett and super spy Lance Sterling in the new animated film.
The preview shows Walter (Holland) turn Lance (Smith) into a pigeon with one of his scientific experiments. The pair must team up to save the world with Lance disguised as the bird.
Spies in Disguise is inspired by Lucas Martell's animated short film Pigeon: Impossible. The movie is directed by Nick Bruno and Troy Quane, and co-stars Ben Mendelsohn, Karen Gillan, Rashida Jones, DJ Khaled and Masi Oka.
"The world's flyest spy. Watch Will Smith and @tomholland1996 in the new trailer for #SpiesInDisguise, in theaters this Christmas," the film tweeted Monday.
Spies in Disguise opens in theaters Dec. 25. Holland will next star in Spider-Man: Far From Home, which opens in theaters Tuesday, while Smith will appear in Gemini Man.