The production on the Bryan Singer-directed film will begin later this year, reported Variety.
While Lee will portray lead guitarist Brian May, Mazzello will take on the role of bass guitarist John Deacon.
Rami Malek stars as frontman Freddie Mercury, with Ben Hardy playing drummer Roger Taylor.
The script has been penned by Justin Haythe, that will follow Queen's journey up to their appearance at the Live Aid concert in 1985.
The movie is slated to release in US on December 25, 2018.