The comedian and Trainwreck star was attached to appear in the famous toy's live-action debut - but has now pulled out, saying she's just too darn busy.
"Sadly, I'm no longer able to commit to Barbie due to scheduling conflicts," the actress told Variety.
"I'm bummed, but look forward to seeing Barbie on the big screen." No replacement has been announced for the film, which is due out next summer.
Film studio Sony said it "respected and supported" Schumer's decision but would press ahead with the movie.
"We look forward to bringing Barbie to the world and sharing updates on casting and film-makers soon," its spokesperson continued.
The plot of the film revolves around a doll who is forced to live in the real world after being deemed not perfect enough for 'Barbieland'.
Schumer, who had faced criticism over her links to the project, said in Decembershe was "honoured... to be considered to play an important and evolving icon".
The first Barbie doll, made by Mattel, was introduced to the world at the New York Toy Fair in 1959.
Since then her screen appearances have been limited to animation, most notably in Toy Story 3 in 2010.
Schumer, 35, will shortly be seen playing Goldie Hawn's daughter in the action comedy Snatched.
According to Variety, she has also signed up to appear alongside Steve Carell and Nicole Kidman in "independent dramedy" She Came to Me.