Cole has been diagnosed with a depressed fracture of the tibia after aggravating the problem prior to Sunday's 1-0 home victory over Manchester United.
"Ashley Cole has not been selected in the England squad for their friendly against Brazil. He has a depressed fracture of the tibia, an injury first picked up in the Atletico Madrid home match last month," a spokesman for Chelsea confirmed.
"He aggravated the injury before the match against Manchester United and required an injection to play.
"However, he further injured it during the game and has therefore been ruled out of this week's England squad."
The defender missed three games after initially sustaining the injury in the Blues' 4-0 home win against Atletico Madrid in the Champions League last month.
He returned against Madrid in the return Group D game last week but required a painkilling injection in order to play against United.