The club had hoped a deal could be reached where it could take over the 80,000-seat venue after the Games and move away from its traditional White Hart Lane ground.
But a spokesman for London 2012 says no such deal will be possible.
They said: "The stadium will continue to have athletics at its core after the Games and it is our job to provide a post-Games legacy for the sport."
English Premier League club Spurs have been told they will not be able to move into the London 2012 Olympic Stadium following the Games.
