Joao Felix scored a match-winner for Atletico as Christian Eriksen made his debut for the Red Devils.
Atletico Madrid defeated Manchester United 1-0 in their pre-season friendly on Saturday.
The Red Devils played without Cristiano Ronaldo, who has stayed out of action amid exit links at Old Trafford.
Erik ten Hag’s side started the game by taking control of possession. However, they were not able to create enough chances during the first half.
The Red Devils doubled down on their efforts to score the opener in the second period. Anthony Martial and Anthony Elanga came the closest but were not lucky enough to find the back of the net.
Ten Hag brought in Christian Eriksen in the 69th minute as the Dane made his Man Utd debut. Playing as an attacking midfielder, Eriksen was central to United’s attacks in the final 20 minutes.
The game seemed to be heading toward a goalless finish when Joao Felix drilled a low shot behind David De Gea to make it 1-0 for the hosts.
United pushed for a late equalizer but Fred’s red card in the 91st minute ended their hopes of escaping the defeat.
Having suffered his first defeat as Man Utd manager, Ten Hag will now prepare his side for their final pre-season match against Rayo Vallecano.
The Red Devils will then face Brighton in their Premier League opener on August 7.