Harry Maguire, Kyle Walker and Sam Johnstone have all withdrawn from an already depleted England ahead of next week's friendly international at home to Belgium, the Football Association announced Sunday.
England suffered their first defeat in 15 months on Saturday as Brazil won 1-0 at Wembley thanks to a late goal from teenager Endrick.
Walker and Maguire started that match but have now left England manager Gareth Southgate's squad, as has backup goalkeeper Johnstone ahead of Tuesday's match against Belgium at Wembley.
Manchester City's Rico Lewis, who made his debut against North Macedonia in November, and uncapped Burnley goalkeeper James Trafford have been moved into the senior squad from the England Under-21 set-up.
Captain Harry Kane missed Saturday's game with an ankle injury and had already been ruled out of facing Belgium.