Ben Chilwell will play no part against Denmark after leaving England’s training camp with a foot injury.
The 23-year-old Chelsea full-back missed England’s previous Nations League clash with Belgium after coming down with an illness and had been training alone at St George’s Park.
Kieran Trippier started at left wing-back in place of Chilwell as Gareth Southgate’s side came from behind to beat Belgium at the weekend but the Atletico Madrid star has also left England’s camp due to a personal matter.
The Blues play host to Southampton when the Premier League gets back underway next weekend before kicking off their Champions League campaign at home to Sevilla.
The absence of Chilwell and Tripper could see Arsenal youngster Bukayo Saka make his first competitive start for the Three Lions.
Saka played 76 minutes of England friendly with Wales last week as goals from Dominic Calvert-Lewin, Conor Coady and Danny Ings sealed a comfortable 3-0 win at Wembley.
England captain Harry Kane is set to return to Southgate’s starting XI after recovering from a muscle injury.
The Tottenham striker started on the bench for England’s 2-1 win over Belgium but came on as a substitute for Calvert-Lewin midway through the second half, shortly after Mason Mount’s winning goal.
‘Well, medically, there’s been conversations, because that would always be the case,’ Southgate replied when asked whether he had spoken to Spurs about Kane’s fitness ahead of England’s clash with Denmark.
‘He reported on the Sunday, we gave players that reported on Sunday, who weren’t involved in the Wales game for us, we gave them two days’ recovery.
‘He started to train on the Wednesday, was a little unhappy with how he felt, so then worked with our medical team the next couple of days, we scanned just to be certain but it’s a muscular fatigue issue rather than an injury, so it just needed a bit more confidence.
‘I think he’ll have gained more confidence from the spell he had against Belgium, and so all being well, we hope he’ll be good to start on Wednesday.’