Liverpool right-back Trent Alexander-Arnold has not been included in England's matchday squad for Monday's Nations League match against Germany.
Alexander-Arnold, who missed last year's Euros through injury, has made just one international appearance this year and was an unused substitute in Friday's defeat by Italy.
Fikayo Tomori, Jarrod Bowen and James Ward-Prowse also miss out.
But Brentford striker Ivan Toney is in line to make his international debut.
Toney, who has scored five goals in the Premier League this season, received his first England call-up this month but was not included for the Italy game.
He has been named in the Three Lions' 23-man squad to face Germany, but Manchester City midfielder Jack Grealish is out through suspension.
The other omissions are squad selections and not due to injury.
How does Alexander-Arnold compare?
Alexander-Arnold tops the standings for chances created by an English full-back so far this season in all club competitions with an average of 2.7 per match.
The 23-year-old has scored two league goals for Liverpool this season, and played 737 minutes - more than any other English full-back.
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Centre-back Tomori also did not feature against Italy despite playing regularly for AC Milan in Serie A.
Instead, manager Gareth Southgate opted to start Harry Maguire, who has made just three appearances in the Premier League for Manchester United this season.
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Monday's match at Wembley is England's final fixture before Southgate announces his squad for the 2022 World Cup, which is being held in Qatar from 20 November to 18 December.
England are on a five-match winless run and Friday's 1-0 defeat in Milan saw them relegated from their Nations League group.Gareth Southgate's squad dilemmas in statistics
BBC Sport's Alex Howell
The omission of Alexander-Arnold shows where he is in the England right-back pecking order and probably the thoughts of England manager Gareth Southgate.
Kyle Walker is the first choice right-back when England play a back four and is one of the back three in Southgate's favoured formation. Reece James has also seemed to nudge ahead of Alexander-Arnold because he offers an attacking threat as well as more defensive solidity, and it can also be argued that Kieran Trippier is ahead of the Liverpool player as well because of his versatility.
There is no doubting the talent of Alexander-Arnold and the fact that he is one of England's best passers of the ball. His attacking output at Liverpool is excellent but with England he doesn't get the same freedom as with his club.
Southgate is criticised for being cautious but his approach has helped England reach a semi-final of a World Cup and a first final in a European Championship, so he has justification for preferring certain players. And that is without mentioning that Alexander-Arnold missed the Euros with injury.
Being left out of the of the last matchday squad before the team heads to a World Cup will be a big blow for him and one that will be felt more because he did not come off of the bench in Italy, so will leave this camp without playing a minute.
There's no doubt that after this international break Alexander-Arnold's place in the 26-man squad for Qatar is uncertain.