The uncapped Tyrone Mings, Aaron Wan-Bissaka, Mason Mount and James Maddison have all received England call-ups as Gareth Southgate surprisingly left out right-back Kyle Walker.
Tottenham midfielder Harry Winks was recalled and Liverpool's Alex Oxlade-Chamberlain was also named in the squad for European Championship qualifiers against Bulgaria and Kosovo, despite making just two starts since suffering a serious knee injury in April 2018.
Southgate has brought Kieran Trippier back into the fold after omitting him from the Nations League squad and there is a maiden call-up for Wan-Bissaka, leaving World Cup semi-finalist Walker conspicuous by his absence from the 25-man squad.
Fellow Manchester City defender John Stones, who has had a thigh problem, also missed out with Aston Villa defender Mings receiving his first call-up.
Chelsea playmaker Mount and Leicester's Maddison have both previously been part of the senior squad and have been included for the upcoming qualifiers.
"We only have eight matches before I name the squad for the European Championships," Southgate said.
"We learned some lessons from last September on the importance of physical fitness at this time of year, also some exciting players we wanted to look at and involve."
Wan-Bissaka joined Manchester United from Crystal Palace in the summer and Southgate explained his inclusion ahead of Manchester City defender Walker.
"We tracked him very closely last year and thought he had an outstanding season last season, but he should stay with the Under-21 group," the Three Lions boss said. "He's adapted very well to a high profile move with the intensity and pressure and we are really looking at him and Trent (Alexander-Arnold).
"We also want to reintegrate Kieran Trippier to know he is still on our radar. It's an area of the pitch where we have a lot of quality. Kyle started the season really well so I explained the situation to him. They all have different strengths."
Southgate said of 26-year-old Mings: "I watched him last year and wanted to see how he adapted to the Premier League.
"He's an interesting case as he entered the game a bit later and hasn't played as much as a centre-back. I see a lot of potential and time for growth given the number of games he's played."
Villa paid an initial £20million - rising to £26.5million - to make Mings' loan from Bournemouth permanent in the summer and, in an inflated market, club manager Dean Smith feels he is a steal.
He said: "I thought it was a tough one to pay but one well worth paying. When you look at the market after that and what some teams have paid, it looks a snip, that's for sure.
"At the time I thought we were paying over the odds but it was a deal worth doing because we knew what a player he could become."
Mount spent last season on loan at Derby under current Chelsea boss Frank Lampard, helping him to hit the ground running this term.
Southgate said: "Mason Mount has been an outstanding player in the league this season.
"He's not in on the back of three games for Chelsea, it's on a year with Derby and the Euro Under-19 Championship with us."
Goalkeepers: Tom Heaton (Aston Villa), Jordan Pickford (Everton), Nick Pope (Burnley)
Defenders: Trent Alexander-Arnold (Liverpool), Ben Chilwell (Leicester), Joe Gomez (Liverpool), Michael Keane (Everton), Harry Maguire (Manchester United), Tyrone Mings (Aston Villa), Danny Rose (Tottenham), Kieran Trippier (Atletico Madrid), Aaron Wan-Bissaka (Manchester United)
Midfielders: Ross Barkley (Chelsea), Jordan Henderson (Liverpool), James Maddison (Leicester), Mason Mount (Chelsea), Alex Oxlade-Chamberlain (Liverpool), Declan Rice (West Ham), Harry Winks (Tottenham)
Forwards: Harry Kane (Tottenham), Jesse Lingard (Manchester United), Marcus Rashford (Manchester United), Jadon Sancho (Borussia Dortmund), Raheem Sterling (Manchester City), Callum Wilson (Bournemouth)