Joe Worsley is set to miss out on the World Cup after being cut from England's training squad.
The Wasps back-row forward, who has won 78 England caps, is one of five players released by manager Martin Johnson.
Leicester's George Chuter and Thomas Waldrom, Saracens wing David Strettle and Gloucester back James Simpson-Daniel have also been let go.
Johnson has retained a 40-man squad in preparation for England's forthcoming back-to-back matches against Wales.
The 34-year-old Worsley made his Test debut in the 1999 World Cup and was part of the victorious squad in 2003. He was a replacement in the 2007 final defeat by South Africa.
New Zealand-born number eight Waldrom has been struggling with a calf injury, meaning the Harlequins captain Chris Robshaw is still pushing for World Cup selection.
Chuter's departure leaves England with three hookers in Dylan Hartley, Steve Thompson and Lee Mears - all of whom are now likely to travel to New Zealand.
Strettle and Simpson-Daniel have been overtaken in the pecking order by Gloucester's Charlie Sharples and Harlequins wing Ugo Monye.
"Thomas had an injury to his calf and he hasn't been able to train that much," said Johnson, who captained England to World Cup victory in 2003.
"He has gone back to Leicester to participate in their pre-season.
"The other decisions have all been tough because everyone has participated and contributed. It is particularly tough on Stretts (Strettle) because he has trained very, very well.
"Simpson-Daniel has had a couple of niggling injuries and a bit of illness. Joe has come back from some injuries last year and worked incredibly hard to get back in but has come up a little bit short right now."
Johnson will select a final squad of 30 on 22 August before flying to New Zealand. The World Cup begins on 9 September with England's first game against Argentina in Dunedin the following day.
England World Cup training squad
Forwards (22): Props: Dan Cole (Leicester Tigers), Alex Corbisiero (London Irish), Paul Doran-Jones (Northampton Saints), Tim Payne (London Wasps), Andrew Sheridan (Sale Sharks), Matt Stevens (Saracens), David Wilson (Bath Rugby). Hookers: Dylan Hartley (Northampton Saints), Lee Mears (Bath Rugby), Steve Thompson (London Wasps). Locks: Mouritz Botha (Saracens), Louis Deacon (Leicester Tigers), Courtney Lawes (Northampton Saints), Tom Palmer (Stade Francais), Simon Shaw (London Wasps). Back row: Tom Croft (Leicester Tigers), Nick Easter (Harlequins), Hendre Fourie (Sale Sharks), James Haskell (Unattached), Lewis Moody (Bath Rugby), Chris Robshaw (Harlequins), Tom Wood (Northampton Saints).
Backs (18): Full-backs: Delon Armitage (London Irish), Ben Foden (Northampton Saints). Wings: Chris Ashton (Northampton Saints), Mark Cueto (Sale Sharks), Ugo Monye (Harlequins), Charlie Sharples (Gloucester Rugby) Centres: Matt Banahan (Bath Rugby), Riki Flutey (London Wasps), Shontayne Hape (London Irish), Mike Tindall (Gloucester Rugby), Manusamoa Tuilagi (Leicester Tigers). Fly-halves: Toby Flood (Leicester Tigers), Charlie Hodgson (Saracens), Jonny Wilkinson (Toulon). Scrum-halves: Danny Care (Harlequins), Joe Simpson (London Wasps), Richard Wigglesworth (Saracens), Ben Youngs (Leicester Tigers).