The National League Cup, a competition between 16 fifth tier sides and 16 Premier League 2 teams, will launch next month – although four non-league clubs have opted out.
The top 14 teams in last season’s National League not to be promoted, along with the two teams relegated from League Two, were invited to take part.
However, Barnet, Eastleigh, Solihull and Southend have decided not to take up their places, meaning only 12 National League sides will be involved.
Manchester United, Newcastle United and Tottenham are among the 16 clubs whose under-21 sides are in the competition.
The tournament will take the format of eight groups of four, with the top two from each group going through to the knockouts.
As in the EFL Trophy, the Premier League 2 sides will play all of their matches away from home, with the hosts keeping all of the gate receipts.
The combined prize fund for the cup is £1m, provided by the Premier League, and all money will be kept by National League clubs.
The first round of games will be played on Tuesday, 1 October and Wednesday, 2 October, with the final scheduled to take place on either Tuesday, 1 April or Wednesday, 2 April.