The Football League insist Southampton's new owners will not be allowed to appeal against the club's ten-point deduction.
The decision means a takeover by the Pinnacle group, which is backed by Matt Le Tissier, may not now take place.
Saints were docked the points after the club's parent company, Southampton Leisure Holdings plc, went into administration earlier this year and they will now start their 2009-10 League One campaign on minus ten points.
A Football League statement said: "The board of the Football League met to consider representations from the Pinnacle Group with regard to their proposed takeover of Southampton Football Club.
"The Football League has the responsibility to ensure all League clubs start the season with certainty as regards the competition they compete in.
"With that in mind, the Football League has responded directly to the Pinnacle Group, and will continue to liaise with their representatives going forward."