Formula 1 hopefuls Stefan GP have had their bid to race in 2010 rejected.
The Serbian team had hoped to race in place of another new team, US F1, which has withdrawn after failing to fulfil its obligations for the new season.
Governing body the FIA said in a statement: "It is not possible for a replacement team to be entered for the championship at this late stage."
The decision means 12 teams will be on the grid when the season starts in Bahrain on 14 March.
Stefan had been campaigning vigorously to be given a place on the grid, and had been given the support of F1 commercial boss Bernie Ecclestone.
The team, run by Serbian businessman Zoran Stefanovich, had obtained the cars that were designed and built by the Toyota team before the Japanese manufacturer quit the sport at the end of last season.
Stefan had stepped up its campaign in recent weeks as it became clear that US F1 would not make the grid.
The FIA said in a statement that US F1 had "indicated that they will not be in a position to participate in the 2010 FIA Formula 1 World Championship.
"Having considered the various options, the FIA confirms that it is not possible for a replacement team to be entered for the Championship at this late stage."
Full 2010 entry list:
1 Jenson Button (GB) McLaren-Mercedes
2 Lewis Hamilton (GB) McLaren-Mercedes
3 Michael Schumacher (Ger) Mercedes
4 Nico Rosberg (Ger) Mercedes
5 Sebastian Vettel (Ger) Red Bull-Renault
6 Mark Webber (Aus) Red Bull-Renault
7 Felipe Massa (Brz) Ferrari
8 Fernando Alonso (Spa) Ferrari
9 Rubens Barrichello (Brz) Williams-Cosworth
10 Nico Hulkenberg (Ger) Williams-Cosworth
11 Robert Kubica (Pol) Renault
12 Vitaly Petrov (Rus) Renault
14 Adrian Sutil (Ger) Force India-Mercedes
15 Vitantonio Luzzi (Ita) Force India-Mercedes
16 Sébastien Buemi (Swi) Toro Rosso-Ferrari
17 Jaime Alguersuari (Spa) Toro Rosso-Ferrari
18 Jarno Trulli (Ita) Lotus-Cosworth
19 Heikki Kovalainen Lotus-Cosworth
20 TBA HRT-Cosworth
21 Bruno Senna (Brz) HRT-Cosworth
22 Pedro de la Rosa (Spa) BMW Sauber-Ferrari
23 Kamui Kobayashi (JPN) BMW Sauber-Ferrari
25 Timo Glock (Ger) Virgin-Cosworth
26 Lucas di Grassi (Brz) Virgin-Cosworth