Juventus, who were demoted to the second division last month with a 17-point penalty for match-fixing, officially lodged their appeal with a civil court on Thursday, a move which could lead to further sanctions against them if unsuccessful.
It follows an unsuccesful appeal to the Italian Olympic Committee (CONI), Italy's highest sports authority.
The civil court that Juve are turning to in a bid to be reinstated in Serie A is the Regional Administrative Court of Lazio, or TAR as it is known in Italy. The TAR has the authority to overturn rulings made by sports federation bodies, but is normally reluctant to intervene in sporting issues.
The FIGC (Italian football federation), who handed the Turin club their punishment, wanted to keep the matter in the hands of sports bodies rather than the criminal courts and fear the move could delay the start of the domestic season, which is due to begin on Sept. 9-10.
Such action is also against the regulations of FIFA, world soccer's governing body.
On Wednesday the FIGC told FIFA they would sanction clubs who have infringed Article 27 of its rules, which forbids clubs taking cases to civil courts.
Article 61-2 of FIFA's rules also states that appealing to civil courts is forbidden and that the only other alternative is the Lausanne-based Court of Arbitration for Sport (CAS).
However, despite the threat of further penalties, Juventus remain defiant.
"We retain that the sanctions are unjustified and our treatment shows a lack of respect when you compare our punishment with those given to the other guilty clubs," said a statement issued by Juventus.
Juventus are also considering a damages claim against the FIGC. The club predict relegation to Serie B will cost them 130 million euros (US$165 million) in lost revenue.
Juventus, whose former general manager Luciano Moggi was the central figure in the match-fixing scandal, were the only club of the five found guilty of match-fixing to be relegated from Italy's top division.
They were also stripped of last season's Serie A title and the one they won in 2004-05.