Juventus have been docked 10 points following a new hearing into the club’s past transfer dealings.
Juve were initially handed a 15-point penalty in January but Italy’s highest sporting court overturned that decision in April and ordered the case to be re-examined.
The new ruling was announced on Monday, before the team’s game away to Empoli.
The penalty drops Juve to seventh place in Serie A, outside the European qualification spots.
Napoli have already secured the Serie A title and, before Monday’s ruling, were 17 points ahead of second-placed Juve.
In April, a tribunal at Italy’s Olympic Committee (CONI) revoked Juve’s 15-point penalty from the Italian Football Federation (FIGC) appeals court.
It asked the appeals court to revise punishments to the club and some key figures, including former vice-chairman Pavel Nedved.
Long bans given to former chairman Andrea Agnelli, ex-chief executive Maurizio Arrivabene and sporting directors Federico Cherubini and Fabio Paratici have now been upheld.
A Juventus statement said that the club “takes note” of the decision and “reserves the right to read the reasons to evaluate a possible appeal” to CONI.
It added: “What was established by the fifth instance of judgment in this matter, which began more than a year ago, arouses great bitterness in the club and in its millions of supporters who, in the absence of clear rules, find themselves extremely penalised with the application of sanctions that seem to take into account the principle of proportionality.
“While not ignoring the need for urgency, which Juventus has never shied away from during the proceedings, it is emphasised that these are facts that still have to be evaluated by a judge.”
If Juve beat Empoli, they will be two points behind AC Milan, who occupy the final Champions League spot, with two games to play.
The club is also involved in criminal proceedings over its accounting practices, with 12 current and former officials, including Agnelli, potentially facing trial.
According to Italian news agencies, a high court hearing on 26 October will decide whether any eventual trial will be held in Turin, where the investigation has taken place, or in either Milan or Rome.