On Dec. 25, a three-judge panel ordered the 76-year-old center-right politician to pay his wife, Veronica Lario, 56, about $46.9 million annually in alimony, Italy's ANSA news agency reported.
The sentence allowed Berlusconi to keep sole possession of his villa outside Milan and his business empire.
In January, Berlusconi alleged the court's decision was biased because the panel was made up of three women. Berlusconi called it an "unreal" verdict from "feminist and Communist judges."
He then filed an appeal, which was recently rejected, ANSA said Thursday.
The former prime minister can now file a final appeal to Italy's high court.
Lario, a former actress, filed for a divorce from Berlusconi after he attended the 18th birthday party of Noemi Letizia, an aspiring starlet and friend, in Naples in 2009, and accused him of socializing with minors.
In an interview with the gossip magazine Chi, Berlusconi said he regretted the end of his "love story"; however, in mid-December he announced his engagement to Naples-born Francesca Pascale, ANSA said.