In 1501, by order of his mother Ginevra, he had several members of the Marescotti family, rivals of the Bentivoglio, killed.
In 1506, when Pope Julius II ordered the Bentivoglio to leave Bologna, he took refuge first in Ferrara and then in Mantua, together with Annibale.
A reward was raised against them by the Pope; in spite of it, the two brothers repeatedly attempted to reconquer their city until they succeeded in 1511, thanks the help of the French.
Annibale was declared new lord of Bologna; the following year, however, once the French left the population rose against them and the Bentivoglio had to leave the city forever.
He is portrayed, as a child, in Lorenzo Costa the Elder's Bentivoglio Altarpiece in San Giacomo Maggiore.