Culture and heritage group Italia Nostra argued that Leonardo's "Vitruvian Man" sketch was too fragile to be transported to the Louvre, where it was expected to be included in an exhibition marking the 500th anniversary of the artist's death.
Italia Nostra launched an appeal with a regional court to halt the sketch's loan, which was upheld by a court ruling on Tuesday.
Italia Nostra argued that the work was too fragile to be transported, and could be damaged if displayed in the Louvre.
It also expressed concern that if the work were exhibited in Paris, Venice's Gallerie dell'Accademia -- where the sketch is held -- would be forced to conceal it for several years on its return to limit the risk of environmental exposure.
"Vitruvian Man" can only be displayed for short periods to limit environmental damage.
The exhibition at the Louvre -- where Leonardo's most famous work, the Mona Lisa, is admired by some 20,000 people a day -- is due to open on Oct. 24.
Alongside its own collection of Leonardo paintings, the museum said it would display "nearly 120 works" from European and American institutions.
In a statement published online, Italia Nostra said it welcomed the court's decision to suspend the loan, and noted that authorities would make a final decision on the loan later in October.
Amid simmering tensions between Rome and Paris earlier this year, Italy's junior culture minister, Lucia Borgonzoni, threatened to cancel the loan, adding, "there are plenty of things in the Louvre that should be returned to us, even above and beyond the old controversy surrounding the Mona Lisa."
Representatives for the Louvre did not respond to CNN's request for comment.
CNN's Melissa Bell and Saskya Vandoorne contributed to this report.