The pair, reported to be a 34-year-old male and his 26-year-old female companion, fell 10 metres from the mediaeval fortifications at the picturesque tourist town.
Local radio station Dubrovacka Televizija reported that the tourists were thought to have been kissing on the ramparts.
A spokesperson for Dubrovnik-Neretva told media they had not climbed the wall, but alcohol may have played a part in the accident.
The incident occurred shortly before 2.30am on Saturday morning, local time.
Blood alcohol readings for the tourists were just over 0.2 for the man and 0.3 for his partner.
A trauma specialist at Dubrovnik General Hospital said the woman remained in a critical condition and the man had undergone surgery for his injuries.
Dr Ivan Bencic said the woman had suffered brain injuries and multiple fractures to her spine. She was receiving care from a neurosurgeon and several CT scans, but remained in a critical condition.
“We are constantly monitoring her and doing everything we can to fight for her life,” he said.
“As far as the male is concerned, he had a far better run than she did.”
The as-yet-unnamed visitors are both thought to be Australians.
The Department of Foreign Affairs and Trade in Croatia said they were providing consular assistance to two injured Australian nationals but would not provide any further information over privacy concerns for the tourists.
The historic quarter of Dubrovnik by the Pile Gate, is known for its impressive 16-century fortifications and Fort Lovrijenac. It received huge tourist attention after appearing in the fantasy television series, Game of Thrones.
The suburb is characterised by 10-metre-high stone walls and the rocky Danče beach, below.