The Brisbane Tigers' previous NRL application, submitted as the Brisbane Firehawks in 2021, was defeated by the Dolphins' Redcliffe-based proposal.
Now, the Queensland Cup club is preparing another attempt to secure the competition's 20th licence for 2029.
The bid would make the Tigers Queensland's fifth NRL team and create a second Brisbane-based side alongside the Broncos.
Chief executive Mitch Cook told News Corp the club remains determined to enter the process.
"Yes, we're absolutely still keen to get the licence," Cook said to News Corp.
He said the Tigers had substantial financial support and were arranging discussions with the Australian Rugby League Commission.
"We want to be recognised as a leading contender because we have great financial backing."
The club, formerly called Easts Tigers, has competed in the state system for 94 years and has established strong Queensland community ties.
Cook believes a team south of the Brisbane River would offer an alternative to the Broncos and strengthen the city's market.
The Dolphins have developed a rivalry with Brisbane despite being based at Redcliffe, he said.
A Toowoomba proposal has recently entered the race, while Christchurch's Southern Orcas remain contenders for the expansion position.