Batgirl's directors say they are "saddened and shocked" after Warner Bros decided to scrap the film before its release date.
Leslie Grace, who was cast as Batgirl, also tweeted to say she was "proud of the love, hard work and intention" put into the film by all involved.
The production was due for release in 2022, but the studio has shelved the project and it will no longer appear in theatres or on the streaming service HBO Max.
Directors Adil El Arbi and Bilall Fallah posted a short statement on Instagram saying they wished fans could have had the "opportunity to see and embrace the final film themselves".
The Bad Boys For Life filmmakers said: "We are saddened and shocked by the news. We still can't believe it.
"As directors, it is critical that our work be shown to audiences, and while the film was far from finished, we wish that fans all over the world would have had the opportunity to see and embrace the final film themselves. Maybe one day they will insha'Allah (if God wills).
"Our amazing cast and crew did a tremendous job and worked so hard to bring Batgirl to life.
"We are forever grateful to have been part of that team. It was a dream to work with such fantastic actors like Michael Keaton, J.K. Simmons, Brendan Fraser, Jacob Scipio, Corey Johnson, Rebecca Front and especially the great Leslie Grace, who portrayed Batgirl with so much passion, dedication and humanity."
They added: "In any case, as huge fans of Batman since we were little kids, it was a privilege and an honor to have been a part of the DCEU, even if it was for a brief moment," and signed off their statement "Batgirl For Life".
A tweet by Grace, pinned to the top of her account, says: "Querida familia! [Dear family!]
"On the heels of the recent news about our movie Batgirl, I am proud of the love, hard work and intention all of our incredible cast and tireless crew put into this film over seven months in Scotland."
Why has Batgirl been shelved?
A spokesperson for Warner Bros said: "The decision to not release Batgirl reflects our leadership's strategic shift as it relates to the DC universe and HBO Max.
"Leslie Grace is an incredibly talented actor and this decision is not a reflection of her performance."
Warner Bros added that they were "incredibly grateful" to the filmmakers and cast, and hoped to collaborate with everyone again in the future.
However, the New York Post reported claims that test screenings for audiences were poorly received by filmgoers and that the studio decided to "cut its losses".
Which Hollywood stars were due to appear in Batgirl?
The film was set to feature Grace in the titular role of Batgirl - also known as Barbara Gordon, the only daughter of Gotham police commissioner Jim Gordon.
The 27-year-old actor is known for her role as Nina Rosario from the musical drama In the Heights. She is also an acclaimed singer-songwriter.
Michael Keaton was to star as Bruce Wayne / Batman, with Brendan Fraser cast as supervillain Firefly, and JK Simmons as Commissioner Gordon.
Where was Batgirl filmed?
The production was being filmed in Glasgow.
In January, the Trongate area of Glasgow was transformed by set-builders into the famous Gotham city, with alterations made to shop fronts and police cars.
Several cast and crew members, including Grace and co star Ethan Kai, as well as directors Adil El Arbi and Bilall Fallah, were pictured filming in Scotland.
Glasgow City Council had agreed to give Warner Bros £150,000 to shoot the production in the city, but confirmed this has not been paid.
A Glasgow City Council spokesman said: "The filming of Batgirl brought a very significant economic benefit to Glasgow when the production was here, with 450 Glasgow-based crew jobs, almost 1,300 Scottish supporting artists, and many subcontractors working as tradespeople and in traffic management and security.
"Any decision on the release of the production is very much a matter for Warner Bros. The £150,000 grant support has not been paid, and discussions continue with the producers."
How much did Batgirl cost to make?
The production reportedly cost an estimated $70 million (£57.6 million) - though some sources put the budget at up to $100m (£83m), or even higher.
Have any other productions been scrapped?
Another Warner Bros film due for release - Scoob! Holiday Haunt - has also been cancelled.
The film was due for release in 2022, with the storyline centered on character Scooby Doo's first Christmas.