Known widely as the Central Park Five, the defendants spent between five and 13 years in prison before they were cleared in 2002 based on new DNA evidence and the confession of another person.
Trump, the Republican nominee for the White House, falsely said at the September 10 debate with Democrat Kamala Harris that the Central Park Five had killed a person and pleaded guilty.
A spokesperson for Trump's campaign did not immediately respond to a request for comment on the lawsuit, filed in federal court in Philadelphia on Monday.
The lawsuit by Yusef Salaam, Raymond Santana, Kevin Richardson, Antron Brown and Korey Wise called Trump's statements "demonstrably false."
A lawyer for the men, Shanin Specter, said in a statement that Trump's remarks "cast them in a harmful false light and intentionally inflicted emotional distress on them."
The plaintiffs are seeking unspecified monetary damages for reputational and emotional harms as well as punitive damages.
Trump has drawn criticism before over his statements about the Central Park Five. After the jogger's assault, he spoke out about the case and took out a full-page ad in several New York newspapers calling for the reinstatement of the death penalty.
Trump asked a US appeals court in September to reject a $US5 million ($A7.5 million) verdict finding him liable for sexually assaulting and defaming the writer E Jean Carroll.
A different jury in January ordered Trump to pay Carroll $US83.3 million ($A125.0 million) for having defamed her and damaging her reputation in June 2019 after she first accused him of rape.Â
