Combs, 55, entered his plea to the new indictment before US District Judge Arun Subramanian at a hearing in Manhattan federal court. A trial remains scheduled for May 5.
With several of his family members looking on from the courtroom audience, Combs was led in to the courtroom sporting grey hair, a grey beard, and a baggy, tan jail outfit.
Combs previously pleaded not guilty to charges of racketeering conspiracy, sex trafficking and transportation to engage in prostitution. While the new indictment added no new charges, it described what prosecutors called the "forced labour" that Combs demanded in connection with the 20-year alleged racketeering conspiracy.
It said Combs and his associates "maintained control" over some employees by forcing them to work long hours with little sleep, through the use of or threats to use physical force, financial harm, psychological harm and reputational harm.
Combs' defence lawyer, Marc Agnifilo, has said his client never forced anyone to engage in sexual acts against their will.
Prosecutors with the Manhattan US Lawyer's Office said Combs used his business empire, including his record label Bad Boy Entertainment, to sexually abuse women between 2004 and 2024.
Combs' alleged abuse included having women take part in recorded sexual performances called "freak offs" with male sex workers, who were sometimes transported across state lines.
Agnifilo has said the freak offs were consensual sexual activity.
Combs also faces dozens of civil lawsuits by women and men who accused him of sexual assault and other misconduct. He has denied all wrongdoing.
Friday's hearing is also expected to discuss jury selection, including the potential difficulty in finding an impartial jury given Combs' fame and the substantial media coverage of his criminal case.
Prosecutors want jury selection to begin on April 21, to help ensure the trial does not run past the July 4 holiday. Defence lawyers want jury selection to begin on May 5, saying an earlier date would be unfair to them.
Combs has been held in a Brooklyn jail awaiting trial since his September 2024 arrest.
Also known during his career as Puff Daddy and P Diddy, Combs founded Bad Boy Records and is credited with helping turn rappers and R&B singers such as Mary J Blige, Faith Evans, Notorious BIG and Usher into stars in the 1990s and 2000s.
His career has been derailed since federal agents raided his Los Angeles and Miami Beach, Florida, homes in early 2024 as part of the sex trafficking investigation that led to his indictment.
