All five victims, four women and another person, were transported to a local hospital; one of the women was pronounced dead there.
The gender of the fifth victim was not immediately announced.
The shooting happened around 3 a.m. ET Thursday at the Days Inn in the 4400 block of Connecticut Avenue NW.
Fox 5 D.C. reported that officers arrived at the hotel to find one female who had been shot in a hallway and another shot in a room. Three other gunshot victims transported themselves to the hospital.
Streets surrounding the hotel remained closed as police continue to investigate.
Fox News' Audrey Conklin contributed to this report.