The North Port Police Department responded to Mile Marker 177 on I-75 South, between Sumter Boulevard and Toledo Blade Boulevard, about a shooting.
The victim, identified as Javaris Miller, had been shot while driving just south of the Toledo Blade exit.
Miller died at the scene.
Detectives believe Miller was targeted by a group of men from the Lee County area.
It is believed that two vehicles are associated with the shooting death.
There is a surviving witness in the case who is cooperating.
Javaris Miller was driving his 2019 Cadillac XTS from "The Hall" nightclub in Palmetto and drove through North Port at approximately 3:30 a.m.
If anyone was driving on I-75 during this time and has any information on either of the two vehicles that were chasing the victim, please contact Crime Stoppers at 1-800-780-TIPS (8477).
Tips may also be made online at www.southwestfloridacrimestoppers.com or on the P3 Tips mobile app.
Crime Stoppers is offering a reward of up to $3,000.