The victims were initially identified only as a woman and man. Their names were not immediately available for release.
The wreck happened around 1:00 p.m. near Yantley, Ala.
"Two vehicles were involved. The driver of each was a fatality," said Trooper Spencer Collier of the Alabama Highway Patrol. "It was a head-on."
Collier said homicide investigators will work to determine the cause of the crash.
The wreck closed the highway for several hours as wreckage was cleared.
Fire units from Lisman, Yantley, Pennington and Butler responded, along with the Alabama Highway Patrol.