The accident took place just a day after 25 people were charred to death when their bus caught fire on the expressway in Buldhana.
The mishap occurred near Bhojade village in Kopargaon at around 11.30am, a Kopargaon police officer said.
The driver lost control of the wheel and the car dashed the divider, killing all three passengers on the spot, the officer said.
The victims were travelling to Mumbai from Nanded, he said, adding that locals rushed the trio to a hospital, but they were declared dead.
A case under section 304-A (causing death by negligence) and other relevant provisions of the Indian Penal Code has been registered against the driver, the official said.
Twenty-five passengers were charred to death when the bus they were travelling in caught fire after dashing into the divider and turning turtle on the expressway in Buldhana district on Saturday.
There were 33 passengers onboard the ill-fated bus and eight survived the tragedy.
(With inputs from PTI)