The deceased was identified as Babu, police said.
The injured persons were identified as Deepak (25) and Balwant (24), they said.
They were referred to PGI Chandigarh where their condition was stated to be critical. All of them were residents of Dhulkot village and nearby areas located on the highway.
The car collided with the bike near Baldev Nagar police station at Ambala city here.
The car driver, who escaped unhurt, has been arrested by the police.