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The Eastern Cape Department of Transport said three people died in a head-on collision between a bakkie and seven-seater vehicle on Monday.
Department spokesperson, Unathi Binqose, said six other people were injured in the accident on the N2 near Butterworth.
He said the deceased were in the seven-seater Toyota vehicle and included a four-year-old boy, the driver and another male.
The driver of the bakkie and his passenger were rushed to hospital with severe injuries.
"A case of culpable homicide has been opened with the Butterworth police station," Binqose said.
He said the MEC Xolile Nqata has extended his condolences to the families of the deceased, while wishing a speedy recovery to those who were injured and in hospital.
Binqose said Nqata also appealed to road users to obey the rules of the road to avoid accidents like head-on collisions.