Gloucestershire Police said the crash happened just after 11:45 GMT near Northleach on the A429 towards Cirencester near the A40 junction.
The driver of a Jaguar, a 56-year-old man from Staffordshire, was taken to hospital in Bristol with serious but not life threatening injuries.
A 46-year-old man from Cheltenham and a female passenger in a Ford Focus suffered minor injuries.
The lorry from Moreton-in-Marsh was carrying a load of Cotswold stone. The driver was uninjured, police said.
The road was closed earlier but has now been reopened to traffic.
A lorry carrying stone shed its load on to a car in a crash in the Cotswolds.