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Pembroke, Maine was settled in 1774 by its first pioneer, Hatevil Leighton, followed by Edmund Meagher (Mahar) and William Clark in 1780 who settled near Cobscook Falls - known today as Reversing Falls.

Incorporated February 4th, 1832, Pembroke was named for a city in Wales.

By 1870 the population had risen to 2,551 compared to the most recent census of 879 full-time residents.

Perhaps one of the most notable persons born in Pembroke, Maine was Dr. Charles Best who co-discovered insulin with Dr. Frederick Banting. In 1923, Dr. Banting and J.J.R. Macleod were awarded the Nobel Prize for Medicine for that discovery. Dr. Banting was so outraged that Dr. Best was overlooked he gave Best half of his prize money. Dr. Best later became an adviser to the medical research committee of the United Nations World Health Organization.