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His family were descended from that of the Frederick baronets: Charles's great-great-grandfather Major-General Marescoe Frederick was a younger brother of Sir John Frederick, 4th Baronet.
His father, William Harcourt Isham Mackworth (1806—1872), a younger son of Sir Digby Mackworth, the 3rd Baronet, took the additional surname Dolben after he married Frances, the heiress of Sir John English Dolben, the 4th Baronet.