| Pope Alexander VI | formerly Cardinal Rodrigo Borgia |
| Cardinal Cesare Borgia | afterwards Duc de Valentinois and Duke of Romagna, the Pope’s son |
| Don Joffré | Duke of Squillace, the Pope’s younger son |
| Louis XII | King of France |
| Don Juan | King of Navarre |
| Cardinal Francesco Borgia | cousin to the Pope |
| Cardinal Ippolito d’Este | son of the Duke of Ferrara |
| Cardinal Giuliano Della Rovere | afterwards Pope Julius II and other Cardinals. |
| Prince Don Alfonso | Duke of Bisceglia, a natural son of the King of Naples, husband to Lucrezia Borgia, after her divorce from Giovanni Sforza |
| Prince Djem | the Sultan’s brother and the Pope’s hostage |
| The Bishop of Venosa | the Pope’s Private Physician |
| Monsignore Bonafede | Bishop of Chiusi |
| Monsignore Burchard | Master of the Ceremonies |
| Monsignore Gaspare Poto | the Pope’s Private Chamberlain |
| Monsignore Gaspare Torella | Cesare Borgia’s Physician |
| Cavaliere Vincenzo Calmeta | a poet |
| Don Pedro de Torpia | Cesare Borgia’s Spanish jailer |
| Don Michelotto Corella | one of Cesare Borgia’s captains |
| Don Federico Altieri | a young Roman gentleman |
| Don Garcilaso de la Vega | Spanish Ambassador |
| Messer Niccolo Macchiavelli | Florentine Envoy |
| Messer Bernardino Betti (Pintoricchio) | a painter |
| Messer Ercole | a goldsmith and metal-worker |
| Messer Cristofero | Lucrezia Borgia’s Secretary |
| Messer Agapito da Amalia | Cesare Borgia’s Secretary |
| Messer Pincione | an apothecary |
| Juanito Grasica | Cesare Borgia’s page |
| Garcia de Magona | a Spanish boy |
| Giorgio | a waterman |
| Donna Lucrezia Borgia | the Pope’s daughter |
| Donna Adriana Borgia | the Pope’s cousin |
| Donna Angela Borgia | Maids of Honour to Lucrezia |
| Donna Hieronyma Borgia | Maids of Honour to Lucrezia |
| Donna Sancia d’Aragon | sister to Don Alfonzo and wife to Don Joffré Borgia |
| Mademoiselle Charlotte d’Albret | afterwards wife to Cesare Borgia |
| Donna Vanozza de’ Catanei | once the Pope’s mistress, and the mother of Cesare and Lucrezia Borgia |
| Donna Giulia Farnese (La Bella) | the Pope’s young mistress |
| Donna Fiammetta | A Roman woman, Cesare Borgia’s mistress |
| Donna Catilena de Valence | Maid of Honour to Lucrezia |
| Suor Lucia | an Anchoress |
| Clarice | Maid to Lucrezia |
| A Mute, Shepherds, Citizens of Rome, Attendants, Bargemen, Girls and Women |
