The Madonna and Child Enthroned with Saints John the Baptist and Sebastian is a painting by the Italian Renaissance artist Pietro Perugino, executed in 1493 and housed in the Uffizi Gallery, Florence.

The work was commissioned by Cornelia Salviati, widow of the Venetian merchant Giovanni Martini, and his son Roberto, for the chapel of the convent of San Domenico, Fiesole, which had been perhaps restored by Giuliano da Sangallo a few years before.

The Virgin sits on a high throne decorated with grotesques at the base, which also includes the signature PETRVS PERVSINVS PINXIT AN[NO] MCCCCLXXXXIII ('Pietro Perugino painted it, 1493').

The composition was one of the first examples in Florence of the new style of sacra conversazione elaborated in Venice by Antonello da Messina and Giovanni Bellini a few years before, with a pyramidal development pivoting on the central figure of the Virgin on a high throne.

The painting is also one of the first by Perugino in which the Madonna is no longer an elegant maid, but a more mature and severe woman, according to the more sober climate introduced in Florence by Girolamo Savonarola.