Fadrique de Ribera, who died 1539, established in his will that a hospital was to be constructed opposite the Macarena Gate. It was to have "such a high quality that it will be regarded as a perpetual work". It was named the Hospital de Las Cinco Llagas and at the time, with more than one thousand beds for both men and women, it was the largest of its kind in Europe. The hospital remained in use until 1972. After several years of neglect, the building was restored and since 1992 has been the seat of Andalusia's Regional Parliament.Typical renaissance hospitals constructed during the times of the Catholic Monarchs had a large rectangular layout with two crossing naves forming four courtyards. Gainza's design from 1546 observed the rules of the time and included a church in the intersection of the naves. This was later changed by Hernán Ruiz II who built a separate church in one of the front courtyards in 1584.

According to Chueca Goitia, this "was the most grandiose design ever for a classic religious building in Spain before the monastery of El Escorial". The monumental main doorway consists of two sections with double columns which support a straight pediment crowned by large vases. In 1564 Juan Bautista Vázquez "the Old" carved the marble reliefs on the arch at the entrance which depicts the theological Virtues. It has a rectangular layout and a single nave. The chapels built between the buttresses gave rise to a new style which was to have great influence in Sevillian Baroque, as single nave functional churches were preferred during the Counter-reformation period because they improved visibility and acoustics for following the Mass.