In early 1503, Leonardo da Vinci returned to Florence, the city of his youth. The Republican government commissioned the artist to create a mural for the Hall of the Great Council in the Palazzo Vecchio. The chosen subject was the Battle of Anghiari, where, in 1440, the Florentines and their allies defeated the army of the Milanese Duke Filippo Maria Visconti, commanded by Niccolò Piccinino.

Leonardo envisioned a multi-figure composition presenting a sweeping panorama of the battle. At its heart was the struggle of horsemen on a bridge as they fought for the standard with unprecedented intensity, what Leonardo himself called "beastly madness" (pazzia bestialissima). On either side of the river, troops were shown preparing for combat. Following the commission in late 1503, Leonardo spent over a year preparing sketches and a full-scale cartoon (approximately 8 by 18 meters) and began the painting in the Palazzo Vecchio in May 1505. He continued his work for a year until his departure for Milan in June 1506. By that time, the central group of horsemen had evidently been completed. However, Leonardo’s experiment with a new binding medium proved unsuccessful, and the paint failed to adhere properly to the wall. Contemporaries could still admire the mural until the mid-16th century; then the hall was remodeled and decorated with new frescoes by Giorgio Vasari. Consequently, Leonardo’s original work was lost, and it can now be studied only through early copies and sketches. The most famous painted version, known as the Tavola Doria, is currently at the Uffizi Gallery in Florence and is considered a 16th-century copy.

The Battle of Anghiari from the Museum of Christian Culture differs significantly from the Uffizi version. The figure of the right-hand rider, which is missing in the Tavola Doria, is rendered in considerable detail on the St. Petersburg panel. This version also features a foreground landscape, terrain, and the lower part of the composition, which is absent from the Florentine painting. X-ray analysis of the panel from the Museum of Christian Culture revealed a hidden, later overpainted portrait of a man in a beret seated in a chair with spheres on the armrests. Infrared reflectography uncovered preparatory charcoal underdrawings of the figures, subsequently refined with ink applied with theleft hand (Leonardo was famously left-handed). This suggests the Master may have reused a previously prepared panel containing an unfinished portrait for the numerous sketches he was producing. Left unfinished, the work was likely completed by one of Leonardo’s pupils. Experts believe the St. Petersburg painting is a rare piece of evidence documenting Leonardo’s work on the Palazzo Vecchio mural—the "experimental panel" mentioned by the Master’s contemporaries.