In 1591, when Kazy-Girey approached Moscow, Tsar Fyodor Ioannovich ordered a procession with the Don Icon of the Mother of God around the city walls and then placed it in the field church of the Russian army. The night before the icon, the tsar had a vision confirming his actions. When the enemy approached the camp, several skirmishes occurred, but the battle never started. Kazy-Girey's army stood for a day and suddenly retreated in panic, leaving behind the wounded and equipment.

The Don Monastery was founded at the site where the Russian troops stood before the battle with the Tatars. A field church with the Don Icon was established in the center of the camp. The Muscovites referred to the monastery as the abode of the Mother of God "that is in the convoy."

With the Don Icon, St. Sergius of Radonezh blessed Prince Dmitry Ioannovich for the Battle of Kulikovo in 1380. After that, the icon was kept in the Assumption Cathedral of Kolomna, then transferred to the Moscow Church of the Nativity of the Mother of God, and later to the Annunciation Cathedral of the Kremlin. Since then, the icon became known as the "Don Icon." Each time Moscow faced danger, prayers for help were offered before it.

With the founding of the monastery, a stone cathedral in honor of the Don Icon was built, which later became the refectory. The Don Monastery became part of the defensive ring of monastery-fortresses around Moscow, protecting it from enemies. It was conceived as a stronghold on the Kaluga road.

In the early 17th century, the monastery was captured and looted by the Poles under the leadership of Khodkevich. After that, it fell into decline and was annexed to the Andronikov Monastery. The monastery was revived in the 1620s thanks to Tsars Mikhail Fyodorovich and Alexei Mikhailovich, becoming a place of pilgrimage.

Construction of the New Cathedral and monastery walls, reminiscent of the walls of the Novodevichy Monastery, began in 1684. In 1705, by order of Peter I, Archimandrite Lavrenty became the abbot, and the monastery became a center of Russian-Georgian relations. Representatives of the Georgian nobility found their resting place here.

In 1745, the monastery received the status of stavropegial. After the plague epidemic of 1771, the monastery cemetery became a burial place for the Moscow nobility. By the end of the 18th century, the monastery became one of the most revered in Moscow.

In 1812, the monastery was looted by Napoleon's army, but the buildings survived. In the 19th century, the architectural ensemble continued to take shape. In 1834, a spiritual school was opened to prepare students for the seminary.

After the October Revolution of 1917, Patriarch Tikhon was located in the monastery. In 1926, the monastery was closed, and its buildings were used for a museum and other needs. In 1935, a museum of Russian architecture opened here.

In 1948, the Small Cathedral was returned to the Moscow Patriarchate. In 1990, the monastery fully returned to the Church's administration. Since then, restoration work has been underway, new churches have been built, and various educational and cultural institutions are in operation.