One of the first churches in Voronezh was dedicated to the Annunciation of the Blessed Virgin Mary. Researchers disagree on the date of its founding. Metropolitan of Kyiv, Eugeniy (Bolkhovitino), believes the church was founded in 1620, while Archpriest Stefan Zverev, Archbishop Dmitriy (Sambikin), and researcher N.I. Polikarpov think it was built along with the founding of Voronezh in 1586.

Initially, the Annunciation Church was wooden and suffered from fires multiple times, after which it was rebuilt and even relocated. In 1682, under the first Bishop of Voronezh, Saint Mitrophan, the church became a Cathedral. On April 19, 1684, at the request of Saint Mitrophan, the Patriarch of Moscow and all Rus, Ioakim, blessed the construction of a stone cathedral, which was completed in 1690. Since 1682, the residence of the ruling archbishops was located on the territory of the Cathedral, and from 1745 to 1822, the Voronezh Spiritual Seminary, established by Bishop Theophilact, was also located there.

After the death of Saint Mitrophan in 1703, his remains were buried in the Cathedral. During the reconstruction of the church, his relics were discovered, and a year later he was canonized by the Russian Orthodox Church. The initiator of the canonization was Archbishop Antoniy (Smirnitskiy), who in 1834 proposed to establish the Annunciation Mitrophan Monastery at the Annunciation Cathedral. The monastery was solemnly opened on September 1, 1836, and the Holy Trinity Cathedral became the Cathedral, while the Annunciation remained an important spiritual center. After the revolution, the church and monastery were closed and destroyed in the 1950s. The main building of Voronezh State University now stands on their site.

The revival of the Annunciation Cathedral began in 1998 at the initiative of Metropolitan Mephodiy in May Day Park, in honor of the 2000th anniversary of the Nativity of Christ. The new Cathedral, like the ancient one, will be consecrated in honor of the Annunciation of the Blessed Virgin Mary, and its exterior will resemble the Vladimir Cathedral, erected in honor of the 900th anniversary of the Baptism of Rus.

On October 15, 1998, Patriarch of Moscow and all Rus Alexiy II consecrated the cornerstone of the new shrine of the diocese during his visit to Voronezh.

The fate of the Annunciation Cathedral and the Mitrophan Monastery is closely linked to Archbishop Tikhon (Nikanorov), a native of Novgorod and a graduate of the Novgorod Spiritual Seminary and St. Petersburg Spiritual Academy. He was appointed to the Voronezh diocese on May 13, 1913, and is remembered as a church builder and the initiator of the canonization of Archbishop Antoniy (Smirnitskiy). Bishop Tikhon participated in the Local Council of the Orthodox Russian Church of 1917-18.

On December 27, 1919 (January 9, 1920), he was hanged on the Royal Gates of the altar of the Annunciation Cathedral. Along with him, 160 people accepted martyrdom. At the Jubilee Archpastoral Council of 2000, he was canonized as a new martyr. His relics rest in the cemetery of the Alexeyev-Akatov Monastery in a common grave of Voronezh archpastors, whose remains were transferred there in 1993.