In the 4th century BCE, Babylon was conquered by Alexander the Great, who used his Staff of Eden given to him by the Order to create one of the largest empires in history. In 323 BCE, the Babylonian proto-assassin Iltani, realizing that she would not be able to take Alexander out through force, set out to find an Achaemenid alchemist, hoping to expand her knowledge of poisons. After a perilous journey, she arrived in Babylon, where she found the alchemist and learned his secrets.
On 13 June, 323 BCE, Iltani infiltrated the palace of Nebuchadnezzar II in Babylon, where Alexander was residing, poisoned him and retrieved the Staff. Soon afterwards, Alexander's empire began to crumble.