In 332 BCE, Alexander the Great of Macedon conquered the region as part of his campaign against the Persian Empire. After his death in 322 BCE, his generals divided the empire and Judea became a frontier region between the Seleucid Empire and Ptolemaic Kingdom in Egypt. Following a century of Ptolemaic rule, Judea was conquered by the Seleucid Empire in 200 BCE at the battle of Panium. Hellenistic rulers generally respected Jewish culture and protected Jewish institutions.^[88] Judea was ruled by the hereditary office of the High Priest of Israel as a Hellenistic vassal. Nevertheless, the region underwent a process of Hellenization, which heightened tensions between Greeks, Hellenized Jews, and observant Jews. These tensions escalated into clashes involving a power struggle for the position of high priest and the character of the holy city of Jerusalem.^[89]

When Antiochus IV Epiphanes consecrated the temple, forbade Jewish practices, and forcibly imposed Hellenistic norms on the Jews, several centuries of religious tolerance under Hellenistic control came to an end. In 167 BCE, the Maccabean revolt erupted after Mattathias, a Jewish priest of the Hasmonean lineage, killed a Hellenized Jew and a Seleucid official who participated in sacrifice to the Greek gods in Modi'in. His son Judas Maccabeus defeated the Seleucids in several battles, and in 164 BCE, he captured Jerusalem and restored temple worship, an event commemorated by the Jewish festival of Hannukah.^[90]

After Judas' death, his brothers Jonathan Apphus and Simon Thassi were able to establish and consolidate a vassal Hasmonean state in Judea, capitalizing on the Seleucid Empire's decline as a result of internal instability and wars with the Parthians, and by forging ties with the rising Roman Republic. Hasmonean leader John Hyrcanus was able to gain independence, doubling Judea's territories. He took control of Idumaea, where he converted the Edomites to Judaism, and invaded Scythopolis and Samaria, where he demolished the Samaritan Temple.^[91] Hyrcanus was also the first Hasmonean leader to mint coins. Under his sons, kings Aristobulus I and Alexander Jannaeus, Hasmonean Judea became a kingdom, and its territories continued to expand, now also covering the coastal plain, Galilee and parts of the Transjordan.^[92]

Under Hasmonean rule, the Pharisees, Sadducees and the mystic Essenes emerged as the principal Jewish social movements. The Pharisee sage Simeon ben Shetach is credited with establishing the first schools based around meeting houses.^[93] This was a key step in the emergence of Rabbinical Judaism. After Jannaeus' widow, queen Salome Alexandra, died in 67 BCE, her sons Hyrcanus II and Aristobulus II engaged in a civil war over succession. The conflicting parties requested Pompey's assistance on their behalf, which paved the way for a Roman takeover of the kingdom.^[94]