The Iroquois Six Nations, Shawnee, Cherokee, and other tribes gave up land claims in what is now West Virginia through various treaties, though not all tribal views were represented.

In 1744, English representatives from Pennsylvania, Maryland, and Virginia met with Iroquois leaders to resolve land disputes. The Iroquois believed they had sold land only in southern Virginia, not most of West Virginia, leading to a conflict that lasted until the 1768 Treaty of Fort Stanwix. The Treaty of Lancaster (1744-1755) allowed over 1,000 settlers into eastern West Virginia without resistance.

The 1768 Treaty of Fort Stanwix forced the Iroquois to give up most of Western Virginia, except for the Northern Panhandle, in exchange for goods and money. The 1768 Treaty of Hard Labor defined a western boundary from Wytheville, Virginia, to Point Pleasant. The Treaty of Lochaber (1770) gave Britain the Cherokees' land in western Virginia, and the 1774 Treaty of Camp Charlotte established the Ohio River as the boundary with Shawnee territory, bringing temporary peace during the early American Revolution.