In a blow to President Donald Trump, Senators voted 63-37 to take forward a motion on ending US support.
Secretary of State Mike Pompeo and Defence Secretary Jim Mattis had urged Senators not to back the motion, saying it would worsen the situation in Yemen.
The vote comes in the wake of the murder of Saudi journalist Jamal Khashoggi, a US resident.
Criticism of Saudi Arabia has grown since the prominent writer was killed while visiting the Saudi consulate in Istanbul in Turkey on 2 October.
The US Senators, who met in a closed session on relations with Saudi Arabia on Wednesday, also strongly criticised the non-appearance of the director of the CIA at their hearing.
Gina Haspel has heard a recording of Khashoggi's murder provided by Turkey and has studied evidence in the case. One senator described her absence as a "cover-up".
The Senate's vote is seen as a rebuke to President Trump who has described Saudi Arabia as a vital ally and resisted calls for sanctions against the kingdom's leadership.
Last week Mr Trump dismissed a reported CIA assessment that Saudi Crown Prince Mohammed bin Salman was to blame for Khashoggi's death.
The vote in the Senate means further debate on US support for Saudi Arabia is expected next week.
However, correspondents say that even if the Senate ultimately passes the bipartisan resolution it has little chance of being approved by the House of Representatives.
The resolution calls for an end to US military involvement in Yemen except for operations targeting Islamist militants. It came up for a Senate vote earlier in the year and failed to pass. But the climate is different now.
Senators are more and more concerned about the way Saudi Arabia is conducting the war, citing its terrible toll on civilians. Defence Secretary Jim Mattis and Secretary of State Mike Pompeo argued that pulling back US intelligence and training assistance for the Saudi campaign would only make a bad situation worse - strengthening Iran which backs the rebels, and undermining peace efforts that are gaining momentum.
But there's been a bipartisan push for a harder line on Riyadh since the Saudi government was linked to the murder of Jamal Khashoggi.