Senators voted 24-67 on the amendment from Paul, which would reduce spending by 2 percent compared to fiscal 2019 levels.
The amendment, had Paul been successful, would have been added to a spending package that includes commerce, science and justice; transportation and housing and urban development; agriculture; and interior.
Sen. Patrick Leahy (D-Vt.), the top Democrat on the Senate Appropriations Committee, urged senators to oppose Paul's amendment.
"His amendment will slash spending below the bipartisan budget act that we all negotiated," Leahy said from the Senate floor ahead of the vote.
It's the latest attempt by Paul, a libertarian-leaning GOP senator, to slash spending, only to be rebuffed by a majority of his Senate colleagues. The Senate previously rejected a balanced budget proposal from Paul in June.