The House rejected the measure, proposed by Congresswoman Marjorie Taylor Greene, by a vote of 147-276-2.
Greene's amendment would have added text to the defense budget that said no cluster munitions or cluster munitions technology shall be sold or transferred to Ukraine.
Greene, in remarks on the House floor, said cluster munitions kill civilians and represent an escalation of the war, not deterrence.
Earlier, the lower chamber rejected an amendment offered by Greene to remove $300 million in funding for Ukraine from the bill.
The House also rejected an amendment from Congressman Matt Gaetz to prohibit federal funds from being made available to provide security assistance to Ukraine.
Last week, the US unveiled a new military assistance package for Ukraine, which included cluster munitions. Russian Defense Minister Sergei Shoigu warned Tuesday that the Russian military would be forced to use similar weapons, which it has plenty of in stocks, against the Ukrainian armed forces if the US supplies cluster munitions to Kiev.
Cluster munitions are banned by the Convention on Cluster Munitions, which has been ratified by 123 countries. The US, Ukraine, Russia, China, India, Pakistan, Israel and South Korea have not signed the convention.