Russian Foreign Ministry spokesman Alexander Lukashevich made the accusation on Thursday a day ahead of talks in Moscow with French President Francois Hollande and German Chancellor Angela Merkel on ways to end the conflict in eastern Ukraine.
Lukashevich told reporters in Moscow: "Washington's actions are purposely driving Russian-American relations into a dead end.
"In the US military budget, $640 million has been pledged ... to Moldova, Georgia and Ukraine in order to, I quote the document; 'oppose Russia'."
He added: "Russia has no doubts about the confrontational attitude of the White House, which started a meaningless game of sanctions.
"But now we are talking about the financing of subversive actions against our country, which puts tension in U.S.-Russian relations to a new level."
U.S. Secretary of State John Kerry arrived in Kiev on Thursday with Hollande and Merkel, but he will not join the European leaders in Moscow.
U.S. and European leaders have accused Russia of supporting the conflict with arms and ammunition, while Russia has said the U.S. and EU have deliberately destabilised Ukraine in order to expand the west's military presence in eastern Europe through NATO.
Observers in Ukraine have also claimed U.S. troops are involved in clandestine operations in the region.
More than 5,000 people have been killed in the Ukrainian conflict between government forces and pro-Russian separatists in the southeast of the country since it began last April.