A group of about 60 pro-regime soldiers moved into the area with a tank, artillery, anti-aircraft weapons and armed technical vehicles, posing a threat to the coalition forces at At-Tanf Garrison near the border with Jordan and Iraq, the coalition said in the statement.
"The coalition issued several warnings via the de-confliction line prior to destroying two artillery pieces, an anti-aircraft weapon, and damaging a tank," it said.
US and British troops are based at At-Tanf, where they train local Syrian fighters for the battle against Islamic State forces.
The coalition said the contingent was "well advanced" into the 55-kilometer (34-mile) de-confliction zone, where any intrusion is considered hostile.
"The coalition is not looking for a fight with the Syrian regime or pro-regime forces but it is ready to defend itself if pro-regime forces refuse to leave the de-confliction zone," the statement said.
It was the second time in less than a month that coalition forces have attacked pro-regime forces as they headed toward the garrison.
Coalition planes on May 18 pounded the front of a convoy that had apparently been headed toward the At-Tanf garrison, in an established "deconfliction zone" northwest of the garrison.
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