The Jordanian army announced on Sunday that its forces shot dead five people who were approaching their position from the Syrian side of the border.
The border guards killed the individuals who were coming from Tanf, a Syrian desert town where US special forces training rebels are based.
The Jordanian army said it destroyed a car and two motorbikes in the incident.
The army statement did not give any details of the identity of the men and whether they were smugglers or militants in the area where Jordan’s northeastern borders meet both Iraq and Syria.
The statement however said that before the shooting, a convoy of nine cars had approached from the Tanf area but fled after the army fired warning shots.
The town has been a flashpoint in recent weeks as militias backed by Iran have tried to get near the US garrison, prompting US coalition jets to strike back.
Tanf lies near the strategic Damascus-Baghdad highway that was once a major weapons supply route for Iranian weapons into Syria.
ISIS militants launched a suicide attack last April on the heavily defended base in which the Pentagon said an estimated 20-30 ISIS fighters were involved. US jets bombed the militants in the hit and run attack.
Staunch US ally Jordan has also been threatened by the militants.
Moscow on Wednesday condemned the US-led coalition strike on pro-regime fighters in Tanf as an “act of aggression” that targeted the most effective forces battling “terrorists” in the war-torn country.
“It was an act of aggression which breaches the territorial sovereignty of Syria and intentionally or not targeted those forces that are the most effective in fighting the terrorists on the ground,” Foreign Minister Sergei Lavrov said.
The US-led coalition on Tuesday said it had destroyed a unit of pro-regime forces in Syria as they advanced near an area where coalition commandos have been training and advising rebels.
A group of about 60 pro-regime forces moved into the area with a tank, artillery, anti-aircraft weapons and armed technical vehicles, the coalition said in a statement. They posed a threat to the coalition forces at the Tanf Garrison, it added.
The assault marks the second time in less than a month that coalition forces have attacked pro-regime forces as they headed toward the garrison inside a supposed “deconfliction zone” claimed by the US.