"Sadly our aid convoy with UN and Syria Red Crescent was refused entry to Daraya, despite being given prior clearance from all sides," the International Committee of the Red Cross said on Twitter.
"We urge the responsible authorities to grant us access to Daraya, so we can return with desperately-needed food & medicines" outside the capital Damascus, it said.
ICRC's Syria chief Marianne Gasser, who was part of the convoy, described the refusal of access as tragic.
"Daraya has been the site of relentless fighting for more than three and a half years, and we know the situation there is desperate," she said.
Daraya had a pre-war population of around 80,000 people but that has dropped by almost 90 per cent, with remaining residents suffering from severe shortages and malnutrition.
A five-truck convoy organised by the ICRC, the United Nations and the Syrian Arab Red Crescent had been due to deliver baby milk and medical and school supplies.
The United Nations says more than 400,000 people are living under siege in Syria, most of them in areas besieged by the regime.
Syria's conflict has killed more than 270,000 and displaced millions since it erupted with the brutal repression of anti-regime protests in 2011.
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