The first lion was darted earlier at a farm along the N4 near the Swaziland border.
"The second lion has been successfully darted. The operation is still continuing and we now have sight of the remaining lions," said SANParks.
The five lions escaped from the KNP on Monday evening, causing panic among residents.
SANParks said the five adult male lions were believed to have been chased out of the park by a dominant pride.
Police, park rangers and a veterinarian were on the trail of the three remaining lions.