Kenseth turned a lap of 186.285 mph around the 2.5-mile oval. Clint Bowyer was second at 186.070 mph.
They were the only two drivers to top 186 mph in practice for Sunday's Crown Royal 400. Brad Keselowski was third with Jimmie Johnson fourth and Kyle Larson fifth. Kurt Busch, Kyle Busch, Joey Logano, Marcos Ambrose and Kevin Harvick rounded out the top 10.
Tony Stewart, a hometown favorite and two-time Indy race winner, was 11th. Series points leader Jeff Gordon was 14th. Danica Patrick, the highest-finishing female in the Indianapolis 500 and Daytona 500, was 16th. Dale Earnhardt Jr. was 24th.
Teams have a final practice session from 9-10:50 a.m. ET Saturday with qualifying set for 2:10 p.m.