World no.1 moved one step closer to his 13th Grand Slam on Tuesday after Jo-Wilfried Tsonga retired in the third set (6-3, 6-2).
Novak booked his spot in the last 8 for the 10th time after dismissing Kyle Edmund 6-2, 6-1, 6-4 on Sunday night at Flushing Meadows.
The world no.1 got pass into the US Open fourth round as Mikhail Youzhny retired injured at 4-2 for the defending champion.
The top seed and defending champion will face Mikhail Youzhny (RUS) in the third round on Friday.
Nole began his quest for a third US Open title with a 6-3, 5-7, 6-2, 6-1 victory over Jerzy Janowicz at Flushing Meadows on Monday night.
Top seed and defending champion will open campaign in his 12th US Open appearance against Jerzy Janowicz (Poland).
The 2016 US Open draw will be led by reigning champion Novak, followed by Olympic gold medalist Murray, Wawrinka and Nadal.
Nole suffered a tough 6-7 (4), 6-7 (2) defeat to Juan Martin del Potro at the Rio Olympics on Sunday evening.
The Serbian duo won their opening round match 6-2, 6-2 against Croatian combination of Marin Cilic and Marin Draganja.
Novak is a top seed and will open his campaign in singles competition against world no.145, Juan Martin del Potro (ARG).