Seven months on from a 2-1 beating of the European champions in Naples - where Kane became England's all-time leading goalscorer - Gareth Southgate's men were out to complete their Euro 2020 revenge mission under the bright lights of Wembley.
With four wins and a draw to boast from their opening five matches in Group C, England only needed to avoid defeat against Luciano Spalletti's side to assure themselves of a place in Germany next summer.
Southgate unsurprisingly reverted to his strongest XI after a nervy friendly win over Australia, but the Wembley crowd were silenced early doors, as an unmarked Gianluca Scamacca met Giovanni Di Lorenzo's cross and found the roof of the net from close range.
However, the Azzurri's lead only lasted for 17 minutes, as a smart link-up between Kane and the effervescent Jude Bellingham ended with the latter being brought down by Di Lorenzo inside the box.
Following a prolonged VAR check, Kane stepped up and calmly sent Gianluigi Donnarumma the wrong way to draw England level, before Marcus Rashford sent the hosts ahead in the 57th minute thanks to a piece of individual brilliance from Bellingham.
Twenty minutes after Rashford picked out the bottom corner, Alessandro Bastoni headed a clearance from Marc Guehi into the path of Kane, who proceeded to poke the ball past Giorgio Scalvini and comfortably finish while one-on-one with Donnarumma.
As well as rubber-stamping England's ticket to next year's tournament in Germany, Kane's double on Tuesday evening saw the former Tottenham Hotspur striker become his nation's all-time highest Wembley scorer.
The 30-year-old has left Sir Bobby Charlton in his wake with 24 goals at England's headquarters, as the Manchester United legend managed 23 at the ground during his distinguished career.
Speaking to Channel 4 after the game, Kane - who now sits on 61 goals for his country - confessed that he 'enjoys' the individual feats, but focus must soon turn to Euro 2024, where England will be regarded as one of the favourites to go all the way.
"I love playing here at Wembley. I love playing for England full stop. To do it here at home is always special," Kane said. "To perform in front of our fans, to wear the badge, to lead the boys out, and of course to score as well is one of the best feelings ever.
"I enjoy the records but we go onto the next one and see how far we can go. This group wants to keep pushing standards. Now we will have to see how far we go next summer."
As well as confirming their place in the pot for December's group-stage draw, England are also guaranteed to finish in first place in Group C thanks to their superior head-to-head results over Ukraine.
Southgate's men round off a smooth qualifying campaign against Malta and North Macedonia next month, hosting the former at Wembley on November 17 before a trip to Skopje to meet the Lynxes on November 20.