The International Rugby Board has confirmed that the North v South clash of the rugby hemispheres will take place at Twickenham on 30 November.
The Millennium Stadium bid for the game when the fixture was initially put out to tender, but the IRB have now opted to go to London for the inaugural fixture.
"The match was put out to tender, and the WRU duly submitted a bid. It was never at any stage formally agreed that it would be in Cardiff, and we're obviously disappointed to have missed out on this occasion," said WRU media manager Lyn Davies.
South Africa's Andre Watson has already been named as the referee for the game and the IRB will shortly announce a match sponsor and TV broadcaster.
Set to become a biennial event in the rugby calendar, the proceeds will go towards the development of rugby in the smaller playing countries around the globe.
IRB chief executive Mike Miller claims there is "tremendous enthusiasm" for the game.
The IRB will also hold their Annual Awards ceremony in London on the same day as the game.
Ebbw Vale captain Damien Hudd picked up the biggest accolade of the night at the 11th Principality Premiership Awards Evening held at Principality Stadium, after leading his side to the title for the first time.
Wales assistant coach Robin McBryde is predicting a tough battle against England on Sunday despite the fact they may not be considered to be at full strength. Players will be out to impress Eddie Jones which makes England a dangerous proposition, he says.
Wales U20 captain Tom Phillips and performance psychologist Dr Rich Neil explain why spending three days at Monmouth RFC has further strengthened the togetherness of the squad ahead of the #WorldRugbyU20s
Alongside Scarlets flanker James Davies and Cardiff Blues centre Cory Allen, three Wales Sevens core players have made the extended training squad for GB Sevens. Wales Head coach Gareth Williams tells WRU TV's Graeme Gillespie why Luke Treharne, Sam Cross and Luke Morgan earned selection in the initial training squad for Rio.
Wales Sevens head coach Gareth Williams tells WRU TV's Graeme Gillespie he has had to overcome a number of injuries before naming a competitive squad for this weekend's penultimate round of the World Series in Paris.