Makfi, Dick Turpin and Canford Cliffs finished in that order at Newmarket last month and a mouthwatering rematch is in prospect.
Makfi, trained in France by Mikel Delzangles, sprang a 33-1 surprise in the British Classic but has not been raced since.
The other two, both trained by Richard Hannon, have covered themselves in glory.
Dick Turpin, who has beaten his stablemate twice this season, went on to finish second to subsequent Prix du Jockey-Club victor Lope de Vega in the French 2000 Guineas.
Canford Cliffs subsequently put up a career-best performance with a brilliant victory in the Irish 2000 Guineas.
Aidan O’Brien bids to make a major impact in the three-year-old division by saddling three runners – Steinbeck, Beethoven and Encompassing.
Alain de Royer-Dupre’s Siyouni, a big disappointment in the French Guineas, also takes his chance.
Pat Eddery, who won the race three times as a jockey, saddles Hearts Of Fire while there is transatlantic interest in American challenger Noble’s Promise, trained by Kenneth McPeek.
Makfi will be is ridden by Olivier Peslier for the first time and the jockey is expecting a big run.
“I think his chances are very good. He won the Guineas. It was a bit of a surprise but it was a good win last time and the trainer is confident,” he told At The Races.
“The horse has been working well but now this is Ascot and he needs to confirm the form.
“I hope I win the race, of course, but it is very strong and will not be easy. The horse is pretty well and I hope he can run a big race.”
Meanwhile Marcus Tregoning is hoping the lack of race fitness will not cost Mawatheeq in the Prince of Wales’s Stakes on Wednesday.
The Lambourn trainer has been unable to get the five-year-old on the track this year but reports Sheikh Hamdan Al Maktoum’s entire to be in great shape.
“He obviously had a bit of a problem over the winter which has got sorted out and it will be his first run of the year,” Tregoning told At The Races.
“I hope it doesn’t come too quick for him but he’s in good form and the race looks all right.
“The ground should be perfect, I think, by Wednesday so I’m looking forward to it.
“He’s got the experience now and he’s a lovely horse physically, he’s very strong and he’s a willing work horse.
“The problem we’ve had is with his soundness. Obviously that did hold us up. He had a seizure in his fore fetlock joint and had to have a screw put in it, but he’s moving fantastically well now.
“Whether the race is a bit too early for him having had that problem we’ll have to see, but I’m happy with him.
“He’s moving great and he looks a picture so we just have to hope for the best.
“He should be thereabouts provided fitness doesn’t tell. We wouldn’t run him unless we thought he was ready for a race.
“In reality we could have done with another couple of weeks but we haven’t had that and Royal Ascot comes and goes very quickly. He’s in good form.”