Jordan Henderson will miss the title of Eredivisie after Ajax had suffered the ultimate choke.
The Dutch giants saw the victory against Groningen left in the 99th minute.
Ajax had taken the lead by Anton Gauei, but was fed back in the second half by Thom van Bergen.
Former Manchester United and Burnley striker Wout Wout Weghorst would have thought that his 68th minute goal would be enough for a vital three points.
But it was the ten men of the hosts who trigger the heartbreaking goal through Thijmen Blokzijl in the dying seconds of added time.
In the result, Ajax falls to second place and one point behind the title Rivals PSV.
Henderson and Co were the favorites for their 37th title with five more games left of the season.
Ajax only had to pick up six points to be confirmed as competition champions.
However, they have encountered defeats against Utrecht and NEC, as well as a draw with NEC and Groningen before the last game of the season.
The side of Peter Bosz fully benefited from the briefs of Ajax to eradicate the nine-point gap, after they have won their previous six games.
This means that one game is to go, the title PSVs is to throw away.
PSV Face 11th placed Sparta Rotterdam while Ajax takes on FC Twente.
Ajax will have to win and hope that PSV is slipping away because their target difference is considerably worse.
After Blokzijl's Last-Gasp delayers were left on social media stunned by Ajax's drop-off.
A posted: “How does Ajax give away a 9 points lead in 4 games? I have no words for it.”
A second wrote: “Ajax has bottled what is happening in the Netherlands.”
A third said, “We call that stifling in rap fights.”
A fourth said, “How did Ajax bottle the competition title?”
Another added: “As a fan, my head would be on Pluto until the latest games BC How?”
Images were recorded from the PSV match of the supporters and coaches of the club who waited to hear the result of the Ajax match.
Many seemed stressed and expected that the Amsterdam -based club would win, but were soon overjoyed when the late goal entered.
However, the race is not completely over, because Ajax manager Francesco Farioli has published a rally call.
He said: “It is never easy to speak after admitting a last-minute equalizer.
“It's not over yet. We still have a game to play: let's go.
“We have given it all, but unfortunately it wasn't enough. Yet football teaches us that something can happen.”
Henderson, 34, joined Ajax in January 2024 after leaving the Saudi Pro League side Al-Intifaq.
He has received the captain's bracelet, but has to lift another trophy with the club.