The news will come as a huge blow to Liverpool manager Jurgen Klopp and Manchester City boss Pep Guardiola, who have been very vocal in their view that the measure should be reintroduced to the English top flight amid a growing number of injuries to players.
The latest vote at the meeting of the 20 shareholder clubs was deadlocked at 10 votes to 10, falling well short of the 14-6 majority needed.
In a bid to appease some clubs, teams will be allowed to name up to nine substitutes on the bench.
The Premier League also announced it would apply to the game's law-making body, the International Football Association Board, to take part in a concussion substitute trial.
The protocol the Premier League has expressed an interest in allows each team to make up to two additional permanent substitutions in the event of a concussion, with the opposition allowed to make the same number of extra changes.
It could be introduced into the competition as early as next month, the league said. The league's medical working group will meet on Friday to further consider the protocols.
The Football Association plans to introduce the same concussion protocols into the men's and women's FA Cups, the Women's Super League and the Women's Championship.
Premier League clubs have rejected a move to allow five substitutions for a THIRD time.