The bus carrying players and officials of the Ghana Premier League team was returning to its base in Kumasi after a game against Inter Allies in Accra.
The incident took place in the southern town of Nkawkaw, where the bus is said to have hit a stationary truck.
The team's deputy equipment officer Kofi Asare has been confirmed dead.
"Management has been in shock and grief following Wednesday's unfortunate accident involving our team bus," Kotoko said in a statement on Thursday.
"We regret to announce that Kofi Asare could not make it."
"Meanwhile, head coach Steven Polack, midfielder Ollennu Ashitey and bus driver Nana Berkye are receiving treatment."
The statement added that players and officials who had sustained minor injuries have been treated and discharged, with a follow-up examination scheduled for Saturday.
Reports suggest there were around 35 people on board - nine of whom have already been treated and discharged from the hospital in Nkawkaw.
In a statement, the Ghanaian FA said it was 'devastated' by the news.
"Our thoughts and prayers are with Kumasi Asante Kotoko, particularly the injured officials and players, who are currently receiving treatment in Nkawkaw," it added.
Kotoko's league game against Accra Hearts of Oak, which was scheduled for Sunday, has been indefinitely postponed.
Asante Kotoko is a famous football club based in Kumasi, Ghana's second-largest city.
The news has been met with an outpouring of grief on social media, says the BBC's Thomas Naadi in Accra.
Ghana's President Nana Akufo-Addo says he's saddened by the incident and has wished the injured a speedy recovery.
Asante Kotoko had lost to Inter Allies, so ending a 10-game run without a defeat to leave the two-time African champions fourth in the league.
One person has been killed and three others injured after the team bus of Ghana's Asante Kotoko was involved in an accident on Wednesday night.