Just under 1,000 tickets were sold to customers for Don't Look Now, a stage adaptation of the Daphne du Maurier novel, starring Rula Lenska.
The curtain was due to rise on the play on Monday for six evening performances and two matinees.
However, Ian Dickens Productions has been forced to cancel the show due to issues 'surrounding the procurement of the set'.
The production, also starring James Gaddas and Nicola Bryant, was due to begin its national tour in Eastbourne.
Theatre managers say they were informed by the producer on Friday of potential problems involved in opening the show on the original date, August 4.
Another opening date was agreed for today (Friday), and existing ticket holders were informed and offered seats for the end of the week.
However, an e-mail was received from Ian Dickens on Monday evening stating that the show would not be going ahead in Eastbourne at all, even though some of the company had actually been at the theatre during the day.
Eastbourne Theatres artistic director Chris Jordan said, "This has never happened before in my time at Eastbourne Theatres.
"I spoke to Mr Dickens on Friday when he informed me that the set would not be ready for him to open by Monday.
"He assured me that the show could open on the following Friday, so reluctantly we set about trying to contact and transfer patrons.
"We are extremely frustrated that the show is now not going to happen at all and that our theatregoers have been so inconvenienced."
Councillor Margaret Bannister added, "I am very disappointed that an external theatre producer has let down our theatre patrons in this way."
Mr Dickens said the Devonshire Park is 'one of our company's regular and favourite venues' and the show was cancelled 'with deep regret'.
"The main reason for this was the fact that our very complicated set, with nine different settings, was not ready in time and on Monday it was unfortunate that further set complications prevented the show opening on the Friday and Saturday," he explained.
"Although the set was the main issue there are a number of other issues which Eastbourne Theatres have conveniently forgotten to mention.
"For instance the fact that contract for this production was only issued from Eastbourne on July 24, reaching my office on July 28 which is seven days before the production was due to open.
"The contract had many clauses which at that late date could not be adhered to and therefore a number of issues needed to be resolved.
"We are sorry for any inconvenience to patrons and I am sure that they will realise that we would never taken this action unless it was absolutely unavoidable."
A theatre spokesman added that the contract was the same as the contract which had been approved by the production company 26 times before over the last few years.
Any customers who purchased a ticket for the show were notified by phone by Eastbourne Theatres, where possible, and all monies are in the process of being refunded.
For more information, or if you have purchased tickets for this production, call the box office on 412000.