Staff at the Dolphin in Old Portsmouth announced it would be closing the pub for the last time at 10pm on Easter Monday after failing to agree a lease renewal with an unnamed pub company.
Writing on the pub's Facebook page they said: "The Dolphin lease had ended and, unfortunately, it has not been possible to agree terms for an extension that are realistic. Maintenance of the Grade II-listed building has become increasingly onerous in cost.
"The landlord required a 25% increase in rent with further increases every year for both rent and the supply of beer, wine and spirits. These factors have made it financially unrealistic for us to continue the business. We have therefore been served with a notice to vacate the premises.
"We have been advised by solicitors that we would not be successful in making an application to the courts to extend the lease at a fair rent under the Landlord and Tenant Act. We understand that the Dolphin will remain closed for the foreseeable future while the landlord plans a 'chain pub' makeover.
"We are all deeply sad and disappointed at this outcome. Alex, Ian and Alan would like to thank customers for making the Dolphin one of the best pubs in the city."
The Publican’s Morning Advertiser approached the pub for further comment but received no response.
Local Claire Moody commented: "Absolutely gutted. Loved every visit we made to you. Great food, great staff and wonderful atmosphere."
On Twitter, beer writer Roger Protz added: "Tragic loss of 250-year-old pub the Dolphin. Served great characters including Captain Cook."