Bakers Wine Merchants in Bishops Waltham will make five members of staff redundant when it shuts later.
The shop has sold alcohol since 1617 and was owned by two families for 244 years.
The off-licence said it was one in a series of closures partly due to fears of a proposed Sainsbury's development.
The store's five-year lease came up for renewal at the end of March.
Managing director Colin Baker felt he could not commit "with Sainsbury's looming".
He also said there was a lack of customers in the village and "the overheads are just too high".
"We had to look at the potential to break even and it just wasn't there", he added.
Shops in Bishops Waltham remain concerned about the impact that the planned Sainsbury's could have on their businesses.
The Liberal Democrat-led Winchester City Council is set to decide on the plans at a meeting on 7 April.
Sainsbury's said as part of the development it would help finance a £1.6m relocation of the village's surgery to the site and create more than 300 jobs for the area.
The chain also said the store would "address an identified lack of main food shopping choice in Bishops Waltham" and help attract more shoppers to the village.