The environmental organisation and fellow green campaigners ReCommon filed the suit in a Rome court along with 12 citizens from areas of Italy severely affected by extreme weather events, Greenpeace said in a statement.
It said it was bringing legal action "for past and potential future damages resulting from its contribution to climate change, of which Eni was well aware but chose to ignore for decades".
In a statement, Eni described the suit as "groundless".
The lawsuit also includes Eni's two main shareholders, Italy's economy ministry and state lender Cassa Depositi e Prestiti (CDP), both of which "strongly influence the company's policies", Greenpeace said.
The group accused Eni's policies of being "in blatant violation of the Paris Agreement", the landmark 2015 deal aimed at limiting global warming, which it said had direct implications for the climate commitments of companies like Eni.
"Eni scored record profits in 2022, yet it continues to invest in expanding its fossil fuels business, dismissing climate impacts and ignoring local communities," said Chiara Campione from Greenpeace.
The plaintiffs have asked the Rome court to rule on whether Eni has caused damages through "violations of their human rights to life, health, and private and family life".
Eni said it would explain in court its "decarbonisation strategy", which it said balanced three aims: "sustainability, energy security and the country's competitiveness".
The suit is the first of its kind against a private company in Italy.
In recent years, a growing number of organisations and citizens have turned to the courts to criticise what they say is government inaction on the climate.
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