BRUSSELS-European Union regulators filed formal charges Tuesday against Honeywell International Inc. and DuPont Co. over their agreements to produce the only car coolant that meets new EU rules on greenhouse-gas emissions.

The European Commission, the EU's top antitrust regulator, said deals between the two companies to produce the chemical, known as R-1234yf, "may have limited its availability and technical development, in breach of EU antitrust rules."

The move steps up a probe opened in 2011 that targeted Honeywell and DuPont's pact to produce a new refrigerant for vehicle air-conditioning systems, which was developed to replace a refrigerant that had fallen foul of new EU anti-global warming rules.

If the companies are found guilty of illegal collusion, they could face fines of up to 10% of annual sales.

The decision to send a so-called statement of objections doesn't prejudge the final outcome of its investigation, the commission said in a statement.

Honeywell said the EU's allegations were "baseless and conflict with the EU's own laws that encourage collaboration on development." The company has said it is targeting $1.5 billion of revenue from the chemical over five years.

DuPont said it had "complied at all times with applicable laws" in the development of the chemical, and would defend itself vigorously.

The EU had also been examining whether Honeywell engaged in "deceptive conduct" during the evaluation of 1234yf between 2007 and 2009. When it opened its probe in 2011, the commission said Honeywell may have failed to disclose its patents while the refrigerant was being assessed and failed to grant licenses on "fair and reasonable terms."

Write to Tom Fairless at tom.fairless@wsj.com

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