Croatia vice-captain Ivan Rakitic and forward Ante Rebic have not been included in the squad to face Wales and Tunisia in June.
Coach Zlatko Dalic confirmed that Barcelona's Rakitic was suffering with a hamstring injury, while Rebic of Frankfurt had not recovered from a knee injury.
Croatia face Wales in their European Qualifier on June 8 having won one and lost one so far in Group E.
They then host a friendly against Tunisia on June 11, a game which the pair will also miss.
Dalic said: "We made the decision together that Rakitic will not be part of this action. He has had a problem with his hamstring, and it flared up again in the final of the King's Cup. He is not fit for the Wales match and will not take part in the Tunisia match either.
"The match with Wales is extremely important and demanding because it comes at the end of the club season, but as it is for us, so is it for our opponents. As far as the result is concerned, we do not have many choices."
Wales go into the match against the World Cup 2018 finalists looking to try to qualify for their second consecutive European Championships, having made it to the semi-finals in 2016. They won their opening game 1-0 against Slovakia thanks to an early goal from Daniel James.
After the Croatia match they face a trip to Hungary on 11 June.
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