Just hours before the opening ceremony, the Court of Arbitration for Sport rejected the cases of 47 Russians who wanted to overturn a decision not to invite them to take part.
The court decision was quickly welcomed by the International Olympic Committee, which said it "brings clarity for all athletes", and the World Anti-Doping Agency.
However, the Kremlin expressed regret over the rejection of the late appeal.
"We regret this. We are taking this decision into consideration... and undoubtedly we will continue to help athletes to stand up for their rights," Russian President Vladimir Putin's spokesman Dmitry Peskov told journalists.
The Russians, including Korean-born star speed skater Victor An, had asked CAS to reverse an IOC decision not to invite them to compete as neutrals.
The Russians also lodged a further case with a Swiss civil court in Lausanne, where the IOC is headquartered, in a final bid to compete in South Korea.
But in a dramatic turnaround, the athletes withdrew that appeal, although the court gave no reason why.
Russia's team is banned after the emergence of systemic drug cheating. But the IOC allowed a large group of Russians deemed clean to take part as "Olympic Athletes from Russia".
A spokesman for the Olympic Athletes from Russia told AFP: "It's a pity the Russian delegation is not bigger."
Russian figure skater Evgenia Tarasova said: "It is sad, of course."
- 'Welcome news' -
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However, the IOC hailed the CAS decision and WADA president Craig Reedie called it "welcome news for WADA... for athletes and all others worldwide that care for clean sport".
The Russian saga has proved highly contentious in the build-up to Pyeongchang. On Thursday, reports emerged of an altercation between a Canadian and a Russian at the athletes' village.
"The message we're sending is the decision has been made and you should be satisfied that every athlete, including the Olympic Athletes from Russia, have had to clear the same high hurdle," she added.
Fifteen of the 47 who lost their bids on Friday were among a group of 28 who controversially had life bans from the Olympics overturned last week by CAS.
The other 32, including three-time Games gold medallist An, were also left off the list of Russians invited to Pyeongchang.
Investigations revealed an elaborate plot where tainted Russian urine samples were switched with clean ones using a "mousehole" in the wall of the Sochi anti-doping laboratory.
Russia has denied any government links to the conspiracy. But the IOC has banned the former sports minister Mutko for life.
"It's clearly disappointing we're still talking about this on the eve of the Winter Games," said Hugh Robertson, chairman of the British Olympic Association.
"It's reassuring the IOC's decision has been upheld and we can get on with the Games and forget about doping."
"I hope IOC president Thomas Bach is listening. For the sake of the Olympic ideal, he needs to resign," Walden said in a statement.
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