A High Court Judge rejected Tottenham and Leyton Orient's appeal for a judicial review.
Spurs and Leyton Orient started the legal battle after the Olympic Park Legacy Company (OPLC) board voted 14-0 in February to make the Hammers the first choice to move into the ?486million stadium.
Battle: Tottenham are refusing to back down after West Ham were awarded the Olympic Stadium
Mr Justice Davis rejected their application to seek a judicial review, but both clubs could still make another legal challenge.
A statement from Tottenham read: 'The club now has the option of renewing its application at an oral hearing at the High Court and we shall give consideration to this in the next few days.
'As previously reported, the club continues to hold discussions with both local and national government bodies in order to seek to determine a feasible stadium solution.'
A spokesman for the OPLC, which is in charge of securing the future of the Olympic Park after the Games, had earlier said: 'The court has decided to refuse both Tottenham Hotspur and Leyton Orient permission to pursue a judicial review challenge in relation to the Legacy Company's decision to select a preferred bidder for the Olympic Stadium.
'We are pleased with the ruling and continue to make good progress in our negotiations with the preferred bidder in order to be in a position to agree the final terms for the stadium's lease.'
Vision: West Ham's depiction of how the Olympic Stadium will look when they take it over
Tottenham lost a head-to-head contest with West Ham to become the new tenant of the stadium in Stratford, east London.
West Ham, in a joint bid with Newham Council, intend to convert the 80,000-seater stadium into a 60,000-capacity facility which retains an athletics track. The club plans to move from Upton Park in 2014/15 with a 250-year lease, and give a 250-year lease to UK Athletics (UKA).
Newham Council has arranged a ?40m loan to finance West Ham's move to the stadium after the 2012 Games.
Tottenham's plans, part of a joint bid with AEG sport and entertainment group, had been to create a football-only stadium without the track and redevelop Crystal Palace for athletics.
The OPLC's choice of West Ham as preferred bidder has also been rubber-stamped by the Government and London mayor Boris Johnson.
Time to move on: West Ham's owners were pleased with the High Court's ruling
A Newham Council spokesman said: 'We are pleased with the judge's decisions.
'We are continuing to focus on securing a lasting legacy for the stadium and, together with West Ham United, are progressing well with the Olympic Park Legacy Company on bringing the matter to a commercial and financial close.'
West Ham vice-chairman Karren Brady wrote on Twitter: 'We welcome today's news as a further endorsement of our strong and viable legacy vision.
'We hope we can now focus all our energy and passion on delivering a fantastic multi-use Olympic Stadium for the whole nation.'
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Source: Daily Mail
Source: Daily Mail