HS2 will continue as planned after the Court of Appeal rejected attempts to derail the plan.
Opponents including Bucks County Council wanted further assessment of the £43 billion scheme.
However, judges dismissed all seven grounds of the challenge, although a final appeal could be made to the Supreme Court.
High Speed Rail Minister Simon Burns said: “I urge opponents not to waste any more taxpayers’ money on expensive litigation and instead work with us on making HS2 the very best it can be.
“We continue to move forward apace with the crucial business of getting the scheme ready for construction in 2017 and delivering enormous benefits for the country.”
The appeals were brought by 15 councils, the HS2 Action Alliance and Heathrow Hub Limited.
Challenges included whether the Government had properly considered alternatives to HS2 and if it should have carried out a strategic environment assessment of the project.
Councillor Martin Tett, chairman of the 51m alliance and Bucks County Council leader said they were disappointed with the result,
However, he was encouraged by the fact there was a split decision on the environmental assessment, with one judge, Lord Justice Sullivan, siding with the objectors.
Mr Tett said: “Lord Justice Sullivan gives a very strong steer that HS2 Ltd has failed both in its obligation to fully assess the environmental implications of the project and vitally to assess these against the alternative we have put forward.
“His comments in his judgement are damning of the Department for Transport’s approach.
“This is another example of the Department for Transport and HS2 Ltd riding roughshod over public opinion, ploughing ahead regardless of what local communities want and ignoring the environmental merits of the alternatives.
“We have evidence that our alternative to HS2 would provide all of the capacity required, far more quickly, at a fraction of the cost and would be less damaging to the environment.”
Hilary Wharf, HS2 Action Alliance director, said it will continue to fight to ensure the Government does not ‘duck its environmental responsibilities’ over the project.
She said: “We can see from the Phase 1 draft environmental statement that cost savings are the top priority rather than protecting irreplaceable landscapes for future generations.
“The Government must properly take the environment into account.
“There is little case law on the specific point of law on which we lost.
“We are confident that our position is a strong one and we are pleased that the court has allowed our appeal to the Supreme Court.
“It’s concerning however that we have to go to the highest court in the land to make the Government give the environment the respect it deserves.”
UKIP councillor Chris Adams, county councillor for Wendover, Stoke Mandeville & Halton, said: “We take heart of course from the Appeal Court’s decision to allow a further appeal to the Supreme Court on the environmental case but are bitterly disappointed that another battle in the Courts against HS2 has been lost.
“Today’s arrogant gloating from Tory Transport Minister Simon Burns MP adds insult to the recent injury of Parliamentarians remaining stuck in their Westminster fact-free bubble, voting 12-1 in favour for HS2, and against the overwhelming majority of people in Bucks.
“This second setback in the courts is of course very disappointing but even an Appeal Court win and almost certain government appeal to the Supreme Court would not in itself have changed the political realities or the need for a political response to kill off HS2.
“Only yesterday Conservative Prime Minister David Cameron was quoted in The Evening Standard as saying that HS2 is essential.
“He and the national Conservative party in particular need to keep getting the message that they are going to be hit hard over this issue at the next general election.”