The Blades took their case to the High Court, following an independent arbitration's panel's decision not to over-turn the Premier League decision to only fine West Ham over the Tevez transfer wrangle.
United felt West Ham should have been deducted points and were looking to be reinstated to the Premier League.
However, Justice Andrew Smith ruled against United and also denied their request to take the case further to the Court of Appeal.
Speaking outside the courts, the club's solicitor Paul Stothard insisted the fight may not be over - but first issued a statement on behalf of the club.
"Sheffield United have not been granted leave to appeal aspects of the arbitration panel's decision earlier this month.
"This application was made on very limited grounds and it has also been denied on very limited grounds.
"Sheffield United are not precluded from taking further action whether against the Premier League or West Ham.
"We will consider our position with our professional advisors and make further statements in due course."
Speaking to Sky Sports News, Stothard added: "We can't go to the Court of Appeal on this particular issue and that's all that we've been precluded from doing. Obviously now we have to go away and think about what we can do.
"As Kevin (McCabe, United chairman) said recently we've got a long agenda of things that we can do with regards to this. We're still on the first page and I think you can be fairly confident that the issue won't rest there."
On possible reinstatement, he said: "Time's been ticking since the last game of the season. We have to go away, look at what's been said today and we have to make our minds up. We will issue a further statement on that one.
"I think you've always got an uphill battle when you're trying to overturn a decision which has already been made. We've seen that, we've seen in the first instance with the arbitral tribunal that they disagreed with that first decision, the decision not to dock points from West Ham, but even that wasn't enough legally.
"The legal tests are very stringent, they are very hard. Many aspects of this case have been completely unprecedented and certainly Sheffield United cannot be criticised for trying these.
"As Kevin said it shows Sheffield United is not a club to be pushed around, they may be a small club but it's a club with good advisors, a professional approach, with good long-term plans and if it won't be playing in the Premier League next season it will certainly be looking to gain promotion and put forward its business plan for the next few years which is to establish itself as a Premier League force."
He also admitted that the club would at the very least look to gain compensation.
"Certainly if we cannot get reinstated then compensation would be top of the agenda because obviously there are very significant consequences financially for being relegated from the Premier League wrongfully as we believe we have been," he said.