Almost 30 conservative MPs so far have signed a motion that wants to stop the Bill regarding the investment into the HS2 project.
The group of MPs, led by former Cabinet minister Cheryl Gillan, aims at derailing the legislation before it reaches the second reading, killing it at the first parliamentary hurdle.
"This House declines to give a second reading to a Bill which authorises preparatory expenditure on a railway without specifying further detail of the route and a limit on expenditure," reads the rebel motion.
The rebels are ready to voice their opinion during Wednesday’s key first votes on the High Speed Rail Preparation Bill.
If passed into law, the Bill would give the Transport Secretary the green light to spend money developing the new High Speed 2 line set to link London to Birmingham by 2026. Two second phase branches to Manchester and Leeds, via Sheffield, are planned by 2032.
Apart from 28 conservative MPs, the motion is supported by Labour representatives Frank Dobson, Natascha Engel, Geoffrey Robinson and Jim Cunningham, Green Caroline Lucas, Plaid Cymru's Hywel Williams and independent Patrick Mercer.