The 178-mile walk - in its second year - will see a core team of over 30 ex-footballers take part including Steph Houghton, Matt Holland and Jay Spearing.
The walk starts at Wembley Stadium on Friday ending at Ipswich Town's Portman Road stadium on Sunday evening passing through numerous National League clubs.
National League Head of Player Development Gary Owers is taking part alongside Hornchurch manager Daryl McMahon and their assistant manager Steve Gritt in the event that has backing from the PFA.
Speaking ahead of the event Owers said: "I'm looking forward to spending some time with a number of my former teammates this weekend but the main reason we're here is to raise money and awareness for the brilliant work done by the Darby Rimmer MND Foundation."
Last year's walk raised £200,000 for the Darby Rimmer MND Foundation with the charity targeting the quarter of a million pounds barrier this time around.
Click here to see more information about the incredible work of the Darby Rimmer MND Foundation and see how you can donate and help support their work to #AttackMND.