The challenge is a 55-mile hike around the county's white horses, and has raised £16,000 over the past two years.
It is organised by a group of Wiltshire businesses, and supported by the Wiltshire Times.
Organisers are calling on walkers to don their boots and take the challenge on the weekend of June 7-8.
Trowbridge and Chippenham-based accountancy firm Monahans are among the organisers.
Marketing Director Richard Edwards said: "By nature of us as a business we can get the corporate teams in.
"But we want groups of friends to do it who might see it in the paper or talk about it down the pub."
25-year-old barrister Ben Jenkins completed the challenge in 2006.
He said: "The full challenge is fairly gruelling. It's not the sort of event where you get out of breath, but it's really mentally tiring.
"But there's great camaraderie and we got a cracking breakfast on the Sunday morning."
The 55-mile hike is spread over the two days, but there are three shorter walks for people who don't fancy the full length.
Walkers can cover half of the course, which covers seven white horses around Devises, Marlbourough and Pewsey, or do the challenge in a team relay.
There is also a seven-mile route on the Sunday for a lighter day's walking.
Mr Edwards said: "Anyone can take part, because of the different lengths you don't need to be an Olympic athlete."
Monahans set up the White Horse Challenge with gardening company Lawntender, truck dealers Rygor Commercials and gym Premier Global.
For more information call Monahans on 01225 785520, or visit www.whitehorsechallenge.co.u
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