The UK-based charity has added its voice to an online petition against the ad, which has attracted 8,300 signatures internationally.
Its intervention carries weight because it has previously benefitted from BrewDog’s marketing; in 2014 it was offered a share of the profits from a special edition beer that BrewDog made in protest against Russia’s discrimination against gay people.
The online ad, aimed at potential investors in the company, features the founders in a variety of comic scenarios, including posing as women in a ‘red light’ district window, that they hope they won’t have to turn to in order to raise money.
However, it has met with a backlash in the form of an online petition that accuses it of "offensive caricatures of people, many of whom already suffer discrimination every day".
Over 8,000 people have signed the petition, which calls on Brewdog to pull the ad, apologise and donate to charities working for the rights of homeless people, trans women and sex workers.
A Stonewall spokesperson said: "Belittling, caricaturing and mocking people who suffer daily discrimination in an advert only serves to deepen and encourage discriminatory attitudes.
"We hope BrewDog will demonstrate their purported commitment to equity by listening to those who started the petition, taking this advert down and having a rethink."
BrewDog, which could not be reached for comment, has previously defended the ad against the petition.
James Watt, co-founder of BrewDog, said: "The video was made in the spirit of fun and sending ourselves up – it’s a shame that some people have taken offence where none was intended. We have a history of supporting and championing the LGBT community, and will continue doing so. Watch this space."