"President Trump got the message from the many Londoners who love and admire America and Americans but find his policies and action the polar opposite of our city's values," Khan said in a statement released on Twitter.
Khan said Trump's visit would "without doubt have been met with mass peaceful protests" and that it was a mistake for Prime Minister Theresa May to invite him on a state visit.
British Foreign Secretary Boris Johnson fired back on Twitter, saying Friday that he would "not allow US-UK relations to be endangered" by Khan, whom he referred to as a "pompous puffed up popinjay in City Hall."
A UK official said on Friday that the invitation to Trump for a state visit, first extended by May during her visit to the US early in Trump's presidency, still stands: "The invitation for a State Visit has been extended and accepted," the official said.
The official drew a distinction between the state visit, which would include a visit with Queen Elizabeth II and royal trappings like a horse parade, and a working visit, that would include a meeting with May and other diplomatic formalities like opening the embassy.
After Trump declared his disapproval with London's new US embassy as the reason for canceling his visit to the city, Madame Tussauds placed a waxwork of the President outside the building.
Khan, London's first elected Muslim mayor, has previously criticized the US President for his proposed travel ban on several Muslim-majority countries.
Trump has also hit out at Khan for his handling of terror attacks carried out in London, seizing on them in his call for the travel ban.
Khan has pushed back, saying he doesn't need advice from the US President.
Khan called Britain First, the far-right party Trump retweeted, "a vile, extremist group that exists solely to sow division and hatred in our country."
He added that the videos make it "increasingly clear that any official visit at all from President Trump to Britain would not be welcomed."
Khan reiterated this sentiment on Friday, saying he hoped that Trump "revisits the pursuit of his divisive agenda."