The Russian president scorned claims that the US president-elect took part in a hotel sex party with two Russian prostitutes in 2013.
It is alleged that the incident took place as secret agents collected “kompromat” material that could be used for blackmail purposes.
Mr Putin said: “The masterminds of fakes, the nature of which are presently being spread against the US president-elect, [are] worse than prostitutes —they have no moral limits.”
The allegations were revealed last week in a dossier compiled by former MI6 officer Christopher Steele.
The documents claimed that Mr Trump had asked prostitutes to urinate on a bed in his suite at the Ritz Carlton, believing it was once used by President Obama and his wife, Michelle.
He was in the city for a Miss Universe pageant.
The allegations were denied by Mr Trump, who also insisted that he was a germaphobe.
Mr Putin said prostitutes were women of “low social responsibility” and he doubted Mr Trump “took the bait”.
He said: “When Trump came to Moscow...he wasn’t any kind of political figure, we didn’t even know about his political ambitions.
“He was just a businessman, one of the richest people in America. Does someone think the special services run around after every American billionaire? Of course not, that’s total nonsense.
“Secondly - [the idea that] Trump arrived and straight away ran to meet up with some Moscow prostitutes. This is an adult person and a person who has spent many years organising beauty contests.
“He spent time with the most beautiful women in the world.
“I find it hard to imagine that he ran to a hotel to meet with our girls ‘of low social responsibility’. They are without question the best in the world but I doubt Trump took the bait.”