In a statement yesterday, Sadiq Khan accused Brokenshire of "hypocrisy" by "blocking", or threatening to block, several residential schemes in the capital after the secretary of state had previously called for a "step change in housing delivery".
The Labour mayor claimed the government had recently intervened in three housing schemes in just one week.
Plans for the redevelopment of Newcombe House were refused by the Royal Borough of Kensington & Chelsea (RBKC) in March, prompting the mayor to call-in the application.
Khan approved the plans in September but said Brokenshire has since issued a holding direction - a precursor to the decision being called-in.
Plans to rebuild Purley Baptist Church alongside 220 homes were approved by Croydon Council in December 2016 and then by Khan in March 2017.
The decision was subsequently called in by the secretary of state and permission was refused earlier this month.
Meanwhile, plans to redevelop the Kensington Forum Hotel alongside 46 homes were refused by RBKC in September then called in by the mayor last month.
Khan said the secretary of state has threatened to issue a holding direction preventing the London mayor from taking over the application.
Khan said: "By blocking these three schemes, this government appears to have bowed to lobbying in what can only been seen as a case of hypocrisy.
"Londoners cannot take this government seriously when the secretary of state for housing goes out of his way to stop new homes - including social housing - being built in the capital.
"Instead of giving in to lobbying, ministers need to show they are serious about supporting me in building the new social rented and other genuinely affordable homes Londoners need.
"Particularly with the two applications in Kensington and Chelsea, and in light of the chronic shortage of affordable homes seen after the terrible Grenfell Tower fire, this intervention by the minister is unhelpful and unnecessary."
MHCLG has been approached for comment but it had yet to respond at time of publication.
The latest row follows a dispute between Khan and Brokenshire earlier this year which was sparked by the housing secretary’s criticism of the emerging London Plan.
Brokenshire wrote to the London mayor to warn he was "not convinced" that the draft plan's "assessment of need reflects the full extent of housing need in London to tackle affordability problems".
The exchange led to fears that the London Plan could be delayed while the mayor’s team sought to address Brokenshire’s criticisms.
Earlier this week, Brokenshire refused plans for the 366-home redevelopment of a Kensington and Chelsea housing estate, after he concluded that the resulting net loss of social housing would be unacceptable.