Parliamentary private secretaries Chris Green and Conor Burns both announced they were resigning on Twitter over Brexit.
And Mr Green's announcement was met by fellow Tory MP – Simon Hart – tweeting "nobody gives a f***".
It comes as the Conservatives leading body the 1922 Committee met to decide the Prime Ministers fate.
Mrs May has been embattled today as Cabinet heavyweights David Davis and Boris Johnson both quit over Brexit.
Tory divisions have been brought to the fore as they disagree over the the so-called Chequers agreement.
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“Brexit must mean Brexit”
Chris Green MP
Mr Green, who worked for transport secretary Chris Grayling, tweeted his resignation letter – saying “Brexit must mean Brexit”.
He wrote: "It must be clear to the electorate that, even if we do not fully leave at the end of March 2019, our full independence is gained in time for them to cast their judgment at the following general election."
He continued: "The direction the negotiations had been taking have suggested that we would not really leave the EU and the conclusion and statements following the Chequers summit confirmed my fears.
"I recognise that delivering Brexit is challenging, however I had hoped at tonight's meeting that there would be some certainty that my fears were unfounded, but, instead they have been confirmed.”
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Meanwhile, Mr Burns said: "I've enjoyed a fantastic year as PPS to Boris Johnson and six years as PPS to five different Ministers.
"I've decided it's time to have greater freedom.
"I want to see the referendum result respected. And there are others areas of policy I want to speak more openly on.”
Junior minister Steve Baker MP also resigned las night, bringing the total to five who quit on Mrs May.
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Mr Hart responded to the resignations in a surprising outburst on Twitter as the divisions in the Tory government continue to emerge over Brexit.
He said: "You're a PPS Chris. It is not relevant and nobody gives a f***. Apart from me obviously."
The MP added "I'm thinking of stepping down myself" and earlier in the day described Mr Davis' resignation as a "s*** show".
And he warned the infighting has "hugely increased chances of Brexit never happening" and branded the Brexit secretary as "failure".
In her reply to Mr Johnson's letter of resignation, Mrs May said she was "sorry – and a little surprised" that he had chosen to leave following their "productive discussions" at Chequers.
However, she said that if he was unable to support the Government's agreed position "it is right that you step down".
"As I outlined at Chequers, the agreement we reached requires the full collective support of Her Majesty's Government," she said.
"During the EU referendum campaign, collective responsibility on EU policy was temporarily suspended. As we developed our policy on Brexit, I have allowed Cabinet colleagues considerable latitude to express their views.
"But the agreement we reached on Friday marks the point where that is no longer the case, and if you are not able to provide the support we need to secure this deal in the interests of the United Kingdom, it is right that you should step down."