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7 year oldThierry Solere's resignation adds to a slew of notable departures, including the campaign treasurer on Thursday.
The conservative candidate has slipped down the opinion polls amid an inquiry into political payments to his wife.
His woes have raised speculation that ex-PM Alain Juppe could return to the race if Mr Fillon were to pull out.
Mr Juppe was overwhelmingly defeated by Mr Fillon in the Republicans' primary in November, securing only 33% of the vote to Mr Fillon's 66%.
Sources close to Mr Juppe said he would be prepared to step in, but only with the unanimous support of the party and only if Mr Fillon were to go voluntarily.
A new opinion poll suggests Mr Fillon would be eliminated in the first round of presidential election voting on 23 April, and that far-right candidate Marine Le Pen would challenge independent centrist Emmanuel Macron in the two-candidate run-off on 7 May. Mr Macron would win that contest, a number of opinion polls have suggested.
Mr Fillon has so far said he has no intention of stepping down despite the continuing haemorrhage of allies.
More than 60 politicians have said they can no longer support him.
The latest, Mr Solere, said on Twitter: "I've decided to end my duties as Francois Fillon's spokesman."
He joins campaign treasurer Gilles Boyer, two deputy directors and Mr Fillon's foreign affairs spokesman, among others.
Mr Fillon's Paris home was raided by investigators on Thursday as part of the inquiry into the payments to his Welsh-born wife, Penelope.
The Le Canard Enchaine newspaper alleges she was paid €831,400 (£710,000; $900,000) over several years for working as a parliamentary assistant for Mr Fillon and his successor, but had no parliamentary pass - raising questions over whether she did the work she was paid for.
Mr Fillon, 62, denies any wrongdoing and says he is the victim of a "political assassination".
He is planning a major rally near the Eiffel Tower in Paris on Sunday.
Mr Juppe, 71, has kept a low profile as the mayor of Bordeaux since his defeat in the primary.
Quoting a close source, Le Parisien said Mr Juppe would "remain loyal" but "not inactive" in a situation resembling a "collective suicide". Mr Juppe indicated that the call for him to return would have to come not just from his own supporters but from a far wider field.
Republican lawmaker Georges Fenech was among those appealing for Mr Juppe to return, telling BFM TV: "I don't think we have a single minute to lose in showing we have the ability to react to what has happened. I think Alain Juppe offers an alternative."
The deadline for candidates to declare they are running is 17 March, two days after Mr Fillon is scheduled to appear before a judge overseeing the investigation.
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