The changes were intended to separate interactions with friends from branded content - including that of celebrities and influencers.
Snapchat founder Evan Spiegel wrote in a blog post that he believed blurring the two had contributed to the rise of fake news.
However, thousands of Snapchat users say that the new layout is hard to use.
Nic Rumsey, who set up the petition, wrote that some are using Virtual Private Network (VPN) apps - which use servers abroad to mask the location of a device - in order to access the older version of the platform: "That's how annoying this update has become," he said.
"Many 'new features' are useless or defeat the original purposes Snapchat has had for the past years."
The petition, posted on the change.org website, is one of several appealing to Snapchat to revert to its previous state.
Users now swipe left to see and interact with their friends, and swipe right to see branded and celebrity content.
Model Chrissie Teigen tweeted that she didn't like feeling that her followers no longer felt like "friends" as a result.
"How many people have to hate an update for it to be reconsidered?" she asked.
Kylie Jenner asked her Twitter followers what they thought of the changes, and then agreed with a follower who replied that they hated it.
Snapchat reiterated its statement that updates "can take a little getting used to".
"We hope the community will enjoy it once they settle in," the firm said.
Rohan Midha, managing director of influencer marketing agency PMYB, told the BBC that Snapchat had made a mistake in removing celebrities and influencers - high profile individuals with lots of followers - from the "friends" section.
"They hadn't realised the value that influencers add to the app," he said.
"People want to follow influencers because they really like them, whereas Snapchat just sees them as brands."
However, Mr Midha added that he didn't think the petition would force Snapchat into a u-turn.
"It doesn't surprise me that so many people have signed it," he said.
"But I don't think this large Silicon Valley company is going to take notice of a petition."
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