Staff at Dempseys Bar and Club have written an open letter to the force, accusing them of letting down the LGBTQ+ community and saying that their appeals for help have been ignored.
Colin Humphreys said the situation is the worst he has known since he opened the nightclub 22 years ago.
He said:"I feel angry about it because we've always been good citizens. I think they think we're an easy target.
"We get homophobia every day and we put up with most of it as we can't have the police running backwards and forwards but when we think it is getting dangerous that's when we ring the police."
Over the past two years there has been a 'massive' increase in incidents of hate crime.
According to Mr Humphreys it took police three hours to respond to the most recent assault.
"When we ring the police we need help. What I would like to see is the police response," he said.
He said he accepted that cutbacks had put pressure on police numbers and resources, but that police had a duty of care.
LGBTQ-plus charity, Sayit said hate crimes have always happened but the over the past two years there has been a 'massive' increase in incidents.
South Yorkshire Chief Constable Lauren Poultney admitted she was concerned about the issues raised and would make contact with the bar's owners.
She said she understood the last two years had presented "numerous challenges" for all of Sheffield's bar, pubs and clubs including Dempseys.
She added: "We do already have the venue factored in to our patrol plans with a view to preventing crime in the city centre in addition to ensuring the door staff feel more supported and there is an increased officer presence at the venue.
"We are committed to working with all our licensed premises in Sheffield, including Dempseys, to provide a safe and welcoming night time economy."