As Iranian delegates looked on in seeming displeasure, Mr Trump claimed one of the greatest security threats the world faces is "the repressive regime in Iran".
The president accused Iran of being the number one state sponsor of terrorism, and fuelling both the wars in Syria and Yemen.
"Iran citizens deserve a government that cares about reducing poverty, ending corruption and increasing jobs, not stealing their money to fund and massacre abroad and at home," he said.
“The regime is squandering the nation's wealth and future in a fanatic quest for nuclear weapons and the means to deliver them.
“To stop Iran’s path to nuclear weapons and missiles, I withdrew the US from the terrible Iran nuclear deal.
“Hoping to free itself from sanctions, the regime has escalated its violent and unprovoked aggression.
“In response to Iran’s recent attack on Saudi Arabia’s oil facilities, we just imposed the world’s highest level of sanctions on Iran’s central bank and sovereign wealth fund.”
President Trump also vowed the sanctions will be tightened until Iran is ready to cooperate with the US, and warned it was a cautionary tale for other countries.
“As long as Iran’s menacing behaviour continues, sanctions will not be lifted,” he said.
“They will be tightened.
“Iran’s leaders will have turned a proud nation into just another cautionary tale of what happens when a ruling class abandons its people and embarks on a crusade for personal power and riches.”
He urged other nations to act, stating: “No responsible government should subsidise Iran’s blood lust."
The President insisted the US desires "peace, cooperation and mutual gain with all”, but will always protect the interests of America first.
This comes as the UK, France and Germany joined the US on Monday in blaming Iran for recent strikes against oil facilities in Saudi Arabia.
On Monday, Boris Johnson called for a “Trump deal” with Iran, claiming that the US president is the “one guy” who can re-negotiate a nuclear deal.
The PM said the current deal with Iran was "bad" and had "many defects" from previous UK policy.
Iran wasn’t Donald Trump's only target in his UN address
Mr Trump has delivered a roaring defence of nationalism and American sovereignty as he addressed the UN General Assembly, imploring other nations to do the same.
“The future does not belong to globalists. The future belongs to patriots. The future belongs to strong,independent nations,” he told a murmuring crowd at the General Assembly in New York.
“Globalism exerted a religious pullover past leaders, causing them to ignore their own national interests. Those days are over.”
Almost in contradiction with his calls for nationalism over globalism, President Trump insisted he and Boris Johnson were working "very closely" to form a "magnificent trade deal".
"As the UK makes preparations to exit the European Union, I have made clear that we stand ready to complete an exceptional new trade agreement that will bring tremendous benefits to both of our countries," he said.
US-China trade wars
President Trump also hit out at China, and said it was “wrong” for world leaders to admit the country into the World Trade Organisation in 2001.
“Our leaders argued that this decision would compel china to liberalise their economy and strengthen protections to provide things that were unacceptable to us and for private property and for the rule of law,” he said.
“Two decades later, this theory has been tested and proven wrong.”
In his statement, Mr Trump asserted: "Not only has China declined adopt promised reforms, it has embraced any economic model dependent on massive market barriers, state subsidies, currency manipulation, product dumping, forced technology transfers and the theft of intellectual property, and also trade secrets on a grand scale.
"The second-largest economy all over the world should not be permitted to declare itself a developing country in order to game the system at others’ expense.
He added, "those days are over" and he will not accept a "bad deal" for the American people, at the same time, justifying his massive tariffs on imported Chinese goods.
"We’re seeking justice," he said.
He added he is currently monitoring the situation in Hong Kong, and how China handles the situation will be reflective of its future role in world trade.
The president also used the platform to implore countries to take human trafficking, and border control seriously.
"Many of the countries here today are coping with the challenge of uncontrolled migration. Each of you has the right to protect your borders and so if course, so does our country," he said.
He then continued to attack his opponents, who he labelled "activists" who have a "false sense of virtue" in promoting illegal immigration.
He stated: "I have a message for activists who cloak themselves in the rhetoric of social justice."
"Your policies are not just. Your policies are cruel and evil. You are empowering criminal organisations who prey on men, women and children."
He also had a hardline message for anyone hoping to cross the US-Mexico border - they are unwelcome to the US.
"To anyone contemplating crossing our borders illegally, please hear these words: Do not pay the smugglers, do not pay the coyotes," he said.
"Do not put yourself in danger. Do not put your children in danger, because if you make it here you will not be allowed in.
"You will be promptly returned home. You will not be released into our country.
As long as I am president of the United states, we will enforce our laws and protect our borders."