Sandusky had asked the Pennsylvania Supreme Court to take up his 45-count conviction, arguing that his lawyers were rushed to trial in 2012 and that prosecutors improperly made reference to his decision not to testify.
He also said the trial judge should have issued a jury instruction about how long it took his alleged victims to report the abuse and that jurors should not have been told to weigh evidence of his good character against all other evidence.
Sandusky defence attorney Norris Gelman said he was disappointed by the court's decision, which was issued in the form of a one-sentence order. Sandusky can file a new appeal.
"I'm sure he will," Gelman said.
Attorney General Kathleen Kane, whose office prosecuted Sandusky, issued a statement saying she was pleased with the decision.
"Protecting Pennsylvania's children is one of my top priorities, and I remain committed to seeking justice for all victims of sexual abuse," Kane said.
The prosecutor's office argued that Sandusky did not provide sufficient basis for the Supreme Court to take up the matter and that decisions made by the trial judge did not violate his rights.
Michael Boni, a lawyer who represents Aaron Fisher and other Sandusky victims, said the Supreme Court made the right call.
"Hopefully this will, once and for all, put to bed any lingering hopes that Jerry will have his sentence reversed, his convictions reversed," Boni said. "It's a happy day for the victims."
Sandusky, 70, is serving a 30- to 60-year prison sentence for sexual abuse of 10 boys.
His wife, Dottie, said in an emailed statement: "All I have to say is Jerry is an innocent man in prison from crimes he did not commit and I hope that the legal system will eventually rectify this injustice."
Gelman said Sandusky can file a new appeal under the state's Post Conviction Relief Act. That appeal could address any newly discovered evidence as well as any claims that Sandusky's lawyers were not effective, Gelman said.
Sandusky also could eventually take his case to federal court.
Eight of his victims testified at trial, describing a range of abuse from grooming and fondling to oral and anal sex, including attacks in the basement of Sandusky's home outside State College. Another witness, a graduate assistant for the team who had been a quarterback for the Nittany Lions, testified he saw Sandusky having sexual contact with a boy inside a team shower late on a Friday night.
Sandusky did not testify on his own behalf but has maintained his innocence. His lawyer has said the victims' testimony was motivated by a desire to cash in. Penn State announced last year it was paying $59.7 million to 26 people who had raised claims of abuse at Sandusky's hands.
His defence lawyers repeatedly sought delays before trial, saying they were swamped by an enormous amount of material from prosecutors and needed more time to examine the background of his accusers.
During a post-sentencing hearing, however, defence attorney Joe Amendola acknowledged that he had not discovered anything afterward that would have changed his trial strategy.
Sandusky's 2011 arrest led to the firing of Hall of Fame football coach Joe Paterno and significant penalties levied against the school by the NCAA. Paterno was stripped of 111 of his 409 career wins while the school was fined $60 million, banned from bowl games for four years and faced steep scholarship cuts. He has since died.
Three other high-ranking school officials, including the then-president, face charges they covered up complaints about Sandusky. Their case has not yet gone to trial.