The hospital wanted the court to quash government orders that brought medical treatment to the late leader under the Terms of Reference of the Justice A Arumughaswamy Commission of Inquiry (CoI).
The plea comes a month after a counsel for the CoI alleged in a petition before the Commission that Tamil Nadu Health Secretary J Radhakrishnan colluded and conspired with Apollo Hospitals and "inappropriate treatment" was provided to her.
Both had denied the allegations.
Citing the accusation, the hospital sought barring the CoI from going into aspects like correctness, adequacy, or inadequacy of the medical treatment provided to Jayalalithaa.
The CoI was a fact finding body and not meant to be "adversarial or accusatory", the hospital said in its plea.
In the present case, however, the commission has stepped into the shoes of an "accuser," and falsely accused it of "conspiracy, collusion, inaction and inappropriate treatment," the hospital said.
Such accusations "impute unsubstantiated serious criminal motives to the bonafide medical treatment" provided to the late leader, the plea said.
There was a "real danger of prejudice and bias" in the manner in which the panel has been conducting examination of witnesses, the hospital alleged.
The nature of questions framed for witnesses and recording of their depositions disclosed a predetermined mindset against it, the hospital where Jayalalithaa was treated for 75 days in 2016 claimed.
The accusation of "collusion and conspiracy," between it and the health secretary and CoIs repeated questioning of the top official during the recording of his deposition on January 4 on the same lines evidenced that the panel was predetermined against it, the hospital submitted.
The petitioner hospital, while referring to dismissal of its application for appointment of a medical board, contended that the CoI has erroneously concluded that it was its duty to analyse complex medical facts without the aid of medical experts.
Last month, the panel had rejected the hospital's plea seeking a medical board to go into voluminous medical records by citing the assistance it received from government doctors to decipher such documents.