Lab's focus is on the Drug Discovery against HEV at Transcription and Post-translational level:
The four enzymes of HEV, Methyl Transferase, Cysteine Protease, Helicase and RNA dependent RNA Polymerase, predicted through computational biology remain to be validated for their presence and functionality. Our current focus is on the enzymes Methyl Trnsferase and Cysteine Protease against which we plan to identify the inhibitors that will stop their activity and effectively, the HEV propagation. The process involves Baculoviral expression of these proteins to ensure their structural characterization in their native forms. Based upon the crystal structure and other characterisation parameters , docking studies will be performed using virtual library of the inhibitors. Having developed and validated the assays of these enzymes, a set of shortlisted inhibitors will be tested in the wet lab. Around 10 inhibitors based upon their IC50 values and the binding strength on the active site will be selected and taken further for pre-clinical studies.
Our lab’s second interest is to identify the molecules that can inhibit the HEV replication so that they can be developed as putative therapeutic molecules in future and used in preventing the propagation of the virus. This has so far remained obscure for the lack of an efficient culture system to grow HEV in vitro or in vivo. Our lab is working on creating the virus culture using novel technology of BacMam, not used earlier and the expertise of our lab, where Baculovirus is used to encapsidate HEV genome and safely deliver in mammalian cells by endocytosis without causing injury to the cells, To our perception, the delivery of higher copy number of the viral genome in a cell in a non-injurious manner, should generate the HEV particles which can replicate in the mammalian cells due to the CMV promoter present in BacMam system. In case we are able to produce and purify the virus , we plan to create a small animal model of HEV culture by injecting them in nude or Balb/C mice. Once done, we will be studying the ex vivo/in vivo processing of the polyprotein, as compared to the in vitro, which we have earlier shown in our lab. Having established an efficient culture system with quantifiable replicative RNA forms, we will test various plant extract components already known for hepatic cures in the traditional science besides testing interferons or other anti-inflammatory drugs against liver available in the market.