High-throughout screening assay for identification of several high-risk HPV inhibitors

Several types of human papillomaviruses (HPVs) may induce transformation of infected cells. Two main genera of HPVs are classified as mucosotropic α and cutaneous β HPVs, both including high-risk cancer-associated species. Regardless of being extensively studied for decades, treatment for HPV-related infections/diseases remain challenging. Three vaccines against clinically relevant HPV types have been developed with their efficacy approved. However, various social, economic and religious concerns limit their effect. Moreover, as the vaccines are not effective against established infection, it needs to administered at a very early age. Therefore no effective and specific cure against an ongoing HPV infection has not yet been developed.

The absence of antiviral drugs has driven investigations into the details of the molecular mechanisms of the HPV life cycle, which is tightly linked to epithelial differentation. Viral helicase E1 and transcription factor E2 are phosporoteins, which play a pivotal role in the viral life cycle. For effective HPV genome replication, two viral proteins E1 and E2 form a complex. This protein-protein interaction has been an attractive target for developing anti-HPV drugs.

In addition to targeting HPV proteins, the viral genome itself could be target as well. Several sequence-specific DNA binding compounds have been developed, that bind to AT-rich regions near the E1 and E2 binding sites in the HPV replication origin and effictively reduce the stably maintained episomal viral genomes.

There are number of systems for conducting high-throughput screening (HTS) of available chemical libraries that is a widely used technique to identify new inhibitors of various diseases. Most of the work has been done in human primary epithelial keratinocytes. However, using these cells is relatively time consuming and expensive, especially for high-throughput screening.