Compounds and methods associated with differential methylation of human papilloma virus genomes in epithelial cells

Abstract

The invention relates to compounds and methods useful in detection and therapy of HPV-associated diseases. The invention is based on the elucidation of a mechanism by which replication of HPVs occurs in naturally infected tissues and cells. Moreover it is based on the identification of distinct epigenetic changes of the viral genome in infected cells that allows promotion of the affected cells to precancerous and cancerous cells. The invention therefore provides methods of diagnosing neoplasias and their precursor lesions as well as methods of preventing the development of malignancies or inhibiting tumor growth.

1 . An isolated nucleic acid comprising a sequence selected from the group consisting of SEQ ID NOs: 1-16. 
     
     
         2 . A demethylating agent as a medicament for topical application for treatment of an HPV related lesion. 
     
     
         3 . The demethylating agents of  claim 2  being selected from a group consisting of 5-Azacytidine, 5-Aza-20-deoxycytidine, Arabinosyl-5-azacytidine, 5-6-Dihydro-5-azacytidine, 5-Fluoro-20-deoxycytidine, Epigallocatechin-3-gallate, Hydralazine, Procainamide, Procaine and Zebularine. 
     
     
         4 . The demethylating agent of  claim 2 , wherein the HPV related lesion is selected from a group consisting of warts, exophytic growing papillomas, condylomata, inverted papillomas, pre-neoplastic or neoplastic HPV-induced lesions, cervical neoplasias, rectal neoplasias, neoplasias of the anal epithelium, neoplasias of the oropharynx or neoplasias of the tonsils. 
     
     
         5 . A pharmaceutical composition consisting of a) a demethylating agent and b) a pharmaceutically acceptable carrier. 
     
     
         6 . The pharmaceutical composition of  claim 5 , comprising in addition active ingredients selected from a group comprising buffers, humectants, excipients, preservatives and antiviral substances. 
     
     
         7 . The pharmaceutical composition of  claim 5 , wherein the demethylating agent is selected from a group consisting of 5-Azacytidine, 5-Aza-20-deoxycytidine, Arabinosyl-5-azacytidine, 5-6-Dihydro-5-azacytidine, 5-Fluoro-20-deoxycytidine, Epigallocatechin-3-gallate, Hydralazine, Procainamide, Procaine and Zebularine. 
     
     
         8 . The pharmaceutical composition of  claim 5 , wherein the demethylating agent is present in a concentration of 0.001 to 50%. 
     
     
         9 . The pharmaceutical composition of  claim 5 , wherein the pharmaceutically acceptable carrier comprises at least one component selected from stearic acid, Aquaphor®, isopropyl palmitate, polyoxyl 40 stearate, propylene glycol, potassium sorbate, sorbic acid, triethanolamine lauryl sulfate, DMSO, betadex, edetate disodium, hydroxyethyl cellulose, methylparaben, niacinamide, phenoxyethanol, propylparaben and purified water. 
     
     
         10 . The pharmaceutical composition of  claim 5 , being provided on a polymer carrier consisting of cotton, cellulose, dextran, polyethylene, polypropylene, poly(dimethylsiloxane), ethylene vinyl acetate copolymers, polycarophil, hydroxypropyl methylcellulose and polyacrylic acids. 
     
     
         11 . The pharmaceutical composition of  claim 5 , wherein the polymer carrier is a cervical cap, a swab, a tampon, vaginal tampon, vaginal ring, vaginal strip, vaginal capsule, bioadhesive film, sponge or a brush.