Phenyl 1,2,3-triazole-thymidine ligands stabilize G-quadruplex DNA, inhibit DNA synthesis and potentially reduce tumor cell proliferation over 3'-azido deoxythymidine

• Published in: PloS one 2013-08, Vol.8 (8), p.e70798
• Main Authors: Mahesh Kumar, Jerald, Idris, Mohammed M, Srinivas, Gunda, Vinay Kumar, Pallerla, Meghah, Vuppalapaty, Kavitha, Mitta, Reddy, Chada Raji, Mainkar, Prathama S, Pal, Biswajit, Chandrasekar, Srivari, Nagesh, Narayana
• Format: Article
• Language: English
• Subjects: Affinity; Animal experimentation; Animal tissues; Animals; Antineoplastic Agents - chemical synthesis; Antineoplastic Agents - pharmacology; Apoptosis; Biology; Cancer therapies; Cell cycle; Cell growth; Cell Line, Tumor; Cell proliferation; Cell Proliferation - drug effects; Chemical synthesis; Chemistry; Click Chemistry; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA biosynthesis; DNA fragmentation; DNA Fragmentation - drug effects; DNA polymerase; DNA synthesis; DNA, Neoplasm - antagonists & inhibitors; DNA, Neoplasm - biosynthesis; Electronegativity; Forming; G-Quadruplexes - drug effects; Gene expression; Genetic research; Genomes; Health aspects; High temperature; Humans; Kinases; Ligands; Medicine; Melanoma, Experimental - chemistry; Melanoma, Experimental - drug therapy; Melanoma, Experimental - pathology; Mice; Mice, Inbred C57BL; Molecular biology; Nitrogen; Nucleoside analogs; Nucleosides; Skin Neoplasms - chemistry; Skin Neoplasms - drug therapy; Skin Neoplasms - pathology; Stability; Studies; Telomerase; Thymidine; Thymidine - chemistry; Toxicity; Triazoles; Triazoles - chemistry; Tumor Burden - drug effects; Tumor cells; Tumors; Zidovudine - chemistry
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0070798

Triazoles are known for their non-toxicity, higher stability and therapeutic activity. Few nucleoside (L1, L2 and L3) and non-nucleoside 1,2,3-triazoles (L4-L14) were synthesised using click chemistry and they were screened for tumor cell cytotoxicity and proliferation. Among these triazole ligands studied, nucleoside ligands exhibited higher potential than non-nucleoside ligands. The nucleoside triazole analogues, 3'-Phenyl-1,2,3- triazole-thymidine (L2) and 3'-4-Chlorophenyl-1,2,3-triazole-thymidine (L3), demonstrated higher cytotoxicity in tumor cells than in normal cells. The IC₅₀ value for L3 was lowest (50 µM) among the ligands studied. L3 terminated cell cycle at S, G2/M phases and enhanced sub-G1 populations, manifesting induction of apoptosis in tumor cells. Confocal studies indicated that nucleoside triazole ligands (L2/L3) cause higher DNA fragmentation than other ligands. Preclinical experiments with tumor-induced mice showed greater reduction in tumor size with L3. In vitro DNA synthesis reaction with L3 exhibited higher DNA synthesis inhibition with quadruplex forming DNA (QF DNA) than non quadruplex forming DNA (NQF DNA). T(m) of quadruplex DNA increased in the presence of L3, indicating its ability to enhance stability of quadruplex DNA at elevated temperature and the results indicate that it had higher affinity towards quadruplex DNA than the other forms of DNA (like dsDNA and ssDNA). From western blot experiment, it was noticed that telomerase expression levels in the tissues of tumor-induced mice were found to be reduced on L3 treatment. Microcalorimetry results emphasise that two nucleoside triazole ligands (L2/L3) interact with quadruplex DNA with significantly higher affinity (K(d)≈10⁻⁷ M). Interestingly the addition of an electronegative moiety to the phenyl group of L2 enhanced its anti-proliferative activity. Though IC₅₀ values are not significantly low with L3, the studies on series of synthetic 1,2,3-triazole ligands are useful for improving and building potential pro-apoptotic ligands.