NSC746364, NSC746365, and NSC746366: the spectra of cytotoxicity and molecular correlates of response to telomerase activity.

NSC746364, NSC746365, and NSC746366 are structurally novel 2,7-diamidoanthraquinone derivatives compared with other clinically used anticancer agents and have exhibited a unique multilog differential pattern of activity in our earlier studies. To systematically evaluate their potential anticancer activity, three selected compounds were tested for their cytotoxicity in vitro against 60 human cancer lines in the National Cancer Institute's anticancer drug screen as well as for dose response curves and telomerase activity. Cell growth was analyzed by the MTT assay, with differences between dose-response curves analyzed nonparametrically. Telomerase activity was detected by a modified version of the PCR-based assay and telomere repeat amplification protocol assay. To elucidate the structure-activity relationships and in-vitro anticancer activity, we correlated their activity profile [GI(50), total growth inhibition (TGI), and LC(50)] in the screening system and also their effects on telomerase activity, human telomerase reverse transcriptase expression, cell proliferations, and cytotoxicity. As a result we found that NSC746364, NSC746365, and NSC746366 have potent activity with 50% net growth inhibition conferred by 0.23-16.0 micromol/l (2.08 micromol/l mean); 0.78-15.9 micromol/l (2.57 micromol/l mean); 1.38-63.1 micromol/l (3.89 micromol/l mean), respectively. Sensitive cell lines exhibit TGI and 50% lethality to NSC746364, exhibited an LC(50) with as little as 2.82 micromol/l and TGI with as little as 0.95 micromol/l; NSC746365, exhibited an LC(50) with as little as 3.30 micromol/l, and TGI with as little as 1.65 micromol/l; NSC746366, exhibited an LC(50) with as little as 8.80 micromol/l; and TGI with as little as 4.06 micromol/l, respectively. Results of the study extend the initial in-vitro observation reported in the data above and confirm the importance of anticancer activity and telomerase inhibition. The unique molecular characterization, cytotoxicity, and telomerase activity profiles warrant further investigation and indicate a potential novel mechanism of anticancer action involved.

Synthesis, human telomerase inhibition and anti-proliferative studies of a series of 2,7-bis-substituted amido-anthraquinone derivatives.

Telomerase is important in tumor initiation and cellular immortalization. Given the striking correlations between telomerase activity and proliferation capacity in tumor cells, telomerase had been considered as a potentially important molecular target in cancer therapeutics. A series of 2,7-diamidoanthraquinone were designed and synthesized. They were evaluated for their effects on telomerase activity, hTERT expression, cell proliferations, and cytotoxicity. In the series, compounds (6, 10, 13, 16, 18, 19, 20-22, and 24) showed potent telomerase inhibitory activity, while compounds 19, 21, and 22 activated hTERT expression in normal human fibroblasts. The results indicated that 2,7-diamidoanthraquinones represent an important class of compounds for telomerase-related drug developments. Compounds 8, 16, 18, 26, and 32 were also selected by the NCI for Screening Program and demonstrated high anti-proliferative activity against 60 human cancer cell lines. Structure-activity relationships (SAR) study revealed that the test compounds with side chains two carbon spacer between amido and amine are important structural moiety for telomerase inhibition. Although the exact mechanism of how this amine group contributes to its activity is still unclear, however, the amine group in the extended arm of the bis-substituted anthraquinone might contribute to proper binding to the residues within the grove of G-quadruplex structure. Our results indicated that the 2,7-disubstituted amido-anthraquinones are potent telomerase inhibitors that have the potential to be further developed into novel anticancer chemotherapeutic agents.