Novel Anthra[1,2-c][1,2,5]Thiadiazole-6,11-Diones as Promising Anticancer Lead Compounds: Biological Evaluation, Characterization & Molecular Targets Determination

Abstract

The novel compounds NSC745885 and NSC757963 developed at our laboratory were tested against a panel of 60 cancer cell lines at the National Cancer Institute, USA, and a panel of 39 cancer cell lines at the Japanese Foundation of Cancer Research. Both compounds demonstrated selective unique multi-log differential patterns of activity, with GI50 values in the sub-micro molar range against cancer cells rather than normal cardiac cells. NSC757963 showed high selectivity towards the leukemia subpanel. Activities of both compounds strongly correlated to expression of NFKB1 and CSNK2B genes, implying that they may inhibit the NF-κB pathway. Immunocytochemical microscopy of OVCAR-3 cells showed clear cytosolic accumulation of the NF-κB p65 subunit following treatment. Western blotting showed dose dependent inhibition of the nuclear expression of the NF-κB p65 subunit with subsequent accumulation in the cytosol following treatment. Docking experiments showed binding of both compounds to the NF-κB activator IKKβ subunit preventing its translocation to the nucleus. Collectively, these results confirm the ability of our compounds to inhibit the constitutively active NF-κB pathway of OVCAR-3 cells. Furthermore, COMPARE analysis indicated that the activity of NSC757963 is similar to the antituberculosis agent rifamycin SV, this was confirmed by testing the antimycobacterial activity of NSC757963 against Mycobacterium tuberculosis, results revealed potent activity suitable for use in clinical practice. Molecular properties and Lipinski’s parameters predicted acceptable bioavailability properties with no indication of mutagenicity, tumorigenicity, irritability and reproductive effects. Oral absorption experiments using the human Caco-2 model showed high intestinal absorption of NSC745885 by passive transport mechanism with no intestinal efflux or active transport mechanisms. The unique molecular characterization as well as the illustrated anticancer spectra of activity and bioavailability properties warrant further development of our compounds and present a foundation brick in the pre-clinical investigations to implement such compounds in clinical practice.

Erlotinib-Conjugated Iron Oxide Nanoparticles as a Smart Cancer-Targeted Theranostic Probe for MRI

“…Viability of cells was tested by the WST-1 assay48. Briefly, either 3 × 10 3 CL1-5/F4 cells or 2 × 10 3 Jurkat cells were seeded into 96-well plates and incubated for 24 hours.…”

“…Inhibition of NF-κB was detected using the immunocytochemical fluorescent imaging according to previous reports4851. Briefly, treated or untreated cells seeded on coverslips were fixed, permeabilized with Triton X-100, treated with anti-NF-κB p65 rabbit monoclonal primary antibody Abcam #ab32536, followed by Alexa Fluor 488 Donkey anti-rabbit IgG secondary antibody Biolegend #406416.…”

NO-Donor Nitrosyl Iron Complex with 2-Aminophenolyl Ligand Induces Apoptosis and Inhibits NF-κB Function in HeLa Cells

“…NF-κB signaling is well recognized as an important molecular crossway that contributes to various diseases, including cancer [67][68][69][70][71][72][73]. Chemical compounds that inhibit NF-κB, either directly or via its upstream regulators, cause cell death and suppress tumor growth [74][75][76][77][78][79][80][81][82][83] or sensitize cancer cells to radiation or chemotherapeutic agents [84][85][86].…”

<p>Efficient Labeling Of Mesenchymal Stem Cells For High Sensitivity Long-Term MRI Monitoring In Live Mice Brains</p>

“…We used the WST-1 cell proliferation reagent (Roche Diagnostics, Germany) to test the viability of FGF21 MSCs according to our previous publications 25,26 with slight modification. We seeded the cells in 96-well plates and incubated them at the optimum growth conditions to reach a confluency of about 80%.…”

“…Extracted total proteins of cells were denatured, subjected to electrophoresis separation in SDS-PAGE gels, and transferred to polyvinylidene difluoride polyacrylamide (PVDF) membranes according to previous protocols. [25][26][27] Rabbit anti-mouse FGF21 antibody (Aviscera Bioscience, Code: A00145-03-100) and beta-actin antibody (C4) (Santa Cruz Biotechnology, Code: sc-47778) were used to probe the FGF21 and beta-actin proteins, respectively (stripping and reprobing were performed). Horseradish peroxidase (HRP) secondary antibodies and Immobilon Western Chemiluminescent HRP Substrate kit were used to visualize the protein bands.…”