Column Chromatography-Free Synthesis of Spirooxindole and Spiroindanone-Based Naphthalimides as Potent c-MYC G4 Stabilizers and HSA Binders for Elevating Anticancer Potential.

Abstract

G-quadruplex (G4) DNA plays a pivotal regulatory role in fundamental biological processes, integral for governing cellular functions such as replication, transcription, and repair in living cells. Within cancer cells, G4 DNA exerts an impact on the expression of crucial genes such as c-MYC, effectively repressing its activity when structured within its promoter region. Therefore, employing molecular scaffolds to target these structures offers an attractive strategy for altering their functions. In our pursuit of potent and selective G-quadruplex binders, herein we report a series of spironaphthalimide-pyrrolidine analogues that demonstrate the ability to stabilize c-MYC G4 formation and subsequently inhibit c-MYC expression. These analogues are evaluated for their anticancer activity against 60 human cancer cell lines at 10 μM. The most potent analogues 8j and 21c underwent additional testing at five dose concentrations (10^-4-10^-8 M) where low MG-MID GI₅₀ values are observed for both the analogues 8j (9.98 μM) and 21c (2.49 μM). To correlate with the antiproliferative activity, the mechanism is explored in vitro by performing Pu27 DNA binding studies through multispectroscopic techniques, and the results are compared with Pu22, human telomere, and calf thymus DNA. Additionally, insights from molecular docking suggested stacking over the G-tetrad of G4 structures of both analogues, with quantum mechanical studies further reinforcing the rationale for the stability of this quadruplex secondary structure. The analogues are also evaluated for their binding affinity to human serum albumin, revealing their robust capability to effectively bind and potentially facilitate targeted delivery to specific sites. Amidst the abundance of G4s across the human genome, the above findings underscore the significance of spiro analogues, with potent multitargeting anticancer attributes, marking a transformative leap forward in G4-ligand innovation, promising frontiers in the quest for effective anticancer modalities.