Novel Isochromene-Fused CF3-Substituted Pyrazole Compounds Induce Cytotoxicity through Caspase Mediated Apoptosis

AUTOR(I) / AUTHOR(S): Milica Marković, Nađa Grozdanić, Ana Andrijević, Milica Selaković, Andrea Nikolić, Igor Opsenica, Tatjana Stanojković

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DOI:  10.46793/ICCBIKG25.469M

SAŽETAK / ABSTRACT:

Pyrazoles, particularly their fluorinated derivatives, have emerged as valuable heterocyclic moieties in medicinal chemistry due to their broad spectrum of biological activities, including anticancer potential. In this study, we applied our previously established one-pot, two- step synthetic protocol for pyrazolo[3,4-c]isochromenes to generate a focused library of 1- (trifluoromethyl)-3,5-dihydroisochromeno[3,4-c]pyrazole derivatives via late-stage functionalization with aryldiazonium tetrafluoroborate salts. The selected derivatives, containing biphenyl-4-carbaldehyde, 4-chlorophenyl, 4-phenol, 4-benzenesulfonyl fluoride, or 4- benzenesulfonamide substituents, demonstrated pronounced cytotoxic effects against HeLa cells. To elucidate the apoptotic pathways involved, we assessed cytotoxicity in the presence of selective inhibitors of caspase-3, caspase-8, and caspase-9. Caspase-3 inhibition allowed us to confirm the requirement for executioner caspase activation, indicating apoptosis as the dominant mode of cell death. The use of caspase-8 and caspase-9 inhibitors enabled discrimination between extrinsic and intrinsic apoptotic pathways. Our results indicate subtle distinctions among the examined compounds, with certain compounds primarily activating the intrinsic apoptotic pathway, whereas others induce a comparatively greater involvement of extrinsic signaling, thereby suggesting refined structure–activity relationship patterns. These results broaden the understanding of the molecular mechanisms underlying the anticancer activity of investigated compounds, while also highlighting the potential of the compounds for the development of agents whose primary mechanism of action is apoptosis.

KLJUČNE REČI / KEYWORDS:

Diazo compounds, Heterocycles, Medicinal Chemistry, Cancer, Caspase Inhibitors

PROJEKAT / ACKNOWLEDGEMENT:

This research was financially supported by the Ministry of Technological Development and Innovation of Republic of Serbia (Contract numbers: 451-03-136/2025- 03/200168 and 451-03-136/2025 03/200288 and 451-03-136/2025-03/200043).

LITERATURA / REFERENCES:

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