In Vitro and In Vivo Anti-Cancer Activity Assessment, X-ray Single-Crystal Analysis, and Synthesis of Strong Indolyl-Hydrazones as Kinase Inhibitors for Breast Cancer

Abstract

According to statistics from the World Health Organisation (WHO), 2.3 million women worldwide were diagnosed with
breast cancer in 2020; 685,000 of these cases ended in death.In the continuous fight against breast cancer, it is critical to investigate
innovative approaches as the disease's incidence numbers continue to grow. As a result, ethyl 3-formyl-1H-indole-2-carboxylate 1 was
reacted with thiosemicarbazide and semicarbazide to create a variety of novel indolyl-hydrazones.The new hit compounds were
obtained by combining HCl, 4-nitrophenylhydrazine, 2-dinitrophenylhydrazine, and 4-amino-5-(1H-indol-2-yl)-1,2,4-triazole-3-
thione. By using NMR, elemental analysis, and X-ray single-crystal analysis, these compounds were given chemical structures
asthiosemicarbazone3, bis(hydrazinederivative)5, semicarbzone6, Schiff base 8, and the corresponding hydrazones 10 and 12.The
compounds' cytotoxicity against breast cancer cells (MCF-7) was examined using the MTT test.In contrast to staurosproine (IC50 =
8.32±0.43μM), cytotoxicity data showed strong IC50 values against MCF-7, particularly compounds 5, 8, and 12, with IC50 values of
2.73±0.14, 4.38±0.23, and 7.03±0.37μM, respectively. Thus, the wound-healing test was used to examine the activities of compounds
5, 8, and 12 in connection to cell migration. With corresponding percentages of wound closure recorded at 48.8%, 60.7%, and 51.8%,
the results demonstrated noteworthy wound-healing effectiveness.By looking at the compounds' ability to induce apoptosis, the effect
of the hit compounds on proliferation was evaluated. Remarkably, compound 5 significantly increased cell death in MCF-7 cells,
showing a noteworthy 39.26% increase compared to the untreated control group, which showed just 1.27% cell death. Additionally,
compound 5's mode of action was examined by testing it against kinase receptors. When compared to conventional medications that
target these receptors, the findings showed promising efficacy and considerable kinase inhibition, especially against PI3K-α, PI3K-β,
PI3K-δ, CDK2, AKT-1, and EGFR. Using an in vivo experiment, compound 5 showed a significant decrease in tumour volume in the
concluding phase, going from 106 mm3 in the untreated control to 56.4 mm3. Additionally, it greatly reduced tumour growth by
46.9%. Because they successfully prevent both cell migration and proliferation, the found leads show promise for possible
development into future drugs for the treatment of breast cancer in light of these results