In vitro antiproliferative and apoptotic effects of thiosemicarbazones based on (-)-camphene and R-(+)-limonene in human melanoma cells

PLoS One. 2023 Nov 30;18(11):e0295012. doi: 10.1371/journal.pone.0295012. eCollection 2023.

Abstract

A series of 38 thiosemicarbazone derivatives based on camphene and limonene were evaluated for their antiproliferative activity. Among them, 19 were synthesized and characterized using proton and carbon-13 nuclear magnetic resonance (1H and 13C NMR). For initial compound selection, human melanoma cells (SK-MEL-37) were exposed to a single concentration of a compound (100 μM) for 24, 48, and 72 hours, and cell detachment was visually observed. Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Nineteen compounds (4, 6, 8, 11, 13, 14, 15, 16, 17, 18, 20, 22, 25, 26, 31, 3', 4', 6', and 9') yielded cell viability below 20%. Subsequently, IC50 values for these compounds were determined, ranging from 11.56 to 55.38 μM, after 72 hours of treatment. Compound 17 (o-hydroxybenzaldehyde (-)-camphene-based thiosemicarbazone) demonstrated the lowest IC50 value, followed by compound 4 (benzaldehyde (-) camphene-based thiosemicarbazone) at 12.84 μM. Regarding compound 4, we observed the induction of a characteristic ladder pattern of DNA fragmentation through gel electrophoresis. Furthermore, fluorescence, flow cytometry and scanning microscopy assays revealed morphological changes consistent with apoptosis induction. Additionally, the measurement of caspase 6 and 8 activity in cellular extracts after treatment for 2, 4, 6, and 24 hours suggested the potential involvement of the extrinsic apoptosis pathway in the mechanism of action of compound 4. Further investigations, including molecular docking studies, are required to fully explore the potential of compound 4 and the other selected compounds, highlighting their promising role in future melanoma therapy research.

MeSH terms

  • Antineoplastic Agents* / chemistry
  • Antineoplastic Agents* / pharmacology
  • Apoptosis
  • Cell Line, Tumor
  • Cell Proliferation
  • Drug Screening Assays, Antitumor
  • Humans
  • Limonene / pharmacology
  • Melanoma* / drug therapy
  • Melanoma* / pathology
  • Molecular Docking Simulation
  • Structure-Activity Relationship
  • Thiosemicarbazones* / chemistry
  • Thiosemicarbazones* / pharmacology

Substances

  • Limonene
  • Thiosemicarbazones
  • Antineoplastic Agents

Grants and funding

This study was made possible through the research fellowships awarded to PROS and DCSP by the National Council for Scientific and Technological Development (CNPq) of Brazil. These fellowships supported PROS during her master's studies and DCSP during her doctoral studies, covering personal expenses during the research period, and they did not include direct salaries. CNPq played no additional role in the study design, data collection and analysis, decision to publish, or manuscript preparation. No additional external finding was received for this study.