Lupane derivatives containing various aryl substituents in the position 3 have selective cytostatic effect in leukemic cancer cells including resistant phenotypes

Eur J Med Chem. 2022 Dec 15:244:114850. doi: 10.1016/j.ejmech.2022.114850. Epub 2022 Oct 20.


In this work, a large set of betulinic acid derivatives modified with various aromatic substituents at the position C-3 were prepared via Suzuki-Myiaura cross-coupling. All compounds were tested for their in vitro cytotoxic activity in 8 cancer and 2 healthy cell lines. Derivatives 6h, 6i, and 6o had the lowest IC50 in the CCRF-CEM cell line (0.69-4.0 μM) and had high selectivity. In addition, 6h and 6i also showed significant activity in daunorubicin-resistant CEM and taxol-resistant K562 cell lines; therefore, they were selected for the evaluation of the mechanism of action. First, the effect of 6h, 6i, and 6o on cell death induction was studied. To our surprise, we have not detected almost any apoptotic cells, even following a long-time exposure of CCRF-CEM cells to the compounds. On the other hand, a dramatic cell number decrease was observed, proportional to the time of the compound's exposure. Based on this data it was concluded that the effect of compounds is cytostatic rather than cytotoxic, which was further confirmed by subsequent studies of the impact of 6h, 6i, and 6o on the cell cycle. Detailed cell cycle analysis revealed a block in the G1 phase accompanied by reduced expression of phosphorylated forms of the RB protein as well as cyclin A protein. Evaluation of the pharmacological properties of the most promising compounds revealed their high stability in the presence of phosphate buffer, human plasma, and microsomes and limited permeability determined using permeability through artificial membrane (PAMPA) and cell permeability assay: Caco-2 and MDCK-MDR1 cell lines. Compounds 6h, 6i, and 6o were selected for further drug development; their cytostatic effect may be advantageous in this process since we expect fewer non-specific interactions and toxicity than in highly cytotoxic compounds. In addition, the activity of 6h and 6i against resistant CEM-DNR and K562-TAX leukemic cell lines makes them promising as a possible future alternative to currently used therapies.

Keywords: ADME parameters; Anticancer; Betulinic acid; Cell cycle; Cytostatic; Lupane; Suzuki- myiaura coupling; Triterpenes.

MeSH terms

  • Antineoplastic Agents* / pharmacology
  • Apoptosis
  • Caco-2 Cells
  • Cell Line, Tumor
  • Cytostatic Agents* / pharmacology
  • Drug Screening Assays, Antitumor
  • Humans
  • Neoplasms*
  • Pentacyclic Triterpenes / pharmacology
  • Phenotype


  • Cytostatic Agents
  • Pentacyclic Triterpenes
  • Antineoplastic Agents