Targeting MAP kinase for inhibiting Leishmania promastigotes and amastigotes stages by L-alanine derived molecules and their anticancer potential against PANC-1 cancer cell lines.

Home to a quarter of the global population, the World Health Organization's South-East Asia Region bears a substantial burden of Neglected Tropical Diseases (NTDs). Identifying means to eliminate NTDs in these regions has been a flagship priority since 2014, with India, Nepal, and Bangladesh committed to eliminating Kala-azar or Visceral Leishmaniasis (VL) sooner or by the end of 2026 (WHO, 2023a). In this context, vanillin-derived molecules are well established in inhibiting several kinases present in cancer cells and have high LD values along with broad therapeutic window. In view of the demand of suitable oral candidate for controlling Leishmania parasitic proliferation in human body here, we are reporting ortho vanillin and l-Alanine amino acid derived ligand molecule t2-{(E)-[(2-hydroxy-3-methoxyphenyl) methylidene] amino} propanoate (L) and its Cu(II) conjugates L & L as water-soluble, and nontoxic drug candidates for the treatment of Visceral Leishmaniasis. In silico binding study against potential chemotherapeutic target MAP kinase (Mitogen-Activated Protein Kinase, (PDB id: 4QNY) was undertaken and significant binding was shown by these molecules. Both compounds demonstrated significant inhibition of amastigote proliferation. For L. donovani, L exhibited an IC of 1.544 μgml and L an IC of 1.691 μgml. Against L. mexicana, the IC values were 1.750 μgml for L and 1.748 μgml for L. These results were comparable to the standard reference drug Amphotericin B, which exhibited an IC of 1.450 μgml for L. donovani and 1.532 μgml for L. mexicana. The toxicity studies were undertaken against macrophages THP-1 cell lines where the IC value is quite higher in comparison to Amphotericin-B. Additionally, the potential of molecules was checked against PANC-1 cancer cell lines in comparison to normal HPNE cell lines. Both complexes caused double-strand DNA nicks, consistent with the higher cytotoxic activities observed in Cancer cell lines. Furthermore, the cell cycle results also confirmed that L arrested (PANC-1) cells in the G1-S phase of the cell cycle followed by an increase in the sub-G1 phase. L, has shown that the population of late/secondary cellular apoptotic cells increased from 2.35 % to 93.56 %, whereas the population of early/primary apoptotic cells increased from 0.13 % (control) to 12.08 %. These findings suggest that L causes apoptosis, or cell death that is programmed.