Bioconjugates of Co(III) complexes with Schiff base ligands and cell penetrating peptides: Solid phase synthesis, characterization and antiproliferative activity

Abstract

In this work we synthesized a chelating Schiff base by a single condensation of salicylaldehyde with 3,4-diamino benzoic acid (1). This ligand was used further for complexation to CoCl₂·6H₂O under nitrogen. In the next step, three six-coordinate Co(III) complexes were synthesized by coordinating this complex with imidazole (2), 2-methyimidazole (3) and N-Boc-l-histidine methyl ester (4) (Boc: tert.-butoxycarbonyl) in axial positions with simultaneous oxidation of Co(II) to Co(III) under ambient environment. All Co(III) complexes were characterized by multinuclear NMR spectroscopy (¹H, ¹³C and ⁵⁹Co NMR), FT-IR, mass spectrometry and HPLC. The Co(III) complexes were conjugated to three different cell penetrating peptides: FFFF (P1), RRRRRRRRRGAL (P2) and FFFFRRRRRRRRRGAL (P3). Standard solid-phase peptide chemistry was used for the synthesis of cell penetrating peptides. Coupling of N-terminal peptides with the cobalt complexes, possessing a carboxylic group on the tetradentate Schiff base ligand, afforded Co(III)-peptide bioconjugates, which were purified by semi-preparative HPLC and characterized by analytical HPLC and mass spectrometry. The antiproliferative activity of the synthesized compounds was studied against different human tumour cell lines: lung cancer A549, liver cancer HepG2 and normal human fibroblasts GM5657T, in comparison with the activity of cisplatin as a reference drug. The bioconjugate 21 containing the Co complex 4 and the combined phenylalanine and polyarginine cell penetrating sequence P3 shows better activity against the liver cancer line HepG2 than the parent Co(III) complex 4.

Keywords: Bioconjugates; Cell targeting peptides; Cobalt(III); Medicinal inorganic chemistry; Schiff base ligands; Solid phase peptide synthesis.