Morpholine
motif is an important pharmacophore and, depending on
the molecular design, may localize in cellular acidic vesicles. To
understand the importance of the presence of pendant morpholine in
a metal complex, six bidentate N,O-donor ligands with or without a
pendant morpholine unit and their corresponding ruthenium(II) p-cymene complexes (1–6) are synthesized, purified, and structurally characterized by various
analytical methods including X-ray diffraction. Complexes 2–4 crystallized in the P21/c space group, whereas 5 and 6 crystallized in the P1̅ space group.
The solution stability studies using 1H NMR support instantaneous
hydrolysis of the native complexes to form monoaquated species in
a solution of 3:7 (v/v) dimethyl sulfoxide-d
6 and 20 mM phosphate buffer (pH* 7.4, containing 4 mM NaCl).
The monoaquated complexes are stable for at least up to 24 h. The
complexes display excellent in vitro antiproliferative activity (IC50 ca. 1–14 μM) in various cancer cell lines,
viz., MDA-MB-231, MiaPaCa2, and Hep-G2. The presence of the pendant
morpholine does not improve the dose efficacy, but rather, with 2-[[(2,6-dimethylphenyl)imino]methyl]phenol
(HL1) and its pendant morpholine analogue (HL3) giving complexes 1 and 3, respectively, the antiproliferative
activity was poorer with 3. MDA-MB-231 cells treated
with the complexes show that the acidic vesicles remain acidic, but
the population of acidic vesicles increases or decreases with time
of exposure, as observed from the dispersed red puncta, depending
on the complex used. The presence of the 2,6-disubstituted aniline
and the naphthyl group seems to improve the antiproliferative dose.
The complex treated MDA-MB-231 cells show that cathepsin D, which
is otherwise present in the cytosolic lysosomes, translocates to the
nucleus as a result of exposure to the complexes. Irrespective of
the presence of a morpholine motif, the complexes do not activate
caspase-3 to induce apoptosis and seem to favor the necrotic pathway
of cell killing.