Abstract:
BACKGROUND : Novel, in silico-designed anticancer compounds were synthesized in our laboratory namely, 2-ethyl-3-Osulphamoyl-
estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) and 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16).
These compounds were designed to have improved bioavailability when compared to their source compound,
2-methoxyestradiol. This theoretically would be due to their increased binding affinity to carbonic anhydrase II,
present in erythrocytes. Since the novel compounds under investigation are proposed to be transported within
erythrocytes bound to carbonic anhydrase II, the morphological effect which they may exert on whole blood
and erythrocytes is of great significance. A secondary outcome included revision of previously reported procedures for
the handling of the whole blood sample.
The purpose of this study was twofold. Firstly, the ultrastructural morphology of a healthy female’s erythrocytes was
examined via scanning electron microscopy (SEM) after exposure to the newly in silico-designed compounds.
Morphology of erythrocytes following exposure to ESE-15-ol and ESE-16 for 3 minutes and 24 hours at 22°C were
described with the use of SEM. The haemolytic activity of the compounds after 24 hours exposure were also determined
with the ex vivo haemolysis assay. Secondly, storage conditions of the whole blood sample were investigated by
determining morphological changes after a 24 hour storage period at 22°C and 37°C.
RESULTS : No significant morphological changes were observed in the erythrocyte morphology after exposure to
the novel anticancer compounds. Storage of the whole blood samples at 37°C for 24 hours resulted in visible
morphological stress in the erythrocytes. Erythrocytes incubated at 22°C for 24 hours showed no structural
deformity or distress.
CONCLUSIONS : From this research the optimal temperature for ex vivo exposure of whole blood samples to
ESE-15-ol and ESE-16 for 24 hours was determined to be 22°C. Data from this study revealed the potential of
these compounds to be applied to ex vivo study techniques, since no damage occurred to erythrocytes ultrastructure
under these conditions. As no structural changes were observed in erythrocytes exposed to ESE-15-ol and ESE-16, further
ex vivo experiments will be conducted into the potential effects of these compounds on whole blood. Optimal
incubation conditions up to 24 hours for whole blood were established as a secondary outcome.
