背景:小说,在我们的实验室合成了硅设计的抗癌化合物,即,2-乙基-3-O-磺酰基-雌胺-1,3,5(10),15-四烯-17-醇(ESE-15-醇)和2-乙基-3-O-磺酰基-雌胺-1,3,5(10)16-四烯(ESE-16)。与它们的来源化合物相比,这些化合物被设计成具有改善的生物利用度,2-甲氧基雌二醇。从理论上讲,这是因为它们与碳酸酐酶II的结合亲和力增加,存在于红细胞中。由于研究中的新化合物被提议在与碳酸酐酶II结合的红细胞内运输,它们对全血和红细胞的形态学作用具有重要意义。次要结果包括修订先前报道的处理全血样品的程序。这项研究的目的是双重的。首先,在暴露于新设计的化合物后,通过扫描电子显微镜(SEM)检查了健康女性红细胞的超微结构形态。使用SEM描述了在22°C下暴露于ESE-15-ol和ESE-163分钟和24小时后的红细胞形态。在24小时暴露后,化合物的溶血活性也用离体溶血测定法测定。其次,通过测定在22°C和37°C下24小时储存期后的形态变化来研究全血样品的储存条件。
结果:在暴露于新的抗癌化合物后,在红细胞形态中没有观察到显著的形态变化。将全血样品在37°C下储存24小时导致红细胞中可见的形态应力。在22°C下孵育24小时的红细胞显示没有结构畸形或不适。
结论:从这项研究中,确定全血样品离体暴露于ESE-15-ol和ESE-1624小时的最佳温度为22°C。这项研究的数据揭示了这些化合物应用于离体研究技术的潜力,因为在这些条件下对红细胞超微结构没有损害。由于在暴露于ESE-15-ol和ESE-16的红细胞中没有观察到结构变化,因此将进行进一步的离体实验来研究这些化合物对全血的潜在作用。作为次要结果,建立了对于全血直至24小时的最佳孵育条件。
BACKGROUND: Novel, in silico-designed anticancer compounds were synthesized in our laboratory namely, 2-ethyl-3-O-sulphamoyl-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.