Department of Medical Biophysics, Institute of Biophysics, Faculty of Biology and Environmental Protection, University of Lodz, 141/143 Pomorska Street, 90-236 Lodz, Poland.
Department of Pharmacy, Cosmetic Chemistry and Biotechnology, Team of Chemistry, Łukasiewicz Research Network-Industrial Chemistry Institute, 8 Rydygiera Street, 01-793 Warsaw, Poland.
Int J Mol Sci. 2022 Feb 3;23(3):1760. doi: 10.3390/ijms23031760.
Despite the continuous developments in pharmacology and the high therapeutic effect of new treatment options for patients with hematological malignancies, these diseases remain a major health issue. Our study aimed to synthesize, analyze in silico, and determine the biological properties of new melphalan derivatives. We obtained three methyl esters of melphalan having in their structures amidine moieties substituted with thiomorpholine (EM-T-MEL), indoline (EM-I-MEL), or 4-(4-morpholinyl) piperidine (EM-MORPIP-MEL). These have not yet been described in the literature. The in vitro anticancer properties of the analogs were determined against THP1, HL60, and RPMI8226 cells. Melphalan derivatives were evaluated for cytotoxicity (resazurin viability assay), genotoxicity (alkaline comet assay), and their ability to induce apoptosis (Hoechst33342/propidium iodide double staining method; phosphatidylserine translocation; and caspase 3/7, 8, and 9 activity measurements). Changes in mitochondrial membrane potential were examined using the specific fluorescence probe JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazol carbocyanine). The EM-T-MEL derivative had the highest biological activity, showing higher cytotoxic and genotoxic properties than the parent drug. Moreover, it showed a high ability to induce apoptosis in the tested cancer cells. This compound also had a beneficial effect in peripheral blood mononuclear cells (PBMC). In conclusion, we verified and confirmed the hypothesis that chemical modifications of the melphalan structure improved its anticancer properties. The conducted study allowed the selection of the compound with the highest biological activity and provided a basis for chemical structure-biological activity analyses.
尽管在药理学方面不断发展，并且新的治疗选择对血液系统恶性肿瘤患者具有很高的治疗效果，但这些疾病仍然是一个主要的健康问题。我们的研究旨在合成、计算机分析并确定新美法仑衍生物的生物学特性。我们获得了三种美法仑的甲酯，其结构中的脒基部分被硫代吗啉（EM-T-MEL）、吲哚啉（EM-I-MEL）或 4-（4-吗啉基）哌啶（EM-MORPIP-MEL）取代。这些在文献中尚未描述过。对类似物的体外抗癌特性进行了测定，针对 THP1、HL60 和 RPMI8226 细胞。对美法仑衍生物进行了细胞毒性（resazurin 活力测定）、遗传毒性（碱性彗星试验）以及诱导细胞凋亡的能力（Hoechst33342/碘化丙啶双重染色法；磷脂酰丝氨酸易位；以及 caspase 3/7、8 和 9 活性测量）的评估。使用特异性荧光探针 JC-1（5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑碳菁）检查线粒体膜电位的变化。EM-T-MEL 衍生物具有最高的生物学活性，表现出比母体药物更高的细胞毒性和遗传毒性特性。此外，它在测试的癌细胞中具有很高的诱导细胞凋亡的能力。该化合物对外周血单核细胞（PBMC）也有有益的作用。总之，我们验证并证实了这样的假设，即美法仑结构的化学修饰改善了其抗癌特性。进行的研究允许选择具有最高生物学活性的化合物，并为化学结构-生物学活性分析提供了基础。
