Nikoleousakos Nikolaos, Dalezis Panagiotis, Polonifi Aikaterini, Geromichalou Elena G, Sagredou Sofia, Alifieris Constantinos E, Deligiorgi Maria V, Sarli Vasiliki, Trafalis Dimitrios T
Laboratory of Pharmacology, Faculty of Medicine, National and Kapodistrian University of Athens, 11527 Athens, Greece.
Department of Chemistry, Aristotle University of Thessaloniki, University Campus, 54124 Thessaloniki, Greece.
Biomedicines. 2021 Aug 17;9(8):1028. doi: 10.3390/biomedicines9081028.
We evaluated three newly synthesized B-lactam hybrid homo-aza-steroidal alkylators (ASA-A, ASA-B and ASA-C) for their PARP1/2 inhibition activity and their DNA damaging effect against human ovarian carcinoma cells. These agents are conjugated with an alkylating component (POPA), which also served as a reference molecule (positive control), and were tested against four human ovarian cell lines in vitro (UWB1.289 + BRCA1, UWB1.289, SKOV-3 and OVCAR-3). The studied compounds were thereafter compared to 3-AB, a known PARP inhibitor, as well as to Olaparib, a standard third-generation PARP inhibitor, on a PARP assay investigating their inhibitory potential. Finally, a PARP1 and PARP2 mRNA expression analysis by qRT-PCR was produced in order to measure the absolute and the relative gene expression (in mRNA transcripts) between treated and untreated cells. All the investigated hybrid steroid alkylators and POPA decreased in vitro cell growth differentially, according to the sensitivity and different gene characteristics of each cell line, while ASA-A and ASA-B presented the most significant anticancer activity. Both these compounds induced PARP1/2 enzyme inhibition, DNA damage (alkylation) and upregulation of PARP mRNA expression, for all tested cell lines. However, ASA-C underperformed on average in the above tasks, while the compound ASA-B induced synthetic lethality effects on the ovarian cancer cells. Nevertheless, the overall outcome, leading to a drug-like potential, provides strong evidence toward further evaluation.
我们评估了三种新合成的β-内酰胺杂化高氮甾体烷基化剂（ASA-A、ASA-B和ASA-C）对PARP1/2的抑制活性以及它们对人卵巢癌细胞的DNA损伤作用。这些试剂与一种烷基化成分（POPA）共轭，POPA也用作参考分子（阳性对照），并在体外对四种人卵巢癌细胞系（UWB1.289 + BRCA1、UWB1.289、SKOV-3和OVCAR-3）进行了测试。此后，在一项研究其抑制潜力的PARP测定中，将所研究的化合物与已知的PARP抑制剂3-AB以及标准的第三代PARP抑制剂奥拉帕尼进行了比较。最后，通过qRT-PCR进行了PARP1和PARP2 mRNA表达分析，以测量处理细胞和未处理细胞之间的绝对和相对基因表达（以mRNA转录本计）。根据每个细胞系的敏感性和不同基因特征，所有研究的杂化甾体烷基化剂和POPA均不同程度地降低了体外细胞生长，而ASA-A和ASA-B表现出最显著的抗癌活性。对于所有测试的细胞系，这两种化合物均诱导了PARP1/2酶抑制、DNA损伤（烷基化）和PARP mRNA表达上调。然而，ASA-C在上述任务中平均表现较差，而化合物ASA-B对卵巢癌细胞诱导了合成致死效应。尽管如此，总体结果显示出具有类药物潜力，为进一步评估提供了有力证据。
