Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106, Warsaw, Poland.
Medical University of Warsaw, Nowogrodzka 59, 02-006, Warsaw, Poland.
Chem Biodivers. 2021 Jan;18(1):e2000733. doi: 10.1002/cbdv.202000733. Epub 2020 Dec 30.
Currently available chemotherapeutic treatments for blood cancers (leukemia) usually have strong side effects. More selective, efficient, and less toxic anticancer agents are needed. We synthesized seven, new, optically pure (12aS)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dione derivatives and examined their cytotoxicity towards eight cancer cell lines, including urinary bladder (TCC-SUP, UM-UC-3, KU-19-9), colon (LoVo), and breast (MCF-7, MDA-MB-231) cancer representatives, as well as two leukemic cell lines (MV-4-11, CCRF-CEM) and normal murine fibroblasts (Balb/3T3) as reference cell line. Three of the seven newly-obtained compounds ((12aS)-8-bromo-2-(3-phenylbenzoyl)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dione, (12aS)-8,9-dimethoxy-2-(4-phenylbenzoyl)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dione and (12aS)-8-nitro-2-(4-phenylbenzoyl)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dione, showed enhanced activity and selectivity toward the leukemic MV-4-11 cell lines when compared to our previously reported compounds, with IC values in the range of 2.9-5.6 μM. Additionally, (12aS)-9-nitro-2-(4-phenylbenzoyl)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dione exhibited a strong cytotoxic effect against the leukemic CCRF-CEM (IC =6.1 μM) and MV-4-11 (IC =11.0 μM) cell lines, a moderate cytotoxic effect toward other tumor lines (IC =31.8-55.0 μM) and very weak cytotoxic effect toward the Balb/3T3 reference cell lines. Selected compounds were further evaluated for their potential to induce apoptotic cell death in MV-4-11 cells by measuring caspase-3 activity. We also established the crystal structure of three products and investigated the effect of 22 derivatives of 1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dione on the activity of the cancer-associated enzyme autotaxin. All compounds proved to be weak inhibitors of autotaxin, although some (R) and (S) enantiomers had K values of 10-19 μM. The obtained results showed that the tested compounds exhibited a selective antileukemic effect, which appeared not to be related directly to autotaxin. Molecular targets responsible for this effect remain to be identified. The newly obtained compounds can be used in the search for new, selective anticancer therapies.
目前用于血液癌症（白血病）的化疗药物通常具有很强的副作用。需要更具选择性、更有效和毒性更低的抗癌药物。我们合成了七种新的光学纯（12aS）-1,3,4,12a-四氢吡嗪并[2,1-c][1,4],12(2H,11H)-二酮衍生物，并研究了它们对八种癌细胞系的细胞毒性，包括膀胱癌（TCC-SUP、UM-UC-3、KU-19-9）、结肠癌（LoVo）和乳腺癌（MCF-7、MDA-MB-231）代表细胞系，以及两种白血病细胞系（MV-4-11、CCRF-CEM）和正常鼠成纤维细胞（Balb/3T3）作为参考细胞系。七种新获得的化合物中的三种（（12aS）-8-溴-2-（3-苯甲酰基）-1,3,4,12a-四氢吡嗪并[2,1-c][1,4],12(2H,11H)-二酮、（12aS）-8,9-二甲氧基-2-（4-苯甲酰基）-1,3,4,12a-四氢吡嗪并[2,1-c][1,4],12(2H,11H)-二酮和（12aS）-8-硝基-2-（4-苯甲酰基）-1,3,4,12a-四氢吡嗪并[2,1-c][1,4],12(2H,11H)-二酮对白血病 MV-4-11 细胞系表现出增强的活性和选择性，与我们之前报道的化合物相比，IC 值在 2.9-5.6μM 范围内。此外，（12aS）-9-硝基-2-（4-苯甲酰基）-1,3,4,12a-四氢吡嗪并[2,1-c][1,4],12(2H,11H)-二酮对白血病 CCRF-CEM（IC=6.1μM）和 MV-4-11（IC=11.0μM）细胞系表现出强烈的细胞毒性作用，对其他肿瘤细胞系（IC=31.8-55.0μM）具有中等细胞毒性作用，对 Balb/3T3 参考细胞系具有非常弱的细胞毒性作用。选择的化合物进一步通过测量半胱天冬酶-3 的活性来评估它们在 MV-4-11 细胞中诱导细胞凋亡的潜力。我们还建立了三种产物的晶体结构，并研究了 1,3,4,12a-四氢吡嗪并[2,1-c][1,4],12(2H,11H)-二酮的 22 种衍生物对癌症相关酶自分泌酶的活性的影响。所有化合物均被证明是自分泌酶的弱抑制剂，尽管一些（R）和（S）对映体的 K 值为 10-19μM。所得结果表明，所测试的化合物表现出选择性抗白血病作用，这似乎与自分泌酶没有直接关系。负责这种作用的分子靶标仍有待确定。新获得的化合物可用于寻找新的、选择性的抗癌疗法。
