Hata Machi, Ueno Jin, Hitomi Yutaka, Kodera Masahito
Molecular Chemistry and Biochemistry, Doshisha University, Tatara-Miyakodani 1-3, Kyotanabe 610-0321, Japan.
ACS Omega. 2023 Jul 27;8(31):28690-28701. doi: 10.1021/acsomega.3c03387. eCollection 2023 Aug 8.
The DNA target/ligand conjugates (HL, X = P and M, = 1-3) were synthesized where various lengths of -CONH(CHCHO)CHCHNHCO- linkers with a 9-phenanthrenyl (P) or methyl (M) terminal as DNA targets replace the methyl group of 2,6-di(amide-tether cyclen)-cresol ligand (HL). DNA binding, DNA cleavage, cellular uptake, and cytotoxicity of Cu(μ-OH)(L) () are examined and compared with those of Cu(μ-OH)(L) () to clarify roles of DNA targets. Upon reaction of with HO, μ-1,1-OH complexes are formed for DNA cleavage. , , and are 22-, 11-, 3-fold more active for conversion of Form II to III in the cleavage of supercoiled plasmid DNA with HO than , where the short P-linker may fix a dicopper moiety within a small number of base pairs to facilitate DNA double-strand breaks (dsb). This enhances the proapoptotic activity of , , and , which are 30-, 12-, and 9.9-fold cytotoxic against HeLa cells than . DNA dsb and cytotoxicity are 44% correlated in but 5% in , suggesting specific DNA binding of P-linkers and nonspecific binding of M-linkers in biological cells. exert cancer cell-selective cytotoxicity against lung and pancreas cancer and normal cells where the short P-linker enhances the selectivity, but do not. Intracellular visualization, apoptosis assay, and caspase activity assay clarify mitochondrial apoptosis caused by . The highest cancer cell selectivity of may be enabled by the short P-linker promoting dsb of mitochondrial DNA with HO increased by mitochondrial dysfunction in cancer cells.
合成了DNA靶标/配体缀合物（HL，X = P和M，n = 1 - 3），其中以9 - 菲基（P）或甲基（M）为末端的不同长度的 -CONH(CH₂CHO)CH₂CH₂NHCO- 连接体作为DNA靶标，取代了2,6 - 二（酰胺 - 连接环糊精） - 甲酚配体（HL）的甲基。研究了Cu(μ-OH)(L)₂（n）的DNA结合、DNA切割、细胞摄取和细胞毒性，并与Cu(μ-OH)(L)₂（m）进行比较，以阐明DNA靶标的作用。n与H₂O₂反应时，形成μ - 1,1 - OH配合物用于DNA切割。在超螺旋质粒DNA与H₂O₂的切割反应中，n₁、n₂和n₃将形式II转化为III的活性分别比m高22倍、11倍和3倍，其中短的P - 连接体可能将双铜部分固定在少数碱基对内，以促进DNA双链断裂（dsb）。这增强了n₁、n₂和n₃的促凋亡活性，它们对HeLa细胞的细胞毒性分别是m的30倍、12倍和9.9倍。DNA dsb与细胞毒性在n中相关性为44%，而在m中为5%，表明P - 连接体在生物细胞中具有特异性DNA结合，而M - 连接体为非特异性结合。n对肺癌和胰腺癌及正常细胞具有癌细胞选择性细胞毒性，其中短的P - 连接体增强了选择性，但m没有。细胞内可视化、凋亡测定和半胱天冬酶活性测定阐明了n引起的线粒体凋亡。n的最高癌细胞选择性可能是由于短的P - 连接体促进了癌细胞中线粒体功能障碍导致的线粒体DNA与H₂O₂的dsb。
