Esterase-sensitive nitric oxide donors of the diazeniumdiolate family: In vitro antileukemic activity
Esterase-sensitive nitric oxide donors of the diazeniumdiolate family: In vitro antileukemic activity
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DOI：
10.1021/jm9903850
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发表时间：
2000-01-27
影响因子：
7.3
通讯作者：
Keefer, LK
中科院分区：
文献类型：
--
作者：
Saavedra, JE;Shami, PJ;Keefer, LK
我们设计了一种新的前药，它在中性水介质中稳定，但通过酯酶代谢释放生物活性一氧化氮（NO）。将结构为R2N- n (O)=N-OR‘， R’ = Na的重氮双酸酯与BrCH2OAc反应，将自发释放NO的盐1a （R2N =二乙基氨基）和1b （R2N =吡咯里迪诺）分别转化为前药2a （AcOM-DEA/NO）和2b (AcOM-PYRRO/NO)，其中R' = CH2OAc。与阴离子la和Ib（在pH 7.4的磷酸盐中，37℃下的半衰期分别为2分钟和3秒）相比，在这些条件下，2a和2b在16小时后仅表现出最小的分解。然而，在猪肝酯酶存在的情况下，水解发生得非常快，在酯酶诱导的2a生成NO的过程中，观察到自由阴离子1a是一种中间体。通过比较1和2在no敏感的人白血病细胞系HL-60和U937中作为抗增殖药物的作用，说明了这类前药的潜在效用。在HL-60细胞培养中，1a和1b（在整个培养基中产生NO）的72 h IC50值为60 μ M，而2a和2b的IC50值分别为8.3和6.4 μ M。这一结果与我们的假设一致，即2在细胞内被选择性地水解为1，然后被水解为NO。对于U937细胞，2a和2b的72小时IC50均为53 μ M。相比之下，R' = CH2CH2SC(O)Me的类似物具有较高的抗增殖IC50（在U937细胞中为100 μ M），其中乙酰基和2-巯基必须依次裂解才能释放no释放的重氮二酸酯功能。在初始浓度为50 μ M的24 h内，2a和2b分别诱导50%和57%的HL-60细胞凋亡（U937细胞分别为35%和40%）。这些数据揭示了这些新化合物在体外显著的抗白血病活性。此外，它们比阴离子重氮二酸盐更容易处理的优点表明它们可以作为探测NO的生物学作用的方便试剂。
We have designed a novel prodrug class that is stable in neutral aqueous media but releases bioactive nitric oxide (NO) on metabolism by esterase. Diazeniumdiolates of structure R2N-N(O)=N-OR', in which R' = Na, were reacted with BrCH2OAc to convert the spontaneously NO-releasing salts 1a (R2N = diethylamino) and 1b (R2N = pyrrolidino) to prodrugs 2a (AcOM-DEA/NO) and 2b (AcOM-PYRRO/NO), respectively, where R' = CH2OAc. In contrast to anions la and Ib (half-lives in pH 7.4 phosphate at 37 degrees C of 2 min and 3 s, respectively), 2a and 2b showed only minimal decomposition after 16 h under these conditions. Very rapid hydrolysis occurred in the presence of porcine liver esterase, however, with free anion 1a being observed as an intermediate in the esterase-induced generation of NO from 2a. The potential utility of this prodrug class is illustrated with a comparison of 1 and 2 as antiproliferative agents in NO-sensitive human leukemia cell lines HL-60 and U937. While the 72-h IC50's for 1a and 1b (which generate NO throughout the medium) in HL-60 cell cultures were >600 mu M, those of 2a and 2b were 8.3 and 6.4 mu M, respectively. This result is consistent with our hypothesis that 2 is selectively hydrolyzed to 1 and thence to NO intracellularly. For U937 cells, the 72-h IC50 for both 2a and 2b was 53 mu M. By contrast, relatively high antiproliferative IC50's (>100 mu M in U937 cells) were observed for analogues in which R' = CH2CH2SC(O)Me, from which acetyl and 2-mercaptoethyl groups must be successively cleaved to free the NO-releasing diazeniumdiolate function. Within 24 h at initial concentrations of 50 mu M, 2a and 2b induced apoptosis in 50% and 57% of the HL-60 cells, respectively (35% and 40% of the U937 cells, respectively). The data reveal significant in vitro antileukemic activity on the part of these novel compounds. Moreover, their substantial ease-of-handling advantages over the anionic diazeniumdiolates from which they are derived suggest their use as convenient agents for probing the biological roles of NO.