改善药物的体外稳定性酯化合物在大鼠和狗血清:抑制特定的酯酶和影响他们的身份。

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Koitka M, Hochel J, Gieschen H, Borchert HH

改善药物的体外稳定性酯化合物在大鼠和狗血清:抑制特定的酯酶和影响他们的身份。

J制药生物医学肛门。2010年2月5日,51 (3):664 - 78。doi: 10.1016 / j.jpba.2009.09.023。Epub 2009年9月23日。

PubMed ID

19850433 (在PubMed

]

文摘

在药物开发中,已经发现一些药物化合物,特别是酯在体外血清样本是不稳定的。这可能导致大幅低估实际的药物浓度。老鼠和狗,代表一种啮齿动物和non-rodent物种,分别在临床前研究中被广泛使用。我们研究了三种结构不同的降解药物血清酯在大鼠和狗。此外,选择酶抑制剂,以防止这些退化的效率进行了研究。此外,我们发现的迹象的身份还酯酶的抑制剂的敏感性以及蛋白质纯化和鉴定。研究药物sagopilone、屈和甲强龙aceponate (MPA) (pre -)中使用的所有的临床药物开发。的sagopilone-cleaving酯酶在大鼠血清丝氨酸水解酶抑制剂所抑制。我们部分纯化酯酶导致活动率为5%,净化472倍。使用matrix-assisted激光解吸电离(MALDI)飞行时间质谱分析(MS) (TOF),河鼠羧酸酯酶同工酶ES-1被确定在这些分数,从而指出其参与sagopilone乳沟。 Drospirenone cleavage in rat serum was effected by butyrylcholinesterase (BChE) and paraoxonase 1 (PON1) as we deduced from the high efficacy of certain serine hydrolase and metallohydrolase inhibitors, respectively. Likewise, some inhibition characteristics implied that MPA was cleaved in rat serum by BChE and serine proteases. Partial purification of the MPA-specific esterases resulted in activity yields of 1-2%, exhibiting up to 10,000-fold purification. In dog serum, we found that sagopilone was not degraded which was in contrast to MPA and drospirenone. MPA degradation was mainly prevented by serine hydrolase inhibitors. We used a three-step purification to isolate the esterases cleaving MPA. This procedure resulted in an activity yield of 12% and 645-fold purification. By protein identification using liquid chromatography (LC)-electrospray ionization (ESI)-MS, we identified alpha(2)-macroglobulin (alpha(2)M) in the active fractions. We therefore assumed that serine hydrolases, probably butyrylcholinesterase, known to form esteratically active complexes with alpha(2)M, were responsible for MPA cleavage. In contrast, PON1 was assumed to be involved in drospirenone cleavage due to the high efficiency of metallohydrolase inhibitors. This indication was supported by the presence of PON1 in drospirenone-cleaving fractions as we found by affinity chromatography and Western immunoblotting for isolation and detection of PON1, respectively. The identity of the assumed cleaving enzymes remains, however, to be further studied. The inhibitors we found can serve as a tool for stabilizing drug ester compounds in biological samples ex vivo.

DrugBank数据引用了这篇文章

药物酶

药物	酶	类	生物	药理作用	行动
屈	血清paraoxonase /芳香酯1	蛋白质	人类	未知的	底物	细节