采用平衡透析和 LC/MS/MS 研究新型 2-氨基-1-丙醇化合物与牛血清白蛋白、α1-酸性糖蛋白和大鼠血清的结合。

Binding of new aminopropan-2-ol compounds to bovine serum albumin, α1-acid glycoprotein and rat serum using equilibrium dialysis and LC/MS/MS.

机构信息

Department of Pharmacokinetics and Physical Pharmacy, Jagiellonian University, Medical College, Medyczna 9, PL 30-688 Kraków, Poland.

出版信息

Pharmacol Rep. 2013;65(5):1294-303. doi: 10.1016/s1734-1140(13)71487-7.

DOI:10.1016/s1734-1140(13)71487-7

PMID:24399725

Abstract

BACKGROUND

The binding of three new aminopropan-2-ol compounds briefly called 2F109, ANBL and TWo8 with potential cardiovascular activity to bovine serum albumin (BSA), α1-acid glycoprotein (AGP) and to rat serum was studied. The chemical structures of these compounds are related to carvedilol. They possess an antiarrhythmic and hypotensive activity, and β- and α-adrenolytic mechanism of action. All analogues are weak bases with pKa values 8.65, 8.85 and 8.26 for 2F109, ANBL and TWo8, respectively, and they possess lipophilic character (log P > 1.9584).

METHODS

The extent of protein binding was determined using equilibrium dialysis in the range 2.5 - 900 μM, and 2.5 - 300 μM for binding of investigated compounds to BSA and AGP, respectively, and the quantitative measurement was done by LC/ESI-MS/MS assay.

RESULTS

The studied compounds bound to a single class of binding sites on BSA which was characterized by low affinity (Kd for 2F109 = 8.49 x 10(-5) M, for ANBL= 1.92 x 10(-5) M, and for TWo8 = 1.71 x 10(-5) M) and low capacity (n = 0.53 for 2F109, 0.132 for ANBL and 0.13 for TWo8). The binding of 2F109, ANBL and TWo8 to AGP revealed one class of binding sites, with moderate affinity (Kd for 2F109 = 4.67 x 10(-6) M, for ANBL = 3.48 x 10(-5) M, and for TWo8 = 1.13 x 10(-5) M) and higher capacity (n = 2.21 for 2F109, 2.76 for ANBL and 2.28 for TWo8).

CONCLUSION

The obtained data indicate that 2F109, ANBL and TWo8 moderately bind to BSA (34.2 - 71.2%) with low capacity (Ka = 6.21 x 10(3) - 7.61 x 10(3)M(-1)) and strongly bind to AGP (71.5 - 85.5%) with moderate affinity (Ka = 7.94 x 10(4) - 4.73 x 10(5)M(-1)).

摘要

背景

研究了三种新型氨基丙-2-醇化合物（简称 2F109、ANBL 和 TWo8）与潜在心血管活性与牛血清白蛋白（BSA）、α1-酸性糖蛋白（AGP）和大鼠血清的结合。这些化合物的化学结构与卡维地洛有关。它们具有抗心律失常和降压作用，以及β-和α-肾上腺素能作用机制。所有类似物均为弱碱，pKa 值分别为 2F109 的 8.65、ANBL 的 8.85 和 TWo8 的 8.26，具有亲脂性（log P > 1.9584）。

方法

在 2.5-900 μM 范围内，使用平衡透析法测定蛋白质结合程度，分别在 2.5-300 μM 范围内测定研究化合物与 BSA 和 AGP 的结合程度，并用 LC/ESI-MS/MS 法进行定量测定。

结果

研究化合物与 BSA 上的单一类结合位点结合，其特点是亲和力低（2F109 的 Kd 为 8.49 x 10(-5) M，ANBL 的 Kd 为 1.92 x 10(-5) M，TWo8 的 Kd 为 1.71 x 10(-5) M）和结合容量低（2F109 的 n = 0.53，ANBL 的 n = 0.132，TWo8 的 n = 0.13）。2F109、ANBL 和 TWo8 与 AGP 的结合显示出单一的结合位点，具有中等亲和力（2F109 的 Kd 为 4.67 x 10(-6) M，ANBL 的 Kd 为 3.48 x 10(-5) M，TWo8 的 Kd 为 1.13 x 10(-5) M）和更高的结合容量（2F109 的 n = 2.21，ANBL 的 n = 2.76，TWo8 的 n = 2.28）。

结论

研究结果表明，2F109、ANBL 和 TWo8 与 BSA 结合程度适中（34.2-71.2%），结合容量较低（Ka = 6.21 x 10(3) - 7.61 x 10(3)M(-1)），与 AGP 结合较强（71.5-85.5%），亲和力中等（Ka = 7.94 x 10(4) - 4.73 x 10(5)M(-1)）。