Jakaria Sardar M, Budil David E, Murtagh James, Revilla Graham
Hikma Pharmaceuticals, Bedford, OH 44146, USA.
Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, USA.
Pharmaceutics. 2024 Nov 22;16(12):1503. doi: 10.3390/pharmaceutics16121503.
The effect of 2-hydroxpropyl-β-cyclodextrin (2HPβCD) with or without divalent metal ions (Ca, Mg, and Zn) on the stability of dalbavancin in acetate buffer was investigated. Dalbavancin recovery from formulations with 2HPβCD and divalent metal ions after four weeks of storage at 5 °C and 55 °C was measured by RP-HPLC and HP-SEC; a longer-term study was carried out over six months at 5 °C, 25 °C, and 40 °C. Binding of 2HPβCD was characterized by isothermal titration calorimetry (ITC) and nuclear magnetic resonance (NMR). The stability of the dalbavancin formulations after 4 weeks at 55 °C in 10 mM acetate buffer was significantly improved with 0.6 mM, 5.5 mM, and 55 mM 2HPβCD relative to without 2HPβCD. No further improvement was observed with the addition of any of the divalent cations. Dalbavancin in a 1:10 molar ratio with 2HPβCD was more stable at a concentration of 1 mg/mL than at 20 mg/mL under accelerated conditions at 40 °C for six months. ITC revealed two 2HPβCD binding sites to dalbavancin in 10 mM acetate: one with a 1:1 stoichiometry and thermodynamics consistent with known cyclodextrin-drug interactions, and a second with 0.1:1 stoichiometry, a positive binding enthalpy, and an unusually large entropy of binding. NMR spectroscopy indicates that dalbavancin exhibits aggregation in acetate buffer that is disrupted by 2HPβCD binding. 2HPβCD significantly improves the short- and long-term heat stability of dalbavancin in pH 4.5 acetate buffer at and above molar ratios of 1:1. The strong 1:1 binding of 2HPβCD to dalbavancin demonstrated by ITC confirms that this stability is conferred by the formation of a stable complex. This observation, combined with the NMR results, points to the aliphatic hydrocarbon chain of the glycone moiety as the most likely site of 2HPβCD-dalbavancin interaction.
研究了含或不含二价金属离子（钙、镁和锌）的2-羟丙基-β-环糊精（2HPβCD）对达巴万星在醋酸盐缓冲液中稳定性的影响。通过反相高效液相色谱（RP-HPLC）和高效体积排阻色谱（HP-SEC）测定了在5℃和55℃储存四周后含2HPβCD和二价金属离子的制剂中达巴万星的回收率；在5℃、25℃和40℃进行了为期六个月的长期研究。通过等温滴定量热法（ITC）和核磁共振（NMR）对2HPβCD的结合进行了表征。相对于不含2HPβCD的情况，在10 mM醋酸盐缓冲液中，55℃下4周后，0.6 mM、5.5 mM和55 mM的2HPβCD显著提高了达巴万星制剂的稳定性。添加任何二价阳离子均未观察到进一步的改善。在40℃加速条件下六个月，与2HPβCD摩尔比为1:10的达巴万星在浓度为1 mg/mL时比在20 mg/mL时更稳定。ITC显示在10 mM醋酸盐中2HPβCD与达巴万星有两个结合位点：一个化学计量比为1:1，其热力学与已知的环糊精-药物相互作用一致，另一个化学计量比为0.1:1，结合焓为正，结合熵异常大。核磁共振光谱表明达巴万星在醋酸盐缓冲液中表现出聚集，而2HPβCD的结合会破坏这种聚集。在pH 4.5醋酸盐缓冲液中，当2HPβCD与达巴万星的摩尔比为1:1及以上时，2HPβCD显著提高了达巴万星的短期和长期热稳定性。ITC证明2HPβCD与达巴万星有很强的1:1结合，这证实了这种稳定性是由形成稳定复合物赋予的。这一观察结果与核磁共振结果相结合，表明糖苷部分的脂肪烃链是2HPβCD与达巴万星相互作用最可能的位点。
