Lyophilization and stability of antibody-conjugated mesoporous silica nanoparticle with cationic polymer and PEG for siRNA delivery.
Lyophilization and stability of antibody-conjugated mesoporous silica nanoparticle with cationic polymer and PEG for siRNA delivery.
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DOI：
10.2147/ijn.s164393
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发表时间：
2018
影响因子：
8
通讯作者：
Yantasee W
中科院分区：
文献类型：
--
作者：
Ngamcherdtrakul W;Sangvanich T;Reda M;Gu S;Bejan D;Yantasee W
候选治疗药物的长期稳定性对其临床应用是必要的。对于大多数在水溶液中配制的纳米颗粒系统，冻干或冷冻干燥是确保长期稳定性的常用方法。虽然已经研究了脂质、聚合物或无机纳米颗粒的冻干，但关于由聚合物、无机颗粒和抗体组成的杂化纳米颗粒系统的冻干和稳定性的报道很少。复杂纳米颗粒系统的冻干在保存材料的物理化学性质和生物活性方面具有挑战性。我们最近报道了一种有效的小干扰RNA （siRNA）纳米颗粒载体，由50纳米介孔二氧化硅纳米颗粒和聚乙烯亚胺和聚乙烯乙二醇共聚物修饰而成，以及抗体组成。为了实现未来的个性化医疗，纳米颗粒载体被单独冻干，并在磷酸盐缓冲盐水中简单混合几分钟后重组后装载siRNA。在此，我们从缓冲液、冷冻保护剂、重构、冷冻和干燥步骤的时间和温度等方面对纳米颗粒的冻干进行了优化，并监测了材料在不同温度和时间下储存后的物理和化学性质（重构、流体动力学大小、电荷和siRNA装载）和生物活性（基因沉默、癌细胞杀伤）。在Tris-HCl和5% w/w海藻糖的缓冲液中配制该材料效果最佳。冷冻步骤在- 55°C下进行3小时，然后在- 40°C（100µBar）下进行一次干燥步骤24小时，在20°C（20µBar）下进行二次干燥步骤12小时。冻干后的材料可以在4°C下稳定保存2个月，在- 20°C下至少保存6个月。我们成功地开发了冻干工艺，该工艺应适用于其他类似的纳米颗粒系统，该系统由阳离子聚合物、PEG和抗体修饰的无机纳米颗粒核心组成。
Long-term stability of therapeutic candidates is necessary toward their clinical applications. For most nanoparticle systems formulated in aqueous solutions, lyophilization or freeze-drying is a common method to ensure long-term stability. While lyophilization of lipid, polymeric, or inorganic nanoparticles have been studied, little has been reported on lyophilization and stability of hybrid nanoparticle systems, consisting of polymers, inorganic particles, and antibody. Lyophilization of complex nanoparticle systems can be challenging with respect to preserving physicochemical properties and the biological activities of the materials. We recently reported an effective small-interfering RNA (siRNA) nanoparticle carrier consisting of 50-nm mesoporous silica nanoparticles decorated with a copolymer of polyethylenimine and polyethyleneglycol, and antibody. Toward future personalized medicine, the nanoparticle carriers were lyophilized alone and loaded with siRNA upon reconstitution by a few minutes of simple mixing in phosphate-buffered saline. Herein, we optimize the lyophilization of the nanoparticles in terms of buffers, lyoprotectants, reconstitution, and time and temperature of freezing and drying steps, and monitor the physical and chemical properties (reconstitution, hydrodynamic size, charge, and siRNA loading) and biological activities (gene silencing, cancer cell killing) of the materials after storing at various temperatures and times. The material was best formulated in Tris-HCl buffer with 5% w/w trehalose. Freezing step was performed at −55°C for 3 h, followed by a primary drying step at −40°C (100 µBar) for 24 h and a secondary drying step at 20°C (20 µBar) for 12 h. The lyophilized material can be stored stably for 2 months at 4°C and at least 6 months at −20°C. We successfully developed the lyophilization process that should be applicable to other similar nanoparticle systems consisting of inorganic nanoparticle cores modified with cationic polymers, PEG, and antibodies.