Achieving long-term stability of lipid nanoparticles: examining the effect of pH, temperature, and lyophilization.
Achieving long-term stability of lipid nanoparticles: examining the effect of pH, temperature, and lyophilization.
复制标题
达到脂质纳米颗粒的长期稳定性：检查pH，温度和冻干的影响。
DOI：
10.2147/ijn.s123062
复制
发表时间：
2017
影响因子：
8
通讯作者：
Whitehead KA
中科院分区：
文献类型：
--
作者：
Ball RL;Bajaj P;Whitehead KA
siRNA疗法的广泛临床应用将由在环境条件下长时间保持稳定性和效力的载药递送系统促进。在目前的研究中，我们试图更好地了解脂质纳米颗粒（LNPs）的稳定性和储存条件的影响，我们的团队和其他研究人员已经证明，在体外和体内，LNPs都能有效地将RNA传递到各种细胞和器官靶点。具体来说，本研究评估了pH、温度和冻干对LNP在HeLa细胞中的作用的影响。当LNPs在水条件下储存时，我们发现冷藏（2°C）比在- 20°C冷冻室或室温下储存的LNPs在150天内保持最稳定。由于没有发现储存缓冲液的pH值影响稳定性，因此建议将LNPs储存在生理适宜的条件下（pH值为7），以便于使用。尽管LNPs在冻融循环中会发生聚集和丧失效力，但在使用冷冻保护剂、海藻糖和蔗糖后，LNPs的稳定性得以保持。最初，LNPs冻干后在水缓冲液中重构也会导致功效降低，这很可能是由于重构时的聚集。虽然在重组缓冲液中加入乙醇可以恢复其功效，但这种方法并不理想，因为LNP溶液在使用前需要透析。幸运的是，我们发现在LNP溶液中加入海藻糖或蔗糖，在冻干之前，可以促进室温储存和在水缓冲液中重构，而不会降低递送效力。
The broadest clinical application of siRNA therapeutics will be facilitated by drug-loaded delivery systems that maintain stability and potency for long times under ambient conditions. In the present study, we seek to better understand the stability and effect of storage conditions on lipidoid nanoparticles (LNPs), which have been previously shown by our group and others to potently deliver RNA to various cell and organ targets both in vitro and in vivo. Specifically, this study evaluates the influence of pH, temperature, and lyophilization on LNP efficacy in HeLa cells. When stored under aqueous conditions, we found that refrigeration (2°C) kept LNPs the most stable over 150 days compared to storage in the −20°C freezer or at room temperature. Because the pH of the storage buffer was not found to influence stability, it is suggested that the LNPs be stored under physiologically appropriate conditions (pH 7) for ease of use. Although aggregation and loss of efficacy were observed when LNPs were subjected to freeze–thaw cycles, their stability was retained with the use of the cryoprotectants, trehalose, and sucrose. Initially, lyophilization of the LNPs followed by reconstitution in aqueous buffer also led to reductions in efficacy, most likely due to aggregation upon reconstitution. Although the addition of ethanol to the reconstitution buffer restored efficacy, this approach is not ideal, as LNP solutions would require dialysis prior to use. Fortunately, we found that the addition of trehalose or sucrose to LNP solutions prior to lyophilization facilitated room temperature storage and reconstitution in aqueous buffer without diminishing delivery potency.