Roy Shouvik, Henderson Ian, Nayar Rajiv, Randolph Theodore W, Carpenter John F
Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA.
J Pharm Sci. 2008 Dec;97(12):5132-46. doi: 10.1002/jps.21409.
The purpose of this study was to evaluate the importance of prelyophilization solution pH on the stability of botulinum neurotoxin, serotype A (rBoNTA(H(c))). This understanding is of significant importance for proteins such as rBoNTA(H(c)), a potential constituent of a multivalent vaccine product. For multivalent vaccines it may be difficult to identify a liquid formulation satisfying the stability requirements for all constituent protein antigens. Consequently, a lyophilized multivalent vaccine formulation may be a more viable alternative. Therefore evaluating the effect of prelyophilization pH (may be suboptimal) on the stability of antigens such as rBoNTA(H(c)) during lyophilization/storage becomes important. We hypothesize that when rBoNTA(H(c)) is lyophilized from a suboptimal pH, using the appropriate stabilizers can provide adequate physicochemical stability during lyophilization and long-term storage. We identified pH 5 and 8 in which the protein was stable and unstable against aggregation. Excipients were identified that could stabilize rBoNTA(H(c)) during lyophilization and storage in a stable solution of pH 5. These excipients were 7.5% (w/v) trehalose and 2.5% (w/v) trehalose with 2.5% (w/v) HES, with and without 0.01% (w/v) polysorbate 20. In support of our hypothesis, these excipients were found to provide adequate physicochemical stability to rBoNTA(H(c)) during lyophilization/storage, when freeze-dried from a prelyophilized solution of pH 8.
本研究的目的是评估冻干前溶液pH值对A型肉毒杆菌神经毒素（rBoNTA(H(c))）稳定性的重要性。对于rBoNTA(H(c))这类蛋白质（一种多价疫苗产品的潜在成分）而言，这种认识具有重要意义。对于多价疫苗来说，可能难以确定一种能满足所有组成蛋白抗原稳定性要求的液体制剂。因此，冻干多价疫苗制剂可能是更可行的选择。所以，评估冻干前pH值（可能并非最佳）对rBoNTA(H(c))等抗原在冻干/储存过程中稳定性的影响就变得很重要。我们假设，当rBoNTA(H(c))从非最佳pH值进行冻干时，使用合适的稳定剂可在冻干和长期储存过程中提供足够的物理化学稳定性。我们确定了pH 5和pH 8，在这两个pH值下蛋白质对聚集分别是稳定和不稳定的。我们还确定了一些辅料，它们能在pH 5的稳定溶液中冻干和储存期间稳定rBoNTA(H(c))。这些辅料是7.5%（w/v）海藻糖以及2.5%（w/v）海藻糖与2.5%（w/v）羟乙基淀粉的组合，添加或不添加0.01%（w/v）聚山梨酯20。为支持我们的假设，当从pH 8的冻干前溶液进行冻干时，发现这些辅料能为rBoNTA(H(c))在冻干/储存过程中提供足够的物理化学稳定性。
