Schmidt Signe Tandrup, Christensen Dennis, Perrie Yvonne
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, UK.
Department of Infectious Disease Immunology, Center for Vaccine Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark.
Pharmaceutics. 2020 Dec 19;12(12):1237. doi: 10.3390/pharmaceutics12121237.
Subunit vaccines require particulate adjuvants to induce the desired immune responses. Pre-clinical manufacturing methods of adjuvants are often batch dependent, which complicates scale-up for large-scale good manufacturing practice (GMP) production. The cationic liposomal adjuvant CAF09b, composed of dioctadecyldimethylammonium bromide (DDA), monomycoloyl glycerol analogue 1 (MMG) and polyinosinic:polycytidylic acid [poly(I:C)], is currently being clinically evaluated in therapeutic cancer vaccines. Microfluidics is a promising new method for large-scale manufacturing of particle-based medicals, which is scalable from laboratory to GMP production, and a protocol for production of CAF09b by this method was therefore validated. The influence of the manufacture parameters [Ethanol] (20-40% ), [Lipid] (DDA and MMG, 6-12 mg/mL) and dimethyl sulfoxide [DMSO] (0-10% ) on the resulting particle size, colloidal stability and adsorption of poly(I:C) was evaluated in a design-of-experiments study. [Ethanol] and [DMSO] affected the resulting particle sizes, while [Lipid] and [DMSO] affected the colloidal stability. In all samples, poly(I:C) was encapsulated within the liposomes. At [Ethanol] 30% , most formulations were stable at 21 days of manufacture with particle sizes <100 nm. An in vivo comparison in mice of the immunogenicity to the cervical cancer peptide antigen HPV-16 E7 adjuvanted with CAF09b prepared by lipid film rehydration or microfluidics showed no difference between the formulations, indicating adjuvant activity is intact. Thus, it is possible to prepare suitable formulations of CAF09b by microfluidics.
亚单位疫苗需要颗粒佐剂来诱导所需的免疫反应。佐剂的临床前制造方法通常依赖批次，这使得大规模良好生产规范（GMP）生产的扩大规模变得复杂。由二辛基二甲基溴化铵（DDA）、单霉菌酰甘油类似物1（MMG）和聚肌苷酸：聚胞苷酸[聚（I:C）]组成的阳离子脂质体佐剂CAF09b目前正在治疗性癌症疫苗中进行临床评估。微流体技术是一种用于大规模制造基于颗粒的药物的有前途的新方法，它可以从实验室规模扩大到GMP生产，因此验证了通过该方法生产CAF09b的方案。在一项实验设计研究中，评估了制造参数[乙醇]（20 - 40%）、[脂质]（DDA和MMG，6 - 12 mg/mL）和二甲基亚砜[DMSO]（0 - 10%）对所得粒径、胶体稳定性和聚（I:C）吸附的影响。[乙醇]和[DMSO]影响所得粒径，而[脂质]和[DMSO]影响胶体稳定性。在所有样品中，聚（I:C）被包裹在脂质体内。在[乙醇]为30%时，大多数制剂在制造21天时稳定，粒径<100 nm。在小鼠体内对脂质膜复水或微流体技术制备的CAF09b佐剂的宫颈癌肽抗原HPV - 16 E7免疫原性进行比较，结果表明不同制剂之间没有差异，表明佐剂活性完好。因此，通过微流体技术制备合适的CAF09b制剂是可能的。
