Biscaia-Caleiras Mariana, Lopes Diana, Henriques Carolina, Lourenço Ana Sofia, Nunes António, Bañobre Manuel, Moreira João Nuno, Simões Sérgio
CNC-UC - Center for Neurosciences and Cell Biology, Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, Faculty of Medicine (Polo 1), Rua Larga, 3004-504 Coimbra, Portugal; Bluepharma-Indústria Farmacêutica, S.A., São Martinho do Bispo, 3045-016 Coimbra, Portugal; Univ Coimbra-University of Coimbra, CIBB, Faculty of Pharmacy, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
Bluepharma-Indústria Farmacêutica, S.A., São Martinho do Bispo, 3045-016 Coimbra, Portugal.
Int J Pharm. 2025 Sep 15;682:125973. doi: 10.1016/j.ijpharm.2025.125973. Epub 2025 Jul 18.
The industrial manufacturing of liposomal injectables faces significant technical challenges. Traditional batch manufacturing methods, like ethanol injection and extrusion, are time-consuming, prompting a shift towards continuous manufacturing. To improve process efficiency, this study tested microfluidics, a technique widely used in lipid nanoparticle (LNP) production for nucleic acid delivery, to optimize doxorubicin liposome manufacturing, focusing on vesicle formation and remote drug loading. The formulation consisted of neutral phospholipids with minimal DSPE-PEG content and no negatively charged lipids, components typically considered critical for liposome self-assembly and colloidal stability, demonstrating that microfluidics can effectively produce stable liposomes under these challenging conditions. Detailed characterization by cryo-TEM confirmed the formation of unilamellar vesicles with internal drug nanocrystals by microfluidics. In addition, it yielded fewer multilamellar liposomes than the conventional process (18 % vs. 35 %), indicating better control of vesicle structure and lamellarity. Importantly, employing microfluidics instead of ethanol injection and extrusion reduced liposome formation time by 70 % while ensuring consistent particle size distribution, relative to ethanol injection and extrusion. Additionally, lowering the temperature during drug loading (45 °C vs. 65 °C) shortened this step by 20 %, due to faster heating and cooling. Consequently, the optimized process was at least 25 % faster and reduced cost by 15 %. Although conducted at a 100 mL scale, these improvements are expected to be amplified on an industrial scale. Hence, these findings highlight the relevance of decreasing process temperatures and underscore the potential of microfluidics to enhance the efficiency and scalability of continuous manufacturing of liposomes for small molecular weight drug delivery, a field in which this technology has been underexplored.
脂质体注射剂的工业化生产面临重大技术挑战。传统的批次生产方法，如乙醇注入法和挤压法，耗时较长，促使人们转向连续生产。为提高工艺效率，本研究测试了微流控技术，该技术在用于核酸递送的脂质纳米颗粒（LNP）生产中广泛应用，以优化阿霉素脂质体的生产，重点关注囊泡形成和远程载药。该制剂由DSPE-PEG含量极低且无带负电荷脂质的中性磷脂组成，这些成分通常被认为对脂质体自组装和胶体稳定性至关重要，这表明微流控技术能够在这些具有挑战性的条件下有效生产稳定的脂质体。通过冷冻透射电子显微镜（cryo-TEM）进行的详细表征证实了微流控技术形成了含有内部药物纳米晶体的单层囊泡。此外，与传统工艺相比，它产生的多层脂质体更少（18%对35%），表明对囊泡结构和层数的控制更好。重要的是，相对于乙醇注入法和挤压法，采用微流控技术而非乙醇注入法和挤压法可将脂质体形成时间缩短70%，同时确保粒径分布一致。此外，在载药过程中降低温度（45°C对65°C）可使该步骤缩短20%，这是因为加热和冷却更快。因此，优化后的工艺速度至少提高了25%，成本降低了15%。尽管是在100 mL规模上进行的，但预计这些改进在工业规模上会得到放大。因此，这些发现凸显了降低工艺温度的重要性，并强调了微流控技术在提高用于小分子药物递送的脂质体连续生产的效率和可扩展性方面的潜力，而该技术在这一领域的探索还不够充分。
