PO.ET02.08 · 实验与分子治疗
长春瑞滨与多柔比星在游离溶液和脂质体中于多细胞球体模型的联合效应
Combination effect of vinorelbine and doxorubicin in free solutions and liposomes in a multicellular spheroids model
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
目的：联合药物治疗是治疗癌症的基石性方法，然而递送药物组合的策略，包括最佳药物比例和药物制剂，仍然充满挑战。多柔比星（DOX）和长春瑞滨（VNR）是成熟的抗癌药物，可抑制细胞分裂的不同阶段，因而成为联合治疗的有前景候选药物。本研究通过细胞活力测定和数学建模，评估了DOX和VNR在各种制剂（游离溶液、pH敏感脂质体和非pH敏感脂质体）中以不同药物比例对A549细胞（NSCLC）多细胞球体（MCSs）的联合效应。
方法：非pH敏感脂质体（NonFlip）由胆固醇、POPC和PEG-Ceramide组成。pH敏感脂质体（Flip）由POPC、PEG-Ceramide以及具有pH敏感构象开关的脂质（flipids）组成。A549 MCSs在96孔超低吸附微孔板中用RPMI-1640和0.3%胶原蛋白构建。将MCSs（约500 μm）用游离或制剂化的DOX、VNR或不同比例的DOX:VNR组合处理。使用CellTiter-Glo 3D测定法评估细胞活力。使用GraphPad Prism 10.6.0软件通过三种数学模型（Greco模型、统一理论模型和Bliss独立模型）评估药物联合效应。
结果：对于单药，游离DOX、NonFlip DOX和Flip DOX表现出相似的IC50值，约为5 μM。游离VNR和Flip VNR显示IC50约为44 μM，而NonFlip VNR的IC50超过200 μM。对于组合，DOX:VNR比例为5:1、1:1和1:5时，在游离药物组和Flip组中分别产生约5、8和16 μM的IC50值。NonFlip组在5:1和1:1时显示的IC50值约为5和8 μM，但NonFlip DOX:VNR 1:5的IC50超过200 μM。与单药相比，游离药物溶液、Flip制剂和NonFlip制剂中的大多数DOX:VNR组合在Greco模型和统一理论模型中均展现出协同作用（R²>0.95）。为优化协同作用，Greco模型倾向于DOX:VNR 5:1的比例，而统一理论倾向于1:5。相比之下，Bliss独立模型对细胞活力数据拟合较差（R²<0.9），且未显示协同效应。
结论：对剂量依赖性细胞活力数据的Greco模型和统一理论建模证明了DOX和VNR在各种制剂中联合的协同效应。Bliss独立模型对细胞活力数据的拟合较差，提示DOX和VNR可能并非独立地抑制癌细胞生长。需要进一步研究以证实这些发现。
查看英文原文 English abstract
Purpose: Combination drug therapy is a cornerstone approach to treat cancers, yet strategies of delivering drug combinations, including optimal drug ratios and drug formulations, remain challenging. Doxorubicin (DOX) and vinorelbine (VNR) are well established anticancer drugs that inhibit different phases of cell division, thus making them promising candidates for combination therapy. This study evaluated the combination effect of DOX and VNR in various formulations (free solution, pH-sensitive liposomes, and non-pH-sensitive liposomes) at various drug ratios against multicellular spheroids (MCSs) of A549 cells (NSCLC) by cell viability assay and mathematical modeling.
Methods: Non-pH-sensitive liposomes (NonFlip) are composed of cholesterol, POPC and PEG-Ceramide. pH sensitive liposomes (Flip) are composed of POPC, PEG-Ceramide and lipids with a pH-sensitive conformational switch (flipids). A549 MCSs were constructed in 96-well ultra-low attachment microplates with RPMI-1640 and 0.3% collagen. MCSs (~500 μm) were treated with free or formulated DOX, VNR, or DOX:VNR combinations at various ratios. Cell viability was assessed using CellTiter-Glo 3D assay. The effects of drug combination were evaluated by three mathematical models (Greco's, unified theory, and Bliss independent) using the GraphPad Prism 10.6.0 software.
Results: For individual drugs, free DOX, NonFlip DOX, and Flip DOX exhibited similar IC50 values around 5 μM. Free VNR and Flip VNR showed IC50 around 44 μM, while NonFlip VNR displayed IC50 exceeding 200 μM. For combinations, DOX:VNR ratios of 5:1, 1:1, and 1:5 yielded IC50 values of approximately 5, 8, and 16 μM, respectively, in free drug and Flip groups. NonFlip groups at 5:1 and 1:1 showed IC50 values of approximately 5 and 8 μM, but the IC50 of NonFlip DOX:VNR 1:5 exceeded 200 μM. Compared to individual drugs, most of the DOX:VNR combinations in Free drug solution, Flip formulations, and NonFlip formulations demonstrated synergism in Greco's and unified theory models (R²>0.95). To optimize the synergism, Greco's model favored the DOX:VNR ratio of 5:1 while unified theory favored 1:5. In contrast, the Bliss independent model fitted the cell viability data poorly (R²<0.9) and did not show synergistic effects.
Conclusions: Greco's and unified theory modeling of the dose dependent cell viability data demonstrated the synergistic effect of combining DOX and VNR in various formulations. The poor fitting of the cell viability data by the Bliss independent model suggests DOX and VNR may not suppress the cancer cell growth independently. Further studies are needed to confirm these findings.
利益披露 Disclosure
Y. Zhu, None..
Z. Xu, None..
Y. Lu, None..
S. Zhao, None..
M. Hossain, None..
X. Guo, None.