本研究旨在制定和统计优化盐酸二甲双胍（MTF）的胶束制剂，以增强其对乳腺癌的抗肿瘤活性。采用Design-Expert®软件使用Box Behnken设计。配方变量包括甘油单油酸酯浓度（GMO，w/w%），Pluronic F-127浓度（PF127，w/w%）和Tween 80浓度（w/w%），而包封效率（EE%），囊泡大小（VS）和Zeta电位（ZP）则被设定为依赖响应变量。设计专家软件用于进行数值优化过程。采用X射线衍射（XRD），透射电子显微镜（TEM），体外释放研究，短期稳定性研究以及在MDA-MB-231乳腺癌和LOVO结肠癌细胞系上进行的体外细胞增殖实验来验证优化的胶束制剂。优化的配方的成分为4.35616（w/w%）GMO，5（w/w%）PF127和7.444E-6（w/w%）Tween 80，所得的适应度为0.733。EE%，VS和ZP的预测值分别为78.0592%，307.273 nm和-26.8275 mV。在优化公式上进行的验证过程表明，与预测的响应相比，差异小于5%。XRD热图显示MTF被包封在胶束囊泡内。优化的MTF胶束制剂的TEM图像显示为球形非聚集纳米囊泡。此外，与MTF溶液相比，它显示出MTF的缓释特性。稳定性研究显示，最佳胶束制剂在30天内保持稳定。与MTF溶液相比，优化的胶束制剂对MDA-MB-231乳腺癌和LOVO结肠癌细胞系的细胞毒性增强，即使在较低浓度下也是如此。然而，对于乳腺癌细胞系，它显示出更强的细胞毒性作用。因此，胶束可以被视为治疗乳腺和结肠癌的有前途的MTF载体。© 2023 The Authors. Published by Elsevier B.V.

This study aimed to formulate and statistically optimize cubosomal formulations of metformin (MTF) to enhance its breast anticancer activity. A Box Behnken design was employed using Design-Expert® software. The formulation variables were glyceryl monooleate concentration (GMO) w/w%, Pluronic F-127 concentration (PF127) w/w% and Tween 80 concentration w/w% whereas Entrapment efficiency (EE%), Vesicles' size (VS) and Zeta potential (ZP) were set as the dependent responses. The design expert software was used to perform the process of optimization numerically. X ray diffraction (XRD), Transmission electron microscope (TEM), in-vitro release study, short-term stability study, and in in-vitro cell proliferation assay on the MDA-MB-231 breast cancer and LOVO cancer cell lines were used to validate the optimized cubosomal formulation. The optimized formulation had a composition of 4.35616 (w/w%) GMO, 5 (w/w%) PF127 and 7.444E-6 (w/w%) Tween 80 with a desirability of 0.733. The predicted values for EE%, VS and ZP were 78.0592%, 307.273 nm and - 26.8275 mV, respectively. The validation process carried out on the optimized formula revealed that there were less than a 5% variance from the predicted responses. The XRD thermograms showed that MTF was encapsulated inside the cubosomal vesicles. TEM images of the optimized MTF cubosomal formulation showed spherical non-aggregated nanovesicles. Moreover, it revealed a sustained release profile of MTF in comparison to the MTF solution. Stability studies indicated that optimum cubosomal formulation was stable for thirty days. Cytotoxicity of the optimized cubosomal formulation was enhanced on the MDA-MB-231 breast and LOVO cancer cell lines compared to MTF solution even at lower concentrations. However, it showed superior cytotoxic effect on breast cancer cell line. So, cubosomes could be considered a promising carrier of MTF to treat breast and colon cancers.© 2023 The Authors. Published by Elsevier B.V.