Pegylated liposomal encapsulation improves the antitumor efficacy of combretastatin A4 in murine 4T1 triple-negative breast cancer model.

Combretastatin A4 (CA4), a vascular disrupting agent has been recently proposed as an anticancer agent. However, its low water solubility and low bioavailability limited its clinical efficacy. Overcomingthis issue requires developing new delivery strategies to enhance its anticancer effects. Here, we prepared various PEGylated liposomal formulations containing CA4 composed of different molar ratios of HSPC/DSPE-mPEG2000/Cholesterol/CA4 (F1: 80:5:10:5; F2: 75:5:15:5; F3: 70:5:20:5; F4: 60:5:30:5 and F5: 50:5:40:5) by the thin-film hydration method plus sonication and extrusion. All formulations had a particle diameter of 100-150 nm, a monomodal distribution with low polydispersity index and a negative zeta potential. Among the formulations only F1, F2, and F3 showed a high CA4 encapsulation efficiency; so their anticancer effects on triple-negative breast cancer (TNBC) were investigated in vitro and in vivo. The release study showed that F3 liposomes had significantly lower CA4 release compared to the F1 and F2 liposomes in different pH of 5.5, 6.5, and 7.4. We found that, CA4-loaded liposomes effectively inhibited both proliferation and migration of 4T1 and MDA-MB-231 TNBC cell lines by inducing cell cycle arrest at the G2/M phase and decreasing MMP-2 and MMP-9 expression and activity. In vivo studies revealed that F3 liposomes were highly accumulated at the tumor site and more effectively delayed tumor growth andprolonged the overall survival than other groups in 4T1 breast tumor-bearing mice. Taken together, encapsulation of CA4 in PEGylated F3 liposomes enhances its anti-tumor activity and may be serve as a promising approach for TNBC treatment and merits further investigation.