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The perhaps unexpected safety proﬁle in mice may be due to the resistance of quiescent endothelia to the effects of combretastatin—likely due to the discriminating dependence of nascent endothelia on the tubulin cytoskeleton (Galbraith et al. 2001; Young and Chaplin 2004). In support of this, a major change that occurs after EC exposure to CA4 is an alteration in shape and morphology, which is profoundly less than that which occurs in quiescent ECs, offering combretastatin, and tubulin inhibitors with similar mechanism of action, an exquisite level of selectivity for ECs in the pathological milieu (Galbraith et al.
Cytotoxic to ECs than would be expected for a tubulin inhibitor and at doses signiﬁcantly lower than the maximum tolerated dose (MTD) in mice. In fact, in vitro studies using human umbilical vein endothelial cells (HUVEC) by several groups demonstrated that CA4 is cytotoxic if the cells are proliferating, resulting in cell rounding, retraction, membrane blebbing, and substratum detachment, but not if they are quiescent (Dark et al. 1997; Galbraith et al. 2001). CA4 has also been shown to inhibit active angiogenesis in a HUVEC tube formation assay (Grosios et al.
Br J Cancer 55(2):159–163 Bibby MC, Double JA et al (1989) Reduction of tumor blood ﬂow by ﬂavone acetic acid: a possible component of therapy. J Natl Cancer Inst 81(3):216–220 Brooks AC, Kanthou C et al (2003) The vascular targeting agent combretastatin A-4-phosphate induces neutrophil recruitment to endothelial cells in vitro. Anticancer Res 23(4):3199–3206 Carmeliet P, Jain RK (2011a) Molecular mechanisms and clinical applications of angiogenesis. Nature 473(7347):298–307 Carmeliet P, Jain RK (2011b) Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases.