title: Vascular disrupting activity of combretastatin analogues.
abstract: Tubulin binding agents (TBAs) are drugs commonly used in cancer therapy as antimitotics. In the last years it has been described that TBAs, like combretastatin A-4 (CA-4), present also vascular disrupting activity and among its derivatives we identified three analogues endowed with potent microtubule depolymerizing activity, higher than that of the lead compound. In this paper we have investigated the anti-vascular activity of these derivatives. We tested the anti-angiogenic effects in human umbilical endothelial cells (HUVEC) and in vivo in chick chorioallantoic membrane assay (CAM), and in a syngeneic tumor mouse model. The three molecules, compound 1: 1-(3,4,5-trimethoxyphenyl)-5-(4-ethoxyphenyl)-1H-1,2,4-triazole; compound 2: (1-(3,4,5-trimethoxyphenyl)-5-(4-ethoxyphenyl)-1H-tetrazole, compound-3 (4-amino-2-p-tolylaminothiazol-5-yl)-(3,4,5-trimethoxyphenyl)-methanone) showed a moderate effect on the growth of HUVEC cells at concentrations below 200nM. At lower concentrations (5-20nM), in particular compound 2, they induced inhibition of capillary tube formation, inhibition of endothelial cell migration and affected endothelial cell morphology as demonstrated by the alteration of the microfilaments network. Moreover, they also increased permeability of HUVEC cells in a time dependent manner. In addition, compounds 1 and 3, as well as the reference compound CA-4, inhibited VEGF-induced phosphorylation of VE-cadherin and in addition compound 3 prevented the VEGF-induced phosphorylation of FAK. In CAM assay, both compounds 2 and 3 efficiently counteracted the strong angiogenic response induced by bFGF, even at the lowest concentration used (1pmol/egg). Moreover in a syngenic mouse model, compounds 1-3 after a single i.p. injection (30mg/kg), showed a stronger reduction of microvascular density. Altogether our results identified these derivatives as potential new vascular disrupting agents candidates.
authors.affiliation: Dipartimento di Salute della Donna e del Bambino, Laboratorio di Oncoematologia, Università di Padova, 35131 Padova, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Porcù
authors.affiliation: Dipartimento di Scienze del Farmaco, Università di Padova, Padova, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Salvador
authors.affiliation: Dipartimento di Scienze del Farmaco, Università di Padova, Padova, Italy.
authors.forename: Irina
authors.initials: I
authors.lastname: Primac
authors.affiliation: Dipartimento di Medicina molecolare e traslazionale, Unità di oncologia sperimentale ed immunologia, Università di Brescia, 25123 Brescia, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Mitola
authors.affiliation: Dipartimento di Medicina molecolare e traslazionale, Unità di oncologia sperimentale ed immunologia, Università di Brescia, 25123 Brescia, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Ronca
authors.affiliation: Dipartimento di Medicina molecolare e traslazionale, Unità di oncologia sperimentale ed immunologia, Università di Brescia, 25123 Brescia, Italy.
authors.forename: Cosetta
authors.initials: C
authors.lastname: Ravelli
authors.affiliation: Dipartimento di Salute della Donna e del Bambino, Laboratorio di Oncoematologia, Università di Padova, 35131 Padova, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Bortolozzi
authors.affiliation: Dipartimento di Scienze del Farmaco, Università di Padova, Padova, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Vedaldi
authors.affiliation: Dipartimento di Scienze Chimiche e Farmaceutiche, Università di Ferrara, 44121 Ferrara, Italy.
authors.forename: Romeo
authors.initials: R
authors.lastname: Romagnoli
authors.affiliation: Dipartimento di Salute della Donna e del Bambino, Laboratorio di Oncoematologia, Università di Padova, 35131 Padova, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Basso
authors.affiliation: Dipartimento di Salute della Donna e del Bambino, Laboratorio di Oncoematologia, Università di Padova, 35131 Padova, Italy. Electronic address: giampietro.viola.1@unipd.it.
authors.forename: Giampietro
authors.initials: G
authors.lastname: Viola
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000494:pharmacology; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001632:Bibenzyls / Q000494:pharmacology*; D015820:Cadherins / Q000378:metabolism; D002199:Capillary Permeability / Q000187:drug effects; D024722:Cardiac Myosins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply*; D004305:Dose-Response Relationship, Drug; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008546:Melanoma, Experimental / Q000098:blood supply / Q000188:drug therapy* / Q000473:pathology; D008810:Mice, Inbred C57BL; D018994:Myosin Light Chains / Q000378:metabolism; D009389:Neovascularization, Pathologic; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010766:Phosphorylation; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D000099152:Cadherin 5
chemical_list: D043925:Angiogenesis Inducing Agents; D020533:Angiogenesis Inhibitors; D015703:Antigens, CD; D001632:Bibenzyls; D015820:Cadherins; D024722:Cardiac Myosins; D051417:Focal Adhesion Kinase 1; D018994:Myosin Light Chains; D000099152:Cadherin 5; C107184:myosin light chain 2; C040105:combretastatin; C496050:PTK2 protein, human
keywords: Angiogenesis; Combretastatin; HUVEC; Tubulin binding agents; Vascular disrupting agents