The novel alkylating prodrug melflufen (J1) inhibits angiogenesis in vitro and in vivo.
Aminopeptidase N (APN) has been reported to have a functional role in tumor angiogenesis and repeatedly reported to be over-expressed in human tumors. The melphalan-derived prodrug melphalan-flufenamide (melflufen, previously designated J1) can be activated by APN. This suggests that this alkylating prodrug may exert anti-angiogenic properties, which will possibly contribute to the anti-tumoral activity in vivo. This work presents a series of experiments designed to investigate this effect of melflufen. In a cytotoxicity assay we show that bovine endothelial cells were more than 200 times more sensitive to melflufen than to melphalan, in HUVEC cells the difference was more than 30-fold and accompanied by aminopetidase-mediated accumulation of intracellular melphalan. In the chicken embryo chorioallantoic membrane (CAM) assay it is indicated that both melflufen and melphalan inhibit vessel ingrowth. Two commercially available assays with human endothelial cells co-cultured with fibroblasts (TCS Cellworks AngioKit, and Essen GFP-AngioKit) also illustrate the superior anti-angiogenic effect of melflufen compared to melphalan. Finally, in a commercially available in vivo assay in mice (Cultrex DIVAA angio-reactor assay) melflufen displayed an anti-angiogenic effect, comparable to bevacizumab. In conclusion, this study demonstrates through all methods used, that melphalan-flufenamide besides being an alkylating agent also reveals anti-angiogenic effects in different preclinical models in vitro and in vivo.
Clinical Pharmacology, Institution of Medical Sciences, Uppsala University, Akademiska Sjukhuset, 751 85 Uppsala, Sweden.
Sara
S
Strese
Malin
M
Wickström
Peder Fredlund
PF
Fuchs
Mårten
M
Fryknäs
Pär
P
Gerwins
Tim
T
Dale
Rolf
R
Larsson
Joachim
J
Gullbo
D000477:Alkylating Agents / Q000494:pharmacology*; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D018826:CD13 Antigens / Q000378:metabolism; D002417:Cattle; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects; D008558:Melphalan / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000494:pharmacology; D051379:Mice; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011355:Prodrugs / Q000494:pharmacology
D000477:Alkylating Agents; D020533:Angiogenesis Inhibitors; D011355:Prodrugs; C585069:melflufen; D010649:Phenylalanine; D018826:CD13 Antigens; D008558:Melphalan
Aminopeptidase N; Angiogenesis; Antiangiogenic; Cancer; Prodrug