Open AccessDevelopment of a Novel Recombinant Disintegrin, Contortrostatin, as an Effective Anti-Tumor and Anti-Angiogenic Agent
Author(s) -
Radu Minea,
Stephen D. Swenson,
Fritz Costa,
Thomas C. Chen,
Francis S. Markland
Publication year - 2005
Publication title -
pathophysiology of haemostasis and thrombosis
Language(s) - English
Resource type - Journals
eISSN - 1424-8840
pISSN - 1424-8832
DOI - 10.1159/000092419
Subject(s) - disintegrin , recombinant dna , snake venom , chemistry , angiogenesis , in vitro , rgd motif , cytotoxicity , metalloproteinase , cancer research , biochemistry , venom , microbiology and biotechnology , integrin , biology , matrix metalloproteinase , cell , gene
Contortrostatin (CN) (Mr 13,500 Da) is a novel homodimeric disintegrin isolated from the venom of Agkistrodon contortrix contortrix (Southern Copperhead) snake and displays two RGD motifs (one on each chain), which modulate its interaction with integrins on tumor cells and angiogenic vascular endothelial cells. In previous studies, we have shown that native CN administered in a liposomal formulation exhibits potent anti-angiogenic and tumor growth inhibitory activities. Current isolation of the protein from crude venom is difficult and prohibitively expensive for translation into the clinic. In this report, we describe a method amenable to large-scale production of a soluble monomeric form of recombinant CN with biologic activity; the protein is expressed directly in the cytoplasm of an engineered bacterial system with an expression yield of approximately 20 mg/l of culture. We present here the in vitro assays as well as the anti-tumor and anti-angiogenic evaluation of liposomal recombinant CN in an orthotopic, xenograft model of human breast cancer.