A bispecific
ligand-directed toxin (BLT) consisting of human
interleukin-13, epithelial
growth factor, and the first 389
amino acids of
diphtheria toxin was assembled in order to target human
glioblastoma. In vitro, DTEGF13 selectively killed the human
glioblastoma cell line U87-luc as well as other human
glioblastomas. DTEGF13 fulfilled the requirement of a successful BLT by having greater activity than either of its monospecific counterparts or their mixture proving it necessary to have both
ligands on the same single chain molecule. Aggressive
brain tumors established intracranially (IC) in nude rats with U87
glioma genetically marked with a
firefly luciferase reporter gene were treated with two
injections of DTEGF13 using convection enhanced delivery resulting in
tumor eradication in 50% of the rats which survived with
tumor free status at least 110 days post
tumor inoculation. An irrelevant BLT control did not protect establishing specificity. The bispecific DTEGF13 MTD dose was measured at 2 microg/injection or 0.5 microg/kg and toxicity studies indicated safety in this dose. Combination of monospecific DTEGF and DTIL13 did not inhibit
tumor growth. ELISA assay indicated that anti-DT
antibodies were not generated in normal immunocompetent rats given identical intracranial DTEGF13
therapy. Thus, DTEGF13 is safe and efficacious as an alternative drug for
glioblastoma therapy and warrants further study.