Introduction: Glioblastoma multiforme (GBM), also known as Grade IV astrocytoma, is most aggressive malignant tumor in human beings. In this study, we newly developed an orally absorbable heparin that can show antiangiogenic activity via binding with growth factors around tumor tissue. It is well known that lactoferrin (Lf) can be orally absorbed via lactoferrin receptor (Lf-R) that is expressed on the intestine, blood-brain barrier (BBB) and GBM. The effect of bovine lactoferrin-conjugated heparin (bLf-heparin) was evaluated in vitro and in vivo.
Materials and Methods: Heparin, typically enoxaparin, was gifted by Prof. YR Byun (Seoul National Univ., Korea). Bovine Lactoferrin was chemically conjugated with heparin (bLf-heparin). Its anticancer ad antiangiogenic activity was evaluated by using U87MG and HUVEC. Its oral absorption mechanism was evaluated with Caco-2 monolayer in vitro. Also therapeutic effect of bLf-heparin was evaluated orthotopic GBM mouse model.
Results and Discussion: The chemical conjugation between Lf and heparin was confirmed via GPC, FT-IR, SDS PAGE and Western blotting. bLf-heparin was could attenuate U87MG cell’s proliferation up to 37.5% with no cytotoxicity. Also, it could block angiogenesis procedure via aorta ring assay. bLf-heparin could be absorbed via transcytosis, which was confirmed by TEER transport system. When Lf-heparin was orally treated in the mice, it could be absorbed in the small intestine via Lf receptor-mediated endocytosis. In addition, it could be targeted to the brain tissue due to the existence of Lf receptor on the blood-brain barrier.
Conclusion: This study demonstrated that bLf-Heparin conjugate could act as an anti-angiogenic drug to the Glioblastoma Multiform, effectively and systemically. And it could be absorbed orally and transported to the brain via lactoferrin receptor. Collectively, bLf-heparin could be used to treat glioblastoma multiforme as a new oral medication.
Korea Health Technology R&D Project