Genetic engineering, expression, and activity of a fusion protein of a human neurotrophin and a molecular Trojan horse for delivery across the human blood-brain barrier.
Neurotrophins, such as brain derived neurotrophic factor (BDNF), do not cross the blood-brain barrier (BBB). Certain monoclonal antibodies (MAb) to the human insulin receptor (HIR) do cross the BBB via receptor-mediated transport, and can act as a molecular Trojan horse to ferry across the BBB an attached drug. A genetically engineered fusion protein was produced whereby the amino terminus of human BDNF is fused to the carboxyl terminus of the heavy chain of a chimeric HIRMAb. The HIRMAb-BDNF fusion protein reacted equally with antibodies to human IgG and BDNF. The bi-functionality of the fusion protein was retained as the affinity of the fusion protein for the HIR was identical to that of the chimeric HIRMAb, and the affinity of the fusion protein for the trkB receptor was identical to that of BDNF. The fusion protein was equi-potent with BDNF in a neuroprotection assay in human neural cells. The pharmacokinetics (PK) of the fusion protein was examined in the adult Rhesus monkey. The mean residence time (MRT) of the fusion protein in blood was >100-fold longer than the MRT of BDNF. Therapeutic levels of BDNF were produced in primate brain following the intravenous administration of the fusion protein. A fusion protein tandem vector was engineered that allowed for isolation of a CHO cell line that produced the fusion protein at high levels in serum free medium. Neurotrophins, such as BDNF, can be re-formulated to enable these molecules to cross the human BBB, and such fusion proteins represent a new class of human neurotherapeutics.
Boado
Ruben J
RJ
ArmaGen Technologies Inc., Santa Monica, California, USA.
Zhang
Yufeng
Y
Zhang
Yun
Y
Pardridge
William M
WM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000493:pharmacokinetics*; D001812:Blood-Brain Barrier / Q000378:metabolism*; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism* / Q000493:pharmacokinetics*; D016503:Drug Delivery Systems / Q000379:methods*; D005260:Female; D000882:Haplorhini; D006801:Humans; D008253:Macaca mulatta; D008657:Metabolic Clearance Rate; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000493:pharmacokinetics
D000911:Antibodies, Monoclonal; D019208:Brain-Derived Neurotrophic Factor; D011993:Recombinant Fusion Proteins