| Autor: |
Georgieva JV; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA., Katt M; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA., Ye Z; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA., Umlauf BJ; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA., Wenthur CJ; Divisions of Pharmaceutical Sciences and Pharmacy Practice, School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA., Shusta EV; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.; Department of Neurological Surgery, University of Wisconsin-Madison, Madison, WI 53792, USA. |
| Abstrakt: |
Central nervous system (CNS) exposure to blood-borne biotherapeutics is limited by the restrictive nature of the brain vasculature. In particular, tightly sealed endothelial cells of the blood-brain barrier (BBB) prevent the uptake of protein and gene medicines. An approach to increase the bioavailability of such therapeutics is harnessing the BBB endothelial cells' own receptor-mediated transcytosis (RMT) mechanisms. Key to this process is a targeting ligand that can engage a BBB-resident RMT receptor. We recently identified an antibody, named 46.1, that accumulates in the mouse brain after intravenous injection. To further characterize the brain targeting and penetrating properties of clone 46.1, we conjugated neurotensin (NT) to an scFv-Fc form of the antibody (46.1-scFv-Fc- L ong L inker-NT). While centrally administered NT decreases the core body temperature and locomotor activity, effects attributed to two spatially segregated brain areas, systemically administered NT has limited effects. Hence, NT can be used as a model therapeutic payload to evaluate the brain penetration of BBB-targeting antibodies and their capability to accumulate in discrete brain areas. We demonstrate that intravenously administered 46.1-scFv-Fc-LL-NT can elicit transient hypothermia and reduce drug-induced hyperlocomotion, confirming that 46.1 can deliver drug cargo to the CNS at pharmacologically relevant doses. Interestingly, when two intravenous administration routes in mice, retro-orbital and tail vein, were compared, only retro-orbital administration led to transient hypothermia. We further explored the retro-orbital route and demonstrated that the 46.1-scFv-Fc-LL-NT could enter the brain arterial blood supply directly from the retro-orbital/cavernous sinus. Taken together, the 46.1 antibody is capable of transporting drug cargo into the CNS, and at least of a portion of its CNS accumulation occurs via the cavernous sinus-arterial route. |
