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Discovery of an Orally Effective Factor IX-Transferrin Fusion Protein for Hemophilia B

Hemophilia B is a severe blood clotting disorder caused by the deficiency of factor IX (FIX). FIX is not bioavailable when given orally due to poor stability and permeability in the gastrointestinal tract. The feasibility of fusing FIX with transferrin (Tf) to enhance the oral bioavailability of FIX...

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Autores principales: Xie, Chen, Wang, Zhijun, Su, Yang, Wang, Jeffrey, Shen, Wei-Chiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6981973/
https://www.ncbi.nlm.nih.gov/pubmed/31861459
http://dx.doi.org/10.3390/ijms21010021
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author Xie, Chen
Wang, Zhijun
Su, Yang
Wang, Jeffrey
Shen, Wei-Chiang
author_facet Xie, Chen
Wang, Zhijun
Su, Yang
Wang, Jeffrey
Shen, Wei-Chiang
author_sort Xie, Chen
collection PubMed
description Hemophilia B is a severe blood clotting disorder caused by the deficiency of factor IX (FIX). FIX is not bioavailable when given orally due to poor stability and permeability in the gastrointestinal tract. The feasibility of fusing FIX with transferrin (Tf) to enhance the oral bioavailability of FIX is explored. Seven recombinant fusion proteins (rFIX-Tf) with different linkers were constructed and expressed in HEK293 cells and characterized by in vitro transcytosis and transferrin receptor (TfR) binding assay in Caco-2 cells and a one-stage clotting assay. The in vivo efficacy study was performed using a tail-bleeding model in hemophilia B mice. Fusion proteins rFIX-Tf/G(2) and rFIX-Tf/SVSQ were most permeable and showed a specific binding ability to TfR in Caco-2 cells. Both proteins retained FIX activity in clotting generation. The in vivo efficacy study showed that both proteins by intravenous injection significantly reduced blood loss. Most significantly, rFIX-Tf/G(2) demonstrated anti-bleeding activity when administered orally. Our results showed that the fusion protein technique with Tf could be potentially used for oral delivery of FIX and the linker between FIX and Tf in the fusion protein is crucial. rFIX-Tf/G(2) appears to be the most promising fusion protein as potential oral therapeutics for hemophilia B.
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spelling pubmed-69819732020-02-07 Discovery of an Orally Effective Factor IX-Transferrin Fusion Protein for Hemophilia B Xie, Chen Wang, Zhijun Su, Yang Wang, Jeffrey Shen, Wei-Chiang Int J Mol Sci Article Hemophilia B is a severe blood clotting disorder caused by the deficiency of factor IX (FIX). FIX is not bioavailable when given orally due to poor stability and permeability in the gastrointestinal tract. The feasibility of fusing FIX with transferrin (Tf) to enhance the oral bioavailability of FIX is explored. Seven recombinant fusion proteins (rFIX-Tf) with different linkers were constructed and expressed in HEK293 cells and characterized by in vitro transcytosis and transferrin receptor (TfR) binding assay in Caco-2 cells and a one-stage clotting assay. The in vivo efficacy study was performed using a tail-bleeding model in hemophilia B mice. Fusion proteins rFIX-Tf/G(2) and rFIX-Tf/SVSQ were most permeable and showed a specific binding ability to TfR in Caco-2 cells. Both proteins retained FIX activity in clotting generation. The in vivo efficacy study showed that both proteins by intravenous injection significantly reduced blood loss. Most significantly, rFIX-Tf/G(2) demonstrated anti-bleeding activity when administered orally. Our results showed that the fusion protein technique with Tf could be potentially used for oral delivery of FIX and the linker between FIX and Tf in the fusion protein is crucial. rFIX-Tf/G(2) appears to be the most promising fusion protein as potential oral therapeutics for hemophilia B. MDPI 2019-12-18 /pmc/articles/PMC6981973/ /pubmed/31861459 http://dx.doi.org/10.3390/ijms21010021 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Xie, Chen
Wang, Zhijun
Su, Yang
Wang, Jeffrey
Shen, Wei-Chiang
Discovery of an Orally Effective Factor IX-Transferrin Fusion Protein for Hemophilia B
title Discovery of an Orally Effective Factor IX-Transferrin Fusion Protein for Hemophilia B
title_full Discovery of an Orally Effective Factor IX-Transferrin Fusion Protein for Hemophilia B
title_fullStr Discovery of an Orally Effective Factor IX-Transferrin Fusion Protein for Hemophilia B
title_full_unstemmed Discovery of an Orally Effective Factor IX-Transferrin Fusion Protein for Hemophilia B
title_short Discovery of an Orally Effective Factor IX-Transferrin Fusion Protein for Hemophilia B
title_sort discovery of an orally effective factor ix-transferrin fusion protein for hemophilia b
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6981973/
https://www.ncbi.nlm.nih.gov/pubmed/31861459
http://dx.doi.org/10.3390/ijms21010021
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