Transferrin Conjugates for Oral Protein Drug Delivery
Principal Investigator: Wei Chiang Shen
Affiliation: University of Southern California
Abstract: Project Summary: Gastrointestinal (Gl) absorption is a desirable route for administration of drugs including therapeutic peptides and proteins, because it can reduce the inconvenience, cost, and complications associated with injection, and increase patient compliance especially in chronic administration. However, no acceptable oral formulation is currently available for peptide and protein drugs. In this application, recombinant fusion proteins with a transferrin (Tf) domain will be designed and developed as a new class of protein drugs that can be orally administered. This novel protein drug design is based on our recent findings on the recombinant fusion protein between human Tf and granulocyte colony stimulating factor (G-CSF), which demonstrated an effective myelopoietic activity when orally administered to BDF1 mice. In this application, G-CSF-Tf will be studied, with a multidisciplinary approach including molecular biology, computer modeling, cell biology, and protein chemistry, to optimize the biological activity, as well as the applicability, of the recombinant products. In addition, the pharmacokinetics and biodistribution of the fusion protein will be investigated in order to elucidate the underlying mechanism of TfR-mediated protein absorption from the Gl tract. Thus, the specific aims during the next 4 years are first to prepare G-CSF-Tf constructs with different spacers and to determine their in vitro biological activities, i.e., Tf receptor binding and NFS-60 cell proliferation. These peptide spacers will be designed either to maintain a distance between the two domains to minimize the mutual interference, or to allow for separation of the two domains in the blood to regenerate the intact and fully bioactive protein drug. Subsequently, the selected fusion proteins with optimal linkers will be compared for in vivo myelopoietic activity with both subcutaneous and oral administration. Finally, pharmacokinetics and biodistribution of the selected fusion proteins will be investigated in BDF1 mice using a specially designed ELISA method. The potential Gl alteration and toxicity from repeated oral adminsitration in mice will be studied. The previously observed prolonged in vivo effect of orally administered G-CSF-Tf fusion protein will also be examined in correlation with dietary iron levels. These data will provide information regarding the deposition, bioavailability, and transport mechanism. Our long term goal is to develop Tf-fusion proteins into a new generation of protein drugs that can be administered orally to treat human diseases. Relevance: Proteins and peptides are an important class of drugs for treating various human diseases; however, they are presently administered only by injection. The objective of this application is to develop recombinant proteins by linking the protein drug with transferrin to promote oral absorption. Such a new generation of protein drugs for effective oral delivery will overcome patient incompliance, cost, and side effects currently associated with injectable dosage forms.
Funding Period: 2002-04-01 - 2011-03-31
more information: NIH RePORT
- Improving the oral efficacy of recombinant granulocyte colony-stimulating factor and transferrin fusion protein by spacer optimizationYun Bai
Department of Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, California 90033, USA
Pharm Res 23:2116-21. 2006....
- Recombinant granulocyte colony-stimulating factor-transferrin fusion protein as an oral myelopoietic agentYun Bai
Department of Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA 90033, USA
Proc Natl Acad Sci U S A 102:7292-6. 2005..Taken together, we conclude that the Tf-based recombinant fusion protein technology represents a promising approach for future development of orally effective peptide and protein drugs...
- Comparison of monomeric and oligomeric transferrin as potential carrier in oral delivery of protein drugsChing-Jou Lim
Department of Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA 90089, USA
J Control Release 106:273-86. 2005..The delayed onset and prolonged effect of Agg-Tf-S-S-In in hypoglycemia strongly suggests that the Tf-oligomer can act as a sustained release carrier in the oral delivery of protein and peptide drugs...
- Accumulation of transferrin in Caco-2 cells: a possible mechanism of intestinal transferrin absorptionChing Jou Lim
Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, 1985 Zonal Avenue, Los Angeles, CA 90033, USA
J Control Release 122:393-8. 2007..The Rab11-positive compartment may control the release of apically internalized Tf for either slow recycling to apical membrane or processing to transcytotic compartments...
- Tyrphostin A8 stimulates a novel trafficking pathway of apically endocytosed transferrin through Rab11-enriched compartments in Caco-2 cellsFariba Norouziyan
Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, California 90033, USA
Am J Physiol Cell Physiol 294:C7-21. 2008..These results suggest an effect of AG10 on the later steps of transferrin receptor trafficking, which are involved in subsequent recycling, and possibly transcytosis, of endocytosed transferrin in Caco-2 cells...