Transferrin Conjugates for Oral Protein Drug Delivery

Summary

Principal Investigator: Wei Chiang Shen
Affiliation: University of Southern California
Country: USA
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

Top Publications

  1. ncbi Improving the oral efficacy of recombinant granulocyte colony-stimulating factor and transferrin fusion protein by spacer optimization
    Yun Bai
    Department of Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, California 90033, USA
    Pharm Res 23:2116-21. 2006
  2. pmc Proinsulin-transferrin fusion protein as a novel long-acting insulin analog for the inhibition of hepatic glucose production
    Yan Wang
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
    Diabetes 63:1779-88. 2014
  3. pmc Characterization of transferrin receptor-mediated endocytosis and cellular iron delivery of recombinant human serum transferrin from rice (Oryza sativa L.)
    Deshui Zhang
    Ventria Bioscience, Fort Collins, CO 80524, USA
    BMC Biotechnol 12:92. 2012
  4. pmc Fusion protein linkers: property, design and functionality
    Xiaoying Chen
    Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA 90089, USA
    Adv Drug Deliv Rev 65:1357-69. 2013
  5. pmc Mass spectrometry study of a transferrin-based protein drug reveals the key role of protein aggregation for successful oral delivery
    Cedric E Bobst
    Department of Chemistry, University of Massachusetts, Amherst, MA 01003, USA
    Proc Natl Acad Sci U S A 109:13544-8. 2012
  6. pmc Insertion of the designed helical linker led to increased expression of tf-based fusion proteins
    Nurmamet Amet
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California School of Pharmacy, PSC 404B, 1985 Zonal Avenue, Los Angeles, California, 90089 9121, USA
    Pharm Res 26:523-8. 2009
  7. pmc Conjugation with cationic cell-penetrating peptide increases pulmonary absorption of insulin
    Leena N Patel
    Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, Will Rogers Institute Pulmonary Research Center, Keck School of Medicine, University of Southern California, Los Angeles, California 90089 9121, USA
    Mol Pharm 6:492-503. 2009
  8. pmc Pharmacokinetics of recombinant bifunctional fusion proteins
    Xiaoying Chen
    University of Southern California School of Pharmacy, Department of Pharmacology and Pharmaceutical Sciences, CA 90089 9121, USA
    Expert Opin Drug Metab Toxicol 8:581-95. 2012
  9. pmc Receptor-mediated activation of a proinsulin-transferrin fusion protein in hepatoma cells
    Yan Wang
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, United States
    J Control Release 155:386-92. 2011
  10. pmc Effects of receptor binding on plasma half-life of bifunctional transferrin fusion proteins
    Xiaoying Chen
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California School of Pharmacy, Los Angeles, California 90089 9121, USA
    Mol Pharm 8:457-65. 2011

Detail Information

Publications16

  1. ncbi Improving the oral efficacy of recombinant granulocyte colony-stimulating factor and transferrin fusion protein by spacer optimization
    Yun Bai
    Department of Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, California 90033, USA
    Pharm Res 23:2116-21. 2006
    ..To improve the oral efficacy of the recombinant fusion protein containing granulocyte colony-stimulating factor (G-CSF) and transferrin (Tf) by inserting a linker between the two protein domains...
  2. pmc Proinsulin-transferrin fusion protein as a novel long-acting insulin analog for the inhibition of hepatic glucose production
    Yan Wang
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
    Diabetes 63:1779-88. 2014
    ..Thus, ProINS-Tf fusion protein can potentially be administered as a prodrug with sustained Tf-mediated activation and selectivity in inhibiting HGP. ..
  3. pmc Characterization of transferrin receptor-mediated endocytosis and cellular iron delivery of recombinant human serum transferrin from rice (Oryza sativa L.)
    Deshui Zhang
    Ventria Bioscience, Fort Collins, CO 80524, USA
    BMC Biotechnol 12:92. 2012
    ..Here, we carry out comparative studies of the TFR binding, TFR-mediated endocytosis and cellular iron delivery of recombinant hTF from rice (rhTF), and evaluate its suitability for biopharmaceutical applications...
  4. pmc Fusion protein linkers: property, design and functionality
    Xiaoying Chen
    Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA 90089, USA
    Adv Drug Deliv Rev 65:1357-69. 2013
    ....
  5. pmc Mass spectrometry study of a transferrin-based protein drug reveals the key role of protein aggregation for successful oral delivery
    Cedric E Bobst
    Department of Chemistry, University of Massachusetts, Amherst, MA 01003, USA
    Proc Natl Acad Sci U S A 109:13544-8. 2012
    ..Our results reveal an exceptional stability of GHTx, as well as the preserved ability to bind TfR, a critical first step in crossing the epithelial-intestinal barrier through receptor-mediated transcytosis...
  6. pmc Insertion of the designed helical linker led to increased expression of tf-based fusion proteins
    Nurmamet Amet
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California School of Pharmacy, PSC 404B, 1985 Zonal Avenue, Los Angeles, California, 90089 9121, USA
    Pharm Res 26:523-8. 2009
    ..To demonstrate a high-level expression of transferrin (Tf)-based fusion proteins by inserting a helical linker between two protein domains...
  7. pmc Conjugation with cationic cell-penetrating peptide increases pulmonary absorption of insulin
    Leena N Patel
    Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, Will Rogers Institute Pulmonary Research Center, Keck School of Medicine, University of Southern California, Los Angeles, California 90089 9121, USA
    Mol Pharm 6:492-503. 2009
    ..These results suggest that oligoarginine can be used to increase the alveolar absorption rate of insulin (and potentially other macromolecules as well)...
  8. pmc Pharmacokinetics of recombinant bifunctional fusion proteins
    Xiaoying Chen
    University of Southern California School of Pharmacy, Department of Pharmacology and Pharmaceutical Sciences, CA 90089 9121, USA
    Expert Opin Drug Metab Toxicol 8:581-95. 2012
    ..The lack of general PK models and study guidelines has become a bottleneck for translation of fusion proteins from basic research to the clinic...
  9. pmc Receptor-mediated activation of a proinsulin-transferrin fusion protein in hepatoma cells
    Yan Wang
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, United States
    J Control Release 155:386-92. 2011
    ..Results from this study suggest that the Tf-TfR pathway can be exploited for the design of prohormone-Tf fusion proteins as protein prodrugs for their sustained and targeted activation...
  10. pmc Effects of receptor binding on plasma half-life of bifunctional transferrin fusion proteins
    Xiaoying Chen
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California School of Pharmacy, Los Angeles, California 90089 9121, USA
    Mol Pharm 8:457-65. 2011
    ..Our findings are useful for predicting the plasma half-lives, as well as for improving the pharmacokinetic profiles of therapeutic bifunctional fusion proteins by applying linker technology...
  11. pmc Design of an in vivo cleavable disulfide linker in recombinant fusion proteins
    Xiaoying Chen
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California School of Pharmacy, Los Angeles, CA 90089, USA
    Biotechniques 49:513-8. 2010
    ....
  12. pmc Human growth hormone-transferrin fusion protein for oral delivery in hypophysectomized rats
    Nurmamet Amet
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California School of Pharmacy, PSC 404B, 1985 Zonal Avenue, Los Angeles, CA 90089 9121, USA
    J Control Release 141:177-82. 2010
    ..However, only the GHT, but not GT, fusion protein promoted a modest but statistically significant weight gain after oral dosing with 12.5mg/kg/day...
  13. ncbi Tyrphostin A8 stimulates a novel trafficking pathway of apically endocytosed transferrin through Rab11-enriched compartments in Caco-2 cells
    Fariba 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...
  14. pmc Accumulation of transferrin in Caco-2 cells: a possible mechanism of intestinal transferrin absorption
    Ching 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...
  15. ncbi Comparison of monomeric and oligomeric transferrin as potential carrier in oral delivery of protein drugs
    Ching 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...
  16. pmc Recombinant granulocyte colony-stimulating factor-transferrin fusion protein as an oral myelopoietic agent
    Yun 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...