G-CSF transferrin fusion proteins
| Title: | G-CSF transferrin fusion proteins |
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| Patent Number: | 8,188,032 |
| Publication Date: | May 29, 2012 |
| Appl. No: | 10/575033 |
| Application Filed: | October 08, 2004 |
| Abstract: | A polypeptide comprising a G-CSF domain operably linked to a Tf domain, wherein the ability of the polypeptide to be transported into a cell expressing a TfR gene or the ability of the polypeptide to be transported across a cell expressing a TfR gene via transcytosis is higher than that of the G-CSF domain alone. |
| Inventors: | Shen, Wei-Chiang (San Marino, CA, US); Bai, Yun (Tucker, GA, US); Ann, David (Arcadia, CA, US); Widera, Adam (Long Beach, CA, US) |
| Assignees: | National Institutes of Health (NIH) (Bethesda, MD, US) |
| Claim: | 1. A fusion polypeptide comprising a granulocyte colony stimulating factor (G-CSF) domain operably linked to a transferrin (Tf) domain, wherein the ability of the polypeptide to be transported into a cell expressing a transferrin receptor (TfR) gene or the ability of the polypeptide to be transported across a cell expressing a TfR gene via transcytosis is higher than that of the G-CSF domain alone, wherein the polypeptide is a recombinant polypeptide, and wherein said Tf domain is preloaded with at least one iron molecule. |
| Claim: | 2. The fusion polypeptide of claim 1 further comprising a secretion signal at the N-terminus. |
| Claim: | 3. The fusion polypeptide of claim 1 , wherein the G-CSF domain is N-terminus to the Tf domain. |
| Claim: | 4. A composition comprising a pharmaceutically acceptable carrier and the fusion polypeptide of claim 1 or 3 . |
| Claim: | 5. The composition of claim 4 , further comprising sodium bicarbonate, BSA, casein, or a combination thereof. |
| Claim: | 6. A method of enhancing production of circulating neutrophils in a subject, comprising administering to a subject in need thereof an effective amount of the composition of claim 4 . |
| Claim: | 7. The method of claim 6 , wherein the subject is undergoing chemotherapy for cancer, or is suffering from or at risk for developing severe chronic neutropenia or a bone marrow transplant-related disorder. |
| Claim: | 8. The method of claim 6 , wherein the composition is administered orally. |
| Claim: | 9. The method of claim 6 , wherein the composition is administered subcutaneously. |
| Claim: | 10. The fusion polypeptide of claim 1 , wherein the Tf domain is preloaded with two iron molecules. |
| Claim: | 11. A method of enhancing transport of G-CSF into or across a gastrointestinal (GI) epithelial cell, comprising contacting the GI epithelial cell with the fusion polypeptide of claim 1 under conditions that allow transport of the fusion polypeptide into the cell through TfR or transport of the fusion polypeptide across the cell through TfR via transcytosis. |
| Current U.S. Class: | 514/11 |
| Patent References Cited: | 5254342 October 1993 Shen et al. 5665863 September 1997 Yeh 5672683 September 1997 Friden et al. 7176278 February 2007 Prior 2003/0082679 May 2003 Sun et al. 2003/0144198 July 2003 Collins WO 93/20834 October 1993 |
| Other References: | Widera et al., Pharm. Res. vol. 20: 1231-1238, 2003. cited by examiner Mouz et al., Nutrition 16: 229-230,2000. cited by examiner Deven Shah, et al. “Transcellular Delivery of an Insulin-Transferrin Conjugate in Enterocyte-like Caco-2 Cells” (Journal of Pharmaceutical Sciences, American Pharmaceutical Assoc., Washington, US, vol. 85, No. 12, Dec. 1996, pp. 1306-1311, XP002906594, ISSN: 0022-3549). cited by other Deven Shah, et al. “The Establishment of Polarity and Enhanced Transcytosis of Transferrin Receptors in Enterocyte-like Caco-2 Cells” (Journal of Drug Targeting, vol. 2, No. 2, 1994, pp. 93-99; XP-009094607, Switzerland). cited by other Wendy Halpern, et al. “Albugranin™, a Recombinant Human Granulocyte Colony Stimulating Factor (G-CSF) Genetically Fused to Recombinant Human Albumin Induces Prolonged Myelopoietic Effects in Mice and Monkeys” (Pharmaceutical Research Nov. 2002, vol. 19, No. 11, Nov. 2002, pp. 1720-1729; XP-002464933; ISSN: 0724-8741). cited by other Ophry Pines, et al. “Expression and Secretion of Proteins in E. coli” (Molecular Biotechnology, Totowa, NJ, US, vol. 2, No. 1, 1999, pp. 25-34; XP-000985935; ISSN:1073-6085). cited by other G.E. Grampp, et al. “Use of Regulated Secretion in Protein Production From Animal Cells: An Overview” (Advances in Biochemical Engineering/Biotechnology 1992, vol. 46, 1992, pp. 35-62; XP-009094668; ISSN: 0724-6145). cited by other Lynn McCarroll, et al. “Stable insect cell cultures for recombinant protein production” (Current Opinion in Biotechnology Oct. 1997, vol. 8, No. 5, Oct. 1997, pp. 590-594; XP-002464935; ISSN: 0958-1669). cited by other Richard G. Buckholz, et al. “Yeast Systems for the Commercial Production of Heterologous Proteins” (Bio/Technology, Nature Publishing Co. New York, US, vol. 9, Nov. 1991, pp. 1067-1072; XP-000918924; ISSN: 0733-222X). cited by other S. Hohaus, et al. “Recombinant human granulocyte and granulocyte-macrophage colony-stimulating factor (G-CSF and GM-CSF) administered following cytotoxic chemotherapy have a similar ability to mobilize peripheral blood stems cells” Bone Marrow Transplantation Oct. 1998, vol. 22, No. 7, Oct. 1998, pp. 625-630; XP-002464936 ISSN: 0268-3369). cited by other Mutay Asian, et al. “The effect of recombinant human granulocyte/macrophage-colony-stimulating factor (rHu GM-CSF) and rHu G-CSF administration on neutrophil chemiluminescence assay in patients following cyclic chemotheraphy” (Cancer Immunology, Immunotherapy : CII Nov. 1998, vol. 47, No. 3, pp. 176-181; XP 002464937 ISSN: 0340-7004). cited by other Adam Widera, et al. “The Transepithelial Transport of a G-CSF-Transferrin Conjugate in Caco-2 Cells and Its Myelopoietic Effect in BDF1 Mice” (Pharmaceutical Research Feb. 2004, vol. 21, No. 2, pp. 278-284; XP-002464938; ISSN: 0724-8741). cited by other PCT Search Report for corresponding PCT application No. PCT/US04/33337 lists the references above. cited by other |
| Primary Examiner: | Chandra, Gyan |
| Attorney, Agent or Firm: | DLA Piper LLP (US) |
| Accession Number: | edspgr.08188032 |
| Database: | USPTO Patent Grants |
| Language: | English |
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