title: Stabilization of a recombinant human epidermal growth factor parenteral formulation through freeze-drying.
abstract: Development studies were performed to design a pharmaceutical composition that allows the stabilization of a parenteral rhEGF formulation in a lyophilized dosage form. Unannealed and annealed drying protocols were tested for excipients screening. Freeze-dry microscopy was used as criterion for excipients and formulation selection; as well as to define freeze-drying parameters. Excipients screening were evaluated through their effect on freeze-drying recovery and dried product stability at 50 °C by using a comprehensive set of analytical techniques assessing the chemical stability, protein conformation and bioactivity. The highest stability of rhEGF during freeze-drying was achieved by the addition of sucrose or trehalose. After storing the dried product at 50 °C, the highest stability was achieved by the addition of dextran, sucrose, trehalose or raffinose. The selected formulation mixture of sucrose and dextran could prevent protein degradation during the freeze-drying and delivery processes. The degradation rate assessed by RP-HPLC could decrease 100 times at 37 °C and 70 times at 50 °C in dried with respect to aqueous formulation. These results indicate that the freeze-dried formulation represents an appropriate technical solution for stabilizing rhEGF.
authors.affiliation: Pharmaceutical Development Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box. 6162, Havana 10600, Cuba. Electronic address: hector.santana@cigb.edu.cu.
authors.forename: Héctor
authors.initials: H
authors.lastname: Santana
authors.affiliation: Pharmaceutical Development Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box. 6162, Havana 10600, Cuba.
authors.forename: Jorge
authors.initials: J
authors.lastname: Sotolongo
authors.affiliation: Pharmaceutical Development Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box. 6162, Havana 10600, Cuba.
authors.forename: Yaima
authors.initials: Y
authors.lastname: González
authors.affiliation: Pharmaceutical Development Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box. 6162, Havana 10600, Cuba.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Hernández
authors.affiliation: Chemical Physical Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box. 6162, Havana 10600, Cuba.
authors.forename: Glay
authors.initials: G
authors.lastname: Chinea
authors.affiliation: Quality Control Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box. 6162, Havana 10600, Cuba.
authors.forename: Haydée
authors.initials: H
authors.lastname: Gerónimo
authors.affiliation: Analytical Development Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box. 6162, Havana 10600, Cuba.
authors.forename: Odalys
authors.initials: O
authors.lastname: Amarantes
authors.affiliation: Central Office for Scientific Pole, 198th and 17, n° 1714, Cubanacan, Playa, Havana 11300, Cuba.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Beldarraín
authors.affiliation: Pharmaceutical Development Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box. 6162, Havana 10600, Cuba.
authors.forename: Rolando
authors.initials: R
authors.lastname: Páez
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D004815:Epidermal Growth Factor / Q000737:chemistry*; D005079:Excipients / Q000737:chemistry; D005612:Freeze Drying; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008853:Microscopy; D009994:Osmolar Concentration; D011994:Recombinant Proteins / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013395:Sucrose / Q000737:chemistry; D013696:Temperature; D014199:Trehalose / Q000737:chemistry
chemical_list: D005079:Excipients; D011994:Recombinant Proteins; D013395:Sucrose; D004815:Epidermal Growth Factor; D014199:Trehalose
keywords: Characterization; Epidermal growth factor; Excipients; Lyophilized formulation; Parenteral formulation; Stabilization