title: Development of Calcitonin Salmon Nasal Spray: similarity of peptide formulated in chlorobutanol compared to benzalkonium chloride as preservative.
abstract: The similarity of an intranasal salmon calcitonin (sCT) employing chlorobutanol as preservative (Calcitonin Salmon Nasal Spray) was compared to the reference listed drug (RLD) employing benzalkonium chloride as preservative (Miacalcin Nasal Spray). Various orthogonal methods assessed peptide structuring, dynamics, and aggregation state. Mass spectrometry, amino acid analysis, and N-terminal sequencing all demonstrated similarity in primary structure. Near- and far-UV circular dichroism (CD) data supported similarity in secondary and tertiary sCT structure. Nuclear magnetic resonance studies further supported similarity of three-dimensional structure and molecular dynamics of the peptide. Other methods, such as sedimentation velocity and size exclusion chromatography, demonstrated similarity in peptide aggregation state. These latter methods, in addition to reversed phase chromatography, were also employed for monitoring stability under forced degradation, and at the end of recommended shelf storage and patient use conditions. In all cases and for all methodologies employed, similarity to the RLD was observed with respect to extent of aggregation and other degradation processes. Finally, ELISA and bioassay data demonstrated similarity in biological properties. These investigations comprehensively demonstrate physicochemical similarity of Calcitonin Salmon Nasal Spray and the RLD, and should prove a useful illustration to pharmaceutical scientists developing alternative and/or generic peptide or protein products.
authors.affiliation: Nastech Pharmaceutical Company, Inc, Bothell, Washington 98021, USA.
authors.forename: Henry R
authors.initials: HR
authors.lastname: Costantino
authors.forename: Heather
authors.initials: H
authors.lastname: Culley
authors.forename: Lishan
authors.initials: L
authors.lastname: Chen
authors.forename: Daniel
authors.initials: D
authors.lastname: Morris
authors.forename: Michael
authors.initials: M
authors.lastname: Houston
authors.forename: Sharin
authors.initials: S
authors.lastname: Roth
authors.forename: Mary Jo
authors.initials: MJ
authors.lastname: Phoenix
authors.forename: Chuck
authors.initials: C
authors.lastname: Foerder
authors.forename: John S
authors.initials: JS
authors.lastname: Philo
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Arakawa
authors.forename: Lisa
authors.initials: L
authors.lastname: Eidenschink
authors.forename: Niels H
authors.initials: NH
authors.lastname: Andersen
authors.forename: Gordon
authors.initials: G
authors.lastname: Brandt
authors.forename: Steven C
authors.initials: SC
authors.lastname: Quay
mesh_terms: D000281:Administration, Intranasal; D000336:Aerosols; D000596:Amino Acids / Q000032:analysis; D018927:Anti-Asthmatic Agents / Q000008:administration & dosage* / Q000737:chemistry; D001548:Benzalkonium Compounds / Q000737:chemistry*; D001681:Biological Assay; D002116:Calcitonin / Q000008:administration & dosage* / Q000737:chemistry; D002152:Calorimetry, Differential Scanning; D002626:Chemistry, Pharmaceutical; D002724:Chlorobutanol / Q000737:chemistry*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004356:Drug Storage; D004797:Enzyme-Linked Immunosorbent Assay; D031521:Fractionation, Field Flow; D009682:Magnetic Resonance Spectroscopy; D010455:Peptides / Q000737:chemistry*; D011310:Preservatives, Pharmaceutical / Q000737:chemistry*; D012015:Reference Standards; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013056:Spectrophotometry, Ultraviolet; D014461:Ultracentrifugation
chemical_list: D000336:Aerosols; D000596:Amino Acids; D018927:Anti-Asthmatic Agents; D001548:Benzalkonium Compounds; D010455:Peptides; D011310:Preservatives, Pharmaceutical; C028815:salmon calcitonin; D002116:Calcitonin; D002724:Chlorobutanol