Stabilisation of polypeptides
Inventors
Drew, Jeffrey • Woodward, David Thomas • Ward, Stephen
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-10029007-B2
Publication Date
2018-07-24
Expiration Date
Abstract
A method for preserving a polypeptide comprising: (a) providing an aqueous solution of (i) the polypeptide, (ii) one or more sugars, and (iii) a compound of formula (I) or a physiologically acceptable salt or ester thereof and/or a compound of formula (II) or a physiologically acceptable salt or ester thereof; and (b) drying the solution to form a composition incorporating the polypeptide.
Core Innovation
The invention relates to preserving a polypeptide during freeze-drying by formulating the polypeptide in a buffered aqueous solution and then freeze-drying the solution to form a composition incorporating the polypeptide. The buffered aqueous solution comprises the polypeptide and, as the sole excipients, one or more sugars and a compound of formula (IA) or a physiologically acceptable salt or ester thereof, with the option of including a compound of formula (II) or a physiologically acceptable salt or ester thereof.
In the disclosed embodiment, the polypeptide is an antibody or an antigen-binding fragment of an antibody, and the buffered aqueous solution does not comprise an aluminium salt adjuvant. The resulting dried composition preserves the polypeptide during freeze-drying and is presented as a thermoprotectant or lyoprotectant for protecting polypeptide structure and activity following thermal challenge.
The described examples include freeze-dried antibody formats such as bivalent F(ab')2 and monovalent Fab, as well as an HRP-conjugated mouse IgG and recombinant protective antigen (rPA). Preservation of protein integrity and residual activity or binding is evaluated by ELISA, HPLC-SEC, and DSC, with improved thermal stability relative to liquid storage and improved thermoprotection reported for systems using dimethylglycine and mannitol.
Claims Coverage
The document includes one independent claim directed to a method for preserving a polypeptide during freeze-drying, with dependent claims refining excipient identities, concentrations, ratios, packaging or formats, and antibody types. The independent claim centers on sugars plus a compound of formula (IA), optionally formula (II), as sole excipients, together with exclusion of aluminium salt adjuvant.
Buffering with sugars and formula (IA) as sole excipients
Providing a buffered aqueous solution comprising the polypeptide and, as the sole excipients, one or more sugars and a compound of formula (IA) or a physiologically acceptable salt or ester thereof.
Optional inclusion of formula (II) compounds
Optionally including a compound of formula (II) or a physiologically acceptable salt or ester thereof in the buffered aqueous solution.
Freeze-drying to form a composition incorporating the polypeptide
Freeze-drying the solution to form a composition incorporating the polypeptide.
Antibody or antigen-binding fragment polypeptide with aluminium salt adjuvant exclusion
Wherein the polypeptide is an antibody or antigen-binding fragment of an antibody, and wherein the buffered aqueous solution does not comprise an aluminium salt adjuvant.
Dimethylglycine as the compound of formula (IA)
The method wherein the compound of formula (IA) is dimethylglycine or a physiologically acceptable salt or ester thereof.
Dimethylsulfone as the optional compound of formula (II)
The method wherein the compound of formula (II) is dimethylsulfone.
Quantified excipient concentration ranges for formula and sugars
The method using specific concentration ranges for the compound of formula (IA) and/or its salt or ester, the optional compound of formula (II) and/or its salt or ester, and the sugar or total sugar concentration.
Non-reducing sugars and/or sugar alcohols with sucrose-to-other-sugar ratio constraint
The method using one or more sugars selected as non-reducing sugars and/or sugar alcohols, optionally including sucrose, and when sucrose is combined with other sugars the sucrose-to-other-sugar concentration ratio is from 1:1 to 20:1.
Specific antibody classes for the polypeptide
The method wherein the polypeptide is a monoclonal antibody or fragment, or a chimeric, humanized, or human antibody or a fragment thereof.
Overall, the claim coverage is centered on freeze-drying a polypeptide, specifically an antibody or antigen-binding fragment, after formulating it in a buffered aqueous solution whose only excipients are sugars plus a compound of formula (IA), optionally supplemented with a compound of formula (II), while explicitly avoiding aluminium salt adjuvant. Dependent refinements specify identities for formula (IA) and formula (II), constrain formulation concentrations and sugar selection or ratios, and narrow the polypeptide to particular antibody classes and fragments.
Stated Advantages
Preserves the polypeptide during freeze-drying.
Improved thermal stability for the freeze-dried composition versus liquid storage, as indicated by residual activity or binding and protein integrity measurements.
Higher retained antigenicity or binding and preserved chromatographic peak profiles after heat challenge for disclosed excipient systems using DMG with mannitol and related sugar systems.
DSC-indicated prevention of mannitol recrystallization during freeze-drying and during cooling, supporting an amorphous mannitol state that correlates with better thermoprotection.
Provides protection of polypeptide structure and activity in the dried composition, including maintaining antibody activity and binding after thermal challenge.
Excludes aluminium salt adjuvant from the buffered aqueous solution.
Documented Applications
Formulation and preservation of antibody formats, including bivalent F(ab')2 and monovalent Fab, as freeze-dried compositions, with thermal stress evaluation using ELISA and other analyses.
Formulation and preservation of an HRP-conjugated mouse IgG and evaluation of residual activity or binding after thermal stress in excipient systems.
Formulation and preservation of recombinant protective antigen (rPA) as a freeze-dried composition and evaluation using ELISA-based competition assays and protein integrity analyses.
Thermal stability assessment of freeze-dried polypeptide compositions using assays including ELISA, HPLC-SEC, and DSC, including monitoring protein integrity and residual binding or activity over time and temperature.
Preservation of antibody and antigen-binding fragment polypeptides during freeze-drying, including maintaining antibody activity and binding following thermal challenge.
Use in vaccine immunogens preservation within the polypeptide scope discussed in the document.
Interested in licensing this patent?