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US 2011/0212127 A1Published application
Method for Preserving Polypeptides Using a Sugar and Polyethyleneimine
AssigneeStabilitech Ltd.
InventorsJeffrey Drew, Stephen John Ward
Life of this patent
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Abstract
The invention relates to the preservation of an active agent, such as a polypeptide, by contacting the active agent with a preservation mixture including a sugar and polyethyleneimine.
Claims32
1:
- A method for preserving a polypeptide comprising:
- (i) providing an aqueous solution of one or more sugars, a polyethyleneimine and said polypeptide;
- and
- (ii) drying the solution to form an amorphous solid matrix comprising said polypeptide.
1 more element
2:
- The method according to claim 1 wherein the concentration of polyethyleneimine is 25 μM or less based on the number-average molar mass (Mn) of the polyethyleneimine and the sugar concentration or, if more than one sugar is present, total sugar concentration is greater than 0.1M.
3:
- The method according to claim 2 in which
- (a) the Mn of the polyethyleneimine is between 20 and 1000 kDa and the concentration of the polyethyleneimine is between 0.001 and 100 nM based on the Mn, and/or
- (b) the Mn of the polyethyleneimine is between 1 and 10000 Da and the concentration of the polyethyleneimine is between 0.0001 and 1004 based on the and/or
- (c) the said concentration of polyethyleneimine is 20 μM or less or less than 500 nM, and/or
- (d) the said concentration of polyethyleneimine is 0.025 nM or more or 0.1 nM or more, and/or
- (e) the said concentration of polyethyleneimine is between 0.1 nM and 5 μM or between 0.1 nM and 200 nM.
3 more elements
4:
- The method according to claim 2 in which
- (a) the sugar concentration, or total sugar concentration, is between 0.5 and 2M;
- and/or
- (b) the sugar is sucrose, stachyose, raffinose or a sugar alcohol, or
- (c) two or more sugars are present in said aqueous solution, or
- (d) two or more sugars are present in said aqueous solution and wherein sucrose is present with another sugar;
- the concentration of sucrose relative to the other sugar is at a ratio of molar concentrations of between 3:7 and 9:1;
- and the concentration of polytheyleneimine based on Mn in step (i) is between 0.0025 nM and 5 μM, and/or
- (e) the sugars are sucrose and raffinose.
6 more elements
5:
- The method according to claim 2 in which
- (a) the solution is freeze-dried in step (ii), or
- (b) the polypeptide is a hormone, growth factor, peptide or cytokine, or
- (c) the polypeptide is a tachykinin peptide, a vasoactive intestinal peptide, a pancreatic polypeptide-related peptide, an opioid peptide or a calcintonin peptide, or
- (d) the polypeptide is an antibody or antigen-binding fragment thereof, or
- (e) the polypeptide is an antibody or antigen-binding fragment thereof in which the antibody or antigen-binding fragment is a monoclonal antibody or fragment thereof, or
- (f) the polypeptide is an antibody or antigen-binding fragment thereof in which the antibody or antigen-binding fragment is a chimeric, humanized or human antibody, or fragment thereof, or
- (g) the polypeptide is an antibody or antigen-binding fragment thereof in which the antibody or antigen-binding fragment is a chimeric, humanized or human antibody, or fragment thereof which is an IgG1, IgG2 or IgG4 or antigen-binding fragment thereof, or
- (h) the polypeptide is an antibody or antigen-binding fragment which is capable of binding to:
- (i) tumour necrosis factor α (TNF-α), interleukin-2 (IL-2), interleukin-6 (IL-6), glycoprotein CD33, CD52, CD20, CD11a, CD3, RSV F protein, HER2/neu (erbB2) receptor, vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR), anti-TRAILR2 (anti-tumour necrosis factor-related apoptosis-inducing ligand receptor 2), complement system protein C5, α4 integrin or IgE, or (ii) epithelial cell adhesion molecule (EpCAM), mucin-1 (MUC1/Can-Ag), EGFR, CD20, carcinoembryonic antigen (CEA), HER2, CD22, CD33, Lewis Y or prostate-specific membrane antigen (PMSA).
7 more elements
6:
- The method according to claim 2 in which the polypeptide is
- (a) an enzyme, or
- (b) an enzyme which is an oxidoreductase, a transferase, a hydrolase, a lyase, an isomerase or a ligase, or
- (c) an enzyme selected from an α-galactosidase, β-galactosidase, luciferase, serine proteinase, endopeptidase, caspase, chymase, chymotrypsin, endopeptidase, granzyme, papain, pancreatic elastase, oryzin, plasmin, renin, subtilisin, thrombin, trypsin, tryptase, urokinase, amylase, xylanase, lipase, transglutaminase, cell-wall-degrading enzyme, glucanase, glucoamylase, coagulating enzyme, milk protein hydrolysate, cell-wall degrading enzyme, coagulating enzyme, lysozyme, fibre-degrading enzyme, phytase, cellulase, hemicellulase, protease, mannanase or glucoamylase, or
- (d) a vaccine immunogen, or
- (e) a vaccine immunogen which is a full-length viral or bacterial protein, glycoprotein or lipoprotein;
- or a fragment thereof.
4 more elements
7:
- The method according to claim 2 which further comprises providing the resulting dried amorphous solid matrix in the form of a powder in a sealed vial, ampoule or syringe.
8:
- A method for preserving a vaccine immunogen comprising:
- (i) providing an aqueous solution of one or more sugars, a polyethyleneimine and said vaccine immunogen;
- and
- (ii) drying the solution to form an amorphous solid matrix comprising said vaccine immunogen.
1 more element
9:
- The method according to claim 8 wherein the concentration of polyethyleneimine is 25 μM or less based on the number-average molar mass (Mn) of the polyethyleneimine and the sugar concentration or, if more than one sugar is present, total sugar concentration is greater than 0.1M.
10:
- The method according to claim 9 in which
- (a) the Mn of the polyethyleneimine is between 20 and 1000 kDa and the concentration of the polyethyleneimine is between 0.001 and 100 nM based on the Mn and/or
- (b) the Mn of the polyethyleneimine is between 1 and 10000 Da and the concentration of the polyethyleneimine is between 0.0001 and 10 μM based on the Mn, and/or
- (c) the said concentration of polyethyleneimine is 20 μM or less or less than 500 nM, and/or
- (d) the said concentration of polyethyleneimine is 0.025 nM or more or 0.1 nM or more, and/or
- (e) the said concentration of polyethyleneimine is between 0.1 nM and 5 μM or between 0.1 nM and 200 nM.
3 more elements
11:
- The method according to claim 9 in which
- (a) the sugar concentration, or total sugar concentration, is between 0.5 and 2M, and/or
- (b) the sugar is sucrose, stachyose, raffinose or a sugar alcohol, and/or
- (c) two or more sugars are present in said aqueous solution, and/or
- (d) sucrose is present with another sugar;
- the concentration of sucrose relative to the other sugar is at a ratio of molar concentrations of between 3:7 and 9:1;
- and the concentration of polytheyleneimine based on Mn in step (i) is between 0.0025 nM and 5 μM, and/or
- (e) the sugars are sucrose and raffinose.
5 more elements
12:
- The method according to claim 9 in which the solution is freeze-dried in step (ii).
13:
- The method according to claim 9 in which
- (a) the vaccine immunogen is a subunit vaccine, conjugate vaccine or toxoid, or
- (b) the vaccine immunogen is a subunit vaccine in which the subunit vaccine immunogen is derived from a viral surface protein or viral capsid protein.
14:
- The method according to claim 8 further comprising providing the resulting dried amorphous solid matrix in the form of a powder in a sealed vial, ampoule or syringe.
15:
- A dry powder comprising preserved polypeptide or vaccine immunogen, obtained by the method as defined in claim 1.
16:
- A dry powder comprising preserved polypeptide or vaccine immunogen, obtained by the method as defined in claim 8.
17:
- A preserved product comprising a polypeptide or vaccine immunogen, one or more sugars and polyethylenimine, which product is in the form of an amorphous solid.
18:
- A method of preparing a vaccine comprising a vaccine immunogen, which method comprises:
- (a) providing an aqueous solution of one or more sugars, a polyethyleneimine and said vaccine immunogen wherein the concentration of polyethyleneimine is 15 μM or less based on the number-average molar mass (Mn) of the polyethyleneimine and the sugar concentration or, if more than one sugar is present, total sugar concentration is greater than 0.1M;
- and
- (b) optionally adding an adjuvant, buffer, antibiotic and/or additive to the admixture;
- and
- drying the solution to form an amorphous solid matrix comprising said vaccine immunogen.
3 more elements
19:
- A vaccine comprising a preserved product as defined in claim 15 and optionally an adjuvant.
20:
- A vaccine comprising a preserved product as defined in claim 16 and optionally an adjuvant.
21:
- A vaccine comprising a vaccine obtained by the method of claim 18 and optionally an adjuvant.
22:
- A sealed vial, ampoule or syringe containing a dry powder as defined in claim 15.
23:
- A sealed vial, ampoule or syringe containing a dry powder as defined in claim 16.
24:
- A sealed vial, ampoule or syringe containing a preserved product as defined in claim 17.
25:
- A sealed vial, ampoule or syringe containing a vaccine as defined in claim 19.
26:
- A sealed vial, ampoule or syringe containing a vaccine as defined in claim 20.
27:
- A sealed vial, ampoule or syringe containing a vaccine as defined in claim 21.
28:
- A method for preserving a polypeptide prior to drying comprising:
- (i) providing an aqueous solution of one or more sugars, a polyethyleneimine and said polypeptide;
- and
- (ii) storing the solution for up to five years in a sealed container.
1 more element
29:
- A method according to claim 28, which further comprises:
- (iii) drying the solution to form an amorphous solid matrix comprising said polypeptide.
30:
- A method according to claim 28, in which (a) the solution is stored in a refrigerator, or (b) the solution is stored in a freezer.
31:
- A bulk aqueous solution of one or more sugars, a polyethyleneimine and a polypeptide, which solution is provided in a sealed container and is stored prior to drying in a refrigerator or freezer.
32:
- A solution according to claim 31 in which the bulk aqueous solution has a volume of 0.1 to 100 litres.