WO2010084298A1 - Stable vaccine compositions and methods of use - Google Patents
Stable vaccine compositions and methods of use Download PDFInfo
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- WO2010084298A1 WO2010084298A1 PCT/GB2009/050051 GB2009050051W WO2010084298A1 WO 2010084298 A1 WO2010084298 A1 WO 2010084298A1 GB 2009050051 W GB2009050051 W GB 2009050051W WO 2010084298 A1 WO2010084298 A1 WO 2010084298A1
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- vaccine formulation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/02—Bacterial antigens
- A61K39/07—Bacillus
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55505—Inorganic adjuvants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55561—CpG containing adjuvants; Oligonucleotide containing adjuvants
Definitions
- the present invention relates generally to the field of protein storage, especially following industrial scale preparation and where said proteins have form part of a vaccine composition intended for administration to mammals, especially human beings.
- Vaccines are commonly formulated using an adjuvant.
- a common adjuvant-type is the aluminium based colloids, usually referred to as alum. More specifically these are usually aluminium hydroxide (aluminium oxyhydroxide) (also called alhydrogel) and aluminium phosphate (also called adju-phos).
- aluminium oxyhydroxide also called alhydrogel
- aluminium phosphate also called adju-phos
- vaccines useful against organisms such as anthrax (Bacillus anthracis) are commonly formulated with alhydrogel, which binds the anthrax antigen used in such vaccines (so called subunit vaccines).
- Physical instability can result from, for example, denaturation or aggregation of the protein(s) present in the vaccines while chemical instability can result from chemical reactions that either break the amino acid chain of the protein or serve to modify one or more amino acid side groups present on the protein (which groups may be essential for immunogenic activity).
- Lyophilization is a well known technique for preserving proteins and operates by removing water from the protein composition of interest. It is a process by which the material to be dried is first frozen and then the frozen solvent (ice in the case of water) is removed by sublimation under high vacuum. Additives may be included in pre-lyophilized formulations to enhance stability during the freeze-drying process and/or to improve stability of the lyophilized product upon storage. Unfortunately, such procedures have heretofore proved unreliable for the lyophilization and storage of certain antigenic proteins useful in vaccines, especially for anthrax vaccines, where stability of the lyophilized powder has resulted in significant loss of activity following reconstitution.
- the present invention solves at least some of the problems heretofore encountered with lyophilization of protein formulations, especially of formulations containing recombinant protective antigen (rPA) of anthrax (Bacillus anthracis), by providing a formulation that retains antigenic activity and thus utility as a vaccine even with long storage in the lyophilized state, so that the reconstituted vaccine composition is still highly useful for protection of mammals, such as human being, against infection by anthrax bacilli.
- rPA recombinant protective antigen
- the present invention relates to a stable reconstituted formulation or vaccine composition, which may be isotonic, comprising an antigen or immunogen for use in a vaccine, preferably an anthrax antigen for use in a vaccine against anthrax, more preferably anthrax Protective Antigen, and most preferably a recombinant Protective Antigen (rPA), in an amount of about 50 ⁇ g/mL to about 400 ⁇ g/mL, preferably about 200 ⁇ g/ml, and a pharmaceutically acceptable carrier, wherein this formulation has been reconstituted from a lyophilized mixture or composition of the anthrax, or other, antigen.
- the reconstituted formulation may contain trehalose, such as trehalose in an amount that provides lyoprotection of the lyophilized mixture or composition.
- said formulation contains a salt, such as phosphate or NaCI or both, a detergent, such as Tween, and an adjuvant, such as alhydrogel (alum), chitosan, MPL or CpG.
- a salt such as phosphate or NaCI or both
- a detergent such as Tween
- an adjuvant such as alhydrogel (alum), chitosan, MPL or CpG.
- the adjuvant is CpG1018 or alhydrogel or, especially preferred, both.
- the vaccine according to the present invention may comprise adjuvants additional to alum.
- adjuvants are well known in the art and include oligonucleotides, especially so-called "CpG” oligonucleotides, especially phosphorothioate oligonucleotides, most preferably oligonucleotides which target Toll Like Receptor 9 receptors.
- additional adjuvants that may be present include saponins, PCPP polymer, lipopolysaccharide derivatives, MPL [Monophosphoryl Lipid A], MDP muramyl di-peptide (MDP) , t-MDP , Flagellin and flagellin-fusion proteins, IC31 , OM- 174 and Leishmania elongation factor, ISCOMS, chitosan, SB-AS2, AS02, SB-AS4, non-ionic block copolymers and SAF [Syntex Adjuvant Formulation].
- MPL Monophosphoryl Lipid A
- MDP muramyl di-peptide (MDP) MDP muramyl di-peptide
- t-MDP t-MDP
- Flagellin and flagellin-fusion proteins IC31 , OM- 174 and Leishmania elongation factor, ISCOMS, chitosan, SB-AS2, AS02, SB-AS4, non-ionic block copolymers and SAF
- the additional adjuvant is preferably incorporated with the stabilised sub-unit vaccine although the additional adjuvant may alternatively be present in the separate adjuvant and/or diluent composition.
- the stable sub-unit vaccine composition or formulation may also incorporate an agent to reduce or prevent agglomeration during preservation, for example a small amount of a non-ionic surfactant, especially an ethoxylated sorbitan ester, for example polyoxyethylenesorbilan monolaurate, typically comprising about 20 ethyleneoxy units.
- the present invention relates to a method for preparing a stable lyophilized vaccine formulation, comprising the steps of:
- the mixture or composition to be lyophilized also contains a buffer, such as phosphate, a salt, such as NaCI, a detergent, such as Tween, and an adjuvant, such as alhydrogel (alum), chitosan, mpl or CpG, with CpG1018 and alhydrogel being especially preferred.
- a buffer such as phosphate
- a salt such as NaCI
- a detergent such as Tween
- an adjuvant such as alhydrogel (alum), chitosan, mpl or CpG, with CpG1018 and alhydrogel being especially preferred.
- the lyophilized composition also comprises a bulking agent added in an amount sufficient to provide mass to the lyophilized composition and contribute to the physical structure of the lyophilized bulk material.
- the present invention relates to a kit comprising a container that holds a lyophilized mixture or composition of an anthrax, or other, vaccine containing rPA, trehalose and a pharmaceutically acceptable carrier; and a set of instructions for reconstituting the lyophilized composition with a diluent to an rPA concentration of about 200 ⁇ g/mL.
- a diluent is provided with the kit.
- the diluent contains an adjuvant, such as alhydrogel.
- the present invention also relates to a method of protecting against, or treating, a microbial, especially a bacterial, infection in a mammal, comprising administering to a mammal either at risk of developing or afflicted with such infection a therapeutically-effective amount of the vaccine composition of the invention.
- the bacterial infection is an anthrax (Bacillus anthracis) infection and/or the mammal is a human being.
- Figure 1 shows the amino acid sequence of a recombinant Protective Antigen (rPA) from Bacillus anthracis (anthrax).
- rPA Protective Antigen
- a “stable” formulation is one wherein the antigenic protein, such as rPA, retains its physical and chemical stability and integrity upon storage, such as in the form of a lyophilized powder, solid or cake.
- Analytical techniques for measuring protein stability are known in the art and will not be described in detail herein. See, for example, Peptide and Protein Drug Delivery, 247-301 , Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991 ) and Jones, A. Adv. Drug Delivery Rev. 10: 29-90 (1993).
- a “stable” formulation may be one wherein less than about 10% and preferably less than about 5% of the protein is present as an aggregate in the formulation.
- any increase in aggregate formation following lyophilization and storage of the lyophilized formulation can be determined.
- a "stable" lyophilized formulation may be one wherein the increase in aggregate in the lyophilized formulation is less than about 5% and preferably less than about 3%, when the lyophilized formulation is stored at 2-8 0 C for at least 6 months, preferably 9 months, more preferably up to one year or longer, such as at least about 16 months.
- the term "about” means an amount that does not substantially reduce the effectiveness of the component whose range or amount is being recited by more than 10%.
- a "reconstituted" formulation is one that has been prepared by dissolving a lyophilized protein formulation in a diluent such that the protein is dispersed in the reconstituted formulation.
- the reconstituted formulation in suitable for administration to a patient.
- isotonic refers to a formulation or composition of the invention having substantially the same osmotic pressure as human blood. Isotonic formulations will generally have an osmotic pressure from about 250 to 350mOsm/Kg. lsotonicity can be measured using a vapor pressure or ice- freezing type osmometer, for example.
- lyoprotectant means a molecule that, when combined with an immunogenic protein, such as rPA, of the invention, substantially reduces, if not prevents, chemical and/or physical instability of said protein upon lyophilization and subsequent storage, sufficient to maintain the immunogenic properties of the protein and thereby insure continued utility as a vaccine.
- lyoprotectants include non-reducing sugars, such as sucrose or trehalose, amino acids, such as monosodium glutamate or histidine, methylamines like betaine, lyotropic salts, like magnesium sulfate, polyols, like trihydric or higher sugar alcohols, including glycerin, erythritol, glycerol, arabitol, xylitol, sorbitol, and mannitol, glycols, like propylene glycol and polyethylene glycol, and combinations of any or all of these.
- non-reducing sugars such as sucrose or trehalose
- amino acids such as monosodium glutamate or histidine
- methylamines like betaine
- lyotropic salts like magnesium sulfate
- polyols like trihydric or higher sugar alcohols, including glycerin, erythritol, glycerol, arabitol, xylitol,
- lyoprotecting amount refers to the amount of a lyoprotectant, for example, trehalose, that when added to an immunogen, such as rPA, of the invention, substantially preserves the physical and chemical stability and integrity of the immunogen upon lyophilization and storage so that, once reconstituted, said immunogen retains its immunogenic properties sufficiently to permit its use in a vaccine.
- diluent means a carrier that is pharmaceutically acceptable (safe and non-toxic for administration to a human) and is useful for the preparation of a reconstituted formulation.
- exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), a pH buffered solution (e.g. phosphate-buffered saline), sterile saline solution, Ringer's solution or dextrose solution.
- the term "preservative" means a compound that can be added to the diluent to essentially reduce bacterial action in the reconstituted formulation, thus facilitating the production of a multi-use reconstituted formulation, for example.
- examples include octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride (a mixture of alkylbenzyldimethylammonium chlorides in which the alkyl groups are long-chain compounds), and benzethonium chloride.
- preservatives include aromatic alcohols such as phenol, 2-phenoxyethanol, thimerosal, benzethonium chloride, formaldehyde, butyl and benzyl alcohol, allyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol.
- aromatic alcohols such as phenol, 2-phenoxyethanol, thimerosal, benzethonium chloride, formaldehyde, butyl and benzyl alcohol
- allyl parabens such as methyl or propyl paraben
- catechol resorcinol
- cyclohexanol 3-pentanol
- m-cresol m-cresol
- the term "bulking agent” refers to a compound that adds mass to the lyophilized mixture or composition and contributes to the physical structure of the lyophilized cake (e.g. facilitates the production of an essentially uniform lyophilized cake which maintains an open pore structure).
- Exemplary bulking agents include mannitol, glycine, dextran, polyethylene glycol and xorbitol.
- Treatment refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented.
- the term "effective amount” or “therapeutically effective amount” means a dosage sufficient to treat, inhibit, or alleviate one or more symptoms of a bacterial infection, especially anthrax infection.
- the precise dosage may vary with such factors as the age, immune system status, general health, and environmental circumstances of the patient, the nature and extent of the infection (or anticipated infection) and the availability of subsequent treatment/vaccination.
- the antigen to be formulated is generally prepared using techniques well known in the art, including isolation from the organism as well as synthesis by recombinant technology or direct chemical synthesis of the polypeptide used in the antigen.
- the Protective Antigen used in the present invention was prepared by recombinant technology and is referred to herein as recombinant Protective Antigen (rPA).
- a stable reconstituted formulation comprising a sub-unit vaccine antigen, for example, an anti-microbial antigen, such as an anthrax antigen containing recombinant Protective Antigen (rPA), and a pharmaceutically acceptable carrier, which reconstituted formulation has been prepared from a lyophilized composition of said anthrax antigen and a lyoprotective amount of trehalose.
- an anti-microbial antigen such as an anthrax antigen containing recombinant Protective Antigen (rPA)
- rPA recombinant Protective Antigen
- rPA recombinant Protective Antigen
- the reconstituted anthrax vaccine formulation of the invention may also contain trehalose.
- the trehalose is present in the reconstituted formulation in the range of about 3% to 7%, preferably in the range of about 4% to 6%, more preferably in the range of about 4.5% to 5.5%, and most preferably at about 5% (w/v).
- the stable reconstituted vaccine formulation of the invention is one reconstituted from a lyophilized composition that was maintained at a temperature of at least 5 0 C for a period of at least 1 month prior to reconstitution, or at least 25 0 C for a period of at least 1 month prior to reconstitution, or wherein said temperature is at least 3O 0 C, 35 0 C, 4O 0 C, 45 0 C, 5O 0 C, 55 0 C, 6O 0 C, 65 0 C, 7O 0 C, or even higher, and for periods of at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7, months, 8 months, 9 months, 10 months, or even longer, such as a year, or 16 months or at least 2 years.
- the reconstituted anthrax vaccine formulation of the invention may also contain a buffer.
- the buffer (or salt) is phosphate at a concentration of in the range of 3 mM to 7 mM, preferably in the range of 4 mM to 6 mM, more preferably in the range of 4.5 mM to 5.5 mM, with concentrations of about 4 mM and about 5.25 mM being especially preferred.
- the reconstituted formulation comprises NaCI, for example, at about 0.39%.
- the reconstituted formulation of the invention may also contain a detergent, such as Tween 20. In one specific but non-limiting example thereof Tween 20 is present at about 0.02%.
- the reconstituted anthrax vaccine formulation of the invention may also contain an adjuvant, especially one selected from the group alhydrogel (alum), chitosan, mpl and CpG, with CpG and alhydrogel preferred.
- the adjuvant is CpG1018, for example, present at about 2 mg/ml.
- the adjuvant is alhydrogel, for example, present at about 0.26% (w/v).
- a complete but non-limiting example of such a formulation is one that contains 200 ⁇ g/ml rPA, 2 mg/ml CpG1018, 5% trehalose, 5.25 mM phosphate, 0.39% NaCI, 0.02% Tween 20, 0.26% w/v alhydrogel (see Example 1 ).
- the reconstituted formulation is isotonic.
- the present invention also provides a method for preparing a stable reconstituted formulation, comprising the steps of:
- a sub-unit vaccine antigen for example, an anti-microbial antigen, such as an anthrax antigen containing rPA, and a lyoprotective amount of trehalose; and
- step (b) reconstituting the lyophilized mixture or composition of step (a) in a diluent such that the reconstituted formulation is stable and has an rPA concentration of between about 0.1 and 10 times the rPA concentration in the mixture or composition before lyophilization.
- the diluent used in step (b) for reconstituting the mixture of step (a) comprises an adjuvant, preferably selected from alhydrogel (alum), chitosan, MPL or CpG, most preferably alhydrogel.
- alhydrogel is present in the range of about 0.2% to about 0.3%, or is present in the range of about 0.25% to about 0.3%, or preferably is present at about 0.26%.
- the diluent comprises a salt, such as phosphate and/or NaCI.
- the phosphate is present in the range of about 3 to 7 mM, or is present in the range of about 4 to 6 mM, or is present in the range of about 4.5 to 5.5 mM, or the phosphate is preferably present at about 5 mM.
- the NaCI is present in the range of about 0.3% to about 0.5%, or is present in the range of about 0.35% to about 0.45%, or is preferably present at about 0.39%.
- the diluent has a pH about 7.4.
- the diluent contains 0.26% alhydrogel in 5 mM phosphate and 0.39% NaCI, pH 7.4.
- Sub-unit vaccines containing anti-microbial antigens and which can be employed are usually recombinant protein-based antigens.
- anti-microbial antigens include Hepatitis B protective antigens, Herpes Simplex Virus antigens, Influenza antigens, Congenital cytomegalovirus (CMV) antigens, Tuberculosis antigens, HIV antigens, Diphtheria antigens, Tetanus antigens, Pertussis antigens and Yersinia pestis protective antigens, such as antigens comprising one, two or more antigenic proteins, for example those disclosed in patent application W096/28551 , incorporated herein by reference.
- the antigen is an anthrax protective antigen, such as recombinant protective antigen (rPA), especially that having the amino acid sequence of SEQ ID NO: 1.
- the present invention further provides a method for preparing a stable lyophilized vaccine formulation, comprising the steps of:
- a sub-unit vaccine antigen for example, an anti-microbial antigen, such as anthrax antigen containing rPA, and a lyoprotective amount of a reducing sugar; and
- the composition of step (a) contains rPA in the range of about 200 to 600 ⁇ g/ml, or about 300 to 500 ⁇ g/ml, or about 350 to 450 ⁇ g/ml, or preferably at about 400 ⁇ g/ml.
- the composition of step (a) contains trehalose is present in the range of about 2 to 20%, or in the range of about 4 to 18%, or about 6 to 16%, or about 8 to 14%, or about 8 to 12%, or about 9 to 12%, or preferably at about 10% (w/v).
- the composition or mixture of step (a) contains a salt, preferably phosphate and/or NaCI (or saline).
- the phosphate is present in the range of about 0.3 to 0.7 mM, or in the range of about 0.35 to 0.65 mM, or about 0.4 to 0.6 mM, or about 0.45 to 0.55 mM, or preferably is present at about 0.5 mM.
- the NaCI is present in the range of about 7 to 13 mM, or about 8 to 12 mM, or about 9 to 11 mM, or about 9.5 to 10.5 mM, or preferably is present at about 10 mM.
- the mixture or composition of step (a) contains an adjuvant, preferably wherein the adjuvant is selected from the group consisting of alhydrogel (alum), chitosan, mpl or CpG.
- the adjuvant is CpG, such as CpG1018.
- CpG1018 is present in the range of about 2 mg/ml to about 6 mg/ml, or in the range of about 3 mg/ml to about 5 mg/ml, or in the range of about 3.5 mg/ml to about 4.5 mg/ml, or CpG1018 is preferably present at about 4 mg/ml.
- the stabilized vaccine composition may also incorporate an agent to reduce or prevent agglomeration during preservation, for example a small amount of a non-ionic surfactant, especially an ethoxylated sorbitan ester, for example polyoxyethylenesorbilan monolaurate, typically comprising about 20 ethyleneoxy units.
- a non-ionic surfactant especially an ethoxylated sorbitan ester, for example polyoxyethylenesorbilan monolaurate, typically comprising about 20 ethyleneoxy units.
- the mixture or composition of step (a) further comprises a detergent, for example, Tween 20.
- Tween 20 is present in the range of about 0.3% to about 0.5%, or is present in the range of about 0.35% to about 0.45%, or preferably is present at about 0.04%.
- the mixture or composition in step (a) comprises 400 ⁇ g/ml rPA, 4 mg/ml CpG1018, 10% trehalose, 0.5 mM phosphate, 10 mM NaCI, and 0.04% Tween 20.
- the present invention also provides for a composition suitable for lyophilization, comprising an anthrax antigen containing recombinant Protective Antigen (rPA) and a lyoprotective amount of trehalose.
- rPA is present in the range of about 100 ⁇ g/mL to about 700 ⁇ g/mL, or is present in the range of about 200 to 600 ⁇ g/ml, or is present in the range of about 250 to 550 ⁇ g/ml, or is present in the range of about 300 to 500 ⁇ g/ml, or is present in the range of about 350 to 450 ⁇ g/ml, is preferably present at about 400 ⁇ g/ml.
- the mixture or composition of step (a) contains trehalose is present in the range of about 2 to 20%, or in the range of about 4 to 18%, or about 6 to 16%, or about 8 to 14%, or about 9 to 12%, or preferably at about 10% (w/v).
- the lyoprotectant is added to the pre- lyophilized formulation or composition to be iyophilized.
- this additive is a non-reducing sugar, for example, sucrose or trehalose, the latter being especially preferred.
- the amount of lyoprotectant added is such that, upon reconstitution, the resulting formulation is isotonic.
- the amount of sugar added must be sufficient to prevent the undesirable degradation or aggregation, for example, of the antigenic protein, such as rPA, often occurring on lyophilization of such proteins.
- a surfactant such as polysorbates (e.g. polysorbates 20 (or Tween 20) or 80); poloxamers (e.g.
- poloxamer 188 Triton; sodium dodecyl sulfate (SDS); sodium laurel sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl- or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palnidopropyl-, or isostearamidopropyl-betaine (e.g lauroamidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl cocoy
- Tween 20 is a preferred surfactant or detergent.
- the amount added is sufficient to reduce aggregation of the reconstituted protein and minimizes the formation of particulates after reconstitution.
- the surfactant or detergent may be present in the pre-lyophilized composition in an amount from about 0.001 -0.5%, and preferably from about 0.005-0.05%.
- the lyoprotectant such as trehalose
- a bulking agent e.g. mannitol or glycine
- Use of such bulking agents may facilitate production of a more uniform lyophilized cake (i.e., without excessive pockets and the like).
- the composition contains a salt, preferably phosphate and/or NaCI (or saline).
- the phosphate is present in the range of about 0.3 to 0.7 mM, or in the range of about 0.35 to 0.65 mM, or about 0.4 to 0.6 mM, or about 0.45 to 0.55 mM, or preferably is present at about 0.5 mM.
- the NaCI is present in the range of about 7 to 13 mM, or about 8 to 12 mM, or about 9 to 11 mM, or about 9.5 to 10.5 mM, or preferably is present at about 10 mM.
- the composition contains an adjuvant, preferably wherein the adjuvant is selected from the group consisting of alhydrogel (alum), chitosan, mpl or CpG.
- the adjuvant is CpG, such as CpG1018.
- CpG1018 is present in the range of about 2 mg/ml to about 6 mg/ml, or in the range of about 3 mg/ml to about 5 mg/ml, or in the range of about 3.5 mg/ml to about 4.5 mg/ml, or CpG1018 is preferably present at about 4 mg/ml.
- composition of further comprises a detergent, for example, Tween 20.
- Tween 20 is present in the range of about 0.3% to about 0.5%, or is present in the range of about 0.35% to about 0.45%, or preferably is present at about 0.04%.
- the composition suitable for lyophilization comprises 400 ⁇ g/ml rPA, 4 mg/ml CpG 1018, 10% trehalose, 0.5 mM phosphate, 10 mM NaCI, and 0.04% Tween 20.
- the present invention also provides a method for preparing such a composition, said method comprising essentially the procedure of step (a) recited elsewhere herein for methods of the invention.
- the phosphate present in said composition to be lyophilized can serve the purposes of buffering the composition as well as the vaccine formulation produced after reconstitution.
- phosphate is not intended as a limiting buffer and other buffers may be used either in place or phosphate or in combination with it.
- buffers include histidine, phosphate, Tris, citrate, succinate and other organic acids.
- the buffer concentration can be from about 1 mM to about 20 mM, or from about 3 mM to about 15 mM, depending, for example, on the buffer and the desired isotonicity of the vaccine formulation (e.g. of the reconstituted formulation).
- Such buffers may also provide a lyoprotective effect in addition to that of trehalose.
- the present invention also provides a kit comprising:
- kit may further comprise a container of a diluent suitable for reconstituting the lyophilized composition.
- a diluent suitable for reconstituting the lyophilized composition. Specific embodiments of such diluent are as described elsewhere herein for the diluent used in the methods of the invention.
- kits of the invention commonly contain the lyophilized composition in a vial or other suitable container, including, for example, bottles, vials (e.g. dual chamber vials), syringes (such as dual chamber syringes) and test tubes.
- the container may be formed from a variety of materials such as glass or plastic.
- the container holds the lyophilized formulation and the label on, or associated with, the container may indicate directions for reconstitution and/or use.
- the label may indicate that the lyophilized formulation is reconstituted to protein concentrations as described above.
- the label may further indicate that the formulation is useful or intended for subcutaneous administration.
- the container holding the formulation may be a multi-use vial, which allows for repeat administrations of the reconstituted formulation.
- the kit which is an article of manufacture, may further comprise a second container comprising a suitable diluent of a type as disclosed herein.
- a suitable diluent of a type as disclosed herein.
- the final rPA concentration in the reconstituted formulation will generally be in a range as disclosed herein, but generally at about 200 ⁇ g/ml.
- the kit may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.
- vaccines are prepared as injectables, in the form of aqueous solutions or suspensions. Vaccines in an oil base are also well known such as for inhaling. Solid forms which are dissolved or suspended prior to use may also be formulated. Pharmaceutically acceptable carriers, diluents and excipients are generally added that are compatible with the active ingredients and acceptable for pharmaceutical use.
- compositions and/or formulations useful herein contain a pharmaceutically acceptable carrier, including any suitable diluent or excipient, which includes any pharmaceutical agent that does not itself induce the production of antibodies harmful to the individual receiving the composition, and which may be administered without undue toxicity.
- Pharmaceutically acceptable carriers include, but are not limited to, liquids such as water, saline, glycerol and ethanol, and the like, including carriers useful in forming sprays for nasal and other respiratory tract delivery or for delivery to the ophthalmic system.
- a thorough discussion of pharmaceutically acceptable carriers, diluents, and other excipients is presented in REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Pub. Co., N.J. current edition). Those described elsewhere herein are especially preferred for use in the formulations, composition s, methods and kits of the invention.
- Vaccine compositions may further incorporate additional substances to stabilize pH, or to function as adjuvants, wetting agents, or emulsifying agents, which can serve to improve the effectiveness of the vaccine.
- Vaccines are generally formulated for parenteral administration and are injected either subcutaneously or intramuscularly. Such vaccines can also be formulated as suppositories or for oral administration, using methods known in the art, or for administration through nasal or respiratory routes.
- the amount of vaccine sufficient to confer immunity to pathogenic bacteria, viruses, or other microbes is determined by methods well known to those skilled in the art in view of the guidance provided herein. This quantity will be determined based upon the characteristics of the vaccine recipient and the level of immunity required (as already noted above).
- a range of .5 to 500 ⁇ g purified protein may be given.
- dosages are commonly sufficient to provide about 1 ⁇ g, possibly 10 ⁇ g, even 50 ⁇ g, and as much as 100 ⁇ g, up to 500 ⁇ g of immunogenic protein, or immunogenic polypeptide, or immunogenically active fragments thereof.
- more than one such active material may be present in the vaccine.
- more than one antigenic structure may be used in formulating the vaccine, or vaccine composition to use in the methods disclosed herein.
- This may include two or more individually immunogenic proteins or polypeptides, proteins or polypeptides showing immunogenic activity only when in combination, either quantitatively equal in their respective concentrations or formulated to be present in some ratio, either definite or indefinite.
- a vaccine composition for use in the processes disclosed herein may include one or more immunogenic proteins, one or more immunogenic polypeptides, and/or one or more immunogenically active immunogens comprising antigenic fragments of said immunogenic proteins and polypeptides, the latter fragments being present in any proportions selected by the use of the present invention.
- the exact components, and their respective quantities, making up the vaccines, and vaccine compositions, useful in the methods of the present invention are determined, inter alia, by the nature of the disease to be treated or prevented, the severity of such condition where it already exists, the age, sex, and general health of the recipient, as well the personal and professional experience and inclinations of the researcher and/or clinician utilizing these methods.
- a vaccine of the present invention such as a sub-unit vaccine for use against anthrax
- specific non-limiting examples of this vaccine composition are those wherein said rPA is present at about 10 to 300 ⁇ g/ml, preferably 50 to 300 ⁇ g/ml, or wherein said rPA is present at about 100 to 300 ⁇ g/m!, more preferably about 150 to 250 ⁇ g/ml, and most preferably wherein said rPA is present at about 200 ⁇ g/ml.
- the dose of antigen, preferably rPA, to be administered is at least about 5 ⁇ g, or at least about 10 ⁇ g, or at least about 25 ⁇ g, or at least about 50 ⁇ g, or at least about 75 ⁇ g, or at least about 100 ⁇ g, or at least about 150 ⁇ g, or at least about 200 ⁇ g, with a preferred dose of about 100 ⁇ g.
- a preferred dose volume is about 0.5 ml.
- the present invention further relates to a vaccine comprising a purified antigen, preferably an anthrax antigen, more preferably anthrax protective antigen (PA) and most preferably a recombinant anthrax protective antigen (rPA), such as that described in WO 2007/122373, pub. 1 November 2007, (the disclosure of which is hereby incorporated by reference in its entirety) bound to an adjuvant, preferably an alum-based adjuvant.
- PA anthrax protective antigen
- rPA recombinant anthrax protective antigen
- the present invention relates to a sub-unit vaccine comprising alum-bound rPA, at least about 2 mM phosphate salt and at about pH 7.1.
- the vaccine formulation used for in vivo administration are commonly sterile, which can be accomplished by filtration through sterile filtration membranes, prior to, or following, lyophilization and reconstitution.
- sterility of the entire composition may be accomplished by autoclaving the ingredients, except for protein, at about 120°C for up to about 30 minutes, as one non-limiting example.
- Freeze-drying is accomplished by freezing the formulation and subsequently subliming ice from the frozen content at a temperature suitable for primary drying. Under this condition, the product temperature is below the eutectic point or the collapse temperature of the formulation.
- the shelf temperature for the primary drying will range from about -30 to 25 0 C. (provided the product remains frozen during primary drying) at a suitable pressure, ranging typically from about 50 to 250 mTorr.
- the formulation, size and type of the container holding the sample (e.g., glass vial) and the volume of liquid will mainly dictate the time required for drying, which can range from a few hours to several days (e.g. 40-60 hrs).
- a secondary drying stage may be carried out at about 0-40 0 C, depending primarily on the type and size of container and the type of protein employed. However, it was found herein that a secondary drying step may not be necessary.
- the shelf temperature throughout the entire water removal phase of lyophilization may be from about 15-30 0 C (e.g., about 20 0 C).
- the time and pressure required for secondary drying will be that which produces a suitable lyophilized cake, dependent, e.g., on the temperature and other parameters.
- the secondary drying time is dictated by the desired residual moisture level in the product and typically takes at least about 5 hours (e.g. 10-15 hours).
- the pressure may be the same as that employed during the primary drying step.
- Freeze- drying conditions can be varied depending on the formulation and vial size.
- the resulting lyophilized powder will commonly possess a moisture content thereof that is less than about 5%, and preferably less than about 3%.
- freeze-drying cycles useful in the methods of the invention are shown in Tables 1 and 2:
- This modified cycle of Table 2 was performed using a Christ freeze-dryer.
- the product was stoppered under vacuum before introducing air into the chamber. Aluminium seals were then applied to the vials and crimped, on removal from the freeze dryer.
- the reconstituted formulation is administered to a patient in need of treatment with the protein, preferably a human, in accord with known methods, such as intravenous administration as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes.
- the lyophilized vaccine compositions prepared by the methods of the invention are typically maintained in the freeze-dried state until the time of use.
- the lyophilized formulation is reconstituted with a diluent as described herein.
- Reconstitution commonly occurs at room temperature to ensure complete hydration, although other temperatures may be employed as desired.
- the time required for reconstitution will depend, e.g., on the type of diluent, amount of excipient(s) and protein, such as rPA.
- diluents include sterile water, bacteriostatic water for injection (BWFI), a pH buffered solution (e.g. phosphate-buffered saline is especially preferred), sterile saline solution, Ringer's solution or dextrose solution.
- BWFI bacteriostatic water for injection
- a pH buffered solution e.g. phosphate-buffered saline is especially preferred
- sterile saline solution e.g. phosphate-buffered saline solution
- Ringer's solution e.g. sterile saline solution
- dextrose solution e.g., Ringer's solution or dextrose solution.
- the amount of preservative employed is determined by assessing different preservative concentrations for • compatibility with the protein and preservative efficacy testing.
- the preservative is an aromatic alcohol (such as benzyl alcohol)
- it can be present in an amount from about 0.1-2.0% and preferably from about 0.5- 1.5%, but most preferably about 1.0-1.2%.
- the present invention further provides a method of protecting against, or a method of treating, a microbial infection, preferably a bacterial infection, comprising administering to a mammal at risk of such infection a therapeutically-effective amount of a stable vaccine formulation as described herein.
- the infecting organism may be any one or more of those recited herein and the formulation being one recited herein and/or a formulation reconstituted from a vaccine composition or mixture freeze-dried according to the methods of the invention.
- the bacterial infection is an anthrax infection (i.e., caused by Bacillus anthracis).
- the animal to be protected is a mammal, especially a human patient.
- a preferred formulation for use in such treatment is a vaccine composition containing 200 ⁇ g/ml rPA, 2 mg/ml CpG1018, 5% trehalose, 5.25 mM phosphate, 0.39% NaCI, 0.02% Tween 20, 0.26% w/v alhydrogel.
- mice Female adult A/J mice were immunized on day 0 intra-muscularly (i.m.) with 0.1 ml of the respective samples indicated in Table 3, which were formulated with diluent prior to administration. All the mice were challenged with S. anthracis STI strain spores on day 21 and survival was determined on day 1 1 .
- the results of Table 3 show a much greater stability for a lyophilized vaccine than would have been expected based on previous experience with such vaccines (compare, for example, the results for storage at 7O 0 C with that for 5 0 C.
- the diluent was comprised of: 0.26% (w/w) alhydrogel (0.4% AI(OH) 3 ) in 0.39% (w/v) sodium chloride buffered with 5 mM phosphate, pH 7.4.
- the na ⁇ ve animals were negative controls that had not received any treatment but were challenged at the day 21 with the same dose of spores as the test animals.
Abstract
Description
Claims
Priority Applications (6)
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EP09785211A EP2381959A1 (en) | 2009-01-22 | 2009-01-22 | Stable vaccine compositions and methods of use |
CA2750321A CA2750321A1 (en) | 2009-01-22 | 2009-01-22 | Stable vaccine compositions and methods of use |
AU2009338516A AU2009338516A1 (en) | 2009-01-22 | 2009-01-22 | Stable vaccine compositions and methods of use |
JP2011546933A JP2012515752A (en) | 2009-01-22 | 2009-01-22 | Stable vaccine composition and method of use |
PCT/GB2009/050051 WO2010084298A1 (en) | 2009-01-22 | 2009-01-22 | Stable vaccine compositions and methods of use |
IL214211A IL214211A0 (en) | 2009-01-22 | 2011-07-20 | Stable vaccine compositions and methods for preparing the same |
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JP (1) | JP2012515752A (en) |
AU (1) | AU2009338516A1 (en) |
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Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014004578A1 (en) * | 2012-06-25 | 2014-01-03 | Emergent Product Development Gaithersburg Inc. | Temperature stable vaccine formulations |
US9616117B2 (en) | 2008-10-02 | 2017-04-11 | Pharmathene, Inc. | Anthrax vaccine formulation and uses thereof |
EP3086780A4 (en) * | 2013-12-27 | 2017-08-09 | Emergent Product Development Gaithersburg Inc. | Temperature stable vaccine formulations |
JP2017214357A (en) * | 2011-07-11 | 2017-12-07 | タケダ ワクチン,インコーポレイテッド | Parenteral norovirus vaccine formulations |
US10512682B2 (en) | 2006-09-29 | 2019-12-24 | Takeda Vaccines, Inc. | Norovirus vaccine formulations |
US10688174B2 (en) | 2007-09-18 | 2020-06-23 | Takeda Vaccines, Inc. | Method of conferring a protective immune response to norovirus |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008079464A2 (en) * | 2006-09-08 | 2008-07-03 | Becton, Dickinson And Company | Stable powder formulations of alum-adsorbed vaccines |
EP1972347A1 (en) * | 2007-03-19 | 2008-09-24 | Becton, Dickinson and Company, Wagner, Jaconda | Stable vaccine powder formulations |
WO2009093072A1 (en) * | 2008-01-22 | 2009-07-30 | Pharmathene Uk Limited | Vaccine composition |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ATE503491T1 (en) * | 2003-02-13 | 2011-04-15 | Becton Dickinson Co | IMPROVED ANTHRAX VACCINES AND DELIVERY METHODS |
-
2009
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- 2009-01-22 CA CA2750321A patent/CA2750321A1/en not_active Abandoned
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008079464A2 (en) * | 2006-09-08 | 2008-07-03 | Becton, Dickinson And Company | Stable powder formulations of alum-adsorbed vaccines |
EP1972347A1 (en) * | 2007-03-19 | 2008-09-24 | Becton, Dickinson and Company, Wagner, Jaconda | Stable vaccine powder formulations |
WO2009093072A1 (en) * | 2008-01-22 | 2009-07-30 | Pharmathene Uk Limited | Vaccine composition |
Non-Patent Citations (3)
Title |
---|
IVINS B ET AL: "Experimental anthrax vaccines: efficacy of adjuvants combined with protective antigen against an aerosol Bacillus anthracis spore challenge in guinea pigs", VACCINE, BUTTERWORTH SCIENTIFIC. GUILDFORD, GB, vol. 13, no. 18, 1 January 1995 (1995-01-01), pages 1779 - 1784, XP004057384, ISSN: 0264-410X * |
JIANG GE ET AL: "Anthrax vaccine powder formulations for nasal mucosal delivery", JOURNAL OF PHARMACEUTICAL SCIENCES, AMERICAN PHARMACEUTICAL ASSOCIATION, WASHINGTON, US, vol. 95, no. 1, 1 January 2006 (2006-01-01), pages 80 - 96, XP002489440, ISSN: 0022-3549 * |
MIKSZTA J A ET AL: "PROTECTIVE IMMUNIZATION AGAINST INHALATIONAL ANTHRAX: A COMPARISON OF MINIMALLY INVASIVE DELIVERY PLATFORMS", JOURNAL OF INFECTIOUS DISEASES, UNIVERSITY OF CHICAGO PRESS, CHICAGO, IL, vol. 191, no. 2, 15 January 2005 (2005-01-15), pages 278 - 288, XP009046310, ISSN: 0022-1899 * |
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10512682B2 (en) | 2006-09-29 | 2019-12-24 | Takeda Vaccines, Inc. | Norovirus vaccine formulations |
US11826415B2 (en) | 2007-09-18 | 2023-11-28 | Takeda Vaccines, Inc. | Method of conferring a protective immune response to Norovirus |
US11040097B2 (en) | 2007-09-18 | 2021-06-22 | Takeda Vaccines, Inc. | Method of conferring a protective immune response to norovirus |
US10688174B2 (en) | 2007-09-18 | 2020-06-23 | Takeda Vaccines, Inc. | Method of conferring a protective immune response to norovirus |
US9616117B2 (en) | 2008-10-02 | 2017-04-11 | Pharmathene, Inc. | Anthrax vaccine formulation and uses thereof |
JP2017214357A (en) * | 2011-07-11 | 2017-12-07 | タケダ ワクチン,インコーポレイテッド | Parenteral norovirus vaccine formulations |
US10675341B2 (en) | 2011-07-11 | 2020-06-09 | Takeda Vaccines, Inc. | Parenteral norovirus vaccine formulations |
US11701420B2 (en) | 2011-07-11 | 2023-07-18 | Takeda Vaccines, Inc. | Parenteral norovirus vaccine formulations |
WO2014004578A1 (en) * | 2012-06-25 | 2014-01-03 | Emergent Product Development Gaithersburg Inc. | Temperature stable vaccine formulations |
US20150335752A1 (en) * | 2012-06-25 | 2015-11-26 | Emergent Product Development Gaithersburg Inc. | Temperature Stable Vaccine Formulations |
CN104470506A (en) * | 2012-06-25 | 2015-03-25 | 新兴产品开发盖瑟斯堡有限公司 | Temperature stable vaccine formulations |
US10357559B2 (en) | 2013-12-27 | 2019-07-23 | Emergent Product Development Gaithersburg Inc. | Temperature stable vaccine formulations |
EP3086780A4 (en) * | 2013-12-27 | 2017-08-09 | Emergent Product Development Gaithersburg Inc. | Temperature stable vaccine formulations |
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CA2750321A1 (en) | 2010-07-29 |
IL214211A0 (en) | 2011-08-31 |
EP2381959A1 (en) | 2011-11-02 |
JP2012515752A (en) | 2012-07-12 |
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