[go: up one dir, main page]

WO2013059408A1 - Etanercept formulations stabilized with meglumine - Google Patents

Etanercept formulations stabilized with meglumine Download PDF

Info

Publication number
WO2013059408A1
WO2013059408A1 PCT/US2012/060743 US2012060743W WO2013059408A1 WO 2013059408 A1 WO2013059408 A1 WO 2013059408A1 US 2012060743 W US2012060743 W US 2012060743W WO 2013059408 A1 WO2013059408 A1 WO 2013059408A1
Authority
WO
WIPO (PCT)
Prior art keywords
etanercept
peak
formulation
meglumine
composition
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2012/060743
Other languages
French (fr)
Inventor
Mark Manning
Brian Murphy
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Coherus Oncology Inc
Original Assignee
Coherus Biosciences Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to KR1020147013301A priority Critical patent/KR20140091705A/en
Priority to SG11201401567YA priority patent/SG11201401567YA/en
Priority to CN201280062761.9A priority patent/CN104010658A/en
Priority to EA201490802A priority patent/EA028520B1/en
Priority to AU2012326080A priority patent/AU2012326080B2/en
Priority to EP12841505.6A priority patent/EP2768531A4/en
Priority to JP2014537221A priority patent/JP6199298B2/en
Priority to HK15101258.8A priority patent/HK1200722A1/en
Application filed by Coherus Biosciences Inc filed Critical Coherus Biosciences Inc
Priority to CA2851646A priority patent/CA2851646A1/en
Priority to BR112014009131A priority patent/BR112014009131A8/en
Priority to MX2014004732A priority patent/MX2014004732A/en
Publication of WO2013059408A1 publication Critical patent/WO2013059408A1/en
Priority to IL231827A priority patent/IL231827A0/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39591—Stabilisation, fragmentation
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
    • A61K38/1793—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/191—Tumor necrosis factors [TNF], e.g. lymphotoxin [LT], i.e. TNF-beta
    • 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
    • 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
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/12—Carboxylic acids; Salts or anhydrides thereof
    • 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
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • 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
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • 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
    • A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0021—Intradermal administration, e.g. through microneedle arrays, needleless injectors
    • 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
    • A61K9/08—Solutions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
    • C07K14/7151—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons for tumor necrosis factor [TNF], for lymphotoxin [LT]
    • 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
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • 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
    • 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
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Definitions

  • the present invention relates to aqueous pharmaceutical compositions stabilized with meglumine for long-term storage of etanereept, methods of manufacture of the compositions, methods of their administration, and kits containing the same.
  • the invention includes etanereept formulations that do not require arginine for stabilization.
  • Polypeptides must often be stored prior to their use. When stored for extended periods, polypeptides are frequently unstable in solution (Manning et .al., 1989, Pharm. Res. 6:903-918). To extend their shelf life, additional processing steps have been developed, such as drying, e.g., lyophilization. However, !yophilized pharmaceutical compositions are less convenient to use.
  • Typical practices to improve polypeptide stability can be addressed by varying the concentration of elements with the formulation, or by adding excipients to modify the formulation (See, for example, U.S. Pat. Nos. 5,580,858 and 6,171 ,586).
  • the use of additives can still result in inactive polypeptides, in addition, in the case of lyophilization, the rehydration step can result in inactivation of the polypeptide by, for example, aggregation or denaturation (Hora et a!., 1992, Pharm. Res., 9:33-36; Liu et al., 1991 , Biotechnoi. Bioeng., 37:177-184).
  • Another way to improve polypeptide stability is to use L-arginine at a specific concentration ⁇ U.S. Pat. No. 7,648,702).
  • etanereept (Enbrel ⁇ , immunex Corporation), which is a dimeric fusion polypeptide consisting of the extracellular ligand-binding portion of the human 75 kilodalton (p75) tumor necrosis factor receptor (TNFR) linked to the Fc portion of human IgGl it consists of 934 amino acids and has an apparent molecular weight of approximately 150 kiioda!ions (Physicians Desk Reference, 2002, Medical Economics Company Inc.)
  • the Fc component of etanercept contains the constant heavy 2 (CH2) domain, the constant heavy 3 (CH3) domain and hinge region, but not the constant heavy 1 (CM) domain of human igG1.
  • An Fc domain can contain one or ali of the domains described above.
  • Etanercept is usually produced by recombinant DMA technology in a Chinese hamster ovary (CHO) mammalian cell expression system.
  • the present invention provides novel stable liquid formulations of etanercept that allow its long-term storage.
  • the present invention is an aqueous stabilized formulation of etanercept comprising: etanercept; and stabilizing ingredients to retard instability, aggregation and/or fragmentation of the etanercept in the formulation, said stabilizing ingredients being comprised of at least one of the following: (a) meglumine; or (b) meglumine in combination with sucrose; or (c) meglumine in combination with sodium chloride; or ⁇ (d) meglumine in combination with sodium chioride and sucrose.
  • the stabilized etanercept formulations of the present invention elicit long term storage stability as characterized by at least one of: (1) SEC analysis at M 3 or T 2 or T 4 of: monomer content greater than about 90%: aggregates content of less than about 3 wt%; and fragment 3 content less than about 5 wt%: and (2) H!C analysis at fv1 3 or T 2 , or T wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 3 wt.%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than 80 wt.%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 20 wt.%.
  • the formulations elicit long term storage stability as characterized by: an HIC analysis at M 3 or T 2 or T 4 wherein the amount of the composition represented by peak 2 of the HIC chromatogram is greater than or equal to about 95 wt.%; and wherein, if peak 3 is present on the HIC chromatogram, the amount of the composition represented by peak 3 is less than or equal to about 3 wt.%.
  • the stabilized etanercept formulation as summarized above optionally and preferably, contains no arginine, or are essentially free of arginine.
  • the formulations of the invention have excellent stability as determined by SEC (Size Exclusion Chromatography) and HIC (Hydrophobic Interaction Chromatography) analysis conducted after one, two or three months of storage at 5° C. These formulations are comparable to or better than a commercially available formulation of etanercept, in which arginine is a required component.
  • the present invention is further directed to formulations of stabilized etanercept, as summarized above, which contain no arginine, or are essentially free of arginine, and wherein the composition, evaluated at M 3 or T 2 or T 4 , elicits long term storage stability that meets one or both of the following criteria: (A) stability comparable to or better than commercially available etanercept marketed under the trademark Enbrel®, as measured by (i) SEC analysis of the amounts of aggregate(s), monomer and fragment 3 in the composition (as defined in the specification) and (ii) HIC analysis of amounts of material in the composition corresponding to peaks 1 , 2 and 3 of the HIC chromatogram (as defined in the specification); and (B) an HIC chromatogram in which (i) peak 3 is absent, or essentially absent and (ii) peak 2 represents greater than about 95 wt% of the composition; an SEC chromatogram containing essentially no peak corresponding to aggregate(s); and an SEC chromatogram in which
  • the formulation of the invention comprises about about 25 to 75 mg/ml etanercept, sodium chloride in an amount up to about 150 mM, about 1 to about 30 mM sodium phosphate; and about 0 to 5 wt.% sucrose or trehalose or combination thereof; wherein the composition has a pH of about 6.0 to about 6.6; and wherein the composition is characterized by SEC analysis at 3 or T 2 or T 4 in which: monomer content is greater than about 85 wt.%; aggregate(s) content is less than about 3 wt.%, and fragment 3 content is less than about 8 wt.%. Formulations meeting these analytical criteria do not require arginine for stabilization.
  • the stabilized etanercept formulation is further characterized by (a) an SEC analysis at 3 or T 2 or T 4 of greater than about 90 t% monomer content; and less than about 3 wt% aggregate(s) content; and (b) an HIC analysis at M 3 or T 2 or T 4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 4 wt%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than about 80 wt%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 20 wt%; and wherein the formulation is free or essentially free of arginine.
  • the etanercept compositions of the invention further afford the ability to provide formulations which contain acceptable levels of subvisible particles. Accordingly, the invention is further directed to stabilized etanercept formulations having, at M 3 or T 2 or T 4l no more than, on average, about 10,000 subvisible particles per mL having a size greater than 5 pm.
  • the stabilized etanercept composition of the present invention are further characterized by: (a) an SEC analysis at IV or T 2 or T of greater than about 90 wt% monomer content; and less than about 3 wt.% aggregate(s) content; and (b) an HIC analysis at 3 or T 2 or T 4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 3 wt%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than 80 wt%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 20 wt% and wherein the formulation is free or essentially free of arginine.
  • the stability of the formulations may be further characterized in that the compositions, optionally free or essentiaily of arginine, exhibit an HIC analysis at 3 or T ⁇ or T 4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 1%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than about 95 t%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 1 wt%.
  • Formulations meeting these analytical criteria do not require arginine for stabilization.
  • Preferred stabilized compositions of the invention, preferably free or essentlaliy free of arginine exhibit an HiC analysis at M 3 or T ?
  • the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 2% or preferably less than about 1%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than about 95 wt.% and preferably greater than about 97%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 1 wt% : and preferably 0 to 1%,
  • nstability denotes the tendency of the etanercept monomer to undergo a variety of undesired transformations during storage.
  • transformations include the formation of oligomers and high molecular weight aggregate(s) (hereinafter terms “aggregate(s)” in which multiple copies of the essentially intact etanercept monomer become irreversibl associated with one another through a variety of non-covalent attractions (e.g., electrostatic interactions.)
  • Undesired transformations during storage may also include degradation of the etanercept monomer to smaller fragments and/or clipped species.
  • a formulation of etanercept should minimize, to the greatest extent possible, the tendency of the formulation to result, during storage, in the formation of aggregates, misfolded protein, oligomers and/or fragments of etanercept.
  • An important benefit resulting from the ability to reduce formation of unwanted aggregates or fragments is a reduction in the potential toxicity and/or immunogenicity of the drug.
  • the etanercept formulation of the present invention which is optionally and preferably free, or essentially free of argihine.
  • arginine is intended to mean that arginine, even if present, is not contributing to the stabilization of the etanercept monomer in the formulation to such an extent that a person skilled in the art would judge its presence beneficial or necessary from a stabilization standpoint.
  • etanercepf or "etanercept monomer” or “monomer” is synonymous with Enbre!®. It refers to a polypeptide which is a dimeric fusion polypeptide consisting of the extracellular iigand-binding portion of the human 75 kilodalton (p75) tumor necrosis factor receptor (TNFR) linked to the Fc portion of human IgGl It consists of 934 amino acids and has an apparent molecular weight of approximately 150 kiiodaStons.
  • etanercepf also encompasses etanercepf with minor modifications in the amino acid structure (including deletions, additions, and/or substitutions of amino acids) which do not significantly affect the function, potency, or avidity of etanercepf.
  • etanercepf encompasses ail forms and formulations of Enbrel®, including but not limited to concentrated formulations, injectable ready-to-use formulations; formulations reconstituted with water, alcohol, and/or other ingredients, and others.
  • sugar refers to monosaccharides, disachharides, and polysaccharides.
  • sugars include, but are not limited to, sucrose, trehalose, dextrose, and others.
  • meglumine refers to a compound with chemical formula H 3 NHCH2(CHOH) 4 CH 2 OH, also known as 1-Deoxy ⁇ 1-methylamtnosorbitol; N- ethyl- d-giucamine; and 1-Deoxy-1-methylamino-D-glucitol.
  • polyo refers to an alcohol containing multiple hydroxy! groups.
  • examples of polyois include, but are not limited to, mannitoi, sorbitol, and others.
  • long-term storage is understood to mean that the pharmaceutical composition can be stored for three months or more, for six months or more, and preferably for one year or more. Long-term storage is also understood to mean that the pharmaceutical composition is stored either as a liquid at 2-8° C, or is frozen, e.g., at -2G°C, or colder. It is also contemplated thai the composition can be frozen and thawed more than once.
  • stable or “stabilized” with respect to long-term storage is understood to mean that etanercepf contained in the pharmaceutical compositions does not lose more than 20%, or more preferably 15%, or even more preferably 10%, and most preferably 5% of its activity relative to activity of the composition at the beginning of storage,
  • mammal includes, but is not limited to, a human.
  • pharmaceutically acceptable carrier refers to a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material, formulation auxiliary, or excipient of an conventional type.
  • a pharmaceutically acceptable carrier is non- toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation.
  • composition refers to a mixture that usually contains a carrier, such as a pharmaceutically acceptable carrier or excipient that is conventional in the art and which is suitable for administration into a subject for therapeutic, diagnostic, or prophylactic purposes. It may include a ceil culture in which the polypeptide or polynucleotide is present in the cells or in the culture medium.
  • a carrier such as a pharmaceutically acceptable carrier or excipient that is conventional in the art and which is suitable for administration into a subject for therapeutic, diagnostic, or prophylactic purposes. It may include a ceil culture in which the polypeptide or polynucleotide is present in the cells or in the culture medium.
  • compositions for oral administration can form solutions, suspensions, tablets, pills, capsules, sustained release formulations, oral rinses or powders.
  • treatment refers to any administration or application of remedies for disease in a mamma! and inciudes inhibiting the disease, arresting its development, relieving the disease, for example, by causing regression, or restoring or repairing a lost, missing, or defective function; or stimulating an inefficient process.
  • treatment includes obtaining a desired pharmacologic and/or physiologic effect, covering any treatment of a pathological condition or disorder in a mammal.
  • the effect may be prophylactic in terms of completely or partially preventing a disorder or symptom thereof and/or may be therapeutic in terms of a partial or complete cure for a disorder and/or adverse affect attributable to the disorder.
  • disease refers to any condition, infection, disorder or syndrome that requires medical intervention or for which medical intervention is desirable. Such medical intervention can include treatment, diagnosis and/or prevention.
  • an effective amount of the polypeptide of the invention for administration to the living subject is an amount that prevents and/or treats an integrin av 3-mediated disease.
  • the exact amount will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques. As is known in the art, adjustments for systemic versus localized delivery, age, body weight, general health, sex, diet, time of administration, drug interaction and the severity of the condition may be necessary, and will be ascertainable with routine experimentation by those skilled in the art.
  • Ti refers to a point in time at which an etanercept formulation has been stored for about one week at 40° C
  • T 2 refers to a point in time at which an etanercept formulation has been stored for about two weeks at 40° C
  • M3 refers, collectively, to three points in time, and in particular to an analytical result being observed for an etanercept formulation after duration of either about one, about two or about three months of storage at a storage temperature of 5° C.
  • reference herein to an analysis being conducted at M3 should be understand to mean that such analysis is be done at the point in time at which etanercept formulation has been in storage for a duration selected from about one, about two, or about three months.
  • an etanercept formulation elicit a certain analytical value or measurement at 3 is satisfied if the required value is observed at a point in time corresponding to at least one of the following storage durations: at approximately one month, at approximately two months or at approximately three months of storage at 5°C.
  • Peak 1 Peak 2
  • Peak 3 Peak 3 when used herein in connection with discussion of HIC chromatography results refers to the same peaks 1 , 2 and 3 discussed in US Patent 7,294,481.
  • the present invention provides several embodinients of aqueous formulations of etanercept that allow stable long-term storage of etanercept, so that etanercept is stable over the course of storage either in liquid or frozen states.
  • the provided formulations include, but are not limited to formulations that do not contain arginine and do not require any extra steps such as rehydratlng. These embodiments are explained in a greater detail below.
  • compositions of the present invention comprise etanercept (Enbrel®).
  • etanercept is a dimeric fusion poiypeptide consisting of the extracellular Iigand-binding portion of the human 75 ki!odalton (p75) tumor necrosis factor receptor (TNFR) linked to the Fc portion of human igG .
  • TNFR tumor necrosis factor receptor
  • Etanercept consists of 934 amino acids.
  • the Fc component of etanercept contains the constant heavy 2 (CH2) domain, the constant heavy 3 (CH3) domain and hinge region of human IgG
  • An Fc domain can contain one or all of the domains described above.
  • Etanercept suitable for storage in the present pharmaceutical composition can be produced by living host ceils that express etanercept, such as hybridomas in the case of antibodies, or host celis that that have been genetically engineered to produce the polypeptide in the case of fusion polypeptides or antibodies.
  • Methods of genetically engineering cells to produce polypeptides are well known in the art. See, e.g., Ausubei et a!., eds. (1990), Current Protocols in Molecular Biology (Wiley, New York). Such methods include introducing nucleic acids that encode and aliow expression of the polypeptide Into living host ceils. These host cells can be bacterial cells, fungal cells, or, preferably, animal ceils grown in culture.
  • Bacterial host ceils include, but are not limited to, Escherichia coii cells. Examples of suitable £ coii strains include: HB101 , DH5.alpha, GM2929, JM109, KW251 , NM538, N 539, and any E. coii strain that fails to cleave foreign DNA.
  • Fungal host cells that can be used include, but are not limited to, Saccharomyces cerevisiae, Pichia pastoris and Aspergillus ceils,
  • a few examples of animal cell lines that can be used are CHO, VERO, BHK, HeLa, Cos, MDCK, 293, 3T3, and W138. New animal cell lines can be established using methods weli know by those skiiied in the art (e.g., by transformation, viral infection, and/or selection).
  • etanercept can be secreted by the host celis into the medium.
  • etanercept Purification of the expressed etanercept can be performed by any standard method. When etanercept is produced intracellular ⁇ , the particulate debris is removed, for example, by centrlfugation or ultrafiltration. When etanercept is secreted into the medium, supernatants from such expression systems can be first concentrated using standard polypeptide concentration filters. Protease inhibitors can also be added to inhibit proteolysis and antibiotics can be included to prevent the growth of microorganisms.
  • Etanercept can be purified using, for example, hydroxyapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, and any combination of known or yet to be discovered purification techniques, including but not limited to Protein A chromatography, fractionation on an ion-exchange column, ethanol precipitation, reverse phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSET®, an anion or cation exchange resin chromatography (such as a polyaspartic acid column), chromatofocusing, SDS- PAGE, and ammonium sulfate precipitation.
  • Protein A chromatography fractionation on an ion-exchange column, ethanol precipitation, reverse phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSET®, an anion or cation exchange resin chromatography (such as a polyaspartic acid column), chromatofocusing, SDS- PAGE, and ammonium sulf
  • the invention provides a stable aqueous formulation comprising etanercept and an ionic poiyol derivative excipient, wherein said excipient is selected from the group consisting of meglumine (N-methyi-D-glucamine), mannosylg!ycerate, glucosylglycerate, mannosyllactate, mannosylglycolate, and diglycerolphosphate.
  • excipient is selected from the group consisting of meglumine (N-methyi-D-glucamine), mannosylg!ycerate, glucosylglycerate, mannosyllactate, mannosylglycolate, and diglycerolphosphate.
  • the invention is an aqueous stabilized formulation of etanercept comprising: etanercept; and stabilizing ingredients to retard instability, aggregation and fragmentation of the etanercept in the formulation, said stabilizing ingredients being comprised of (a) meglumine; or (b) meglumine in combination with sucrose; or (c) meglumine in combination with sodium chloride; or (d) meglumine ' in combination with sodium chloride and sucrose.
  • Meglumine is commonl used as a small molecule excipient
  • meglumine is also able to stabilize aqueous pharmaceutical compositions containing a large protein, such as etanercept.
  • meglumine reduces etanercepfs tendency to associate in undesired ternary or quaternary complexes, and therefore, reduces aggregation of etanercept
  • the reduction in aggregation is believed to last for a long period of time, e.g., two years or more.
  • ionic poiyol derivatives, including meglumine are able to stabilize aqueous pharmaceutical compositions containing etanercept by a combination of three different mechanisms. First, they can act as an excluded solute in the same way mannitol, sucrose, and sorbitol increase conformational stability.
  • charged solutes can alter the colloidal stability, thereby reducing the propensity to self-associate, thereby slowing aggregation.
  • these ionic polyoS derivatives being charged near neutral H, can act as salting-in agents, as arginine does, potentially resolubilizing aggregates.
  • the stabilizing effects of meglumine are not limited to reduction in aggregates but may involve other aspects of stabiiization of the etanercept monomer in a formulation containing the monomer.
  • compositions of the invention may be prepared by combining, a purified etanercept and the ionic polyol derivative, preferably meglumine, Further, a buffer, a tonicity modifier and an additional excipient and other commonly used inactive ingredients can be added as needed. For simplicity, these are discussed more fully later in the specification.
  • a buffer can be added first, middle or last
  • the tonicity modifier can also be added first, middle or last.
  • some of these chemicals can be incompatible in certain combinations, and accordingly, are easily substituted with different chemicals that have similar properties but are compatible in the relevant mixture.
  • the concentration of meglumine in the provided formulations is preferably between about 0.1% ⁇ w/v) to 40% (w/v), more preferably about 1% to about 20%, more preferably about 2% to about 10%, even more preferably about 2% to about 5%.
  • Meglumine is available from commercial suppliers.
  • a preferred embodiment comprises about 25 to about 75 mg/ml etanercept, about 1-30 mM of sodium phosphate; up to about 10% meglumine; optionally up to about 5 wt.% sucrose; and optionally up to about 100 mM sodium chloride, wherein the composition has a pH of about 6.0 to 7.0, and preferably about 6.0 to about 6.6 and most preferably about 6.3 to about 6.5.
  • a meglumine stabilized etanercept composition is preferably characterized by SEC analysis at T 2 in which: the monomer content is greater than about 85 wt.%; aggregate(s) content is less than about 3 wt.%; and fragment 3 content is Sess than about 8 wt.%.
  • a particularly preferred etanercept formulation stabilized with meglumine is characterized by HIC analysis at T 4 or T 2 wherein the amount of the composition represented by peak 1 of the H!C chromatogram is less than about 1%; the amount of the composition represented by peak 2 of the H!C chromatogram is greater than about 99 wt.%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 1 wt.%.
  • meglumine can be replaced with another sonic po!yol derivative of sorbitol, glycerol or mannito!, such as mannosylgiycerate, glucosylglycerate, mannosyllactate, mannosyiglycolate, and diglyceroiphosphate (at about 0.1% to about 40%) in the formulation.
  • a preferred meg!umine-siabiiized etanercept formulation free of arginine and exhibiting analytical properties as described above comprises about 25 to about 75 mg/ml etanercept; about 0,5 wt.% megiumine; about 25 mM phosphate; about 1% sucrose; and about 100 mM sodium chloride.
  • a further preferred meglumine-stabilized etanercept formulation free of arginine and exhibiting analytical properties as described above comprises about 50 mg/ml etanercept; about 5 wt.% meglumine; about 25 mM phosphate.
  • the etanercept formulations comprising ionic poiyol derivatives such as meglumine for stabilization according to the present invention are preferably free or essentially free of arginine.
  • the formulations of the invention may also include buffers, tonicity modifiers, excipients, pharmaceutically acceptable carriers and other commonly used inactive ingredients of the pharmaceutical compositions. For simplicity, these are discussed more fully later in the application.
  • Buffers maintain p ' H in a desired range.
  • Suitable buffers include histidine, potassium phosphate, sodium or potassium citrate, maleic acid, ammonium acetate, tris-(hydroxymethyi)-aminomethane (tris), various forms of acetate and diethanolamine.
  • concentration of the buffer in the formulation is preferably between about 1 mM to about 1M, and more preferably about 10 mM to about 200 mM. Buffers are well known in the art and are manufactured by known methods and available from commercial suppliers.
  • buffers examples include phosphate, histidine, citrate, maleate, tartrate, succinate, acetate, tris-(hydroxymethyl)-aminomethane (tris), bicarbonate.
  • the buffer is sodium phosphate.
  • the pH of the pharmaceutical composition is at or near physiological levels.
  • the pH of the provided compositions Is between about 5.8 and about 8.4; and even more preferably, betwee about 6.2 and about 7.4.
  • the pH can be adjusted as necessary to maximize stabilit and solubility of etanercept in a particular formulation.
  • etanercept formulations at a phi outside of physiological ranges, yet tolerable to the patient are also within the scope of the invention.
  • a tonicity modifier is a molecule that contributes to the osmolality of a solution.
  • the osmolality of a pharmaceutical composition is preferably adjusted to maximize the active ingredient's stability and/or to minimize discomfort to the patient upon administration. It is generally preferred that a pharmaceutical composition be isotonic with serum, i.e., having the same or similar osmolality, which is achieved by addition of a tonicity modifier.
  • the osmolality of the provided formulations is from about 180 to about 420 mOs .
  • the osmolality can be either higher or lower as specific conditions require.
  • tonicity modifiers suitable for modifying osmolality include, but are not limited to amino acids (not including arginine) (e.g., cysteine, histidine and glycine), salts (e.g., sodium chloride, potassium chloride and sodium citrate) and/or saccharides (e.g., sucrose, glucose and mannitol).
  • amino acids not including arginine
  • cysteine e.g., cysteine, histidine and glycine
  • salts e.g., sodium chloride, potassium chloride and sodium citrate
  • saccharides e.g., sucrose, glucose and mannitol
  • Preferred tonicity modifiers are glycine, alanine, sodium chloride, potassium chloride, and sodium sulfate.
  • the concentratio of the tonicity modifier in the formulation is preferably between about 1 mM to about 1 , more preferably about 10 mM to about 200 mM.
  • Tonicity modifiers are well known in the art and are manufactured by known methods and available from commercial suppliers.
  • Excipients also referred to as chemical additives, co ⁇ solutes, or co-solvents, that stabilize the polypeptide while in solution (also in dried or frozen forms) can also be added to a pharmaceutical composition.
  • Excipients are well known in the art and are manufactured by known methods and available from commercial suppliers.
  • excipients include but are not limited to sugars/polyols such as: sucrose, lactose, glycerol, xylite!, sorbitol, mannitol, maltose, inositol, trehalose, glucose; polymers such as: serum albumin (bovine serum albumin (BSA), human SA or recombinant HA), dextran, poiy(viny alcohol) PVA, hydroxypropyl methylceliulose (HPfvIC), polyethyleneimine, gelatin, polyvinylpyrrolidone (PVP), hydroxyethylcellu!ose (HEC); non-aqueous solvents such as: polyhydric alcohols, (e.g., PEG, and glycerol) and dlmethylformamide (D F); amino acids such as; proline, L-serine, sodium glutamic acid, alanine, glycine, lysine hydrochloride
  • Preferred excipients are sucrose, lactose, glycerol, xyiitol, sorbitol, mannitoi, maltose, inositol, trehalose, glucose, bovine serum albumin (BSA), human serum albumin (HSA), recombinant albumin, dextran, PVA, hydroxypropyl methylcei!ulose (HP C), pol ethyleneimine, gelatin, polyvinylpyrrolidone (PVP), hydroxyethylceilulose (HEC), polyethylene glycol, ethylene glycol, glycerol, alanine, glycine, lysine hydrochloride, sarcosine, SDS, polysorbate 20, polysorbate 80, poioxamer 188, trimethyiamine N-oxide, betaine, zinc ions, calcium ions, magnesium ions, CHAPS, sucrose monolaurate, and 2-O-beta-manno
  • concentration of one or more excipients in a formulation of the invention is/are preferably between about 0.001 to 5 weight percent, more preferably about 0.1 to 2 weight percent.
  • the invention provides a method of treating a mammal comprising administering a therapeutically effective amount of the pharmaceutical compositions of the invention to a mammal, wherein the mammal has a disease or disorder that can be beneficially treated with etanercept.
  • the etanercept is derived from the same species of mammal as is to be treated with the composition.
  • the mammal is a human.
  • Diseases or disorders that can be treated with the provided compositions include but are not limited to rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Wegener's disease (granulomatosis), Crohn's disease (or inflammatory bowel disease), chronic obstructive pulmonary disease (COPD), Hepatitis C, endometriosis, asthma, cachexia, psoriasis, and atopic dermatitis.
  • Additional diseases or disorders that can be treated with the compositions of the present invention include those described in WO 00/62790, WO 01/62272, U.S. Patent Application No. 2001/0021380, and US Pat. 7,648,702 B2, the relevant portions of which are incorporated herein by reference.
  • compositions may be administered to a subject in need of treatment by injection systemicaily, such as by intravenous injection; or by injection or application to the relevant site, such as by direct injection, or direct application to the site when the site is exposed in surgery; or by topical application.
  • the invention provides a method of treatment and/or prevention of rheumatoid arthritis comprises administering to a mammal in need thereof a therapeutically effective amount of one of the provided etanercept compositions.
  • the therapeutically effective amount of the etanercept in the provided compositions will depend on the condition to be treated, the severity of the condition, prior therapy, and the patient's clinical history and response to the therapeutic agent.
  • the proper dose can be adjusted according to the judgment of the attending physician such that it can be administered to the patient one time or over a series of administrations.
  • the effective etanercept amount per adult dose is from about 1-500 mg/m 2 , or from about 1-200 mg/m 2 , . or from about 1-40 mg/m 2 or about 5-25 mg/m 2 .
  • a flat dose may be administered, whose amount may range from 2-500 mg/dose, 2-100 mg/dose or from about 10-80 mg/dose.
  • an exemplary dose range is the same as the foregoing described dose ranges or lower and preferably administered two or more times per week at a per dose range of 25-100 mg/dose.
  • an acceptable dose for administration by injection contains 80-100 mg/dose, or alternatively, containing 80 mg per dose.
  • the dose can be administered weekly, biweekly, or separated by several weeks (for example 2 to 8).
  • etanercept is administered at 25 to 75 mg/ml by a single subcutaneous (SC) injection.
  • SC subcutaneous
  • an improvement in a patient's condition will be obtained by administering a dose of up to about 100 mg of the pharmaceutical composition one to three times per week over a period of at least three weeks. Treatment for longer periods may be necessary to induce the desired degree of improvement. For incurable chronic conditions the regimen may be continued indefinitely. For pediatric patients (ages 4-17), a suitable regimen may involve administering a dose of 0.4 mg/kg to 5 mg/kg of etanercept, one or more times per week,
  • the pharmaceutical formulations of the invention may be prepared in a bulk formulation, and as such, the components of the pharmaceutical composition are adjusted to be higher than would be required for administration and diluted appropriately prior to administration.
  • the pharmaceutical compositions can be administered as a sole therapeutic or in combination with additional therapies as needed.
  • the provided methods of treatment and/or prevention are used in combination with administering a therapeutically effective amount of another active agent.
  • the other active agent may be administered before, during, or after administering the pharmaceutical compositions of the present invention.
  • Another active agent may be administered either as a part of the provided compositions, or alternatively, as a separate formulation.
  • Administration of the provided pharmaceutical compositions can be achieved in various ways, including parenteral, peroral, buccal, sublingual, nasal, rectal, intraperitoneal, intradermal, transdermal, subcutaneous, intravenous, intra-arterial, intracardiac, intraventricular, intracranial, intratracheal, intrathecal administration, intramuscular injection, intravitreal injection, and topical application.
  • compositions of this invention are particularly useful for parenteral administration, i.e., subcutaneously, intramuscularly, intravenously, intraperitoneal, intracerebrospina!, Intra-articular, intrasynovial, intravitreal, and/or intrathecal.
  • Parenteral administration can be by bolus injection or continuous infusion.
  • Pharmaceutical compositions for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative.
  • compositions of the present invention are suitable for administration using these new methods, e.g., !nject-ease®, Genject®, injector pens such as GenPen® * and needleless devices such as ediJector® and BioJector®.
  • the present pharmaceutical composition can also be adapted for yet to be discovered administration methods. See also Langer, 1990, Science, 249:1527-1533.
  • compositions can also be formulated as a depot preparation.
  • Such long acting formulations may be administered by implantation (for example subcutaneousiy or intramuscuiariy) or by intramuscuiar injection.
  • the formulations may be modified with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
  • compositions may, if desired, be presented in a vial, pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient, in one embodiment the dispenser device can comprise a syringe having a single dose of the liquid formulation ready for injection.
  • the syringe can be accompanied by instructions for administration.
  • the present invention is directed to a kit or container, which contains an aqueous pharmaceutical composition of the invention.
  • concentration of the polypeptide in the aqueous pharmaceutical composition can vary over a wide range, but is generally within the range of from about 0.05 to about 20,000 micrograms per milliliter (Mg/m! of aqueous formulation.
  • the kit can also be accompanied by instructions for use.
  • Etanercept formulations stabilized with meglumine may be prepared and tested using the procedures generally described below.
  • Each solid formulation component is weighed to the amount required for a given volume of formulation buffer. These components are combined into a beaker or vessel capable of carrying and measuring the given volume of formulation buffer. A volume of deionized water equal to approximately 3 ⁇ 4 of the target given formulation buffer is added to the beaker, and the components are then soiublized. The pH of the buffer is adjusted to the target formulation pH using 1 M sodium hydroxide and/or 1 fvl hydrogen chloride. The final formulation buffer volume is then raised to the target volume through the addition of deionized water.
  • Etanercept protein solution is placed in dialysis material housing (such as Thermo Scientific Slide-A-Lyzer MINI Dialysis Unit 10,000 MVVCQ), which is then placed in contact with the desired formulation buffer for 2 hours at 4°C.
  • Formulation buffer volume to protein solution volume ratio should be no less than 1000:1.
  • the dialysis housing and protein solution it contains is then placed in a second, equal volume of formulation buffer for an additional 12 hours at 4°C.
  • Resulting protein solution is removed from the dialysis material housing, and the concentration of protein determined using ultraviolet spectroscopy. Protein concentration is adjusted to the desired level using centrifugation (such as Arnicon Ultra 10,000 MWCO Centrifugal Concentrators) and/or dilution with formulation buffer.
  • centrifugation such as Arnicon Ultra 10,000 MWCO Centrifugal Concentrators
  • compositions can be tested for long-term stability by size exclusion chromatography (SEC), denatured SEC (dSEC), hydrophobic interaction chromatography (HIC), sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), and for binding and bioactivlty at various timepoints.
  • SEC size exclusion chromatography
  • dSEC denatured SEC
  • HIC hydrophobic interaction chromatography
  • SDS-PAGE sodium dodecylsulfate polyacrylamide gel electrophoresis
  • bioactivity can be measured by any number of well-known assays.
  • composition will be stable over the term of two years or more.
  • STEP 1 Cell Expansion.
  • cell expansion necessary to generate a sufficient number of cells for inoculation of a production bioreactor is performed using a clone of CHO celis expressing the etanercept fusion protein.
  • the product of this expression process results in a mixture of correctly folded etanercept, as well as incorrectly folded and/or aggregated etanercept, along with additional impurities.
  • the harvested cell culture fluid comprising such protein mixture is subjected to detergent viral inactivation.
  • Affinity Chromatography is performed on the harvested cell culture obtain in Step 1 above using a conventional Protein A affinity column in a well known manner. Product recovery is approximately 85%.
  • the product obtained is a complex protein mixture comprising correctl folded etanercept, incorrectly folded etanercept, and/or aggregates of correctly and/or Incorrectly folded etanercept, or protein fragments.
  • the product obtained from this Protein A affinity column purification step is adjusted to pH 3,5 and then subjected to a viral inactivation step. Following viral inactivation the product is adjusted to pH 5.5 and then further clarified in a known manner using a commercially obtained capsule filter.
  • the Protein A pool from step 2 above is diluted to ⁇ 8 mS/cm with WFI and applied to a column loading of up to 15 g/L media for each cycle.
  • the column is operated at a linear velocity of 200 cm/h to give a 6 minute residence time.
  • the column is washed with 2 CV of 25 mM acetate pH 5.5.
  • the product is then eluted with an 8.5 CV, 15% to 85% gradient of 25 mM acetate pH 5.5 to 25 mM acetate, 0.7 U NaCi, pH 5.5.
  • Product collection begins at 0.15 OD (A280, 1.0 cm path length) and collection ends at 50% of peak maximum.
  • the eiuate volume is approximately 5 CV.
  • STEP 3B Mixed Mode Anion Exchange Chromatopgraphy.
  • a 27.0 L (45 cm diameter X 17 cm bed height) packed bed GE Healthcare Capto Adhere chromatography column is used to further purify the product obtained in step 3A above.
  • the column Prior to use, the column is equilibrated with 2 CV of 25 mM Tris, pH 8.0 and sanitized with 2 CV/ of 0.1 N NaOH, 1M NaCI and neutralized and equilibrated with 2 CV of 25 mM Tris, pH 8.0.
  • the Capto MMC pool from Step 3A above is adjusted to pH 8.1 with -0.045 kg of 1 M Tris, pH 8.3 per kg of pool.
  • the product from Step 3A above was diluted in-line 1 :3.8 with F! to adjust the conductivity to 12.0 mS/cm and pH 8.0.
  • the resulting material is then applied to a column loading of up to 15 g/L media.
  • the column is operated at a linear velocity of 170 cm/h to give a 6 minute residence time. After loading, the column is washed with 2 CV of 25 mM Tris, pH 8.0.
  • the product is then eluted with a 10 CV gradient (20% to 90%) of 25 mM Tris, pH 8.0 to 10 mM Tris, 1 M NaCI, pH 8.0.
  • Product collection is started at 0.15 OD (A280, 1.0 cm path length) and collection ended at 25% of peak maximum.
  • the eiuate volume is 4-6 CV.
  • the eiuted product is filtered using a commercially available capsule filter and then subjected in a known manner to viral filtration and tangential flow filtration steps.
  • Overall product recovery from step3B (including the final viral and tangential flow filtration steps) was approximately 68%.
  • Product recovery measured before the filtration steps was about 75%.
  • a schematic representation of HIC data obtained on eluation fractions from this step are representing in Figure 12.
  • the final filtered product obtained in this example is found to have greater than about 90 wt % correctly folded etanercept as determined by HIC: less than 5 wt% incorrectly folded etanercept species as determined by HIC; less than about 3 wt% of clipped material by HIC analysis (beiieved to be fragments of etanercept in which the TNFR portion thereof has been truncated) and a combined amount of correctly and incorrectly folded etanercept of greater than 95 wt% as determined by size exclusion chromatography,
  • samples of the etanercept formulations exemplified above were sterile filtered in a bio safety cabinet. Using sterilized pipettes and autoclaved pipette tips, sampies of the etanercept formulations were transferred to pre-labeled and autoclaved 1 mL lyophiiization vials. Vials were stoppered with sterile butyl stoppers and crimped with aluminum caps. All vials were then transferred to thermal stability ovens. Samples were subject to two thermal stability regimes: (1) two weeks at 40 and (2) four weeks at 25 °C. Throughout this specification, these two temperature regimes are denoted "T and T4," respectively.
  • Etanercept formulations disclosed herein were analyzed using the well known technique of Size Exclusion Chromatography (SEC), a high-performance liquid chromatography method in which anaiytes are separated by size (see Rogner, , (2000). Size Exclusion Chromatography. Protein Liquid Chromatography. M. Kastner. Amsterdam, Elsevier. 61 : 89-145.).
  • SEC Size Exclusion Chromatography
  • the mobile phase buffer was prepared to contain 50 mM sodium phosphate monobasic monohydrate and 150 mM arginine. The pH was adjusted to 8.5 using 1 M HCI. All separations were performed using a Tosoh TSK-Gel SWxl 6 mm x 4 cm guard column (cat. no. 8543) attached linearly to a Tosoh TSK-Gel G4000 SWxl 7.8 mm x 30 cm (cat. no. 8542).
  • the columns were brought to room temperature (23°C) and equilibrated with mobiie phase at a fiow rate of 0,5 mL/min. 5 microliters of 50 mg/mL etanercept formulation were injected onto the column using an autosamp!er. The separation was accomplished over 30 minutes at a flow rate of 0.5 mlJminute. Column eluent was monitored at a wavelength of 280 nm during this time.
  • AH integration was performed using Chromeieon software (Dionex). Prior to integration, the SEC chromatogram for a buffer containing no etanercept was subtracted from all chromatograms. A!l integration was performed between retention times of 12 minutes and 26 minutes. Several parameters were used to define a peak. The minimum area for a detected peak was set to 0.05 mAu * min. The two- dimensional sensitivity for peak defection was set to 0.01 mAu and 75 seconds. Peak shoulders were added manually using a manual integration too!. Ail detected peaks were manually adjusted in two steps. First, peak baselines (the bottom boundary of the peak) were adjusted to horizontal. Secondly, the vertical positions of the peak baselines were adjusted to that of the chromatogram baseiine. The chromatogram baseiine value was defined as the signal in absence of ana!yte. The signal i absence of analyte was defined as the absorbance in mAu at 12 minutes retention time.
  • the foiiowing tables shows the relative amounts of Aggregates, Monomer and Fragment 3 determined by SEC analysis as described above.
  • T 0 formulation maintained at 5 C and analyzed within 24 hours of creation.
  • HSC chromatography may be carried out in a manner known in the art and generally described in U.S. Patent 7,294,481 , incorporated herein by reference, Samples are evaluated at t 0 (within 24 hours of preparation at 5°C.) and again after either two weeks of storage at 25°C. (t 2 ) or after 4 weeks of storage at 25°C.
  • Peak 1 in the H!C chro.matogram is believed to be or include "Fragment 3", which is identified and quantified using SEC, as referenced above in the discussion of SEC data; Peak 2 is etanercept monomer as referenced above in the discussion of SEC data; and Peak 3 includes "Aggregate(s)” as referenced above in the discussion of SEC data.
  • Peak 1 peak 1
  • peak 2 peak 2
  • peak 3 includes "Aggregate(s)” as referenced above in the discussion of SEC data.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Dermatology (AREA)
  • Cell Biology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Rheumatology (AREA)
  • Pulmonology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Toxicology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Pain & Pain Management (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Reproductive Health (AREA)
  • Inorganic Chemistry (AREA)
  • Neurosurgery (AREA)
  • Diabetes (AREA)
  • Hematology (AREA)

Abstract

The present invention is an aqueous stabilized formulation of etanercept comprising: etanercept; and stabilizing ingredients to retard instability, aggregation and/or fragmentation of the etanercept in the formulation, said stabilizing ingredients being comprised of at least one of the following: {a) meglumine; or (b) meglumine in combination with sucrose; or {c) meglumine in combination with sodium chloride; or (d) meglumine in combination with sodium chloride and sucrose. Various technical terms used in the following discussion are defined below in the section entitled "Definitions'' and throughout the remainder of the specification. The stabilized etanercept formulations of the present invention elicit long term storage stability.

Description

Etanereept Formulations Stabilized with Meglumine
Fieid of the Invention
The present invention relates to aqueous pharmaceutical compositions stabilized with meglumine for long-term storage of etanereept, methods of manufacture of the compositions, methods of their administration, and kits containing the same. The invention includes etanereept formulations that do not require arginine for stabilization.
Background of the Invention
Polypeptides must often be stored prior to their use. When stored for extended periods, polypeptides are frequently unstable in solution (Manning et .al., 1989, Pharm. Res. 6:903-918). To extend their shelf life, additional processing steps have been developed, such as drying, e.g., lyophilization. However, !yophilized pharmaceutical compositions are less convenient to use.
Typical practices to improve polypeptide stability can be addressed by varying the concentration of elements with the formulation, or by adding excipients to modify the formulation (See, for example, U.S. Pat. Nos. 5,580,858 and 6,171 ,586). However, the use of additives can still result in inactive polypeptides, in addition, in the case of lyophilization, the rehydration step can result in inactivation of the polypeptide by, for example, aggregation or denaturation (Hora et a!., 1992, Pharm. Res., 9:33-36; Liu et al., 1991 , Biotechnoi. Bioeng., 37:177-184). Aggregation of polypeptides is undesirable, as it may result in immunogenicity (Cleland et al., 1993, Chi Rev. Therapeutic Drug Carrier Systems, 10:307-377; and Robbins et al., 1987, Diabetes, 36:838-845).
Another way to improve polypeptide stability is to use L-arginine at a specific concentration {U.S. Pat. No. 7,648,702).
One of the polypeptides that is stored for up to two years prior to use is etanereept (Enbrel©, immunex Corporation), which is a dimeric fusion polypeptide consisting of the extracellular ligand-binding portion of the human 75 kilodalton (p75) tumor necrosis factor receptor (TNFR) linked to the Fc portion of human IgGl it consists of 934 amino acids and has an apparent molecular weight of approximately 150 kiioda!ions (Physicians Desk Reference, 2002, Medical Economics Company Inc.) The Fc component of etanercept contains the constant heavy 2 (CH2) domain, the constant heavy 3 (CH3) domain and hinge region, but not the constant heavy 1 (CM) domain of human igG1. An Fc domain can contain one or ali of the domains described above. Etanercept is usually produced by recombinant DMA technology in a Chinese hamster ovary (CHO) mammalian cell expression system.
The present invention provides novel stable liquid formulations of etanercept that allow its long-term storage.
Summary of the invention
The present invention is an aqueous stabilized formulation of etanercept comprising: etanercept; and stabilizing ingredients to retard instability, aggregation and/or fragmentation of the etanercept in the formulation, said stabilizing ingredients being comprised of at least one of the following: (a) meglumine; or (b) meglumine in combination with sucrose; or (c) meglumine in combination with sodium chloride; or ■(d) meglumine in combination with sodium chioride and sucrose.
Various technical terms used in the following discussion are defined below in the section entitled "Definitions" and throughout the remainder of the specification.
The stabilized etanercept formulations of the present invention elicit long term storage stability as characterized by at least one of: (1) SEC analysis at M3 or T2 or T4 of: monomer content greater than about 90%: aggregates content of less than about 3 wt%; and fragment 3 content less than about 5 wt%: and (2) H!C analysis at fv13 or T2, or T wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 3 wt.%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than 80 wt.%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 20 wt.%.
In preferred aspects of the stabilized formulations, the formulations elicit long term storage stability as characterized by: an HIC analysis at M3 or T2 or T4 wherein the amount of the composition represented by peak 2 of the HIC chromatogram is greater than or equal to about 95 wt.%; and wherein, if peak 3 is present on the HIC chromatogram, the amount of the composition represented by peak 3 is less than or equal to about 3 wt.%.
The stabilized etanercept formulation as summarized above, optionally and preferably, contains no arginine, or are essentially free of arginine.
The formulations of the invention have excellent stability as determined by SEC (Size Exclusion Chromatography) and HIC (Hydrophobic Interaction Chromatography) analysis conducted after one, two or three months of storage at 5° C. These formulations are comparable to or better than a commercially available formulation of etanercept, in which arginine is a required component. Accordingly the present invention is further directed to formulations of stabilized etanercept, as summarized above, which contain no arginine, or are essentially free of arginine, and wherein the composition, evaluated at M3 or T2 or T4, elicits long term storage stability that meets one or both of the following criteria: (A) stability comparable to or better than commercially available etanercept marketed under the trademark Enbrel®, as measured by (i) SEC analysis of the amounts of aggregate(s), monomer and fragment 3 in the composition (as defined in the specification) and (ii) HIC analysis of amounts of material in the composition corresponding to peaks 1 , 2 and 3 of the HIC chromatogram (as defined in the specification); and (B) an HIC chromatogram in which (i) peak 3 is absent, or essentially absent and (ii) peak 2 represents greater than about 95 wt% of the composition; an SEC chromatogram containing essentially no peak corresponding to aggregate(s); and an SEC chromatogram in which the monomer content represents at least about 95 wt% of the composition.
In one preferred aspect, the formulation of the invention comprises about about 25 to 75 mg/ml etanercept, sodium chloride in an amount up to about 150 mM, about 1 to about 30 mM sodium phosphate; and about 0 to 5 wt.% sucrose or trehalose or combination thereof; wherein the composition has a pH of about 6.0 to about 6.6; and wherein the composition is characterized by SEC analysis at 3 or T2 or T4 in which: monomer content is greater than about 85 wt.%; aggregate(s) content is less than about 3 wt.%, and fragment 3 content is less than about 8 wt.%. Formulations meeting these analytical criteria do not require arginine for stabilization.
_ ¾ - In a further preferred embodiment the stabilized etanercept formulation is further characterized by (a) an SEC analysis at 3 or T2 or T4 of greater than about 90 t% monomer content; and less than about 3 wt% aggregate(s) content; and (b) an HIC analysis at M3 or T2 or T4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 4 wt%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than about 80 wt%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 20 wt%; and wherein the formulation is free or essentially free of arginine.
The etanercept compositions of the invention further afford the ability to provide formulations which contain acceptable levels of subvisible particles. Accordingly, the invention is further directed to stabilized etanercept formulations having, at M3 or T2 or T4lno more than, on average, about 10,000 subvisible particles per mL having a size greater than 5 pm.
The stabilized etanercept composition of the present invention are further characterized by: (a) an SEC analysis at IV or T2 or T of greater than about 90 wt% monomer content; and less than about 3 wt.% aggregate(s) content; and (b) an HIC analysis at 3 or T2 or T4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 3 wt%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than 80 wt%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 20 wt% and wherein the formulation is free or essentially free of arginine.
The stability of the formulations may be further characterized in that the compositions, optionally free or essentiaily of arginine, exhibit an HIC analysis at 3 or T≥ or T4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 1%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than about 95 t%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 1 wt%. Formulations meeting these analytical criteria do not require arginine for stabilization. Preferred stabilized compositions of the invention, preferably free or essentlaliy free of arginine, exhibit an HiC analysis at M3 or T?_ or T4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 2% or preferably less than about 1%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than about 95 wt.% and preferably greater than about 97%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 1 wt%: and preferably 0 to 1%,
As differentiated from commercially available etanercept provided in an arginihe-containing formulation, we found it surprising, in light of US. Pat. No. 7,848,702, that the formulation embodiments of etanercept described and exemplified herein do not require arginine for long-term stabilization, although arginine may still be added if desired. The ability to provide etanercept formulations stabilized without arginine represents a potentially significant benefit to the health care system by providing patients and health care providers with alternative formulations of etanercept that may become available at lower cost compared with present commercial etanercept formulation (i.e., Enbrel®) that require arginine for stabilization.
As used herein the term nstability" or like terms denotes the tendency of the etanercept monomer to undergo a variety of undesired transformations during storage. Such transformations include the formation of oligomers and high molecular weight aggregate(s) (hereinafter terms "aggregate(s)" in which multiple copies of the essentially intact etanercept monomer become irreversibl associated with one another through a variety of non-covalent attractions (e.g., electrostatic interactions.) Undesired transformations during storage may also include degradation of the etanercept monomer to smaller fragments and/or clipped species. Ideally, a formulation of etanercept should minimize, to the greatest extent possible, the tendency of the formulation to result, during storage, in the formation of aggregates, misfolded protein, oligomers and/or fragments of etanercept. An important benefit resulting from the ability to reduce formation of unwanted aggregates or fragments is a reduction in the potential toxicity and/or immunogenicity of the drug. The etanercept formulation of the present invention which is optionally and preferably free, or essentially free of argihine. The term "essentially free of arginine" is intended to mean that arginine, even if present, is not contributing to the stabilization of the etanercept monomer in the formulation to such an extent that a person skilled in the art would judge its presence beneficial or necessary from a stabilization standpoint.
These and other aspects will become apparent from the following description although variations and modifications therein may be affected without departing from the spirit and scope of the novel concepts of the disclosure.
It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.
DETAILED' DESCRIPTION OF THE INVENTION
Various embodiments of the invention are now described in detail. As used in the description and throughout the claims, the meaning of "a", "an", and "the" includes plural reference unless the context clearly dictates otherwise. Also, as used in the description and throughout the claims, the meaning of "in" includes "in" and "on" unless the context clearly dictates otherwise. Additionally, some terms used in this specification are more specifically defined below.
DEFINITIONS
The terms used in this specification generally have their ordinary meanings in the art, within the context of the invention, and in the specific context where each term is used. Certain terms that are used to describe the invention are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the invention. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in this specification including examples of any terms discussed herein is illustrative only, and in no way limits the scope and meaning of the invention or of any exemplified term. The invention is not limited to the various embodiments given in this specification. Unless otherwise defined, ail technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains, in the case of conflict, the present document, including definitions will control,
"Around," "about8 or "approximately" shall generally mean within 20 percent, within 10 percent, within 5, 4, 3, 2 or 1 percent of a given value or range. Numerical quantities given are approximate, meaning that the term "around," "about" or "approximately" can be inferred if not expressly stated.
The term "etanercepf or "etanercept monomer" or "monomer" is synonymous with Enbre!®. It refers to a polypeptide which is a dimeric fusion polypeptide consisting of the extracellular iigand-binding portion of the human 75 kilodalton (p75) tumor necrosis factor receptor (TNFR) linked to the Fc portion of human IgGl It consists of 934 amino acids and has an apparent molecular weight of approximately 150 kiiodaStons. For the purposes of the present application, the term "etanercepf also encompasses etanercepf with minor modifications in the amino acid structure (including deletions, additions, and/or substitutions of amino acids) which do not significantly affect the function, potency, or avidity of etanercepf. The term "etanercepf encompasses ail forms and formulations of Enbrel®, including but not limited to concentrated formulations, injectable ready-to-use formulations; formulations reconstituted with water, alcohol, and/or other ingredients, and others.
The term "sugar" refers to monosaccharides, disachharides, and polysaccharides. Examples of sugars include, but are not limited to, sucrose, trehalose, dextrose, and others.
The term "meglumine" refers to a compound with chemical formula H3NHCH2(CHOH)4CH2OH, also known as 1-Deoxy~1-methylamtnosorbitol; N- ethyl- d-giucamine; and 1-Deoxy-1-methylamino-D-glucitol.
The terms "mannosyiglycerate," "m.annosyilactate," "mannosy!glycolate", and "diglycerolphosphate" are well known in the art and have their commonly accepted meanings. The following references describe these compounds in some detail: Faria et a!., Carbohydrate Res, 2008, 343: 3025-3033; Borges et al., Extremophi!es 2002, 8: 209-218: Faria et at, ChemBioChem 2003, 4; 734-741 ; Sawangwan et al., Biotechno!. J. 2010, 5: 187-191 ; and Pais et al., J. Mot: Biol. 2009, 394: 237-250. The application incorporates by reference the description of these compounds contained in these references.
The term "po!yol" refers to an alcohol containing multiple hydroxy! groups. Examples of polyois include, but are not limited to, mannitoi, sorbitol, and others.
The term "long-term storage" is understood to mean that the pharmaceutical composition can be stored for three months or more, for six months or more, and preferably for one year or more. Long-term storage is also understood to mean that the pharmaceutical composition is stored either as a liquid at 2-8° C, or is frozen, e.g., at -2G°C, or colder. It is also contemplated thai the composition can be frozen and thawed more than once.
The term "stable" or "stabilized" with respect to long-term storage is understood to mean that etanercepf contained in the pharmaceutical compositions does not lose more than 20%, or more preferably 15%, or even more preferably 10%, and most preferably 5% of its activity relative to activity of the composition at the beginning of storage,
The term "mammal" includes, but is not limited to, a human.
The term "pharmaceutically acceptable carrier" refers to a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material, formulation auxiliary, or excipient of an conventional type. A pharmaceutically acceptable carrier is non- toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation.
The term "composition" refers to a mixture that usually contains a carrier, such as a pharmaceutically acceptable carrier or excipient that is conventional in the art and which is suitable for administration into a subject for therapeutic, diagnostic, or prophylactic purposes. It may include a ceil culture in which the polypeptide or polynucleotide is present in the cells or in the culture medium. For example, compositions for oral administration can form solutions, suspensions, tablets, pills, capsules, sustained release formulations, oral rinses or powders.
The terms "pharmaceutical composition" and "formulation" are used interchangeably. The term "treatment" refers to any administration or application of remedies for disease in a mamma! and inciudes inhibiting the disease, arresting its development, relieving the disease, for example, by causing regression, or restoring or repairing a lost, missing, or defective function; or stimulating an inefficient process. The term includes obtaining a desired pharmacologic and/or physiologic effect, covering any treatment of a pathological condition or disorder in a mammal. The effect may be prophylactic in terms of completely or partially preventing a disorder or symptom thereof and/or may be therapeutic in terms of a partial or complete cure for a disorder and/or adverse affect attributable to the disorder. It inciudes (1) preventing the disorder from occurring or recurring in a subject who may be predisposed to the disorder but is not yet symptomatic, (2) inhibiting the disorder, such as arresting its development, (3) stopping or terminating the disorder or at least its associated symptoms, so that the host no longer suffers from the disorder or its symptoms, such as causing regression of the disorder or its symptoms, for example, by restoring or repairing a lost, missing or defective function, or stimulating an inefficient process, or (4) relieving, alleviating or ameliorating the disorder, or symptoms associated therewith, where ameliorating is used in a broad sense to refer to at least a reduction in the magnitude of a parameter, such as inflammation, pain and/or tumor size.
The term "disease" refers to any condition, infection, disorder or syndrome that requires medical intervention or for which medical intervention is desirable. Such medical intervention can include treatment, diagnosis and/or prevention.
The term "therapeutically effective amount" refers to an amount which, when administered to a living subject, achieves a desired effect on the living subject. For example, an effective amount of the polypeptide of the invention for administration to the living subject is an amount that prevents and/or treats an integrin av 3-mediated disease. The exact amount will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques. As is known in the art, adjustments for systemic versus localized delivery, age, body weight, general health, sex, diet, time of administration, drug interaction and the severity of the condition may be necessary, and will be ascertainable with routine experimentation by those skilled in the art. The term "Ti"refers to a point in time at which an etanercept formulation has been stored for about one week at 40° C
The term "T2"refers to a point in time at which an etanercept formulation has been stored for about two weeks at 40° C
The term "T/refe.rs to a point in time at which an etanercept formulation has been stored for about four weeks at 40° C,
The term "M3" refers, collectively, to three points in time, and in particular to an analytical result being observed for an etanercept formulation after duration of either about one, about two or about three months of storage at a storage temperature of 5° C. For example, reference herein to an analysis being conducted at M3 should be understand to mean that such analysis is be done at the point in time at which etanercept formulation has been in storage for a duration selected from about one, about two, or about three months. Thus, a requirement herein that an etanercept formulation elicit a certain analytical value or measurement at 3 is satisfied if the required value is observed at a point in time corresponding to at least one of the following storage durations: at approximately one month, at approximately two months or at approximately three months of storage at 5°C.
The terms "Peak 1 ," Peak 2" and "Peak 3" when used herein in connection with discussion of HIC chromatography results refers to the same peaks 1 , 2 and 3 discussed in US Patent 7,294,481.
Embodiments of the Invention
When pharmaceutical compositions containing etanercept (Enbrel®), including aqueous and lyophi!ized formulations of etanercept are stored on a long term basis, the activity of etanercept can be lost or decreased due to instability of the etanercept monomer via aggregation and/or chemical degradation including formation of fragments. Thus, the present invention provides several embodinients of aqueous formulations of etanercept that allow stable long-term storage of etanercept, so that etanercept is stable over the course of storage either in liquid or frozen states. The provided formulations include, but are not limited to formulations that do not contain arginine and do not require any extra steps such as rehydratlng. These embodiments are explained in a greater detail below.
Etanercept
AS! of the compositions of the present invention comprise etanercept (Enbrel®). As explained in the Background section of this application, etanercept is a dimeric fusion poiypeptide consisting of the extracellular Iigand-binding portion of the human 75 ki!odalton (p75) tumor necrosis factor receptor (TNFR) linked to the Fc portion of human igG . Etanercept consists of 934 amino acids. The Fc component of etanercept contains the constant heavy 2 (CH2) domain, the constant heavy 3 (CH3) domain and hinge region of human IgG An Fc domain can contain one or all of the domains described above.
Etanercept suitable for storage in the present pharmaceutical composition can be produced by living host ceils that express etanercept, such as hybridomas in the case of antibodies, or host celis that that have been genetically engineered to produce the polypeptide in the case of fusion polypeptides or antibodies. Methods of genetically engineering cells to produce polypeptides are well known in the art. See, e.g., Ausubei et a!., eds. (1990), Current Protocols in Molecular Biology (Wiley, New York). Such methods include introducing nucleic acids that encode and aliow expression of the polypeptide Into living host ceils. These host cells can be bacterial cells, fungal cells, or, preferably, animal ceils grown in culture. Bacterial host ceils include, but are not limited to, Escherichia coii cells. Examples of suitable £ coii strains include: HB101 , DH5.alpha, GM2929, JM109, KW251 , NM538, N 539, and any E. coii strain that fails to cleave foreign DNA. Fungal host cells that can be used include, but are not limited to, Saccharomyces cerevisiae, Pichia pastoris and Aspergillus ceils, A few examples of animal cell lines that can be used are CHO, VERO, BHK, HeLa, Cos, MDCK, 293, 3T3, and W138. New animal cell lines can be established using methods weli know by those skiiied in the art (e.g., by transformation, viral infection, and/or selection). Optionally, etanercept can be secreted by the host celis into the medium.
Purification of the expressed etanercept can be performed by any standard method. When etanercept is produced intracellular^, the particulate debris is removed, for example, by centrlfugation or ultrafiltration. When etanercept is secreted into the medium, supernatants from such expression systems can be first concentrated using standard polypeptide concentration filters. Protease inhibitors can also be added to inhibit proteolysis and antibiotics can be included to prevent the growth of microorganisms.
Etanercept can be purified using, for example, hydroxyapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, and any combination of known or yet to be discovered purification techniques, including but not limited to Protein A chromatography, fractionation on an ion-exchange column, ethanol precipitation, reverse phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSET®, an anion or cation exchange resin chromatography (such as a polyaspartic acid column), chromatofocusing, SDS- PAGE, and ammonium sulfate precipitation.
Etanercept Stabilized with art Ionic Poiyo! Derivative Excipient
The invention provides a stable aqueous formulation comprising etanercept and an ionic poiyol derivative excipient, wherein said excipient is selected from the group consisting of meglumine (N-methyi-D-glucamine), mannosylg!ycerate, glucosylglycerate, mannosyllactate, mannosylglycolate, and diglycerolphosphate.
Preferably, in this embodiment or aspect, the invention is an aqueous stabilized formulation of etanercept comprising: etanercept; and stabilizing ingredients to retard instability, aggregation and fragmentation of the etanercept in the formulation, said stabilizing ingredients being comprised of (a) meglumine; or (b) meglumine in combination with sucrose; or (c) meglumine in combination with sodium chloride; or (d) meglumine 'in combination with sodium chloride and sucrose.
Meglumine is commonl used as a small molecule excipient We have now surprisingly found that meglumine is also able to stabilize aqueous pharmaceutical compositions containing a large protein, such as etanercept.
it is believed that meglumine reduces etanercepfs tendency to associate in undesired ternary or quaternary complexes, and therefore, reduces aggregation of etanercept The reduction in aggregation is believed to last for a long period of time, e.g., two years or more. Without wishing to be bound to a particular theory, it is believed that ionic poiyol derivatives, including meglumine, are able to stabilize aqueous pharmaceutical compositions containing etanercept by a combination of three different mechanisms. First, they can act as an excluded solute in the same way mannitol, sucrose, and sorbitol increase conformational stability. Second, charged solutes can alter the colloidal stability, thereby reducing the propensity to self-associate, thereby slowing aggregation. Third, these ionic polyoS derivatives, being charged near neutral H, can act as salting-in agents, as arginine does, potentially resolubilizing aggregates. The stabilizing effects of meglumine are not limited to reduction in aggregates but may involve other aspects of stabiiization of the etanercept monomer in a formulation containing the monomer.
The pharmaceutical compositions of the invention may be prepared by combining, a purified etanercept and the ionic polyol derivative, preferably meglumine, Further, a buffer, a tonicity modifier and an additional excipient and other commonly used inactive ingredients can be added as needed. For simplicity, these are discussed more fully later in the specification. A person of ordinary skill in the art will understand that the combining of the various components to be included in the composition can be done in any appropriate order. For example, the buffer can be added first, middle or last, and the tonicity modifier can also be added first, middle or last. A person of ordinary skill in the art will also understand that some of these chemicals can be incompatible in certain combinations, and accordingly, are easily substituted with different chemicals that have similar properties but are compatible in the relevant mixture.
In a preferred embodiment, the concentration of meglumine in the provided formulations is preferably between about 0.1% {w/v) to 40% (w/v), more preferably about 1% to about 20%, more preferably about 2% to about 10%, even more preferably about 2% to about 5%.
Meglumine is available from commercial suppliers.
A preferred embodiment comprises about 25 to about 75 mg/ml etanercept, about 1-30 mM of sodium phosphate; up to about 10% meglumine; optionally up to about 5 wt.% sucrose; and optionally up to about 100 mM sodium chloride, wherein the composition has a pH of about 6.0 to 7.0, and preferably about 6.0 to about 6.6 and most preferably about 6.3 to about 6.5.
A meglumine stabilized etanercept composition is preferably characterized by SEC analysis at T2 in which: the monomer content is greater than about 85 wt.%; aggregate(s) content is less than about 3 wt.%; and fragment 3 content is Sess than about 8 wt.%.
A more preferred formulation of etanercept wherein an ionic poiyoi derivative such as meglumine is present for stabilization is one that is characterized by:
(a) an SEC analysis at T4 of greater than about 90, 91, 92, 93, 94, 95, 96, or 97 wt.% monomer content; and less than about 3, 2 or 1 wt.% aggregate(s) content; and
(b) an HiC analysis at T2 wherein the amount of the composition represented by peak 1 of the HiC chromatogram is less than about 4, 3, 2 or 1 wt.%; the amount of the composition represented by peak 2 of the H!C chromatogram is greater than 80, 81 , 82, 83, 84 or 85 wt.%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 20, 19, 18, 17, 16, 15, 14, or 13 wt.%; and
(c) an HIC analysis at T4 wherein the amount of the composition represented by peak 1 of the HiC chromatogram is less than about 3, 2 or 1 wt.%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than 80, 81 , 82, 83, 84 or 85 wt.%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than..20, 19, 18, 17, 16, 15, 14 or 13 wt.%. A particularly preferred etanercept formulation stabilized with meglumine is characterized by HIC analysis at T4 or T2 wherein the amount of the composition represented by peak 1 of the H!C chromatogram is less than about 1%; the amount of the composition represented by peak 2 of the H!C chromatogram is greater than about 99 wt.%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 1 wt.%.
The terms "SEC", "T2" "T4" "HIC" "monomer content" "aggregate s)" and "fragment 3" "peak 1 ," "peak 2," and "peak 3," are defined in the examples below.
In other embodiments meglumine can be replaced with another sonic po!yol derivative of sorbitol, glycerol or mannito!, such as mannosylgiycerate, glucosylglycerate, mannosyllactate, mannosyiglycolate, and diglyceroiphosphate (at about 0.1% to about 40%) in the formulation. A preferred meg!umine-siabiiized etanercept formulation free of arginine and exhibiting analytical properties as described above comprises about 25 to about 75 mg/ml etanercept; about 0,5 wt.% megiumine; about 25 mM phosphate; about 1% sucrose; and about 100 mM sodium chloride.
A further preferred meglumine-stabilized etanercept formulation free of arginine and exhibiting analytical properties as described above comprises about 50 mg/ml etanercept; about 5 wt.% meglumine; about 25 mM phosphate.
Although the invention does not exclude the use of arginine, the etanercept formulations comprising ionic poiyol derivatives such as meglumine for stabilization according to the present invention are preferably free or essentially free of arginine.
Additional Components of the Provided Pharmaceutical Compositions
The formulations of the invention may also include buffers, tonicity modifiers, excipients, pharmaceutically acceptable carriers and other commonly used inactive ingredients of the pharmaceutical compositions. For simplicity, these are discussed more fully later in the application.
Buffers maintain p'H in a desired range. Suitable buffers include histidine, potassium phosphate, sodium or potassium citrate, maleic acid, ammonium acetate, tris-(hydroxymethyi)-aminomethane (tris), various forms of acetate and diethanolamine. The concentration of the buffer in the formulation is preferably between about 1 mM to about 1M, and more preferably about 10 mM to about 200 mM. Buffers are well known in the art and are manufactured by known methods and available from commercial suppliers.
Examples of suitable buffers are phosphate, histidine, citrate, maleate, tartrate, succinate, acetate, tris-(hydroxymethyl)-aminomethane (tris), bicarbonate.
In a preferred embodiment, the buffer is sodium phosphate.
In a preferred embodiment, the pH of the pharmaceutical composition is at or near physiological levels. Thus, preferably, the pH of the provided compositions Is between about 5.8 and about 8.4; and even more preferably, betwee about 6.2 and about 7.4. A person of ordinary skill in the art will understand that the pH can be adjusted as necessary to maximize stabilit and solubility of etanercept in a particular formulation. Thus, etanercept formulations at a phi outside of physiological ranges, yet tolerable to the patient, are also within the scope of the invention.
A tonicity modifier is a molecule that contributes to the osmolality of a solution. The osmolality of a pharmaceutical composition is preferably adjusted to maximize the active ingredient's stability and/or to minimize discomfort to the patient upon administration. It is generally preferred that a pharmaceutical composition be isotonic with serum, i.e., having the same or similar osmolality, which is achieved by addition of a tonicity modifier.
In a preferred embodiment, the osmolality of the provided formulations is from about 180 to about 420 mOs .. However, it is to be understood that the osmolality can be either higher or lower as specific conditions require.
Examples of tonicity modifiers suitable for modifying osmolality include, but are not limited to amino acids (not including arginine) (e.g., cysteine, histidine and glycine), salts (e.g., sodium chloride, potassium chloride and sodium citrate) and/or saccharides (e.g., sucrose, glucose and mannitol).
Preferred tonicity modifiers are glycine, alanine, sodium chloride, potassium chloride, and sodium sulfate.
In a preferred embodiment, the concentratio of the tonicity modifier in the formulation is preferably between about 1 mM to about 1 , more preferably about 10 mM to about 200 mM. Tonicity modifiers are well known in the art and are manufactured by known methods and available from commercial suppliers.
Excipients, also referred to as chemical additives, co~solutes, or co-solvents, that stabilize the polypeptide while in solution (also in dried or frozen forms) can also be added to a pharmaceutical composition. Excipients are well known in the art and are manufactured by known methods and available from commercial suppliers.
Examples of suitable excipients include but are not limited to sugars/polyols such as: sucrose, lactose, glycerol, xylite!, sorbitol, mannitol, maltose, inositol, trehalose, glucose; polymers such as: serum albumin (bovine serum albumin (BSA), human SA or recombinant HA), dextran, poiy(viny alcohol) PVA, hydroxypropyl methylceliulose (HPfvIC), polyethyleneimine, gelatin, polyvinylpyrrolidone (PVP), hydroxyethylcellu!ose (HEC); non-aqueous solvents such as: polyhydric alcohols, (e.g., PEG, and glycerol) and dlmethylformamide (D F); amino acids such as; proline, L-serine, sodium glutamic acid, alanine, glycine, lysine hydrochloride, sarcosine and gamma-aminobutyric acid; surfactants such as: Tween®-80 (polysorbate 80), Tween©-20 (polysorbate 20), SDS, polysorbate, poloxamers; and miscellaneous excipients such as: potassium phosphate, sodium acetate, ammonium sulfate, magnesium sulfate, sodium sulfate, trimethyiamine N-oxide, betaine, metal ions (e.g., zinc, calcium, and magnesium), CHAPS, monolaurate, 2- O-beta-mannog!ycerate or any combination of the above.
Preferred excipients are sucrose, lactose, glycerol, xyiitol, sorbitol, mannitoi, maltose, inositol, trehalose, glucose, bovine serum albumin (BSA), human serum albumin (HSA), recombinant albumin, dextran, PVA, hydroxypropyl methylcei!ulose (HP C), pol ethyleneimine, gelatin, polyvinylpyrrolidone (PVP), hydroxyethylceilulose (HEC), polyethylene glycol, ethylene glycol, glycerol, alanine, glycine, lysine hydrochloride, sarcosine, SDS, polysorbate 20, polysorbate 80, poioxamer 188, trimethyiamine N-oxide, betaine, zinc ions, calcium ions, magnesium ions, CHAPS, sucrose monolaurate, and 2-O-beta-mannoglycerate.
The concentration of one or more excipients in a formulation of the invention is/are preferably between about 0.001 to 5 weight percent, more preferably about 0.1 to 2 weight percent.
Methods of Treatment
In another embodiment, the invention provides a method of treating a mammal comprising administering a therapeutically effective amount of the pharmaceutical compositions of the invention to a mammal, wherein the mammal has a disease or disorder that can be beneficially treated with etanercept.
In a preferred embodiment, the etanercept is derived from the same species of mammal as is to be treated with the composition.
In a preferred embodiment, the mammal is a human.
Diseases or disorders that can be treated with the provided compositions include but are not limited to rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, Wegener's disease (granulomatosis), Crohn's disease (or inflammatory bowel disease), chronic obstructive pulmonary disease (COPD), Hepatitis C, endometriosis, asthma, cachexia, psoriasis, and atopic dermatitis. Additional diseases or disorders that can be treated with the compositions of the present invention include those described in WO 00/62790, WO 01/62272, U.S. Patent Application No. 2001/0021380, and US Pat. 7,648,702 B2, the relevant portions of which are incorporated herein by reference.
The provided pharmaceutical compositions may be administered to a subject in need of treatment by injection systemicaily, such as by intravenous injection; or by injection or application to the relevant site, such as by direct injection, or direct application to the site when the site is exposed in surgery; or by topical application.
!n one embodiment, the invention provides a method of treatment and/or prevention of rheumatoid arthritis comprises administering to a mammal in need thereof a therapeutically effective amount of one of the provided etanercept compositions.
The therapeutically effective amount of the etanercept in the provided compositions will depend on the condition to be treated, the severity of the condition, prior therapy, and the patient's clinical history and response to the therapeutic agent. The proper dose can be adjusted according to the judgment of the attending physician such that it can be administered to the patient one time or over a series of administrations.
in one embodiment, the effective etanercept amount per adult dose is from about 1-500 mg/m2, or from about 1-200 mg/m2,. or from about 1-40 mg/m2 or about 5-25 mg/m2.
Alternatively, a flat dose may be administered, whose amount may range from 2-500 mg/dose, 2-100 mg/dose or from about 10-80 mg/dose.
If the dose is to be administered more than one time per week, an exemplary dose range is the same as the foregoing described dose ranges or lower and preferably administered two or more times per week at a per dose range of 25-100 mg/dose.
In another embodiment, an acceptable dose for administration by injection contains 80-100 mg/dose, or alternatively, containing 80 mg per dose.
The dose can be administered weekly, biweekly, or separated by several weeks (for example 2 to 8). in one embodiment, etanercept is administered at 25 to 75 mg/ml by a single subcutaneous (SC) injection.
In some instances, an improvement in a patient's condition will be obtained by administering a dose of up to about 100 mg of the pharmaceutical composition one to three times per week over a period of at least three weeks. Treatment for longer periods may be necessary to induce the desired degree of improvement. For incurable chronic conditions the regimen may be continued indefinitely. For pediatric patients (ages 4-17), a suitable regimen may involve administering a dose of 0.4 mg/kg to 5 mg/kg of etanercept, one or more times per week,
In another embodiment, the pharmaceutical formulations of the invention may be prepared in a bulk formulation, and as such, the components of the pharmaceutical composition are adjusted to be higher than would be required for administration and diluted appropriately prior to administration.
The pharmaceutical compositions can be administered as a sole therapeutic or in combination with additional therapies as needed. Thus, in one embodiment, the provided methods of treatment and/or prevention are used in combination with administering a therapeutically effective amount of another active agent. The other active agent may be administered before, during, or after administering the pharmaceutical compositions of the present invention. Another active agent may be administered either as a part of the provided compositions, or alternatively, as a separate formulation.
Administration of the provided pharmaceutical compositions can be achieved in various ways, including parenteral, peroral, buccal, sublingual, nasal, rectal, intraperitoneal, intradermal, transdermal, subcutaneous, intravenous, intra-arterial, intracardiac, intraventricular, intracranial, intratracheal, intrathecal administration, intramuscular injection, intravitreal injection, and topical application.
The pharmaceutical compositions of this invention are particularly useful for parenteral administration, i.e., subcutaneously, intramuscularly, intravenously, intraperitoneal, intracerebrospina!, Intra-articular, intrasynovial, intravitreal, and/or intrathecal. Parenteral administration can be by bolus injection or continuous infusion. Pharmaceutical compositions for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. In addition, a number of recent drug delivery approaches have been developed and the pharmaceutical compositions of the present invention are suitable for administration using these new methods, e.g., !nject-ease®, Genject®, injector pens such as GenPen®* and needleless devices such as ediJector® and BioJector®. The present pharmaceutical composition can also be adapted for yet to be discovered administration methods. See also Langer, 1990, Science, 249:1527-1533.
The provided pharmaceutical compositions can also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneousiy or intramuscuiariy) or by intramuscuiar injection. Thus, for example, the formulations may be modified with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
The pharmaceutical compositions may, if desired, be presented in a vial, pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient, in one embodiment the dispenser device can comprise a syringe having a single dose of the liquid formulation ready for injection. The syringe can be accompanied by instructions for administration.
Sn another embodiment, the present invention is directed to a kit or container, which contains an aqueous pharmaceutical composition of the invention. The concentration of the polypeptide in the aqueous pharmaceutical composition can vary over a wide range, but is generally within the range of from about 0.05 to about 20,000 micrograms per milliliter (Mg/m!) of aqueous formulation. The kit can also be accompanied by instructions for use.
The present invention is more particularly described in the following examples that are intended as illustrative only, since many modifications and variations therein will be apparent to those skilled in the art. EXAMPLE
Etanercept Stabilized with Meglumine
Etanercept formulations stabilized with meglumine may be prepared and tested using the procedures generally described below.
Each solid formulation component is weighed to the amount required for a given volume of formulation buffer. These components are combined into a beaker or vessel capable of carrying and measuring the given volume of formulation buffer. A volume of deionized water equal to approximately ¾ of the target given formulation buffer is added to the beaker, and the components are then soiublized. The pH of the buffer is adjusted to the target formulation pH using 1 M sodium hydroxide and/or 1 fvl hydrogen chloride. The final formulation buffer volume is then raised to the target volume through the addition of deionized water. Etanercept protein solution is placed in dialysis material housing (such as Thermo Scientific Slide-A-Lyzer MINI Dialysis Unit 10,000 MVVCQ), which is then placed in contact with the desired formulation buffer for 2 hours at 4°C. Formulation buffer volume to protein solution volume ratio should be no less than 1000:1. The dialysis housing and protein solution it contains is then placed in a second, equal volume of formulation buffer for an additional 12 hours at 4°C. Resulting protein solution is removed from the dialysis material housing, and the concentration of protein determined using ultraviolet spectroscopy. Protein concentration is adjusted to the desired level using centrifugation (such as Arnicon Ultra 10,000 MWCO Centrifugal Concentrators) and/or dilution with formulation buffer.
The compositions can be tested for long-term stability by size exclusion chromatography (SEC), denatured SEC (dSEC), hydrophobic interaction chromatography (HIC), sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), and for binding and bioactivlty at various timepoints. The bioactivity can be measured by any number of well-known assays.
For example, the techniques of Size Exclusion Chromatography are described in Hawe et al, Phar . Res. 20 1, 28: 2302 and/or van Marrscha!kerweerd et ai.( Eur. J. Pharm. Biopharm. 2011 , 78: 213. Similarly, the techniques of Denatured Size Exclusion Chromatography, Hydrophobic interaction Chromatography, and Sodium DodecylSuliate-PoiyAcryiamide Gel Electrophoresis are also well known to persons having ordinary skill in the art.
It is believed that the composition will be stable over the term of two years or more.
(Formulation 1 :19)
Ingredient I concentration
Etanercept (active ingredient) 50 mg/ml
Meglumine (inactive ingredient) 5 % (w/v)
Sodium phosphate, pH 6.3 (inactive) 25 mM
(Form ulatson 1 :21)
Ingredient concentration
Etanercept (active ingredient) 50 mg/ml
Meglumine (inactive ingredient) 0.49 % (w/v)
Sodium phosphate, pH 6.3 (inactive) 25 mM
Sucrose (inactive) 1 % (w/v)
NaCI (inactive) 100 mM
!V!annosyigSy cerate Formulation
Inqredient % by weight [
Etanercept (active ingredient) 50 rng/m!
Mannosyiglycerate (inactive ingredient) 4% (w/v) EXAMPLE 2
Preparation of Etanercept
STEP 1. Cell Expansion. In a manner known in the art, cell expansion necessary to generate a sufficient number of cells for inoculation of a production bioreactor is performed using a clone of CHO celis expressing the etanercept fusion protein. The product of this expression process (a harvested cell culture fluid) results in a mixture of correctly folded etanercept, as well as incorrectly folded and/or aggregated etanercept, along with additional impurities. The harvested cell culture fluid comprising such protein mixture is subjected to detergent viral inactivation.
STEP 2. Affinity Chromatography. Affinity chromatography is performed on the harvested cell culture obtain in Step 1 above using a conventional Protein A affinity column in a well known manner. Product recovery is approximately 85%. The product obtained is a complex protein mixture comprising correctl folded etanercept, incorrectly folded etanercept, and/or aggregates of correctly and/or Incorrectly folded etanercept, or protein fragments. The product obtained from this Protein A affinity column purification step is adjusted to pH 3,5 and then subjected to a viral inactivation step. Following viral inactivation the product is adjusted to pH 5.5 and then further clarified in a known manner using a commercially obtained capsule filter.
STEP 3A. Mixed-Mode Cation Exchange Chromatography. A 31.8 L
(45 cm diameter X 20 cm bed height) packed bed GE Healthcare Capto M C chromatograph column is used to purify the product obtained in Step 2 above. Prior to use, the column is equilibrated with 2 CV of 25 m acetate pH 5.5 and sanitized with 2 CV of 0.1 N NaOH, 1 NaCS and neutralized with 2 CV of 25 mM acetate, 0.7 M NaCI, pH 5.5. The column is then equilibrated with 8-10 CV of 25 mM acetate pH 5.5 until the effluent is pH 5.5 and 3,5 mS/cm. The Protein A pool from step 2 above is diluted to < 8 mS/cm with WFI and applied to a column loading of up to 15 g/L media for each cycle. The column is operated at a linear velocity of 200 cm/h to give a 6 minute residence time. After loading, the column is washed with 2 CV of 25 mM acetate pH 5.5. The product is then eluted with an 8.5 CV, 15% to 85% gradient of 25 mM acetate pH 5.5 to 25 mM acetate, 0.7 U NaCi, pH 5.5. Product collection begins at 0.15 OD (A280, 1.0 cm path length) and collection ends at 50% of peak maximum. The eiuate volume is approximately 5 CV. Residual product and contaminants are stripped from the column with 2 CV of 10 mM Tris, 1 M NaC!, pH 8.0 and discarded. The product obtained from the mixed mode column is filtered using a illipore Opticap XL10, 0.22 urn Durapore capsule filter, (0.69 m2). The product obtained from this step represents a recovery of about 70% of the Protein A material obtained in Step 2
STEP 3B. Mixed Mode Anion Exchange Chromatopgraphy. A 27.0 L (45 cm diameter X 17 cm bed height) packed bed GE Healthcare Capto Adhere chromatography column is used to further purify the product obtained in step 3A above. Prior to use, the column is equilibrated with 2 CV of 25 mM Tris, pH 8.0 and sanitized with 2 CV/ of 0.1 N NaOH, 1M NaCI and neutralized and equilibrated with 2 CV of 25 mM Tris, pH 8.0. Prior to product loading, the column is equilibrated with 3 CV of 10 mM Tris, pH 8.0. The Capto MMC pool from Step 3A above is adjusted to pH 8.1 with -0.045 kg of 1 M Tris, pH 8.3 per kg of pool. The product from Step 3A above was diluted in-line 1 :3.8 with F! to adjust the conductivity to 12.0 mS/cm and pH 8.0. The resulting material is then applied to a column loading of up to 15 g/L media. The column is operated at a linear velocity of 170 cm/h to give a 6 minute residence time. After loading, the column is washed with 2 CV of 25 mM Tris, pH 8.0. The product is then eluted with a 10 CV gradient (20% to 90%) of 25 mM Tris, pH 8.0 to 10 mM Tris, 1 M NaCI, pH 8.0. Product collection is started at 0.15 OD (A280, 1.0 cm path length) and collection ended at 25% of peak maximum. The eiuate volume is 4-6 CV. The eiuted product is filtered using a commercially available capsule filter and then subjected in a known manner to viral filtration and tangential flow filtration steps. Overall product recovery from step3B (including the final viral and tangential flow filtration steps) was approximately 68%. Product recovery measured before the filtration steps was about 75%. A schematic representation of HIC data obtained on eluation fractions from this step are representing in Figure 12.
Analysis: The final filtered product obtained in this example is found to have greater than about 90 wt % correctly folded etanercept as determined by HIC: less than 5 wt% incorrectly folded etanercept species as determined by HIC; less than about 3 wt% of clipped material by HIC analysis (beiieved to be fragments of etanercept in which the TNFR portion thereof has been truncated) and a combined amount of correctly and incorrectly folded etanercept of greater than 95 wt% as determined by size exclusion chromatography,
Analysis of Etanercept Formulations A. Thermal Stability Storage
Following dialysis and concentration, samples of the etanercept formulations exemplified above were sterile filtered in a bio safety cabinet. Using sterilized pipettes and autoclaved pipette tips, sampies of the etanercept formulations were transferred to pre-labeled and autoclaved 1 mL lyophiiization vials. Vials were stoppered with sterile butyl stoppers and crimped with aluminum caps. All vials were then transferred to thermal stability ovens. Samples were subject to two thermal stability regimes: (1) two weeks at 40 and (2) four weeks at 25 °C. Throughout this specification, these two temperature regimes are denoted "T and T4," respectively.
B. Size Exclusion Chromatography (SEC)
Etanercept formulations disclosed herein were analyzed using the well known technique of Size Exclusion Chromatography (SEC), a high-performance liquid chromatography method in which anaiytes are separated by size (see Rogner, , (2000). Size Exclusion Chromatography. Protein Liquid Chromatography. M. Kastner. Amsterdam, Elsevier. 61 : 89-145.). I order to evaluate thermal stability of the Etanercept samples decribed above, the samples were examined by a SEC method based on the literature (van Maarscha!keaveerd, A., G. J. Wolbink, et al. (201 1). "Comparison of analytical methods to detect instability of etanercept during thermal stress testing." European Journal of Pharmaceutics and Biopharmaceutics 78(2): 213-221.) The mobile phase buffer was prepared to contain 50 mM sodium phosphate monobasic monohydrate and 150 mM arginine. The pH was adjusted to 8.5 using 1 M HCI. All separations were performed using a Tosoh TSK-Gel SWxl 6 mm x 4 cm guard column (cat. no. 8543) attached linearly to a Tosoh TSK-Gel G4000 SWxl 7.8 mm x 30 cm (cat. no. 8542). To perform a separation, the columns were brought to room temperature (23°C) and equilibrated with mobiie phase at a fiow rate of 0,5 mL/min. 5 microliters of 50 mg/mL etanercept formulation were injected onto the column using an autosamp!er. The separation was accomplished over 30 minutes at a flow rate of 0.5 mlJminute. Column eluent was monitored at a wavelength of 280 nm during this time.
C, Integration of Size Exclusion Chromatography Chromatograms
AH integration was performed using Chromeieon software (Dionex). Prior to integration, the SEC chromatogram for a buffer containing no etanercept was subtracted from all chromatograms. A!l integration was performed between retention times of 12 minutes and 26 minutes. Several parameters were used to define a peak. The minimum area for a detected peak was set to 0.05 mAu * min. The two- dimensional sensitivity for peak defection was set to 0.01 mAu and 75 seconds. Peak shoulders were added manually using a manual integration too!. Ail detected peaks were manually adjusted in two steps. First, peak baselines (the bottom boundary of the peak) were adjusted to horizontal. Secondly, the vertical positions of the peak baselines were adjusted to that of the chromatogram baseiine. The chromatogram baseiine value was defined as the signal in absence of ana!yte. The signal i absence of analyte was defined as the absorbance in mAu at 12 minutes retention time.
D. SEC Fractions of Etanercept Formulations
in the SEC analysis of etanercept formulations described above, three SEC chromatography fractions were identified and studied. The fractions that were analyzed were, in the order of elutlon from the SEC column: (1) a high moiecular weight fraction representing aggregates of the intact etanercept TNFR:FC fusion protein likely assembled via non-covaient electrostatic attraction among intact etanercept molecules (hereinafter "aggregate(s)"' or aggregate(s) content); (2) monomer content, representing the intact etanercept TNFR:Fc fusion protein (hereinafter referred to as "monomer" of "monomer content"); (3) a fraction likely representing one fragment or a population of fragments of the etanercept moiecu!e in which one portion of the TNFR:molecuie fusion protein has become cleaved from the monomer; in the ioss of an arm of the Fab portion of the fusion protein at the hinge region of the moiecule. The most common fragment or clipped species, as measured by SEC, is referred to as Fragment 3. in conducting the SEC analysis, it will be observed that aggregates e!ute first, followed by monomer, foliowed by fragment 3.
The foiiowing tables shows the relative amounts of Aggregates, Monomer and Fragment 3 determined by SEC analysis as described above.
TABLE 1
SEC ANALYSIS OF MONOMER
Note: Amounts reported Tables I, II and III are percentages by weight
T0 = formulation maintained at 5 C and analyzed within 24 hours of creation.
Ti = formulation stored for one week at 40° C
T2 - formulation stored for two weeks at 40 C
Figure imgf000028_0001
TABLE U
SEC ANALYSIS OF AGGREGATES
Figure imgf000029_0001
HIC ANALYSIS OF ETANERCEPT FORMULATIONS
HSC chromatography may be carried out in a manner known in the art and generally described in U.S. Patent 7,294,481 , incorporated herein by reference, Samples are evaluated at t0 (within 24 hours of preparation at 5°C.) and again after either two weeks of storage at 25°C. (t2) or after 4 weeks of storage at 25°C. In the HIC chromatograms of the formulations of the present invention, Peak 1 in the H!C chro.matogram is believed to be or include "Fragment 3", which is identified and quantified using SEC, as referenced above in the discussion of SEC data; Peak 2 is etanercept monomer as referenced above in the discussion of SEC data; and Peak 3 includes "Aggregate(s)" as referenced above in the discussion of SEC data. It should further be understood that the terms "peak 1", "peak 2" and "peak 3 as used here also constitute a reference to the HiC peak 1 , peak 2 and peak 3 referred to and disclosed in Figure 4 of U.S. patent 7,294,481 incorporated herein by reference. Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being Indicated by the following claims.

Claims

WHAT IS CLAIMED IS:
1. An aqueous stabilized formuiation of etanercept comprising: etanercept; and stabilizing ingredients to retard instability, aggregatio and/or fragmentation of the etanercept in the formulation, said stabilizing ingredients being comprised of at least one of the following: (a) meglumine; or (b) meglumine in combination with sucrose; or (c) meglumine in combination with sodium chloride; or (d) meglumine in combination with sodium chloride and sucrose.
2. The formulation of claim 1 , further comprising one or more additional components selected from a buffer, a tonicity modifier and an excipient.
3. The formulation of claim 1 wherein the stabilizing ingredient consists essentially of at least one of the following: (a) meglumine; or (b) meglumine in combination with sucrose; or (c) meglumine in combination with sodium chloride; or (d) meglumine in combination with sodium chloride and sucrose; and wherein, the formulation, optionally, is free or essentially free of arginine, and the formulation elicits long term storage stability as characterized by at least one of:
SEC analysis at M3 or T2 or T4 of: monomer content greater than about 90%; aggregates content of less than about 3 wt%; and fragment 3 content less than about 5 wt%: and
HIC analysis at M3 or T2 or T4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 3 wt.%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than 80 wt.%; and the amount of the composition represented by peak 3 of the HiC chromatogram is less than about 20 wt.%.
4. The formulation of claim 2, optionally free or essentially free of arginine, comprising about 25 to about 75 mg/mS etanercept, about 1-30 mlV! of sodium phosphate; up to about 10% meglumine; optionally up to about 5 wt.% sucrose; and optionally up to about 100 mM sodium chloride, wherein the composition has a pH of about 6.0 to 6.6; and wherein the composition is characterized by SEC analysis at M3 or T2 or T4 in which: the monomer content is greater than about 85 wt.%; aggregate(s) content is less than about 3 wt.%; and fragment 3 content is less than about 8 wt.%.
5. The composition of claim 4 eliciting long term storage stability characterized by:
(a) an SEC analysis at M3 or T2 or T4 of greater than about 90 wt.% monomer content; and less than about 3 wt.% aggregate(s) content; and
■(b) an HIC analysis at 3 or T2 or T wherein the amount of the composition represented by peak 1 of the HIC chromatogram is iess than about 4 wt.%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than about 80 wt.%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 20 wt.%; and
6. The formulation of claim 3 having an HIC analysis at 3 or T2 or T4 wherein the amount of the composition represented by peak 1 of the HIC chromatogram is less than about 1 wt.%; the amount of the composition represented by peak 2 of the HIC chromatogram is greater than about 95 wt.%; and the amount of the composition represented by peak 3 of the HIC chromatogram is less than about 3 wt.%.
7. The formulation of claim 3 comprising: about 50 mg/ml etanercept; about 0.5 wt.% meglumine; about 10 to 30 mM phosphate; about 1% sucrose; and about 100 mM sodium chloride.
8. The formulation of claim 3 comprising: about 50 mg/ml etanercept; about 5 wt.% meglumine; about 10 to 30 mM phosphate; and free or essentially free of sucrose and sodium chloride.
9. The formulation of claim 3 eliciting long term storage stabilit as characterized by: an HIC analysis at 3 or T2 or T4 wherein the amount of the composition represented by peak 2 of the HIC chromatogram is greater than or equal to about 95 wt.%; and wherein, if peak 3 is present on the H!C chromatogram, the amount of the composition represented by peak 3 is less than or equal to about 1 wt.%.
10, The formulation of claim 3 having, at 3 or T2 or T4 no more than, on average, about 10,000 subvisible particles per rrsL having a size greater than 5pm.
11. The formulation of claim 1 , containing no arginine, or essentiall free of arginine, wherein the composition, at M3 o T2 or T4, elicits long term storage stability that meets one or both of the following criteria:
(A) stability comparable to or better than commercially available etanercept marketed under the trademark Enbrei©, as measured by (i) SEC analysis of the amounts of aggregate(s), monomer and fragment 3 in the composition (as defined in the specification) and (ii) H!C analysis of amounts of materia! in the composition corresponding to peaks 1 , 2 and 3 of the HIC chromatogram (as defined in the specification); and
■(B) an H!C chromatogram in which (i) peak 3 is absent, or essentially absent and (ii) peak 2 represents greater than about 95 wt% of the composition; an SEC chromatogram containing essentially no peak corresponding to aggregate(s); and an SEC chromatogram in which the monomer content represents at least about 95 wt% of the composition.
PCT/US2012/060743 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine Ceased WO2013059408A1 (en)

Priority Applications (12)

Application Number Priority Date Filing Date Title
JP2014537221A JP6199298B2 (en) 2011-10-18 2012-10-18 Etanercept formulation stabilized by meglumine
CN201280062761.9A CN104010658A (en) 2011-10-18 2012-10-18 Etanercept Formulations Stabilized With Meglumine
EA201490802A EA028520B1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine
AU2012326080A AU2012326080B2 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine
EP12841505.6A EP2768531A4 (en) 2011-10-18 2012-10-18 MEGANLUMIN STABILIZED STERILE FORMANTIONS
HK15101258.8A HK1200722A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine
CA2851646A CA2851646A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine
KR1020147013301A KR20140091705A (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine
SG11201401567YA SG11201401567YA (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine
BR112014009131A BR112014009131A8 (en) 2011-10-18 2012-10-18 meglumine stabilized etanercept formulations
MX2014004732A MX2014004732A (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine.
IL231827A IL231827A0 (en) 2011-10-18 2014-03-31 Etanercept formulations stabilized with meglumine

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201161548518P 2011-10-18 2011-10-18
US61/548,518 2011-10-18
US201261669480P 2012-07-09 2012-07-09
US61/669,480 2012-07-09

Publications (1)

Publication Number Publication Date
WO2013059408A1 true WO2013059408A1 (en) 2013-04-25

Family

ID=48136153

Family Applications (6)

Application Number Title Priority Date Filing Date
PCT/US2012/060743 Ceased WO2013059408A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with meglumine
PCT/US2012/060739 Ceased WO2013059406A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with metal ions
PCT/US2012/060748 Ceased WO2013059412A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with combinations of sugars and polyols
PCT/US2012/060745 Ceased WO2013059410A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with xylitol
PCT/US2012/060738 Ceased WO2013059405A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with amino acids
PCT/US2012/060741 Ceased WO2013059407A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with sodium chloride

Family Applications After (5)

Application Number Title Priority Date Filing Date
PCT/US2012/060739 Ceased WO2013059406A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with metal ions
PCT/US2012/060748 Ceased WO2013059412A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with combinations of sugars and polyols
PCT/US2012/060745 Ceased WO2013059410A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with xylitol
PCT/US2012/060738 Ceased WO2013059405A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with amino acids
PCT/US2012/060741 Ceased WO2013059407A1 (en) 2011-10-18 2012-10-18 Etanercept formulations stabilized with sodium chloride

Country Status (28)

Country Link
US (21) US10293049B2 (en)
EP (6) EP2768532A4 (en)
JP (6) JP6220788B2 (en)
KR (7) KR20140091707A (en)
CN (6) CN104010657A (en)
AR (6) AR088383A1 (en)
AU (6) AU2012326168B2 (en)
BR (6) BR112014009031A2 (en)
CA (6) CA2851628A1 (en)
CY (1) CY1121843T1 (en)
DK (1) DK2768525T3 (en)
EA (6) EA026410B1 (en)
ES (1) ES2734070T3 (en)
HK (3) HK1200718A1 (en)
HR (1) HRP20191215T1 (en)
HU (1) HUE045624T2 (en)
IL (6) IL231824A0 (en)
IN (3) IN2014CN02527A (en)
LT (1) LT2768525T (en)
MX (7) MX2014004734A (en)
PL (1) PL2768525T3 (en)
PT (1) PT2768525T (en)
RS (1) RS59179B1 (en)
SG (6) SG11201401576WA (en)
SI (1) SI2768525T1 (en)
SM (1) SMT201900422T1 (en)
TW (6) TWI619504B (en)
WO (6) WO2013059408A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015061584A1 (en) * 2013-10-24 2015-04-30 Medimmune, Llc Stable, aqueous antibody formulations

Families Citing this family (33)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008543839A (en) 2005-06-14 2008-12-04 アムジェン インコーポレーテッド Self-buffering protein formulation
US10493151B2 (en) 2011-10-18 2019-12-03 Coherus Biosciences, Inc. Etanercept formulations stabilized with sodium chloride
HK1200718A1 (en) 2011-10-18 2015-08-14 科荣生生物科学公司 Etanercept formulations stabilized with sodium chloride
WO2014011672A1 (en) * 2012-07-09 2014-01-16 Coherus Biosciences, Inc. Etanercept formulations exhibiting marked reduction in sub-visible particles
PE20150996A1 (en) * 2012-09-11 2015-08-01 Coherus Biosciences Inc PROPERLY FOLDED ETANERCEPT FOR HIGH PURITY AND EXCELLENT PERFORMANCE
CN105873601A (en) * 2013-05-02 2016-08-17 马柏克辛斯公司 Alternative formulations for TNFR: Fc fusion polypeptides
WO2015056613A1 (en) * 2013-10-15 2015-04-23 Ｍｅｉｊｉ Ｓｅｉｋａファルマ株式会社 Stabilized polypeptide aqueous preparation
CN106061468B (en) * 2013-11-29 2020-10-02 阿雷斯贸易股份有限公司 Liquid formulations of fusion proteins comprising TNFR and Fc regions
IS3008B (en) 2014-05-14 2018-12-15 Calor ehf A peptide or protein stabilizing formulation
US20160074515A1 (en) 2014-06-20 2016-03-17 Reform Biologics, Llc Viscosity-reducing excipient compounds for protein formulations
US11357857B2 (en) 2014-06-20 2022-06-14 Comera Life Sciences, Inc. Excipient compounds for protein processing
US10478498B2 (en) 2014-06-20 2019-11-19 Reform Biologics, Llc Excipient compounds for biopolymer formulations
WO2016080367A1 (en) * 2014-11-18 2016-05-26 塩野義製薬株式会社 Stable peptide composition
WO2016102328A1 (en) * 2014-12-22 2016-06-30 Ares Trading S.A. Liquid pharmaceutical composition
WO2016103034A1 (en) * 2014-12-23 2016-06-30 Drug Discovery Laboratory As Protein compositions and use thereof
BR112017014224A2 (en) * 2014-12-31 2018-03-06 Lg Chem, Ltd. method for the preparation of fc-tnfr fusion protein containing target content of impurities
CN104694355B (en) * 2015-03-20 2016-09-28 吉林大学 Radix Puerariae liver-protection health-care wine and preparation method thereof
MX389818B (en) * 2015-08-13 2025-03-20 Landsteiner Scient S A De C V ENHANCED STABILITY COMPOSITION OF ETANERCEPT.
JP5938762B1 (en) * 2015-09-01 2016-06-22 日揮株式会社 Microcapsule preparation and production method thereof
CN105748414A (en) * 2016-03-02 2016-07-13 张光泉 Anti-infection micafungin freeze-drying composition and preparation method thereof
WO2017179683A1 (en) * 2016-04-15 2017-10-19 Ｍｅｉｊｉ Ｓｅｉｋａファルマ株式会社 Pharmaceutical composition containing micafungin in stabilized state
WO2018075818A1 (en) * 2016-10-21 2018-04-26 Amgen Inc. Pharmaceutical formulations and methods of making the same
WO2018080196A2 (en) * 2016-10-28 2018-05-03 (주)셀트리온 Stable pharmaceutical formulation
EP3558363A1 (en) 2016-12-21 2019-10-30 Amgen Inc. Anti-tnf alpha antibody formulations
MX2017013995A (en) * 2017-10-31 2019-05-01 Probiomed S A De C V Stable pharmaceutical formulation of a fusion protein.
BR112020010761A2 (en) * 2017-11-30 2020-11-24 Bio-Thera Solutions, Ltd. liquid formulation of humanized antibody for the treatment of il-6 related diseases
CA3189025A1 (en) * 2017-12-22 2019-06-27 Samsung Bioepis Co., Ltd. Liquid composition comprising vegf antagonist
US11253569B2 (en) 2018-05-03 2022-02-22 Seattle Children's Hospital Methods of treating Kawasaki Disease
CN111228225B (en) * 2018-11-28 2022-08-26 鲁南制药集团股份有限公司 Recombinant human tumor necrosis factor receptor-Fc fusion protein freeze-dried preparation
GB201911461D0 (en) 2019-08-09 2019-09-25 Arecor Ltd Novel composition
JP2023502777A (en) * 2019-11-26 2023-01-25 コメラ ライフ サイエンシズ，インコーポレイテッド Excipient compounds for biopolymer formulations
KR20230146579A (en) 2021-02-17 2023-10-19 아레콜 리미티드 Aqueous compositions of engineered protein constructs comprising an Fc domain
CN114788809B (en) * 2022-01-25 2023-04-14 江苏广承药业有限公司 Loratadine liquid preparation

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5580856A (en) 1994-07-15 1996-12-03 Prestrelski; Steven J. Formulation of a reconstituted protein, and method and kit for the production thereof
WO2000062790A2 (en) 1999-04-19 2000-10-26 Immunex Corporation Soluble tumor necrosis factor receptor treatment of medical disorders
US6171586B1 (en) 1997-06-13 2001-01-09 Genentech, Inc. Antibody formulation
WO2001062272A2 (en) 2000-02-25 2001-08-30 Immunex Corporation Soluble tumor necrosis factor receptor and il-4 inhibitor for the treatment of medical disorders
US20030180287A1 (en) * 2002-02-27 2003-09-25 Immunex Corporation Polypeptide formulation
US7294481B1 (en) 1999-01-05 2007-11-13 Immunex Corporation Method for producing recombinant proteins
US20080311119A1 (en) * 2007-06-14 2008-12-18 Biogen Idec Ma Inc. Antibody formulations

Family Cites Families (58)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5605690A (en) 1989-09-05 1997-02-25 Immunex Corporation Methods of lowering active TNF-α levels in mammals using tumor necrosis factor receptor
IT1240314B (en) 1989-09-28 1993-12-07 Immunobiology Research Institutes, Inc. STABILIZED AQUEOUS FORMULATIONS OF SMALL PEPTIDES.
KR100232688B1 (en) 1992-09-15 1999-12-01 스코트 쥐. 홀퀴스트 Pharmaceutical composition for treating TNF-dependent inflammation containing tumor necrosis factor antagonist
US5656730A (en) 1995-04-07 1997-08-12 Enzon, Inc. Stabilized monomeric protein compositions
EP0852951A1 (en) 1996-11-19 1998-07-15 Roche Diagnostics GmbH Stable lyophilized monoclonal or polyclonal antibodies containing pharmaceuticals
US6677326B2 (en) * 1999-03-15 2004-01-13 Arakis, Ltd. Corticosteroid formulation comprising less than 2.5 mg prednisolone for once daily administration
US20040220103A1 (en) * 1999-04-19 2004-11-04 Immunex Corporation Soluble tumor necrosis factor receptor treatment of medical disorders
WO2001058473A1 (en) 2000-02-10 2001-08-16 Wyeth Method of treating or inhibiting cellular injury or cell death
EP2311492B1 (en) 2000-08-11 2017-10-04 Chugai Seiyaku Kabushiki Kaisha Antibody-containing stabilized preparations
WO2003102013A2 (en) * 2001-02-23 2003-12-11 Gonzalez-Villasenor Lucia Iren Methods and compositions for production of recombinant peptides
RS51041B (en) 2003-02-28 2010-10-31 Ares Trading S.A. LIQUID FORMULATIONS OF TBP-1 PROTEIN BINDING TUMOR NECESSITY FACTOR
JPWO2004082715A1 (en) 2003-03-20 2006-06-22 エーザイ株式会社 Concomitant medications as therapeutic agents for inflammatory bowel disease
CN100509847C (en) 2003-08-01 2009-07-08 安美基公司 Crystalline tumor necrosis factor receptor 2 polypeptides
TW201245229A (en) 2003-10-14 2012-11-16 Hoffmann La Roche Macrocyclic carboxylic acids and acylsulfonamides as inhibitors of HCV replication
JP5554466B2 (en) 2004-03-01 2014-07-23 味の素株式会社 Anti-human TNF-α antibody activity decrease inhibitor
AU2005229359C1 (en) 2004-03-05 2011-07-28 Patheon Holdings I B.V. Process for cell culturing by continuous perfusion and alternating tangential flow
US20070196364A1 (en) 2004-07-27 2007-08-23 Human Genome Sciences, Inc. Pharmaceutical Formulation and Process
US7597884B2 (en) * 2004-08-09 2009-10-06 Alios Biopharma, Inc. Hyperglycosylated polypeptide variants and methods of use
US7300773B2 (en) 2004-08-27 2007-11-27 Wyeth Research Ireland Limited Production of TNFR-Ig
JP5339901B2 (en) 2005-05-10 2013-11-13 バイオジェン・アイデック・エムエイ・インコーポレイテッド Treatment and evaluation of inflammatory injury
WO2006132363A1 (en) * 2005-06-10 2006-12-14 Chugai Seiyaku Kabushiki Kaisha Stabilizer for protein preparation comprising meglumine and use thereof
JP2008543839A (en) * 2005-06-14 2008-12-04 アムジェン インコーポレーテッド Self-buffering protein formulation
KR101378194B1 (en) * 2005-12-20 2014-03-27 브리스톨-마이어스 스큅 컴퍼니 Stable protein formulations
WO2007076062A2 (en) 2005-12-21 2007-07-05 Wyeth Protein formulations with reduced viscosity and uses thereof
WO2007092772A2 (en) 2006-02-03 2007-08-16 Medimmune, Inc. Protein formulations
SG170750A1 (en) 2006-03-17 2011-05-30 Biogen Idec Inc Stabilized polypeptide compositions
EP1996076A2 (en) 2006-03-21 2008-12-03 Eugen Oetringer Devices for and methods of analyzing a physiological condition of a physiological subject based on a workload related property
US20080003220A1 (en) 2006-04-21 2008-01-03 Amgen, Inc. Buffering agents for biopharmaceutical formulations
CN103540575A (en) * 2006-08-01 2014-01-29 奇尼塔二有限责任公司 Pharmaceutical manufacturing methods
TW200831129A (en) * 2006-10-06 2008-08-01 Amgen Inc Stable formulations
TW200833357A (en) 2006-10-20 2008-08-16 Amgen Inc Stable polypeptide formulations
ES2541546T3 (en) 2006-11-03 2015-07-21 Wyeth Llc Substances that inhibit glycolysis in cell culture
EP2129401B8 (en) 2006-12-21 2020-01-15 Amgen Inc. Stable buffered formulations containing polypeptides
HUE026403T2 (en) 2007-03-02 2016-06-28 Wyeth Llc Use of copper and glutamate in cell culture for production of polypeptides
EP2014760A1 (en) 2007-06-13 2009-01-14 CMC Biopharmaceuticals A/S A method for producing a biopolymer (e.g. polypeptide) in a continuous fermentation process
TWI543768B (en) * 2007-11-30 2016-08-01 艾伯維生物技術有限責任公司 Protein formulation and method of making same
SG2013054218A (en) * 2008-01-15 2014-10-30 Abbott Gmbh & Co Kg Powdered protein compositions and methods of making same
JP2011514895A (en) 2008-02-29 2011-05-12 バイオジェン・アイデック・エムエイ・インコーポレイテッド Purified immunoglobulin fusion protein and purification method thereof
WO2011015926A1 (en) 2009-08-03 2011-02-10 Avesthagen Limited A process of fermentation, purification and production of recombinant soluble tumour necrosis factor alfa receptor (tnfr) - human igg fc fusion protein
SI2464725T2 (en) 2009-08-11 2025-06-30 F. Hoffmann-La Roche Ag Production of proteins in glutamine-free cell culture media
US20120208986A1 (en) 2009-10-20 2012-08-16 Wenger Marc D Use of mixed mode chromatography for the capture and purification of basic antibody products
WO2011079308A2 (en) * 2009-12-23 2011-06-30 Emergent Product Development Seattle, Llc Compositions comprising tnf-alpha and il-6 antagonists and methods of use thereof
RU2644651C2 (en) 2010-04-26 2018-02-13 Новартис Аг Medium for cells cultivation
WO2011141926A2 (en) * 2010-05-10 2011-11-17 Intas Biopharmaceuticals Limited Liquid formulation of polypeptides containing an fc domain of an immunoglobulin
WO2012013980A1 (en) * 2010-07-30 2012-02-02 Arecor Limited Stabilized aqueous antibody compositions
CA2807607A1 (en) 2010-08-20 2012-02-23 Wyeth Llc Cell culture of growth factor-free adapted cells
JP2013536683A (en) 2010-08-31 2013-09-26 フリエスランド ブランズ ビー．ブイ． Culture medium for eukaryotic cells
MX344268B (en) 2010-10-11 2016-12-09 Abbvie Inc * Processes for purification of proteins.
WO2012143418A1 (en) * 2011-04-20 2012-10-26 Sandoz Ag STABLE PHARMACEUTICAL LIQUID FORMULATIONS OF THE FUSION PROTEIN TNFR:Fc
UY34105A (en) * 2011-06-03 2012-07-31 Lg Life Sciences Ltd STABLE LIQUID FORMULATION OF ETANERCEPT
US20140142055A1 (en) 2011-06-29 2014-05-22 Insite Vision Corporation Methods of treating recurrent meibomian glands disorder and thereby decreasing the frequency of recurrence
IL288509B2 (en) 2011-07-01 2023-04-01 Amgen Inc Mammalian cell culture
AU2012279205B2 (en) 2011-07-01 2017-08-31 Biogen Ma Inc. Arginine - free TNFR : Fc- fusion polypeptide compositions and methods of use
US9781540B2 (en) * 2011-07-07 2017-10-03 Qualcomm Incorporated Application relevance determination based on social context
BR112014000352A2 (en) 2011-07-08 2017-02-14 Merck Sharp & Dohe Corp method of purifying a fc-fc protein, fc proteins containing purified fc, purified tnfr: fc, and highly purified tnfr: fc
EP2753634B1 (en) 2011-08-17 2017-11-01 Ares Trading S.A. Method for preparing active form of tnfr-fc fusion protein
HK1200718A1 (en) * 2011-10-18 2015-08-14 科荣生生物科学公司 Etanercept formulations stabilized with sodium chloride
PE20150996A1 (en) 2012-09-11 2015-08-01 Coherus Biosciences Inc PROPERLY FOLDED ETANERCEPT FOR HIGH PURITY AND EXCELLENT PERFORMANCE

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5580856A (en) 1994-07-15 1996-12-03 Prestrelski; Steven J. Formulation of a reconstituted protein, and method and kit for the production thereof
US6171586B1 (en) 1997-06-13 2001-01-09 Genentech, Inc. Antibody formulation
US7294481B1 (en) 1999-01-05 2007-11-13 Immunex Corporation Method for producing recombinant proteins
WO2000062790A2 (en) 1999-04-19 2000-10-26 Immunex Corporation Soluble tumor necrosis factor receptor treatment of medical disorders
US20010021380A1 (en) 1999-04-19 2001-09-13 Pluenneke John D. Soluble tumor necrosis factor receptor treatment of medical disorders
WO2001062272A2 (en) 2000-02-25 2001-08-30 Immunex Corporation Soluble tumor necrosis factor receptor and il-4 inhibitor for the treatment of medical disorders
US20030180287A1 (en) * 2002-02-27 2003-09-25 Immunex Corporation Polypeptide formulation
US7648702B2 (en) 2002-02-27 2010-01-19 Immunex Corporation Stable aqueous formulation of a soluble TNF receptor and arginine
US20080311119A1 (en) * 2007-06-14 2008-12-18 Biogen Idec Ma Inc. Antibody formulations

Non-Patent Citations (18)

* Cited by examiner, † Cited by third party
Title
"Physicians Desk Reference", 2002, MEDICAL ECONOMICS COMPANY INC.
AUSUBEL ET AL.,: "Current Protocols in Molecular Biology", 1990, WILEY
BORGES ET AL., EXTREMOPHILES, vol. 6, 2002, pages 209 - 216
CLELAND ET AL., CRIT. REV. THERAPEUTIC DRUG CARRIER SYSTEMS, vol. 10, 1993, pages 307 - 377
FARIA ET AL., CARBOHYDRATE RES., vol. 343, 2008, pages 3025 - 3033
FARIA ET AL., CHEMBIOCHEM, vol. 4, 2003, pages 734 - 741
HAWE ET AL., PHARM. RES., vol. 28, 2011, pages 2302
HORA ET AL., PHARM. RES., vol. 9, 1992, pages 33 - 36
LANGER, SCIENCE, vol. 249, 1990, pages 1527 - 1533
LIU ET AL., BIOTECHNOL. BIOENG., vol. 37, 1991, pages 177 - 184
M. KASTNER.: "Protein Liquid Chromatography", vol. 61, 2000, ELSEVIER, article ROGNER, M.: "Size Exclusion Chromatography", pages: 89 - 145
MANNING ET AL., PHARM. RES., vol. 6, 1989, pages 903 - 918
PAIS ET AL., J. MOL. BIOL., vol. 394, 2009, pages 237 - 250
ROBBINS ET AL., DIABETES, vol. 36, 1987, pages 838 - 845
SAWANGWAN ET AL., BIOTECHNOL. J., vol. 5, 2010, pages 187 - 191
TELLEZ C.M. ET AL: "Method for characterization of size-exclusion chromatography media for preparative purification of DNA restriction fragments", BIOTECHNOLOGY TECHNIQUES, vol. 13, June 1999 (1999-06-01), pages 395 - 401, XP003031139, Retrieved from the Internet <URL:http://link.springer.com/article/10.1023%2FA%3A1008932407733?LI=true> [retrieved on 20121203] *
VAN MAARSCHALKERWEERD, A.; G. J. WOLBINK ET AL.: "Comparison of analytical methods to detect instability of etanercept during thermal stress testing", EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS, vol. 78, no. 2, 2011, pages 213 - 221, XP028203395, DOI: doi:10.1016/j.ejpb.2011.01.012
VAN MARRSCHALKERWEERD ET AL., EUR. J. PHARM. BIOPHARM., vol. 78, 2011, pages 213

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015061584A1 (en) * 2013-10-24 2015-04-30 Medimmune, Llc Stable, aqueous antibody formulations
AU2014339984B2 (en) * 2013-10-24 2020-04-09 Astrazeneca Ab Stable, aqueous antibody formulations

Also Published As

Publication number Publication date
IL231828A0 (en) 2014-05-28
TW201325607A (en) 2013-07-01
EA201490802A1 (en) 2014-08-29
TW201325608A (en) 2013-07-01
CN103998061A (en) 2014-08-20
JP6110393B2 (en) 2017-04-05
SG11201401517VA (en) 2014-09-26
AU2012326168B2 (en) 2016-12-15
AU2012326084A1 (en) 2014-04-17
EP2768531A1 (en) 2014-08-27
US9770510B2 (en) 2017-09-26
AR088460A1 (en) 2014-06-11
MX2014004725A (en) 2015-02-05
AU2012326170A1 (en) 2014-04-24
EP2768533A4 (en) 2015-03-11
HK1200720A1 (en) 2015-08-14
CN104011073B (en) 2017-08-25
EA201490817A1 (en) 2014-09-30
AU2012326082A1 (en) 2014-04-24
US9943601B2 (en) 2018-04-17
WO2013059412A1 (en) 2013-04-25
DK2768525T3 (en) 2019-07-22
CA2851639A1 (en) 2013-04-25
IL231829A0 (en) 2014-05-28
AU2012326171A1 (en) 2014-04-24
JP2014530255A (en) 2014-11-17
US20190216930A1 (en) 2019-07-18
BR112014009087A2 (en) 2017-04-18
EA201490815A1 (en) 2014-09-30
US20190290766A1 (en) 2019-09-26
LT2768525T (en) 2019-09-25
CN104010654A (en) 2014-08-27
EP2768535A4 (en) 2015-03-11
AR088380A1 (en) 2014-05-28
US20160317667A1 (en) 2016-11-03
EA025267B1 (en) 2016-12-30
US10980885B2 (en) 2021-04-20
EP2768854A1 (en) 2014-08-27
HK1200721A1 (en) 2015-08-14
EP2768535A1 (en) 2014-08-27
IL231824A0 (en) 2014-05-28
AR088383A1 (en) 2014-05-28
CN104010658A (en) 2014-08-27
MX2014004733A (en) 2015-05-15
JP2014530864A (en) 2014-11-20
IN2014CN02592A (en) 2015-09-04
HK1200851A1 (en) 2015-08-14
US20190290765A1 (en) 2019-09-26
US20190314499A1 (en) 2019-10-17
HK1200719A1 (en) 2015-08-14
IN2014CN02591A (en) 2015-06-26
EA025663B1 (en) 2017-01-30
US20180193463A1 (en) 2018-07-12
US20180125982A1 (en) 2018-05-10
EA028520B1 (en) 2017-11-30
CA2851635A1 (en) 2013-04-25
US20190328875A1 (en) 2019-10-31
HK1200722A1 (en) 2015-08-14
JP6199298B2 (en) 2017-09-20
BR112014009131A2 (en) 2017-06-13
CA2851646A1 (en) 2013-04-25
JP2014530256A (en) 2014-11-17
CN104010654B (en) 2017-10-27
BR112014009146A8 (en) 2017-06-20
US20190298837A1 (en) 2019-10-03
CN104011073A (en) 2014-08-27
AU2012326168A1 (en) 2014-04-24
KR20140091705A (en) 2014-07-22
US9302002B2 (en) 2016-04-05
US20190184017A1 (en) 2019-06-20
US20200405865A1 (en) 2020-12-31
JP2014530862A (en) 2014-11-20
TWI644681B (en) 2018-12-21
BR112014009022A2 (en) 2017-05-02
ES2734070T3 (en) 2019-12-04
US10980884B2 (en) 2021-04-20
AR088381A1 (en) 2014-05-28
EP2768533A1 (en) 2014-08-27
PT2768525T (en) 2019-07-17
EP2768531A4 (en) 2015-03-11
IL231827A0 (en) 2014-05-28
EP2768525A1 (en) 2014-08-27
AU2012326080B2 (en) 2017-02-16
SI2768525T1 (en) 2019-10-30
IL231825A0 (en) 2014-05-28
TWI595883B (en) 2017-08-21
EA201490801A1 (en) 2014-09-30
US10376588B2 (en) 2019-08-13
EA027325B1 (en) 2017-07-31
AR088382A1 (en) 2014-05-28
HRP20191215T1 (en) 2019-10-04
SG11201401563SA (en) 2014-09-26
BR112014009146A2 (en) 2017-06-13
CY1121843T1 (en) 2020-07-31
JP6104922B2 (en) 2017-03-29
AU2012326171B2 (en) 2017-03-09
AU2012326082B2 (en) 2016-12-15
WO2013059407A1 (en) 2013-04-25
MX2014004734A (en) 2015-05-15
SG11201401562RA (en) 2014-09-26
BR112014009073A2 (en) 2017-05-09
KR20140091706A (en) 2014-07-22
JP2014530254A (en) 2014-11-17
US20190314500A1 (en) 2019-10-17
AU2012326170B2 (en) 2016-12-22
EP2768525B1 (en) 2019-06-19
SG11201401519RA (en) 2014-07-30
US10888619B2 (en) 2021-01-12
US10293049B2 (en) 2019-05-21
US9393305B2 (en) 2016-07-19
US20160199489A1 (en) 2016-07-14
CA2851628A1 (en) 2013-04-25
TW201325611A (en) 2013-07-01
HUE045624T2 (en) 2020-01-28
TW201325609A (en) 2013-07-01
EA026410B1 (en) 2017-04-28
US20190314498A1 (en) 2019-10-17
US11129876B2 (en) 2021-09-28
BR112014009031A2 (en) 2017-05-09
AR088379A1 (en) 2014-05-28
US20190290768A1 (en) 2019-09-26
HK1200718A1 (en) 2015-08-14
PL2768525T3 (en) 2019-10-31
MX2014004726A (en) 2015-05-15
US10772963B2 (en) 2020-09-15
JP6220788B2 (en) 2017-10-25
JP6220789B2 (en) 2017-10-25
KR20140079491A (en) 2014-06-26
KR102163150B1 (en) 2020-10-08
EP2768525A4 (en) 2015-07-15
US20130101584A1 (en) 2013-04-25
US9801942B2 (en) 2017-10-31
EA201490804A1 (en) 2014-09-30
TW201325606A (en) 2013-07-01
RS59179B1 (en) 2019-10-31
MX2014004732A (en) 2015-05-15
CA2851651A1 (en) 2013-04-25
CN104010657A (en) 2014-08-27
KR20200008021A (en) 2020-01-22
MX2019009176A (en) 2019-10-07
US20190290767A1 (en) 2019-09-26
US20130108632A1 (en) 2013-05-02
EP2768854A4 (en) 2015-03-11
SG11201401567YA (en) 2014-07-30
CN103998060B (en) 2016-12-21
KR20140091707A (en) 2014-07-22
CA2851639C (en) 2020-12-29
US10987405B2 (en) 2021-04-27
US11135267B2 (en) 2021-10-05
JP2014530863A (en) 2014-11-20
TWI619504B (en) 2018-04-01
CN103998060A (en) 2014-08-20
IL231826A0 (en) 2014-05-28
KR20140083037A (en) 2014-07-03
AU2012326080A1 (en) 2014-04-24
TW201325610A (en) 2013-07-01
CA2851642A1 (en) 2013-04-25
US20130108633A1 (en) 2013-05-02
MX2014004728A (en) 2015-05-15
WO2013059406A1 (en) 2013-04-25
EP2768532A4 (en) 2015-03-11
SG11201401576WA (en) 2014-10-30
EP2768532A1 (en) 2014-08-27
JP6113176B2 (en) 2017-04-12
KR102068462B1 (en) 2020-01-22
US20180028668A1 (en) 2018-02-01
WO2013059410A1 (en) 2013-04-25
SMT201900422T1 (en) 2019-09-09
BR112014009131A8 (en) 2017-06-20
WO2013059405A1 (en) 2013-04-25
KR20140097184A (en) 2014-08-06
AU2012326084B2 (en) 2016-12-15
US10213508B2 (en) 2019-02-26
US20130101640A1 (en) 2013-04-25
MX367054B (en) 2019-08-02
EA201490803A1 (en) 2014-09-30
IN2014CN02527A (en) 2015-06-26

Similar Documents

Publication Publication Date Title
US10772963B2 (en) Etanercept formulations stabilized with xylitol
US11000588B2 (en) Etanercept formulations stabilized with sodium chloride

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 12841505

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2851646

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: MX/A/2014/004732

Country of ref document: MX

ENP Entry into the national phase

Ref document number: 2014537221

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2012326080

Country of ref document: AU

Date of ref document: 20121018

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 2012841505

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 201490802

Country of ref document: EA

ENP Entry into the national phase

Ref document number: 20147013301

Country of ref document: KR

Kind code of ref document: A

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112014009131

Country of ref document: BR

ENP Entry into the national phase

Ref document number: 112014009131

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20140415