WO2021125852A1 - Nouvelle formulation injectable - Google Patents

Nouvelle formulation injectable Download PDF

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Publication number
WO2021125852A1
WO2021125852A1 PCT/KR2020/018595 KR2020018595W WO2021125852A1 WO 2021125852 A1 WO2021125852 A1 WO 2021125852A1 KR 2020018595 W KR2020018595 W KR 2020018595W WO 2021125852 A1 WO2021125852 A1 WO 2021125852A1
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formulation
conditions
analysis
aflibercept
injection
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PCT/KR2020/018595
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English (en)
Korean (ko)
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진현탁
남은주
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주식회사 프로젠
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • 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
    • A61K38/179—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
    • 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
    • A61K47/06—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
    • A61K9/00—Medicinal preparations characterised by special physical form
    • 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
    • A61K9/0048—Eye, e.g. artificial tears
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • 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
    • C07K14/71—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • 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 a novel injection formulation, and more particularly, to an injection formulation containing a VEGFR receptor extracellular domain-IgG Fc domain fusion protein.
  • Angiogenesis is a solid tumor, proliferative retinopathies, age-related macular degeneration (hereinafter, abbreviated as 'AMD'), rheumatoid arthritis, 'RA') is known to be involved in the pathogenesis of various diseases.
  • Vascular endothelial growth factor (hereinafter abbreviated as 'VEGF'), which is one of the factors necessary for angiogenesis, is expressed in human cancer and plays an important role in tumor neoangiogenesis.
  • 'VEGF' Vascular endothelial growth factor
  • the presence of high VEGF concentrations in the eye fluid is highly correlated with the activity of vascular proliferation in diabetic and other ischemia-associated retinopathy patients (Aiello et al ., N. Engl. J. Med .
  • VEGF vascular endothelial growth factor
  • AMD age-related macular degeneration
  • anti-VEGF antibodies such as Bevacizumab, Ranibizumab, or water-soluble Flt Attempts to use VEGF antagonists such as -1 and HuMV-833 or VEGFR antagonists such as SU-5416 and CGP-41251 as therapeutic agents for the above diseases are continuing.
  • VEGF antagonists aflibercept, a recombinant fusion protein in which the Fc portion of human IgG1 immunoglobulin and VEGF binding sites derived from the extracellular domains of human VEGF receptors 1 and 2 are fused
  • EYLEA ® the trade name of human VEGF receptors 1 and 2
  • insoluble particulates are generated or physicochemical denaturation occurs under conditions that are not optimal. These denaturation include oxidation, deamidation, isomerization, and multimerization.
  • Protein denaturation can ultimately reduce the protein's physiological activity by generating protein aggregation, fragments, and isomers.
  • protein aggregation is a major intrinsic factor in the formation of insoluble particles, which can cause side effects such as immune responses.
  • the criteria for insoluble particulates are being kept more stringent (Table 1).
  • Detection criteria for sub-visible insoluble particles Item number of particles ⁇ 10 ⁇ m ⁇ 25 ⁇ m Small injection ( ⁇ 100 mL) ⁇ 6,000/container ⁇ 600/container Bulk injection (> 100 mL) ⁇ 25/mL ⁇ 3/mL ophthalmic preparations ⁇ 50/mL ⁇ 5/mL
  • Korean Patent Registration No. 1406811 discloses an organic cosolvent such as aflibercept and polysorbate, an ionic isotonic agent selected from sodium chloride or potassium chloride, and a stabilizer such as sodium phosphate buffer and sucrose in a specific content.
  • an ionic isotonic agent such as sodium chloride is used, it is difficult to solve the above problems.
  • VEGFR receptor extracellular domain comprising a therapeutically effective amount of VEGFR receptor extracellular domain-IgG Fc domain fusion protein, citrate-sodium citrate buffer, sucrose, two or more amino acids in an aqueous medium -An injection composition containing an IgG Fc domain fusion protein is provided.
  • the injection composition according to an embodiment of the present invention exhibits superior stability compared to when a phosphate buffer is used, and thus has excellent long-term storage.
  • the scope of the present invention is not limited by these effects.
  • 1 is a graph showing the results of 12 weeks of analysis through SEC-HPLC analysis of long-term stability at low temperature of aflibercept-containing injection formulation candidates and control (phosphate formulations) prepared according to an embodiment of the present invention.
  • Figure 2 shows the low-temperature long-term stability of aflibercept-containing injection formulation candidates and controls (phosphate formulation) prepared according to an embodiment of the present invention through SDS-PAGE analysis under non-reducing (top) and reducing conditions (bottom). A series of gel photographs showing the results of weekly analysis.
  • FIG. 4 shows the storage stability under accelerated conditions (25° C.) of aflibercept-containing injection formulation candidates and control (phosphate formulations) prepared according to an embodiment of the present invention in non-reducing (top) and reducing conditions (bottom). It is a series of gel pictures showing the results of 3 months of analysis through SDS-PAGE analysis.
  • FIG. 6 shows the storage stability under severe conditions (40° C.) of aflibercept-containing injection formulation candidates and control (phosphate formulations) prepared according to an embodiment of the present invention in non-reducing (top) and reducing conditions (bottom). It is a series of gel pictures showing the results of analysis for 2 weeks through SDS-PAGE analysis.
  • FIG. 7 is a graph showing the results of 12-week analysis of the low-temperature long-term stability of aflibercept-containing injection formulation candidates and control (phosphate formulations) prepared according to an embodiment of the present invention through SEC-HPLC analysis.
  • Figure 8 shows the low-temperature long-term stability of aflibercept-containing injection formulation candidates and controls (phosphate formulation) prepared according to an embodiment of the present invention through SDS-PAGE analysis under non-reducing (top) and reducing conditions (bottom). A series of gel photographs showing the results of weekly analysis.
  • FIG. 10 shows storage stability under accelerated conditions (25° C.) of aflibercept-containing injection formulation candidates and controls (phosphate formulation) prepared according to an embodiment of the present invention in non-reducing (top) and reducing conditions (bottom). It is a series of gel pictures showing the results of 4 weeks analysis through SDS-PAGE analysis.
  • FIG. 12 shows the freezing conditions (Control, upper left ear) and low temperature conditions (Long-) of aflibercept-containing injection formulation candidates (#1 to #4) and control (phosphate formulation, #5) prepared according to an embodiment of the present invention.
  • term, upper right ear), accelerated condition (Accelerated, lower left ear), and severe condition (Stressed, lower right ear) is a series of graphs showing the results of analysis for 3 months.
  • FIG. 13 shows the freezing conditions (Control, upper left ear) and low temperature conditions (Long-) of aflibercept-containing injection formulation candidates (#1 to #4) and control (phosphate formulation, #5) prepared according to an embodiment of the present invention.
  • term, upper right ear), accelerated condition (Accelerated, lower left ear), and severe condition (Stressed, lower right ear) is a series of graphs showing the results of analysis for 3 months.
  • FIG. 14 shows the freezing conditions (Control, upper left ear) and low temperature conditions (Long-) of aflibercept-containing injection formulation candidates (#1 to #4) and control (phosphate formulation, #5) prepared according to an embodiment of the present invention.
  • term, upper right ear), accelerated condition (Accelerated, lower left ear), and severe condition (Stressed, lower right ear) is a series of graphs showing the results of 6 months of analysis by UV spectroscopy analysis using absorbance at 280 nm.
  • FIG. 15 shows the freezing conditions (Control, upper left ear) and low temperature conditions (Long-) of aflibercept-containing injection formulation candidates (#1 to #4) and control (phosphate formulation, #5) prepared according to an embodiment of the present invention.
  • term, upper right ear), accelerated condition (Accelerated, lower left ear), and severe condition (Stressed, lower right ear) are a series of graphs showing the results of analysis using SEC-HPLC for 6 months.
  • Figure 16a is a non-reducing (left) and reducing the stability of the initial formulation of aflibercept-containing injection formulation candidates (#1 to #4) and control (phosphate formulation, #5) prepared according to an embodiment of the present invention; It is a series of gel pictures showing the results of analysis through SDS-PAGE analysis of the condition (right), and FIG. 16b is an injection formulation candidate containing aflibercept prepared according to an embodiment of the present invention (#1 to #4) and After storing the control (phosphate formulation, #5) for 3 months under freezing conditions (Control) and low temperature conditions (Long-term), the results of analysis through SDS-PAGE analysis under non-reducing (left) and reducing conditions (right) is a series of gel pictures showing, and FIG.
  • 16c is an accelerated condition of the aflibercept-containing injection formulation candidates (#1 to #4) and the control group (phosphate formulation, #5) prepared according to an embodiment of the present invention. And it is a series of gel pictures showing the results of analysis through SDS-PAGE analysis under non-reducing (left) and reducing conditions (right) after storage for 3 months under severe conditions (Stressed), Figure 16d is an embodiment of the present invention After storing the final formulation candidates (#1 to #4) and control (phosphate formulation, #5) containing aflibercept prepared according to the method for 6 months under frozen conditions (Control) and low temperature conditions (Long-term), non-reducing (Left) and a series of gel photographs showing the results of analysis through SDS-PAGE analysis under reducing conditions (right), and FIG.
  • 16E is a candidate for injection formulation containing aflibercept prepared according to an embodiment of the present invention (# 1 to #4) and the control (phosphate formulation, #5) were stored for 6 months under accelerated conditions (Accelerated) and then analyzed through SDS-PAGE analysis under non-reducing (left) and reducing conditions (right). A series of gel pictures.
  • Figure 17a shows the results of analysis by GelIEF analysis of the initial stability of formulation of aflibercept-containing injection formulation candidates (#1 to #4) and control (phosphate formulation, #5) prepared according to an embodiment of the present invention. It is a gel photograph showing, and FIG. 17b is a frozen condition (Control) and low temperature condition of aflibercept-containing injection formulation candidates (#1 to #4) and control (phosphate formulation, #5) prepared according to an embodiment of the present invention. It is a gel photograph showing the result of analyzing the stability of the formulation by GelIEF analysis after storage for 3 months in (long-term), and FIG.
  • FIG. 17c is a candidate for injection formulation containing aflibercept prepared according to an embodiment of the present invention (#1 to #4) and control (phosphate formulation, #5) are gel pictures showing the results of analysis of the stability of the formulation by GelIEF analysis after storage for 3 months in accelerated and stressed conditions
  • FIG. 17d is the present invention Aflibercept-containing injection formulation candidates (#1 to #4) and control (phosphate formulation, #5) prepared according to an example of It is a series of gel pictures showing the results of analyzing the stability of the formulation by GelIEF analysis after storage for 6 months under accelerated conditions (Accelerated, right).
  • aflibercept-containing injection formulation candidates A1-DS-2007-D and A1-DS-2008-D
  • controls phosphate formulation, A1-
  • DS-2006-D in freezing conditions (Control, upper left ear), low temperature conditions (Long-term, upper right ear), accelerated conditions (Accelerated, lower left ear) and harsh conditions (Stressed, lower right ear) for up to 6 months. It is a series of graphs showing the results.
  • FIG. 19 shows aflibercept-containing injection formulation candidates (A1-DS-2007-D and A1-DS-2008-D) and control (phosphate formulation, A1-) according to the type of surfactant prepared according to an embodiment of the present invention; DS-2006-D) under frozen conditions (Control, upper left ear), low temperature conditions (Long-term, right upper ear), accelerated conditions (Accelerated, lower left ear) and severe conditions (Stressed, lower right ear) for up to 6 months It is a series of graphs showing one result.
  • aflibercept-containing injection formulation candidates A1-DS-2007-D and A1-DS-2008-D
  • controls phosphate formulation, A1-
  • DS-2006-D of protein content in freezing conditions (Control, upper left ear), low temperature condition (Long-term, right upper ear), accelerated condition (Accelerated, lower left ear) and severe condition (Stressed, lower right ear) using absorbance at 280 nm
  • FIG. 21 shows aflibercept-containing injection formulation candidates (A1-DS-2007-D and A1-DS-2008-D) and controls (phosphate formulation, A1-) according to the type of surfactant prepared according to an embodiment of the present invention.
  • DS-2006-D the protein content under freezing conditions (Control, upper left ear), low temperature condition (Long-term, right upper ear), accelerated condition (Accelerated, lower left ear), and severe condition (Stressed, lower right ear) using SEC-HPLC This is a series of graphs showing the results of analysis for up to 6 months.
  • Figure 22a shows aflibercept-containing injection formulation candidates (A1-DS-2007-D and A1-DS-2008-D) and control (phosphate formulation, A1-) according to the type of surfactant prepared according to an embodiment of the present invention.
  • DS-2006-D is a series of gel photographs showing the results of analysis of the stability of the initial formulation through SDS-PAGE analysis under non-reducing (left) and reducing conditions (right), and
  • FIG. 22b is an embodiment of the present invention.
  • aflibercept-containing injection formulation candidates (A1-DS-2007-D and A1-DS-2008-D) and control (phosphate formulation, A1-DS-2006-D) were frozen It is a series of gel pictures showing the results of SDS-PAGE analysis under non-reducing (left) and reducing conditions (right) after storage for 3 months under conditions (Control) and low-temperature conditions (Long-term), Fig.
  • 22c is aflibercept-containing injection formulation candidates (A1-DS-2007-D and A1-DS-2008-D) and control (phosphate formulation, A1-DS-) according to the type of surfactant prepared according to an embodiment of the present invention 2006-D) after storage for 3 months under accelerated and stressed conditions, a series of gel photographs showing the results of SDS-PAGE analysis under non-reducing (left) and reducing conditions (right) 22d shows aflibercept-containing injection formulation candidates (A1-DS-2007-D and A1-DS-2008-D) and controls (phosphate formulation, A1-DS-2006-D) was stored for 6 months under freezing conditions (Control), low temperature conditions (Long-term) and accelerated conditions (Accelerated), and then SDS-PAGE analysis of non-reducing (left) and reducing conditions (right) It is a series of gel pictures showing the results of analysis through
  • Figure 23a shows aflibercept-containing injection formulation candidates (A1-DS-2007-D and A1-DS-2008-D) and control (phosphate formulation, A1-) according to the type of surfactant prepared according to an embodiment of the present invention
  • DS-2006-D is a gel photograph showing the results of analysis of the initial stability of formulation by GelIEF analysis
  • FIG. 23b is a candidate for injection formulation containing aflibercept according to the type of surfactant prepared according to an embodiment of the present invention.
  • FIG. 23c is an applique according to the type of surfactant prepared according to an embodiment of the present invention.
  • Injectable formulation candidates including sept (A1-DS-2007-D and A1-DS-2008-D) and control (phosphate formulation, A1-DS-2006-D) were subjected to freezing conditions (Control), low temperature conditions (Long-term) and a series of gel photographs showing the results of analysis of the stability of the formulation by GelIEF analysis after storage for 6 months in accelerated conditions.
  • VEGFR extracellular domain-IgG Fc domain fusion protein is a VEGF receptor VEGFR1 or VEGFR2 extracellular domain that is a VEGF binding site of immunoglobulin Fc (fragment crystalizable region) linked to the N-terminus It is one of the VEGF antagonists that bind to VEGF as a fusion protein and inhibit its function.
  • an ophthalmic injection for the treatment of age-related macular degeneration, etc. with the ingredient name "aflibercept" has been developed and sold.
  • Aflibercept used in this document is the name of an active ingredient included in an ophthalmic injection developed by Regeneron Pharmaceuticals and sold as a treatment for macular degeneration under the trade name EYLEA ®, and the Fc region of human IgG1 Refers to a fusion protein comprising the extracellular domain of VEGFR1 or VEGFR2 fused to.
  • biosimilar refers to a generic drug for a biological drug that can be produced by a third party other than the original patent holder after the patent has expired.
  • OTC drugs are chemically synthesized agents and low molecular weight compounds, so it is possible to chemically synthesize the same ingredients. Therefore, a drug that reproduces it is called a generic drug.
  • biopharmaceuticals are not synthesized chemically, but go through the process of making high-molecular protein products using animal cells, yeast, E. coli, etc., so the same product cannot be duplicated.
  • biopharmaceuticals when making such biopharmaceuticals, they must be manufactured using living cells, but in the sense that it is impossible to make a completely identical product and only similar products can be made, it is not simply called a generic drug, but 'virosimilar'. Therefore, the term 'biosimilar' used in this document has the same amino acid sequence information as that of a patented biological drug, but is not manufactured by the same process, but if it produces almost the same biological effect as a result of a blood test through a clinical test, it is certified for equivalence. It is recognized as a biosimilar, and it is called a biosimilar because it is expressed as a similar in the sense that it is not identical but similar.
  • surfactant refers to a compound having both a water-soluble hydrophilic moiety and an oil-soluble hydrophobic moiety. Due to these properties, it has been used in soaps and detergents, and is also used in cosmetics and pharmaceuticals.
  • Surfactants include ionic surfactants and nonionic surfactants, and ionic surfactants are divided into cationic surfactants and anionic surfactants.
  • Surfactants are also called emulsifiers because they induce emulsification of lipids and aqueous solutions in terms of use.
  • buffer refers to a weak acid or base used to maintain a constant pH of an aqueous solution after addition of another acid or base.
  • pK a Phosphate with a value of 7.2 close to physiological pH is commonly used for biological and medicinal purposes.
  • isotonic agent refers to a solute added to a pharmaceutical formulation to match an osmolality similar to the osmotic pressure of a body fluid in a living body.
  • Tonicity agents include ionic isotonic agents, such as sodium chloride, and nonionic isotonic agents, including sugars, such as sorbitol.
  • VEGFR receptor extracellular domain comprising a therapeutically effective amount of VEGFR receptor extracellular domain-IgG Fc domain fusion protein, citrate-sodium citrate buffer, sucrose, two or more amino acids in an aqueous medium -An injection composition containing an IgG Fc domain fusion protein is provided.
  • the VEGFR may be VEGFR1 or VEGFR2.
  • the fusion protein may be aflibercept or a biosimilar thereof.
  • the amino acid may be arginine and glycine.
  • the injection composition may include 30 to 50 mg/ml of the fusion protein, 5 to 15 mM citrate-sodium citrate buffer, 3 to 7 wt/v% sucrose, 20 to 70 mM arginine, 20 to 70 mM glycine have.
  • the pH of the injection composition may be adjusted to 5.8 to 6.6.
  • the injection composition may or may not further include an isotonic agent.
  • the isotonic agent when the isotonic agent is further included, the isotonic agent may be sorbitol.
  • the injection composition may or may not contain a surfactant.
  • the surfactant may be included in 0.01 to 0.2 wt/v%, and the surfactant may be Polysorbate 20, Polysorbate 80 or poloxamer 188.
  • the injection composition may be used for ophthalmology, and in this case, it may be used for the treatment of angiogenesis-related ophthalmic diseases, and the angiogenesis-related ophthalmic diseases may be diabetic macular edema, diabetic retinopathy, macular edema, or macular degeneration. .
  • the injectable composition of the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions.
  • suitable carriers excipients and diluents commonly used in the preparation of pharmaceutical compositions.
  • liquid formulation additives can be used in the preparation of the injectable composition.
  • the additive for formulation may be either organic or inorganic.
  • the therapeutically effective amount of the fusion protein may vary depending on the disease to be treated. In the case of an intraocular injection composition for the treatment of macular degeneration, the therapeutically effective amount is about 2 mg. In this case, the injection formulation contains 40 mg/ml of the fusion protein. It may be provided at a concentration, and in the case of an injection for treating colon cancer, it may be provided at a concentration of 25 mg/ml at 4 mg/kg body weight.
  • the dosage of the injection composition of the present invention may be administered once a day, or divided into 2-3 times a day at appropriate intervals, or may be administered intermittently at intervals of several days.
  • the present inventors have made intensive efforts to provide an injection formulation containing aflibercept that has high stability without using an ionic isotonic agent and close to physiological pH without using a phosphate buffer.
  • citrate-sodium citrate as a buffer was used.
  • an injection containing aflibercept or a biosimilar thereof with improved stability at low temperature storage and accelerated conditions without isotonicity agents or surfactants was completed.
  • the injection composition according to an embodiment of the present invention is compared with 10 mM sodium phosphate, 5% sucrose, 40 mM sodium chloride, and 0.03% Polysorbate 20, which is a composition of a conventional ophthalmic aflibercept-containing injection (EYLEA ®).
  • EYLEA ® ophthalmic aflibercept-containing injection
  • VEGFR receptor extracellular domain comprising a therapeutically effective amount of VEGFR receptor extracellular domain-IgG Fc domain fusion protein, citrate-sodium citrate buffer, sucrose, two or more amino acids in an aqueous medium -An injection composition containing an IgG Fc domain fusion protein is provided.
  • the VEGFR may be VEGFR1 or VEGFR2.
  • the fusion protein may be aflibercept or a biosimilar thereof.
  • the amino acid may be arginine and glycine.
  • the injection composition may include 30 to 50 mg/ml of the fusion protein, 5 to 15 mM citrate-sodium citrate buffer, 3 to 7 wt/v% sucrose, 20 to 70 mM arginine, 20 to 70 mM glycine have.
  • the pH of the injection composition may be adjusted to 5.8 to 6.6.
  • the injection composition may or may not further include an isotonic agent.
  • the isotonic agent when the isotonic agent is further included, the isotonic agent may be sorbitol.
  • the injection composition may or may not contain a surfactant.
  • the surfactant may be included in 0.01 to 0.2 wt/v%, and the surfactant may be Polysorbate 20, Polysorbate 80 or poloxamer 188.
  • the injection composition may be used for ophthalmology, and in this case, it may be used for the treatment of angiogenesis-related ophthalmic diseases, and the angiogenesis-related ophthalmic diseases may be diabetic macular edema, diabetic retinopathy, macular edema, or macular degeneration. .
  • the injectable composition of the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions.
  • suitable carriers excipients and diluents commonly used in the preparation of pharmaceutical compositions.
  • liquid formulation additives can be used in the preparation of the injectable composition.
  • the additive for formulation may be either organic or inorganic.
  • the therapeutically effective amount of the fusion protein may vary depending on the disease to be treated. In the case of an intraocular injection composition for the treatment of macular degeneration, the therapeutically effective amount is about 2 mg. In this case, the injection formulation contains 40 mg/ml of the fusion protein. It may be provided at a concentration, and in the case of an injection for treating colon cancer, it may be provided at a concentration of 25 mg/ml at 4 mg/kg body weight.
  • the dosage of the injection composition of the present invention may be administered once a day, or divided into 2-3 times a day at appropriate intervals, or may be administered intermittently at intervals of several days.
  • the present inventors have made intensive efforts to provide an injection formulation containing aflibercept that has high stability without using an ionic isotonic agent and close to physiological pH without using a phosphate buffer.
  • citrate-sodium citrate as a buffer was used.
  • an injection containing aflibercept or a biosimilar thereof with improved stability at low temperature storage and accelerated conditions without isotonicity agents or surfactants was completed.
  • the injection composition according to an embodiment of the present invention is compared with 10 mM sodium phosphate, 5% sucrose, 40 mM sodium chloride, and 0.03% Polysorbate 20, which is a composition of a conventional ophthalmic aflibercept-containing injection (EYLEA ®).
  • EYLEA ® ophthalmic aflibercept-containing injection
  • UV-Vis spectrophotometer Ultraviolet-Visible Spectrophotometer
  • aflibercept SEQ ID NO: 1
  • the present inventors used 10 mM citrate-sodium citrate as a basic buffer, and sugars (sucrose and mannitol) as excipients.
  • amino acids arginine, glycine, histidine, methionine, proline, alanine, and glutamic acid
  • isotonicity agents sorbitol, mannitol, sodium chloride
  • surfactants Polysorbate 20, and Polysorbate 80
  • the PROTEOSTAT dye When a protein is unfolded to form a polymer impurity aggregate, the PROTEOSTAT dye binds to the polymer impurity aggregate, and the temperature at which the polymer impurity aggregate is detected is the degree to which the protein shows structural stability ( T agg ). Therefore, the higher T agg compared to the control can be interpreted as showing structural stability when the corresponding excipient was added compared to the control sample, and Ta agg was used as a relative comparative value.
  • the T agg of the control (10 mM citrate-sodium citrate buffer without the addition of an excipient) in 10 mM citrate-sodium citrate buffer was confirmed to be 64.36, and in each combination, excipients with higher T agg values than the control were selected for each of two types.
  • sucrose and trehalose in the sugar group, arginine and glycine in the amino acid group, sodium chloride and sorbitol in the isotonic group, and Polysorbate 80 and Polysorbate 20 as surfactants were selected as candidates.
  • sucrose and trehalose were used as sugars, better values could be confirmed in the DSF analysis results, and in the case of surfactants, there was no significant difference between the two, and amino acids In the case of glycine and arginine were confirmed to be effective, and for isotonic agents, both sorbitol and sodium chloride were confirmed to be suitable.
  • Example 1 From the results of Example 1, the present inventors designed 11 combinations by varying the composition of excipients (Table 3). Analysis was performed by including three control types (10 mM citrate-sodium citrate buffer phosphate formulation and histidine formulation) in the formulation formulated as described above. The 11 combinations consist of 2.5-20% sucrose, 5% trehalose, 40-100 mM sodium chloride, 50-100 mM arginine/Arg-HCl, 40 mM sorbitol, 20-50 mM glycine, 0.03-0.1% Polysorbate 20, 0.03% Polysorbate 80 constitutes the excipient concentration range. The concentration of aflibercept was prepared in the following composition at 37-44 mg/mL, pH 5.9-6.2, and osmolality and DSF analysis results by arbitrarily adjusting the concentration referring to the results of Example 1 was confirmed.
  • Example 2 From the results of Example 2, the present inventors selected 6 candidate formulations as shown in Table 3 to form aflibercept injection formulations (Table 4), low temperature conditions (5°C, 12 weeks), and accelerated conditions (25°C) , 12 weeks) and storage stability under severe conditions (40°C, 2 weeks) were investigated.
  • composition of the formulation candidate group sample number Selected excipient composition control One 10 mM sodium phosphate, 5% sucrose, 40 mM sodium chloride, 0.03% Polysorbate 20 (phosphate formulation) Formulation candidate 1 2 10 mM Citric Acid-Sodium Citrate, 7% Sucrose, 40 mM Sodium Chloride, 0.1% Polysorbate 20 Formulation candidate 2 3 10 mM citrate-sodium citrate, 5% sucrose, 50 mM Arg/Arg-HCl, 50 mM glycine, 0.03% Polysorbate 80 Formulation Candidate 3 4 10 mM citrate-sodium citrate, 5% trehalose, 100 mM sodium chloride, 0.03% Polysorbate 20 Formulation candidate 4 5 10 mM citrate-sodium citrate, 5% trehalose, 40 mM sorbitol, 20 mM glycine, 0.03% Polysorbate 20 Formulation candidate 5 6 10 mM Citric Acid-Sodium Citrate, 5% Suc
  • formulation candidate 2 (10 mM citrate-sodium citrate, 5% sucrose, 50 mM arginine/Arg-HCl, 50 mM glycine, 0.03% Polysorbate 80) was the highest. Therefore, the present inventors fixed 5% sucrose, 50 mM arginine, and 50 mM glycine as amino acids, and designed five formulation candidates in which the type and pH of the surfactant were changed (Table 5).
  • composition of formulation candidates sample sample number
  • Formulation information remark control
  • One 10 mM Sodium Phosphate, 5% Sucrose, 40 mM Sodium Chloride, 0.03% Polysorbate 20 phosphate formulation Formulation candidate 1 2 10 mM citrate-sodium citrate, 5% sucrose, 50 mM arginine/Arg-HCl, 50 mM glycine, pH 6.2 without Polysorbate 80
  • Formulation candidate 2 3 10 mM citrate-sodium citrate, 5% sucrose, 50 mM arginine/Arg-HCl, 50 mM glycine, 0.03% Polysorbate 80 (pH 6.2) Polysorbate 80 Formulation candidate 3 4 10 mM citrate-sodium citrate, 5% sucrose, 50 mM arginine/Arg-HCl, 50 mM glycine, 0.1% poloxamer 188 (pH 6.2) poloxamer 188 Formulation candidate 4 5 10 mM citrate
  • formulation candidate 2 (10 mM citrate-sodium citrate, 5% sucrose, 50 mM arginine/Arg-HCl, 50 mM glycine, 0.03% Polysorbate 80) had the highest overall stability.
  • the present inventors made the formulation candidate 2 as the additional candidate formulation #1, but lowered the concentration of amino acids arginine and glycine to control the osmolality
  • additional candidate formulations #2 to #4 were additionally prepared (Table 6)
  • various items such as stability of these additional candidate formulations were analyzed.
  • Candidate Formulations sample sample number
  • Formulation information remark Additional Candidate Formulation #1
  • Polysorbate 20 Additional Candidate Formulation #2 2 10 mM citrate-sodium citrate, 5% sucrose, 30 mM arginine/Arg-HCl, 30 mM glycine, 0.03% Polysorbate 20 (pH 6.2)
  • the candidate formulations and the control formulation (sodium phosphate formulation) according to an embodiment of the present invention were formulated during storage under freezing conditions or low temperature conditions. It was found that there was no difference between the formulations at the beginning and after storage for 3 months. However, as shown in FIG. 16c , in the case of severe conditions, the bands in the cleaved form were somewhat confirmed, and no difference was observed for each formulation. As confirmed in FIGS. 16D and 16E , this trend was similar even when stored for 6 months.
  • the storage stability of the candidate formulations according to an embodiment of the present invention was also investigated through gel isoelectric intensive analysis (GelIEF), and as shown in FIG. 17a , immediately after formulation, it was confirmed to have an isoelectric point of between 6.0 and 7.8. , the difference in the pattern between the formulations was not confirmed, and exhibited almost the same isoelectric point pattern as in the initial formulation even when stored under frozen or low temperature conditions, and as confirmed in FIGS. 17b and 17c, when stored for 3 months, frozen conditions, The same pattern was observed in the low temperature condition and the accelerated condition, but in the case of the severe condition, a shift of the pI value toward acidity was observed. In addition, in the case of long-term storage for 6 months, as confirmed in FIG. 17d , there was no significant difference in the band pattern in the freezing condition, low temperature condition, and accelerated condition, and no significant difference was confirmed for each formulation.
  • GelIEF gel isoelectric intensive analysis
  • the present inventors confirmed that there is no significant problem in the long-term storage stability of the formulation even if the osmolality of the formulation is lowered by slightly lowering the concentrations of arginine and glycine through the results of Example 5. Accordingly, the present inventors tried to determine the effect on stability when Polysorbate 80 is used instead of Polysorbate 20, which is the surfactant used in Example 4.
  • composition according to change of surfactant in formulation Formulation name
  • Formulation composition A1-DS-2006-D 10 mM Sodium Phosphate, 5% Sucrose, 40 mM Sodium Chloride, 0.03% Polysorbate 20 (pH 6.2) A1-DS-2007-D 10 mM citrate-sodium citrate, 5% sucrose, 30 mM arginine/Arg-HCl, 30 mM glycine, 0.03% Polysorbate 20 (pH 6.2) A1-DS-2008-D 10 mM citrate-sodium citrate, 5% sucrose, 30 mM arginine/Arg-HCl, 30 mM glycine, 0.03% Polysorbate 80 (pH 6.2)
  • the protein content during storage under freezing conditions, low temperature conditions and accelerated conditions was confirmed to be 6 months storage stability in the range of 5% compared to the initial.
  • storage stability for 3 months was confirmed in the range of 12% compared to the initial period during storage under severe conditions.
  • the initial band pattern (Fig. 23a) was almost similar to the initial band pattern (Fig. 23b) when stored under frozen conditions, low temperature conditions, and accelerated conditions for 3 months, whereas under severe conditions, the band pattern tended to shift toward acid. , in this case, the difference by the formulation was not confirmed.
  • the frozen condition and the low temperature condition showed the same pattern, and in the case of the accelerated condition, the band pattern slightly shifted toward the acid side, but the difference was insignificant.
  • no difference was confirmed by the formulation (FIG. 23C).
  • the injectable formulation containing aflibercept or a biosimilar thereof uses a citrate buffer instead of the conventionally used phosphate buffer and does not add isotonic agents or surfactants.
  • the aflibercept-containing injection formulation according to an embodiment of the present invention can be used as a more economical and stable therapeutic agent for angiogenesis-related eye diseases such as age-related macular degeneration.
  • the injectable composition according to an embodiment of the present invention can be used as a therapeutic agent for ophthalmology, in particular, for the treatment of angiogenesis-related ophthalmic diseases such as monolithic retinopathy and age-related macular degeneration.
  • SEQ ID NO: 1 is the amino acid sequence of aflibercept.

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Abstract

La présente invention concerne une nouvelle composition injectable, et fournit une composition injectable contenant une protéine de fusion domaine extracellulaire du VEGFR-domaine Fc de l'IgG, la composition injectable comprenant, dans un milieu aqueux, une quantité thérapeutiquement efficace de la protéine de fusion domaine extracellulaire du VEGFR-domaine Fc de l'IgG, un tampon acide citrique-citrate de sodium, du saccharose et au moins deux acides aminés.
PCT/KR2020/018595 2019-12-17 2020-12-17 Nouvelle formulation injectable WO2021125852A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023031478A1 (fr) * 2021-09-06 2023-03-09 Lek Pharmaceuticals D.D. Formulations pour protéines de fusion du récepteur vegf

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Publication number Priority date Publication date Assignee Title
KR20150033620A (ko) * 2012-06-01 2015-04-01 옵쏘테크 코포레이션 항-pdgf 압타머 및 vegf 길항제를 포함하는 조성물
US20160376342A1 (en) * 2015-06-23 2016-12-29 Alteogen, Inc. STABLE LIQUID FORMULATION OF FUSION PROTEIN WITH IgG Fc DOMAIN
WO2018094316A1 (fr) * 2016-11-21 2018-05-24 Just Biotherapeutics, Inc. Formulations d'aflibercecept et leurs utilisations
KR101861163B1 (ko) * 2017-04-26 2018-05-25 삼천당제약주식회사 안과용 약학 조성물
WO2019173767A1 (fr) * 2018-03-08 2019-09-12 Coherus Biosciences Inc. Formulations aqueuses stables d'aflibercept

Patent Citations (5)

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Publication number Priority date Publication date Assignee Title
KR20150033620A (ko) * 2012-06-01 2015-04-01 옵쏘테크 코포레이션 항-pdgf 압타머 및 vegf 길항제를 포함하는 조성물
US20160376342A1 (en) * 2015-06-23 2016-12-29 Alteogen, Inc. STABLE LIQUID FORMULATION OF FUSION PROTEIN WITH IgG Fc DOMAIN
WO2018094316A1 (fr) * 2016-11-21 2018-05-24 Just Biotherapeutics, Inc. Formulations d'aflibercecept et leurs utilisations
KR101861163B1 (ko) * 2017-04-26 2018-05-25 삼천당제약주식회사 안과용 약학 조성물
WO2019173767A1 (fr) * 2018-03-08 2019-09-12 Coherus Biosciences Inc. Formulations aqueuses stables d'aflibercept

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023031478A1 (fr) * 2021-09-06 2023-03-09 Lek Pharmaceuticals D.D. Formulations pour protéines de fusion du récepteur vegf

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