WO2021105393A1 - Procédés d'obtention de compositions de glp-1 stables - Google Patents

Procédés d'obtention de compositions de glp-1 stables Download PDF

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Publication number
WO2021105393A1
WO2021105393A1 PCT/EP2020/083686 EP2020083686W WO2021105393A1 WO 2021105393 A1 WO2021105393 A1 WO 2021105393A1 EP 2020083686 W EP2020083686 W EP 2020083686W WO 2021105393 A1 WO2021105393 A1 WO 2021105393A1
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WIPO (PCT)
Prior art keywords
alternatively
glp
hmwp
peptide
iron
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PCT/EP2020/083686
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English (en)
Inventor
Jesper Søndergaard PEDERSEN
Arne Staby
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Novo Nordisk A/S
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Publication of WO2021105393A1 publication Critical patent/WO2021105393A1/fr

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    • 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
    • A61K38/22—Hormones
    • A61K38/26—Glucagons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics

Definitions

  • the present invention relates to processes for increasing the stability of compositions comprising a GLP-1 peptide, stable GLP-1 compositions and uses of such compositions.
  • GLP-1 peptides liraglutide and semaglutide have been marketed for use in, for example, treatment of type 2 diabetes.
  • GLP-1 peptides have a limited stability in solution among others due to the formation of high molecular weight proteins (HMWP) which are a highly diverse group of aggregate compounds made by covalent cross-binding between two or more GLP-1 peptides or partly degraded GLP-1 peptides and/or by incorporating tracing impurities into GLP-1 peptides.
  • HMWP high molecular weight proteins
  • EP2209800B1 discloses a shelf-stable pharmaceutical composition for parenteral administration, which comprises liraglutide, a basal insulin peptide, a pharmaceutically acceptable additives and zinc, wherein the zinc content is between 7 and 12 Zn ions per 6 insulin molecules and wherein the pH is between 7.7 and 8.2.
  • the present invention relates to a process for obtaining a stabilised pharmaceutical composition comprising a GLP-1 peptide, said process comprising the steps of: a. Producing said GLP-1 peptide; b. Purifying said GLP-1 peptide; and c. Formulating said GLP-1 peptide into said stabilised pharmaceutical composition, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 2 pg iron/g GLP-1 peptide, such as below 1.5 pg iron/g GLP-1 peptide.
  • the present invention relates to a process for obtaining a stabilised pharmaceutical composition comprising a GLP-1 peptide, said process comprising the steps of: a. Producing said GLP-1 peptide; b. Purifying said GLP-1 peptide; and c. Formulating said GLP-1 peptide into said stabilised pharmaceutical composition, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 1 ug iron/g GLP-1 peptide.
  • the present invention relates to a process for obtaining a GLP-1 peptide with high purity, said process comprising the steps of: a. Producing said GLP-1 peptide; and b. Purifying said GLP-1 peptide, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 2 pg iron/g GLP-1 peptide, such as below 1.5 pg iron/g GLP-1 peptide.
  • the present invention further relates to stabilised pharmaceutical compositions and GLP-1 peptides obtainable by such processes and the use of such stabilised pharmaceutical compositions and GLP-1 peptides.
  • the present invention relates to a process for obtaining a GLP-1 peptide with high purity, said process comprising the steps of: a. Producing said GLP-1 peptide; and b. Purifying said GLP-1 peptide, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 1 ug iron/g GLP-1 peptide.
  • the present invention further relates to stabilised pharmaceutical compositions and GLP-1 peptides obtainable by such processes and the use of such stabilised pharmaceutical compositions and GLP-1 peptides.
  • Fig. 1 shows formation of liraglutide and semaglutide HMWP at 37°C after 19 days as a function of iron (Fe 2+ ) concentration in 10 mM Tris, and 0.6 mM liraglutide or semaglutide, pH 7.4, with 50 mM phenol (A) and without 50 mM phenol (B).
  • Fe 2+ iron
  • Fig. 1 shows formation of liraglutide and semaglutide HMWP at 37°C after 19 days as a function of iron (Fe 2+ ) concentration in 10 mM Tris, and 0.6 mM liraglutide or semaglutide, pH 7.4, with 50 mM phenol (A) and without 50 mM phenol (B).
  • the present invention relates to a process for obtaining a stabilised pharmaceutical composition comprising a GLP-1 peptide, said process comprising the steps of: a. Producing said GLP-1 peptide; b. Purifying said GLP-1 peptide; and c. Formulating said GLP-1 peptide into said stabilised pharmaceutical composition, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 2 pg iron/g GLP-1 peptide, such as below 1.8 pg iron/g GLP-1 peptide, such as below 1.5 pg iron/g GLP-1 peptide.
  • the present invention relates to a process for obtaining a stabilised pharmaceutical composition comprising a GLP-1 peptide, said process comprising the steps of: a. Producing said GLP-1 peptide; b. Purifying said GLP-1 peptide; and c. Formulating said GLP-1 peptide into said stabilised pharmaceutical composition, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 1 ug iron/g GLP-1 peptide.
  • the concentration of iron is kept between 0.001-2, alternatively 0.002-2, alternatively 0.005-2, alternatively 0.01-2, alternatively 0.02-2, alternatively 0.03-2, alternatively 0.04-2, alternatively 0.05-2, alternatively 0.06-2, alternatively 0.07-2, alternatively 0.08-2, alternatively 0.09-2, alternatively 0.1-2, alternatively 0.2-2, alternatively 0.3-2, alternatively 0.4-2, alternatively 0.5- 2, alternatively 0.6-2, alternatively 0.7-2, alternatively 0.8-2, alternatively 0.9-2 pg iron/g GLP- 1 peptide.
  • the concentration of iron is kept between 0.001-1.8, alternatively 0.002-1.8, alternatively 0.005-1.8, alternatively 0.01-1.8, alternatively 0.02-1.8, alternatively 0.03-1.8, alternatively 0.04-1.8, alternatively 0.05-1.8, alternatively 0.06-1.8, alternatively 0.07-1.8, alternatively 0.08-1.8, alternatively 0.09-1.8, alternatively 0.1-1.8, alternatively 0.2-1.8, alternatively 0.3-1.8, alternatively 0.4-1.8, alternatively 0.5-1.8, alternatively 0.6-1.8, alternatively 0.7-1.8, alternatively 0.8-1.8, alternatively 0.9-1.8 pg iron/g GLP-1 peptide.
  • the concentration of iron is kept between 0.001-1.5, alternatively 0.002-1.5, alternatively 0.005-1.5, alternatively 0.01-1.5, alternatively 0.02-1.5, alternatively 0.03-1.5, alternatively 0.04-1.5, alternatively 0.05-1.5, alternatively 0.06-1.5, alternatively 0.07-1.5, alternatively 0.08-1.5, alternatively 0.09-1.5, alternatively 0.1-1.5, alternatively 0.2-1.5, alternatively 0.3-1.5, alternatively 0.4-1.5, alternatively 0.5-1.5, alternatively 0.6-1.5, alternatively 0.7-1.5, alternatively 0.8-1.5, alternatively 0.9-1.5 pg iron/g GLP-1 peptide.
  • the concentration of iron is kept between 0.001-1, alternatively 0.002-1, alternatively 0.005-1, alternatively 0.01-1, alternatively 0.02-1, alternatively 0.03-1, alternatively 0.04-1, alternatively 0.05-1, alternatively 0.06-1, alternatively 0.07-1, alternatively 0.08-1, alternatively 0.09-1, alternatively 0.1-1, alternatively 0.2-1, alternatively 0.3-1, alternatively 0.4-1, alternatively 0.5- 1, alternatively 0.6-1, alternatively 0.7-1, alternatively 0.8-1, alternatively 0.9-1 ug iron/g GLP- 1 peptide.
  • the concentration of iron is kept between 0.001-0.95, alternatively 0.001-0.90, alternatively 0.001-0.85, alternatively 0.001-0.80, alternatively 0.001-0.75, alternatively 0.001-0.70, alternatively 0.001-0.65, alternatively 0.001-0.60, alternatively 0.001-0.55, alternatively 0.001-0.50, alternatively 0.001-0.45, alternatively 0.001-0.40, alternatively 0.001-0.35, alternatively 0.001-0.30, alternatively 0.001-0.25, alternatively 0.001-0.20, alternatively 0.001-0.15, alternatively 0.001-0.10, alternatively 0.001-0.05, alternatively 0.001-0.01 ug iron/g GLP-1 peptide.
  • the concentration of iron is controlled for the entire process. In some embodiments, the concentration of iron is controlled for step a and/or step b and/or step c. In some embodiments, the concentration of iron is controlled in the stabilised pharmaceutical composition.
  • the process further comprises the step of: d. Isolating said stabilised pharmaceutical composition.
  • HMWP is a highly diverse group of aggregate compounds made by covalent cross binding between two or more GLP-1 peptides or partly degraded GLP-1 peptides and/or by incorporating tracing impurities into GLP-1 peptides.
  • HMWP in the drug product may give rise to detrimental effects in vivo, including enhanced immunogenicity, altered subcutaneous absorption kinetics, enhanced injection site toxicity, and diminished biopotency.
  • HMWP formation needs to be kept at a minimum to maximise the stability of solutions comprising GLP-1 peptides.
  • HMWP formation may be managed by identifying and controlling the specific chemical degradation pathways associated with polypeptide reactivity.
  • the present inventors have observed that even very low concentrations of iron can lead to HMWP formation in pharmaceutical compositions comprising liraglutide or semaglutide.
  • the present inventors have further observed a decreased amount of HMWP generation upon controlling the amount of iron present in pharmaceutical compositions comprising liraglutide or semaglutide.
  • Metal impurities may descend from various sources, both for drug substance and drug product, such as exposure to metal surfaces during processing and as trace impurities in raw materials.
  • the stabilised pharmaceutical composition comprises less than or equal to 3% HMWP.
  • HMWP formation is measured after storage for 1 or 2 years at 5°C. In some embodiments, HMWP formation is measured after storage for 3 years at 5°C. Storage conditions may be 2-8°C, such as 5°C, or at least 2.5 years at 5°C. In some embodiments, HMWP formation is measured after storage for 1 month at 37°C. In some embodiments, HMWP formation is measured after storage for 19 or 22 days at 37°C. Alternatively, storage conditions may be at least 4 weeks, such as 6 weeks or 3 months, optionally at 30°C. Alternatively, the conditions of storage may be at 25°C for 24 hours or 1 week.
  • the conditions of storage may be room temperature for two months, such as up to two months.
  • HMWP formation is measured within the in-use time of the drug product.
  • the stabilised pharmaceutical composition comprises less than or equal to 2.9% HMWP, alternatively 2.8% HMWP, alternatively 2.7% HMWP, alternatively 2.6% HMWP, alternatively 2.5% HMWP, alternatively 2.4% HMWP, alternatively 2.3% HMWP, alternatively 2.2% HMWP, alternatively 2.1% HMWP, alternatively 2.0% HMWP, alternatively 1.9% HMWP, alternatively 1.8% HMWP, alternatively 1.7% HMWP, alternatively 1.6% HMWP, alternatively 1.5% HMWP, alternatively 1.4% HMWP, alternatively 1.3% HMWP, alternatively 1.2% HMWP, alternatively 1.1% HMWP, alternatively 1.0% HMWP, alternatively 0.9% HMWP, alternatively 0.8% HMWP, alternatively 0.7% HMWP, alternatively 0.6% HMWP, alternatively 0.
  • the stabilised pharmaceutical composition comprises 0.01-3.0%, alternatively 0.01-2.9%, alternatively 0.01-2.8%, alternatively 0.01-2.7%, alternatively 0.01-2.6%, alternatively 0.01-2.5%, alternatively 0.01-2.4%, alternatively 0.01-2.3%, alternatively 0.01-2.2%, alternatively 0.01- 2.1%, alternatively 0.01-2.0%, alternatively 0.01-1.9%, alternatively 0.01-1.8%, alternatively 0.01-1.7%, alternatively 0.01-1.6%, alternatively 0.01-1.5%, alternatively 0.01-1.4%, alternatively 0.01-1.3%, alternatively 0.01-1.2%, alternatively 0.01-1.1%, alternatively 0.01- 1.0%, alternatively 0.01-0.9%, alternatively 0.01-0.8%, alternatively 0.01-0.7%, alternatively 0.01-0.6%, alternatively 0.01-0.5%, alternatively 0.01-0.4%, alternatively 0.01-0.3%, alternatively 0.01-0.2%, alternatively 0.01-0.1% HMWP.
  • the stabilised pharmaceutical composition comprises 0.01-3.0%, alternatively 0.1-3.0%, alternatively 0.2-3.0%, alternatively 0.3-3.0%, alternatively 0.4-3.0%, alternatively 0.5-3.0%, alternatively 0.6-3.0%, alternatively 0.7-3.0%, alternatively 0.8-3.0%, alternatively 0.9-3.0%, alternatively 1.0-3.0%, alternatively 11-3.0%, alternatively 12-3.0%, alternatively 1.3- 3.0%, alternatively 14-3.0%, alternatively 15-3.0%, alternatively 16-3.0%, alternatively 1.7- 3.0%, alternatively 18-3.0%, alternatively 19-3.0%, alternatively 2.0-3.0%, alternatively 2.1- 3.0%, alternatively 2.2-3.0%, alternatively 2.3-3.0% HMWP.
  • trace metals such as iron may be quantified using Inductively Coupled Plasma - Optical Emission Spectroscopy (ICP-OES) (Ph. Eur. 2.4.20 Metal Catalysts or Metal Reagents. European Pharmacopoeia 10, 2019; USP ⁇ 233> Elemental Impurities - Procedures USP42-NF37 1S. United States Pharmacopeia-National Formulary. 2010; USP ⁇ 730> Plasma Spectrochemistry, USP42- NF37 1S).
  • ICP-OES Inductively Coupled Plasma - Optical Emission Spectroscopy
  • the stabilised pharmaceutical composition is a liquid pharmaceutical composition.
  • the stabilised pharmaceutical composition has a pH between 7 and 10. In some embodiments, the stabilised pharmaceutical composition has a pH between 7 and 9. In some embodiments, the stabilised pharmaceutical composition has a pH between 7.40 and 8.15. In some embodiments, the stabilised pharmaceutical composition has a pH of 7.40 or 8.15.
  • the GLP-1 peptide is liraglutide and the stabilised pharmaceutical composition comprises less than or equal to 2.5% HMWP. In some embodiments, the stabilised pharmaceutical composition comprises less than or equal to 2.4% HMWP. In some embodiments, the stabilised pharmaceutical composition comprises less than or equal to 2.3% HMWP. In some embodiments, the stabilised pharmaceutical composition comprises less than or equal to 1.2% HMWP. In some embodiments, the stabilised pharmaceutical composition comprises less than or equal to 0.9% HMWP. In some embodiments of the present invention, the GLP-1 peptide is semaglutide and the stabilised pharmaceutical composition comprises less than or equal to 2.6% HMWP. In some embodiments, the stabilised pharmaceutical composition comprises less than or equal to 1.8% HMWP.
  • the stabilised pharmaceutical composition comprises less than or equal to 1.5% HMWP. In some embodiments, the stabilised pharmaceutical composition comprises less than or equal to 0.9% HMWP. In some embodiments, the stabilised pharmaceutical composition comprises less than or equal to 0.7% HMWP.
  • the present invention relates to a process for obtaining a GLP-1 peptide with high purity, said process comprising the steps of: a. Producing said GLP-1 peptide; and b. Purifying said GLP-1 peptide, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 2 pg iron/g GLP-1 peptide, such below 1.8 pg iron/g GLP-1 peptide, such as below 1.5 pg iron/g GLP-1 peptide
  • the present invention relates to a process for obtaining a GLP-1 peptide with high purity, said process comprising the steps of: a. Producing said GLP-1 peptide; and b. Purifying said GLP-1 peptide, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 1 ug iron/g GLP-1 peptide.
  • the concentration of iron is kept between 0.001-2, alternatively 0.002-2, alternatively 0.005-2, alternatively 0.01-2, alternatively 0.02-2, alternatively 0.03-2, alternatively 0.04-2, alternatively 0.05-2, alternatively 0.06-2, alternatively 0.07-2, alternatively 0.08-2, alternatively 0.09-2, alternatively 0.1-2, alternatively 0.2-2, alternatively 0.3-2, alternatively 0.4-2, alternatively 0.5- 2, alternatively 0.6-2, alternatively 0.7-2, alternatively 0.8-2, alternatively 0.9-2 pg iron/g GLP- 1 peptide.
  • the concentration of iron is kept between 0.001-1.8, alternatively 0.002-1.8, alternatively 0.005-1.8, alternatively 0.01-1.8, alternatively 0.02-1.8, alternatively 0.03-1.8, alternatively 0.04-1.8, alternatively 0.05-1.8, alternatively 0.06-1.8, alternatively 0.07-1.8, alternatively 0.08-1.8, alternatively 0.09-1.8, alternatively 0.1-1.8, alternatively 0.2-1.8, alternatively 0.3-1.8, alternatively 0.4-1.8, alternatively 0.5-1.8, alternatively 0.6-1.8, alternatively 0.7-1.8, alternatively 0.8-1.8, alternatively 0.9-1.8 qg iron/g GLP-1 peptide.
  • the concentration of iron is kept between 0.001-1.5, alternatively 0.002-1.5, alternatively 0.005-1.5, alternatively 0.01-1.5, alternatively 0.02-1.5, alternatively 0.03-1.5, alternatively 0.04-1.5, alternatively 0.05-1.5, alternatively 0.06-1.5, alternatively 0.07-1.5, alternatively 0.08-1.5, alternatively 0.09-1.5, alternatively 0.1- 1.5, alternatively 0.2-1.5, alternatively 0.3-1.5, alternatively 0.4-1.5, alternatively 0.5-1.5, alternatively 0.6-1.5, alternatively 0.7-1.5, alternatively 0.8-1.5, alternatively 0.9-1.5 pg iron/g GLP-1 peptide.
  • the concentration of iron is kept between 0.001-1, alternatively 0.002-1, alternatively 0.005-1, alternatively 0.01-1, alternatively 0.02-1, alternatively 0.03-1, alternatively 0.04-1, alternatively 0.05-1, alternatively 0.06-1, alternatively 0.07-1, alternatively 0.08-1, alternatively 0.09-1, alternatively 0.1-1, alternatively 0.2-1, alternatively 0.3-1, alternatively 0.4-1, alternatively 0.5- 1, alternatively 0.6-1, alternatively 0.7-1, alternatively 0.8-1, alternatively 0.9-1 ug iron/g GLP- 1 peptide.
  • the concentration of iron is kept between 0.001-0.95, alternatively 0.001-0.90, alternatively 0.001-0.85, alternatively 0.001-0.80, alternatively 0.001-0.75, alternatively 0.001-0.70, alternatively 0.001-0.65, alternatively 0.001-0.60, alternatively 0.001-0.55, alternatively 0.001-0.50, alternatively 0.001-0.45, alternatively 0.001-0.40, alternatively 0.001-0.35, alternatively 0.001-0.30, alternatively 0.001-0.25, alternatively 0.001-0.20, alternatively 0.001-0.15, alternatively 0.001-0.10, alternatively 0.001-0.05, alternatively 0.001-0.01 ug iron/g GLP-1 peptide.
  • the concentration of iron is controlled for the entire process. In some embodiments, the concentration of iron is controlled for step a and/or step b and/or step c.
  • the concentration of HMWP is kept below or equal to 3% HMWP. In some embodiments, the concentration of HMWP is below or equal to 2.9% HMWP, alternatively 2.8% HMWP, alternatively 2.7% HMWP, alternatively 2.6% HMWP, alternatively 2.5% HMWP, alternatively 2.4% HMWP, alternatively 2.3% HMWP, alternatively 2.2% HMWP, alternatively 2.1% HMWP, alternatively 2.0% HMWP, alternatively 1.9% HMWP, alternatively 1.8% HMWP, alternatively 1.7% HMWP, alternatively 1.6% HMWP, alternatively 1.5% HMWP, alternatively 1.4% HMWP, alternatively 1.3% HMWP, alternatively 1.2% HMWP, alternatively 1.1% HMWP, alternatively 1.0% HMWP, alternatively 0.9% HMWP, alternatively 0.8% HMWP, alternatively 0.7% HMWP, alternatively 0.6% HMWP, alternatively 0.5% HMWP, alternatively 0.4% HMWP, alternatively 0.3% HMWP, alternatively 0.3% HMW
  • the concentration of HMWP is between 0.01-3.0%, alternatively 0.01-2.9%, alternatively 0.01- 2.8%, alternatively 0.01-2.7%, alternatively 0.01-2.6%, alternatively 0.01-2.5%, alternatively 0.01-2.4%, alternatively 0.01-2.3%, alternatively 0.01-2.2%, alternatively 0.01-2.1%, alternatively 0.01-2.0%, alternatively 0.01-1.9%, alternatively 0.01-1.8%, alternatively 0.01- 1.7%, alternatively 0.01-1.6%, alternatively 0.01-1.5%, alternatively 0.01-1.4%, alternatively 0.01-1.3%, alternatively 0.01-1.2%, alternatively 0.01-1.1%, alternatively 0.01-1.0%, alternatively 0.01-0.9%, alternatively 0.01-0.8%, alternatively 0.01-0.7%, alternatively 0.01- 0.6%, alternatively 0.01-0.5%, alternatively 0.01-0.4%, alternatively 0.01-0.3%, alternatively 0.01-0.1% HMWP.
  • the concentration of HMWP is between 0.01-3.0%, alternatively 0.1-3.0%, alternatively 0.2-3.0%, alternatively 0.3-3.0%, alternatively 0.4-3.0%, alternatively 0.5-3.0%, alternatively 0.6-3.0%, alternatively 0.7-3.0%, alternatively 0.8-3.0%, alternatively 0.9-3.0%, alternatively 10-3.0%, alternatively 11-3.0%, alternatively 12-3.0%, alternatively 13-3.0%, alternatively 14-3.0%, alternatively 15-3.0%, alternatively 16-3.0%, alternatively 17-3.0%, alternatively 18-3.0%, alternatively 19-3.0%, alternatively 2.0-3.0%, alternatively 2.1-3.0%, alternatively 2.2-3.0%, alternatively 2.3-3.0% HMWP.
  • the concentration of iron is measured as Fe ions. In some embodiments, the concentration of iron is measured as Fe 2+ and/or Fe 3+ ions.
  • the concentration of iron is measured by ICP-OES (Ph. Eur. 2.4.20 Metal Catalysts or Metal Reagents. European Pharmacopoeia 10, 2019; USP ⁇ 233> Elemental Impurities - Procedures USP42-NF37 1S. United States Pharmacopeia-National Formulary. 2010; USP ⁇ 730> Plasma Spectrochemistry, USP42-NF37 1S).
  • ICP-OES Ph. Eur. 2.4.20 Metal Catalysts or Metal Reagents. European Pharmacopoeia 10, 2019; USP ⁇ 233> Elemental Impurities - Procedures USP42-NF37 1S. United States Pharmacopeia-National Formulary. 2010; USP ⁇ 730> Plasma Spectrochemistry, USP42-NF37 1S).
  • the GLP-1 peptide is liraglutide and the concentration of HMWP is kept below or equal to 2.5% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 2.4% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 2.3% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 1.2% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 0.9% HMWP.
  • GLP-1 peptide is semaglutide and wherein the concentration of HMWP is kept below or equal to 2.6% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 1.8% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 1.5% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 0.9% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 0.7% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 0.5% HMWP. In some embodiments, the concentration of HMWP is kept below or equal to 0.3% HMWP.
  • the present invention relates to a stabilised pharmaceutical composition obtainable by the process of the invention.
  • the present invention relates to a method for prevention or treatment of diabetes or obesity comprising administering the stabilised pharmaceutical composition obtainable by the process of the invention in an effective amount to a subject in need thereof.
  • the present invention relates to a stabilised pharmaceutical composition obtainable by the process of the invention for use in medicine.
  • the present invention relates to a stabilised pharmaceutical composition obtainable by the process of the invention for use in the prevention or treatment of diabetes or obesity.
  • the present invention relates to a use of the stabilised pharmaceutical composition obtainable by the process of the invention for the manufacture of a medicament.
  • the present invention relates to a use of the stabilised pharmaceutical composition obtainable by the process of the invention for the manufacture of a medicament for prevention or treatment of diabetes or obesity.
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a GLP-1 peptide, less than 1 ug iron/g GLP-1 peptide and less than or equal to 3.0% HMWP, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide.
  • the GLP-1 peptide is liraglutide and the stabilised pharmaceutical composition comprises less than or equal to 2.5% HMWP, alternatively 2.4% HMWP, alternatively 2.3% HMWP, alternatively 1.2% HMWP, alternatively 0.9% HMWP.
  • the GLP-1 peptide is semaglutide and the stabilised pharmaceutical composition comprises less than or equal to 2.5% HMWP, alternatively 1.8% HMWP, alternatively 1.5% HMWP, alternatively 0.9% HMWP, alternatively 0.7% HMWP.
  • the present invention relates to a GLP-1 peptide obtainable by the process of the invention.
  • the present invention relates to a method for prevention or treatment of diabetes or obesity comprising administering a GLP-1 peptide obtainable by the process of the invention in an effective amount to a subject in need thereof.
  • the present invention relates to a GLP-1 peptide obtainable by the process of the invention for use in medicine.
  • the present invention relates to a GLP-1 peptide obtainable by the process of the invention for use in the prevention or treatment of diabetes or obesity.
  • the present invention relates to use of a GLP-1 peptide obtainable by the process of the invention for the manufacture of a medicament.
  • the present invention relates to use of a GLP-1 peptide obtainable by the process of the invention for the manufacture of a medicament for prevention or treatment of diabetes or obesity.
  • the composition of the invention comprises a GLP-1 peptide, such as liraglutide or semaglutide, and less than 2 pg iron/g GLP-1 peptide. In some embodiments, the composition of the invention comprises a GLP-1 peptide, such as liraglutide or semaglutide, and less than 1.8 pg iron/g GLP-1 peptide. In some embodiments, the composition of the invention comprises a GLP-1 peptide, such as liraglutide or semaglutide, and less than 1.5 pg iron/g GLP-1 peptide.
  • the composition of the invention comprises a GLP-1 peptide, such as liraglutide or semaglutide, and less than 1 ug iron/g GLP-1 peptide.
  • the compositions of the invention may comprise less than 2 pg iron/g GLP-1 peptide and less than or equal to 3.0% HMWP.
  • the compositions of the invention may comprise less than 1.8 pg iron/g GLP-1 peptide and less than or equal to 3.0% HMWP.
  • the compositions of the invention may comprise less than 1.5 pg iron/g GLP-1 peptide and less than or equal to 3.0% HMWP.
  • the compositions of the invention may comprise less than 1 ug iron/g GLP-1 peptide and less than or equal to 3.0% HMWP.
  • pharmaceutical composition and “composition” are used interchangeably herein and both refer to a pharmaceutical composition.
  • the composition of the invention may be a liquid composition.
  • the composition of the invention may be a solid composition.
  • the composition of the invention may be for subcutaneous or oral administration.
  • the composition of the invention may comprise one or more further pharmaceutically acceptable excipients.
  • excipient broadly refers to any component other than the active therapeutic ingredient(s).
  • the excipient may be an inert substance, an inactive substance, and/or a not medicinally active substance.
  • the excipient may serve various purposes, non-limiting example include as a carrier, vehicle, diluent, tablet aid, and/or to improve administration, and/or absorption of the active substance.
  • the formulation of pharmaceutically active ingredients with various excipients is known in the art, see e.g.
  • Non-limiting examples of excipients are: Solvents, diluents, buffers, preservatives, tonicity regulating agents, chelating agents, and stabilisers.
  • the pharmaceutical composition may have a pH in the range of 7.0-10.0, such as 7.0-9.0 or 7.4- 8.15. In some embodiments, the pH of said pharmaceutical composition is 7.40 or 8.15.
  • said excipients are one or more selected from the group consisting of isotonic agent (e.g. propylene glycol), buffer (e.g. phosphate buffer, such as disodium phosphate dihydrate), delivery agent (e.g.
  • sodium N-(8-(2-hydroxybenzoyl)amino)caprylate also referred to as “SNAC”
  • sodium 8-(salicyloylamino)octanoate also known as sodium 8-(salicyloylamino)octanoate
  • a preservative e.g. phenol
  • the processes of the present invention provide a stable pharmaceutical composition.
  • stable pharmaceutical composition when used herein refers to a composition, e.g. a solution or suspension, comprising GLP-1 peptide, and which composition following storage comprises at least 80%(w/v) of said GLP-1 peptide.
  • stability or “stable” as used interchangeably herein refers to stability following quiescent storage. Stability following quiescent storage may also be referred herein as “storage stability”.
  • stability is chemical stability of the GLP-1 peptide (e.g. determined as described under “Chemical stability assay” herein). Storage conditions for stability testing may be 37°C for 22 or 30 days.
  • storage conditions for stability testing may be 3 months, optionally at 25°C.
  • storage conditions for stability testing may be 2-8°C, such as 5°C or at least 2.5 years at 5°C.
  • the conditions of storage for this stable pharmaceutical composition may be at 5°C for 1 or 2 years.
  • the conditions of this storage may be at 5°C for 24 hours or 1 week.
  • the conditions of this storage may room temperature for two months
  • chemical stability of the GLP-1 peptide requires at least 80%(w/v), such as at least 90%(w/v) or at least 95%(w/v), of said GLP-1 peptide remains in solution in said composition at the end of the storage period. In some embodiments chemical stability of the GLP-1 peptide requires at least 95%(w/v), such as at least 97%(w/v) or at least 99%(w/v), of said GLP-1 peptide remains in solution in said composition at the end of the storage period.
  • the GLP-1 peptide is liraglutide.
  • Liraglutide is Arg34,Lys26- (N-epsilon-(gamma-L-glutamyl(N-alfa-hexadecanoyl)))-GLP-1(7-37), also known as N26- (hexadecanoyl-y-glutamyle)-[34-arginine]GLP-1-(7-37)-peptide (WHO Drug Information Vol. 17, No. 2, 2003). Liraglutide may be prepared as described in Example 37 of WO98/08871.
  • the GLP-1 peptide is semaglutide.
  • Semaglutide is also known as N 626 - ⁇ 18-[N-(17-carboxyheptadecanoyl)-L-y-glutamyl]-10-oxo-3,6, 12,15-tetraoxa-9, 18- diazaoctadecanoyl ⁇ -[8-(2-amino-2-propanoic acid),34-L-arginine]human glucagon-like peptide 1(7-37), see WHO Drug Information Vol. 24, No. 1 , 2010.
  • the GLP-1 peptide semaglutide may be prepared as described in W02006/097537, Example 4.
  • the concentration of GLP-1 peptide may be determined using any suitable method.
  • LC-MS Liquid Chromatography Mass Spectroscopy
  • immunoassays such as RIA (Radio Immuno Assay), ELISA (Enzyme-Linked Immuno Sorbent Assay), and LOCI (Luminescence Oxygen Channeling Immunoasssay).
  • RIA Radio Immuno Assay
  • ELISA Enzyme-Linked Immuno Sorbent Assay
  • LOCI Luminescence Oxygen Channeling Immunoasssay
  • GLP-1 peptide refers to a compound comprising a peptide and which, when active, fully or partially activates the human GLP-1 receptor, e.g. with an EC50 on the human GLP-1 receptor below 5000 pM, such as below 1000 pM or below 500 pM.
  • the EC50 is determined in a medium containing membranes expressing the human GLP-1 receptor, and/or in an assay with whole cells expressing the human GLP-1 receptor.
  • the GLP-1 peptide comprises one or more substituents, such as one or more lipophilic moieties.
  • the GLP-1 peptide is a peptide wherein at least one amino acid residue of the peptide has been substituted with another amino acid residue and/or wherein at least one amino acid residue has been deleted from the peptide and/or wherein at least one amino acid residue has been added to the peptide and/or wherein at least one amino acid residue of the peptide has been modified, compared to e.g. GLP-1(7-37).
  • Such addition or deletion of amino acid residues may take place at the N-terminal of the peptide and/or at the C-terminal of the peptide.
  • GLP-1 peptide e.g., [Arg34]
  • GLP-1 (7-37) designates GLP-1 (7-37) wherein the naturally occurring Lys in position 34 has been substituted with Arg. Unless otherwise stated the GLP-1 comprises only L- amino acids.
  • the GLP-1 peptide is human Glucagon-Like Peptide-1 (GLP- 1(7-37)) comprising one or more amino acid substitutions, deletions, additions and/or insertions.
  • GLP-1 (7-37) has the sequence HAEGTFTSDV SSYLEGQAAKEFIAWLVKGRG.
  • the GLP-1 peptide exhibits at least 60%, 65%, 70%, 80% or 90% sequence identity to GLP-1 (7-37) over the entire length of GLP-1 (7-37).
  • sequence identity As an example of a method for determination of sequence identity between the two peptides [Arg34]GLP- 1(7-37) and GLP-1 (7-37) are aligned.
  • the sequence identity of [Arg34]GLP- 1(7-37) relative to GLP-1 (7-37) is given by the number of aligned identical residues minus the number of different residues divided by the total number of residues in GLP-1 (7-37). Accordingly, in said example the sequence identity is (31-1)/31. Processes for production GLP-1 peptides are well-known in the art.
  • the amino acid sequence of the GLP-1 peptide may for instance be produced by classical peptide synthesis, e.g., solid phase peptide synthesis using t-Boc or Fmoc chemistry or other well established techniques, see, e.g., Greene and Wuts, “Protective Groups in Organic Synthesis”, John Wiley & Sons, 1999, Florencio Zaragoza Dorwald, “Organic Synthesis on solid Phase”, Wley-VCH Verlag GmbH, 2000, and “Fmoc Solid Phase Peptide Synthesis”, Edited by W.C. Chan and P.D. White, Oxford University Press, 2000.
  • peptides may be produced by recombinant methods, viz. by culturing a host cell containing a DNA sequence encoding the unbranched amino acid sequence (peptide) and capable of expressing the peptide in a suitable nutrient medium under conditions permitting the expression of the peptide.
  • host cells suitable for expression of these peptides are: Escherichia coli, Saccharomyces cerevisiae, as well as mammalian BHK or CHO cell lines.
  • the GLP-1 peptide of the invention might be recombinantly produced, synthetically produced or a combination of recombinantly and synthetically produced.
  • the GLP-1 peptide may be in the form of a pharmaceutically acceptable salt, amide, or ester.
  • Salts are e.g. formed by a chemical reaction between a base and an acid, e.g.:
  • the salt may be a basic salt, an acid salt, or it may be neither nor (i.e. a neutral salt).
  • Basic salts produce hydroxide ions and acid salts hydronium ions in water.
  • the salts of GLP-1 peptide may be formed with added cations or anions between anionic or cationic groups, respectively. These groups may be situated in the peptide moiety, and/or in the side chain of GLP-1 peptide. Arg34-GLP- 1(7-37) may be regarded as the peptide moiety of GLP-1 peptide.
  • Gamma-L-glutamyl(N-alfa-hexadecanoyl) may be regarded as the side chain of GLP-1 peptide.
  • anionic groups of GLP-1 peptide include free carboxylic groups in the side chain as well as in the peptide moiety.
  • the peptide moiety of GLP-1 peptide includes a free carboxylic acid group at the C-terminus, and it may also include free carboxylic groups at internal acid amino acid residues, such as Asp and Glu.
  • Non-limiting examples of cationic groups in the peptide moiety include the free amino group at the N-terminus as well as any free amino group of internal basic amino acid residues, such as His, Arg, and Lys.
  • the ester of GLP-1 peptide may be formed by the reaction of a free carboxylic acid group with an alcohol or a phenol, which leads to replacement of at least one hydroxyl group by an alkoxy or aryloxy group.
  • the ester formation may involve the free carboxylic group at the C-terminus of the peptide, and/or any free carboxylic group in the side chain.
  • the amide of GLP-1 peptide may be formed by the reaction of a free carboxylic acid group with an amine or a substituted amine, or by reaction of a free or substituted amino group with a carboxylic acid.
  • the amide formation may involve the free carboxylic group at the C-terminus of the peptide, any free carboxylic group in the side chain, the free amino group at the N-terminus of the peptide, and/or any free or substituted amino group of the peptide in the peptide and/or the side chain.
  • GLP-1 peptide is in the form of a pharmaceutically acceptable salt.
  • GLP-1 peptide is in the form of a pharmaceutically acceptable amide, preferably with an amide group at the C-terminus of the peptide.
  • GLP-1 peptide is in the form a pharmaceutically acceptable ester.
  • the pharmaceutical composition of the invention may be for use in medicine.
  • the pharmaceutical composition of the invention may be for use in the treatment and/or prevention of diabetes or obesity.
  • the pharmaceutical composition of the invention is for use in prevention and/or treatment of type 1 diabetes or type 2 diabetes.
  • the pharmaceutical composition of the invention is for use in prevention and/or treatment of NASH. In some embodiments the pharmaceutical composition of the invention is for use in prevention and/or treatment of diabetic complications, such as angiopathy; neuropathy, including peripheral neuropathy; nephropathy; and/or retinopathy. In some embodiments the pharmaceutical composition of the invention is for use in prevention and/or treatment of one or more cardiovascular diseases. In some embodiments the pharmaceutical composition of the invention is for use in prevention and/or treatment of sleep apnoea. In some embodiments the invention provides a method for prevention or treatment of diabetes or obesity comprising administering the composition as defined herein in an effective amount to a subject in need thereof.
  • the term “a” means “one or more”. In some embodiments, and unless otherwise indicated herein, terms presented in singular form also include the plural situation. In some embodiments, the term “about” means the value referred to ⁇ 10%.
  • a process for obtaining a stabilised pharmaceutical composition comprising a GLP-1 peptide, said process comprising the steps of: a. Producing said GLP-1 peptide; b. Purifying said GLP-1 peptide; and c. Formulating said GLP-1 peptide into said stabilised pharmaceutical composition, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 2 pg iron/g GLP-1 peptide, such as below 1.8 pg iron/g GLP-1 peptide, such as below 1.5 pg iron/g GLP-1 peptide.
  • a process for obtaining a stabilised pharmaceutical composition comprising a GLP-1 peptide, said process comprising the steps of: a. Producing said GLP-1 peptide; b. Purifying said GLP-1 peptide; and c. Formulating said GLP-1 peptide into said stabilised pharmaceutical composition, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 1 ug iron/g GLP-1 peptide.
  • said stabilised pharmaceutical composition comprises less than or equal to 3% HMWP.
  • said stabilised pharmaceutical composition comprises less than or equal to 2.9% HMWP, alternatively 2.8% HMWP, alternatively 2.7% HMWP, alternatively 2.6% HMWP, alternatively 2.5% HMWP, alternatively 2.4% HMWP, alternatively 2.3% HMWP, alternatively 2.2% HMWP, alternatively 2.1% HMWP, alternatively 2.0% HMWP, alternatively 1.9% HMWP, alternatively 1.8% HMWP, alternatively 1.7% HMWP, alternatively 1.6% HMWP, alternatively 1.5% HMWP, alternatively 1.4% HMWP, alternatively 1.3% HMWP, alternatively 1.2% HMWP, alternatively 1.1% HMWP, alternatively 1.0% HMWP, alternatively 0.9% HMWP, alternatively
  • said stabilised pharmaceutical composition comprises 0.01-3.0%, alternatively 0.01-2.9%, alternatively 0.01-2.8%, alternatively 0.01-2.7%, alternatively 0.01-2.6%, alternatively 0.01-2.5%, alternatively 0.01-2.4%, alternatively 0.01-2.3%, alternatively 0.01-2.2%, alternatively 0.01-2.1%, alternatively 0.01-2.0%, alternatively 0.01-1.9%, alternatively 0.01-1.8%, alternatively 0.01-1.7%, alternatively 0.01-1.6%, alternatively 0.01-1.5%, alternatively 0.01-1.4%, alternatively 0.01-1.3%, alternatively 0.01-1.2%, alternatively 0.01-1.1%, alternatively 0.01-1.0%, alternatively 0.01-0.9%, alternatively 0.01-0.8%, alternatively 0.01-0.7%, alternatively 0.01-0.6%, alternatively 0.01-0.5%, alternatively 0.01-0.4%, alternatively 0.01-0.3%, alternatively 0.01-0.2%, alternatively 0.01-0.1%, alternatively 0.01-1.0%, alternatively
  • said stabilised pharmaceutical composition comprises 0.01-3.0%, alternatively 0.1-3.0%, alternatively 0.2-3.0%, alternatively 0.3-3.0%, alternatively 0.4-3.0%, alternatively 0.5-3.0%, alternatively 0.6-3.0%, alternatively 0.7-3.0%, alternatively 0.8-3.0%, alternatively 0.9- 3.0%, alternatively 10-3.0%, alternatively 11-3.0%, alternatively 12-3.0%, alternatively 13-3.0%, alternatively 14-3.0%, alternatively 15-3.0%, alternatively 16-3.0%, alternatively 17-3.0%, alternatively 18-3.0%, alternatively 19-3.0%, alternatively 2.0- 3.0%, alternatively 2.13.0%, alternatively 2.2-3.0%, alternatively 2.3-3.0% HMWP.
  • a process for obtaining a GLP-1 peptide with high purity comprising the steps of: a. Producing said GLP-1 peptide; and b. Purifying said GLP-1 peptide, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 2 pg iron/g GLP-1 peptide.
  • a process for obtaining a GLP-1 peptide with high purity comprising the steps of: a. Producing said GLP-1 peptide; and b. Purifying said GLP-1 peptide, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 1.8 pg iron/g GLP-1 peptide.
  • a process for obtaining a GLP-1 peptide with high purity comprising the steps of: a. Producing said GLP-1 peptide; and b. Purifying said GLP-1 peptide, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 1.5 pg iron/g GLP-1 peptide.
  • a process for obtaining a GLP-1 peptide with high purity comprising the steps of: a. Producing said GLP-1 peptide; and b. Purifying said GLP-1 peptide, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide, characterised in that the concentration of iron is kept below 1 ug iron/g GLP-1 peptide.
  • HMWP is kept below or equal 2.9% HMWP, alternatively 2.8% HMWP, alternatively 2.7% HMWP, alternatively 2.6% HMWP, alternatively 2.5% HMWP, alternatively 2.4% HMWP, alternatively 2.3% HMWP, alternatively 2.2% HMWP, alternatively 2.1% HMWP, alternatively 2.0% HMWP, alternatively 1.9% HMWP, alternatively 1.8% HMWP, alternatively 1.7% HMWP, alternatively 1.6% HMWP, alternatively 1.5% HMWP, alternatively 1.4% HMWP, alternatively 1.3% HMWP, alternatively 1.2% HMWP, alternatively 1.1% HMWP, alternatively 1.0% HMWP, alternatively 0.9% HMWP, alternatively 0.8% HMWP, alternatively 0.7% HMWP, alternatively 0.6% HMWP, alternatively 0.5% HMWP, alternatively 0.4% HMWP, alternatively 0.3% HMWP, alternatively 0.2% HMWP, alternatively 0.1% HMWP, alternatively 0.1% HMWP, alternatively 1.0% HMWP,
  • a stabilised pharmaceutical composition obtainable by any of embodiments 1-3835.
  • a GLP-1 peptide obtainable by any of embodiments 42-66.
  • a method for prevention or treatment of diabetes or obesity comprising administering the stabilised pharmaceutical composition obtainable by any of embodiments 1-35 in an effective amount to a subject in need thereof.
  • the stabilised pharmaceutical composition obtainable by any of embodiments 1-35 for use in medicine.
  • the stabilised pharmaceutical composition obtainable by any of embodiments 1-35 for use in the prevention or treatment of diabetes or obesity.
  • the stabilised pharmaceutical composition obtainable by any of embodiments 1-35 for use in the prevention or treatment of NASH.
  • a method for prevention or treatment of diabetes or obesity comprising administering the GLP-1 peptide obtainable by any of embodiments 42-66 in an effective amount to a subject in need thereof.
  • a method for prevention or treatment of NASH comprising administering the GLP-1 peptide obtainable by any of embodiments 42-66 in an effective amount to a subject in need thereof.
  • the GLP-1 peptide obtainable by any of embodiments 42-66 for use in medicine.
  • the GLP-1 peptide obtainable by any of embodiments 42-66 for use in the prevention or treatment of diabetes or obesity.
  • the GLP-1 peptide obtainable by any of embodiments 42-66 for use in the prevention or treatment of NASH.
  • the GLP-1 peptide obtainable by any of embodiments 42-66 for use in the prevention or treatment of NASH.
  • a pharmaceutical composition comprising a GLP-1 peptide, less than 2 pg iron/g GLP-1 peptide and less than or equal to 3.0% HMWP, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide.
  • a pharmaceutical composition comprising a GLP-1 peptide, less than 1.8 pg iron/g GLP-1 peptide and less than or equal to 3.0% HMWP, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide.
  • a pharmaceutical composition comprising a GLP-1 peptide, less than 1.5 pg iron/g GLP-1 peptide and less than or equal to 3.0% HMWP, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide.
  • a pharmaceutical composition comprising a GLP-1 peptide, less than 1 ug iron/g GLP-1 peptide and less than or equal to 3.0% HMWP, wherein said GLP-1 peptide is selected from the group consisting of liraglutide and semaglutide.
  • composition according to embodiment 88, wherein said stabilised pharmaceutical composition comprises less than or equal to 2.3% HMWP.
  • Example 1 Stock solutions of 50mM calcium chloride, copper (II) acetate (hydrate), copper (II) chloride, iron (II) chloride (tetrahydrate), iron (II) sulphate (heptahydrate), iron (iii) chloride (anhydrous) and iron (iii) sulphate (anhydrous) were prepared by weighing out materials and dissolving powder in appropriate volumes of MilliQ water.
  • the metal salt stock solutions were used to prepare two-fold dilution series of each of the metals in 96 deep well plate. Blank reference solutions were also prepared as two-fold dilution with MilliQ water. Target concentrations were ranging from 0.4 mM to 0.0002 mM.
  • the diluted metals stock solutions were diluted a further 4x in a 384 well assay plate and mixed with a 2x liraglutide stock solution containing 3.2 mM liraglutide 16 mM phosphate,
  • the final solutions in the 384 well plate contained 1.6 mM liraglutide, 8 mM phosphate (pH 8.15 or 7.4), 58 mM phenol and 184 mM propylene glycol, and metal concentrations ranging from 0.1 mM to 0.00005 mM in duplicates.
  • Example 2 A freshly prepared 50 mM stock solution of FeSCU was used to make 2- fold dilution series (400 mM to 0.2 mM). Stock solutions of 4 mM liraglutide and semaglutide in MilliQ water, 500 mM phenol and 100 mM tris pH 7.4 were mixed with the 2-fold dilution series of FeSC to create 2 x 100 pL samples for HMWP stability assay containing 0.6 mM liraglutide or semaglutide, 0 or 50 mM phenol and 10 mM tris pH 7.4 and 100 to 0.05pM FeSC in half-area 96 well plates.
  • Quantification of trace metals may be performed by Inductively Coupled Plasma - Optical Emission Spectroscopy (ICP-OES) (Ph. Eur. 2.4.20 Metal Catalysts or Metal Reagents. European Pharmacopoeia 10, 2019; USP ⁇ 233> Elemental Impurities - Procedures USP42-NF37 1S. United States Pharmacopeia-National Formulary. 2010; USP ⁇ 730> Plasma Spectrochemistry, USP42-NF37 1S.) Sample may be diluted in 1% nitric acid containing cesium chloride, extracted at approximately 70 °C for 1 h and centrifuged before analysis with ICP-OES.
  • ICP-OES Inductively Coupled Plasma - Optical Emission Spectroscopy
  • the 96/384 well assay plate was incubated at 37 °C in a Liconic STX plate hotel.
  • An integrated robotic system Freeslate / Unchained Labs
  • After a total cumulated time of 19 or 22 days at 37°C in the incubators the plates were removed and analysed using denaturing SEC UPLC (Waters BEH125A, Isocratic B (HMWP): R-496: 0.15 % v/v TFA in 60 % v/v acetonitrile, UV detection 215nm and 280nm).
  • Example 1 HMWP formation as function of ug metal / g liraglutide at pH 7.40 and 8.15.
  • Samples were prepared as described under General Methods, “Sample Preparation (Example 1)” and HMWP formation was measured as function of ug metal / g liraglutide at pH 7.40 and 8.15 as described under General methods, “Chemical stability assay”.
  • Table 1 Formation of liraglutide HMWP at 37°C after 22 days as a function of metal concentration in formulations at pH 7.4 and 8.15. Data for the effect of Ca 2+ , Cu 2+ , Fe 2+ and Fe 3+ are represented.
  • Table 1 show an increased level of HMWP formation in the presence of iron (II) ions and iron (III) ions.
  • the presence of iron (II) and iron (III) ions clearly affects the HMWP formation down to a concentration of 1 ug metal / g liraglutide.
  • the effect on HMWP formation is more pronounced for iron (II) and iron (III) ions than the effect of copper (II) ions.
  • the presence of calcium ions does not influence the HMWP formation.
  • Example 2 HMWP formation as function of iron concentration in liraqlutide or semaqlutide formulations at pH 7.40
  • Samples were prepared as described under General Methods, “Sample Preparation (Example 2)” and HMWP formation was measured as function of ug metal / L at pH 7.40 as described under General methods, “Chemical stability assay”.
  • Table 2 Formation of liraglutide and semaglutide HMWP at 37°C after 19 days as a function of iron (Fe 2+ ) concentration in 10 mM Tris, and 0.6 mM liraglutide or semaglutide, pH 7.4, with and without 50 mM phenol. Data for the effect of Fe 2+ are represented. * These data points were not reproducible in follow-up experiments and are considered outliers.

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Abstract

La présente invention concerne des procédés d'augmentation de la stabilité de compositions comprenant un peptide GLP-1, des compositions de GLP-1 stables et des utilisations de telles compositions.
PCT/EP2020/083686 2019-11-29 2020-11-27 Procédés d'obtention de compositions de glp-1 stables WO2021105393A1 (fr)

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998008871A1 (fr) 1996-08-30 1998-03-05 Novo Nordisk A/S Derives de glp-1
WO2006097537A2 (fr) 2005-03-18 2006-09-21 Novo Nordisk A/S Composes de glp-1 acyles
WO2009030738A1 (fr) 2007-09-05 2009-03-12 Novo Nordisk A/S Dérivés de glucagon-like peptide-1 et leur utilisation pharmaceutique
EP2209800B1 (fr) 2007-11-16 2013-07-24 Novo Nordisk A/S Composition pharmaceutique durable comprenant du liraglutide et du degludec
WO2018210919A1 (fr) * 2017-05-17 2018-11-22 Novo Nordisk A/S Compositions de glp-1 et leurs utilisations
WO2019038412A1 (fr) * 2017-08-24 2019-02-28 Novo Nordisk A/S Compositions de glp-1 et leurs utilisations
WO2020084126A1 (fr) * 2018-10-26 2020-04-30 Novo Nordisk A/S Compositions de sémaglutide stables et leurs utilisations

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998008871A1 (fr) 1996-08-30 1998-03-05 Novo Nordisk A/S Derives de glp-1
WO2006097537A2 (fr) 2005-03-18 2006-09-21 Novo Nordisk A/S Composes de glp-1 acyles
WO2009030738A1 (fr) 2007-09-05 2009-03-12 Novo Nordisk A/S Dérivés de glucagon-like peptide-1 et leur utilisation pharmaceutique
EP2209800B1 (fr) 2007-11-16 2013-07-24 Novo Nordisk A/S Composition pharmaceutique durable comprenant du liraglutide et du degludec
WO2018210919A1 (fr) * 2017-05-17 2018-11-22 Novo Nordisk A/S Compositions de glp-1 et leurs utilisations
WO2019038412A1 (fr) * 2017-08-24 2019-02-28 Novo Nordisk A/S Compositions de glp-1 et leurs utilisations
WO2020084126A1 (fr) * 2018-10-26 2020-04-30 Novo Nordisk A/S Compositions de sémaglutide stables et leurs utilisations

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
"European Pharmacopoeia", vol. 10, 2019, article "Ph. Eur. 2.4.20 Metal Catalysts or Metal Reagents"
"Remington: The Science and Practice of Pharmacy", 1995
ANONYMOUS: "EMEA - Ozempic - ANNEX I SUMMARY OF PRODUCT CHARACTERISTICS", 5 July 2019 (2019-07-05), pages 1 - 68, XP055710791, Retrieved from the Internet <URL:http://web.archive.org/web/20190705172542if_/https://www.ema.europa.eu/en/documents/product-information/ozempic-epar-product-information_en.pdf> [retrieved on 20200701] *
ANONYMOUS: "EMEA: Assessment report for Victoza", 23 March 2019 (2019-03-23), pages 1 - 49, XP055710793, Retrieved from the Internet <URL:http://web.archive.org/web/20190323191350if_/https://www.ema.europa.eu/en/documents/assessment-report/victoza-epar-public-assessment-report_en.pdf> [retrieved on 20200701] *
FLORENCIO ZARAGOZA DORWALD: "Fmoc Solid Phase Peptide Synthesis", 2000, OXFORD UNIVERSITY PRESS
GREENEWUTS: "Protective Groups in Organic Synthesis", 1999, JOHN WILEY & SONS
LABOUR ET AL: "Pharmaceuticals and Medical Devices Agency Japan : Victoza Subcutaneous Injection 18 mg - Report on the Deliberation Results", PHARMACEUTICAL AND FOOD SAFETY BUREAU, MINISTRY OF HEALTH, 1 January 2009 (2009-01-01), pages 1 - 119, XP055515927, Retrieved from the Internet <URL:https://www.pmda.go.jp/files/000153180.pdf> [retrieved on 20181016] *
STABY ARNE ET AL: "Influence of Production Process and Scale on Quality of Polypeptide Drugs: a Case Study on GLP-1 Analogs", PHARMACEUTICAL RESEARCH, SPRINGER NEW YORK LLC, US, vol. 37, no. 7, 8 June 2020 (2020-06-08), XP037160410, ISSN: 0724-8741, [retrieved on 20200608], DOI: 10.1007/S11095-020-02817-9 *
WHO, DRUG INFORMATION, vol. 17, no. 2, 2003
WHO, DRUG INFORMATION, vol. 24, no. 1, 2010

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