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CA-3134757-C - METHOD FOR PREPARING STABLE PEPTIDE FORMULATIONS

CA3134757CCA 3134757 CCA3134757 CCA 3134757CCA-3134757-C

Abstract

The present invention provides an improved method for preparing a powder formulation containing a peptide. The present invention further provides an improved method for preparing a powder formulation containing glucagon or a glucagon analog, wherein said powder formulation is suitable for nasal administration.

Inventors

  • Gregory Nelson Brown
  • Kurt Gard Van Scoik

Assignees

  • ELI LILLY AND COMPANY

Dates

Publication Date
20260505
Application Date
20200420
Priority Date
20190426

Claims (20)

  1. WE CLAIM: 1. A method for preparing a peptide powder formulation comprising the steps of: a. forming a first mixture of an acid, a phospholipid surfactant, and a cyclodextrin in an aqueous carrier; b. subjecting the first mixture to a first filtration step wherein the filter comprises a membrane with a pore size of about 0.4 µm to about 0.5 µm; c. adding a peptide to the first filtration product to form a second mixture, and subjecting the second mixture to a second filtration step wherein the filter comprises a membrane with a pore size of about 0.4 µm to about 0.5 µm; and d. drying the second filtration product to form a solid formulation and processing the solid formulation to produce the peptide powder formulation.
  2. 2. The method of claim 1 wherein the peptide is glucagon or a glucagon analog.
  3. 3. The method of claim 2 wherein the glucagon analog is [Lys17,18, Glu21]glucagon or a glucagon of SEQ ID NO: 1 in which one amino acid is replaced with an alanine.
  4. 4. The method of any one of claim 1 to 3 wherein the surfactant, the cyclodextrin and the peptide together constitute between about 1.5% and about 3% by weight of the second mixture.
  5. 5. The method of claim 4 wherein the surfactant, the cyclodextrin and the peptide together constitute about 2% by weight of the second mixture.
  6. 6. The method of claim 4 wherein the surfactant, the cyclodextrin and the peptide together constitute about 2.5% by weight of the second mixture.
  7. 7. The method of any one of claims 1 to 6 wherein the membrane in both the first and the second filtration steps comprise a polyvinylidene difluoride (PVDF) membrane.
  8. 8. The method of any one of claims 1 to 7 wherein the membrane in both the first and the second filtration steps comprises a pore size of about 0.45 µm.
  9. 9. The method of any one of claims 1 to 8 wherein the acid is citric acid or acetic acid.
  10. 10. The method of claim 9 wherein the acid is acetic acid.
  11. 11. The method of claim 10 wherein the acetic acid is at a concentration of 1M.
  12. 12. The method of any one of claims 1 to 11 wherein the surfactant is dodecylphosphocholine, didecylphosphatidylcholine, lysolauroylphosphatidylcholine, dioctanoylphosphatidylcholine, or dilauroylphosphatidylglycerol. 18
  13. 13. The method of claim 12 wherein the surfactant is dodecylphosphocholine.
  14. 14. The method of any one of claims 1 to 13 wherein the cyclodextrin is α-cyclodextrin, β-cyclodextrin, hydroxypropyl β-cyclodextrin, or γ-cyclodextrin.
  15. 15. The method of claim 14 wherein the cyclodextrin is β-cyclodextrin.
  16. 16. A method for preparing a glucagon powder formulation comprising the steps of: a. forming a first mixture of acetic acid, dodecylphosphocholine, and β cyclodextrin in an aqueous carrier; b. subjecting the first mixture to a first filtration step to generate a first filtration product wherein the filter comprises a membrane with a pore size of about 0.4 µm to about 0.5 µm; c. adding glucagon to the first filtration product to form a second mixture, and subjecting the second mixture to a second filtration step to generate a second filtration product, wherein the filter comprises a membrane with a pore size of about 0.4 µm to about 0.5 µm; and d. drying the second filtration product to form a solid formulation and processing the solid formulation to produce the glucagon powder formulation.
  17. 17. The method of claim 16 wherein the dodecylphosphocholine, the β-cyclodextrin and the glucagon together constitute about 2.5% by weight of the second mixture.
  18. 18. The method of claim 16 or claim 17 wherein the membrane in both the first and the second filtration steps comprises a PVDF membrane.
  19. 19. The method of any one of claims 16 to 18 wherein the membrane in both the first and the second filtration steps comprises a pore size of about 0.45 µm.
  20. 20. The method of any one of claims 16 to 19 wherein the acetic acid is at a concentration of 1M.

Description

Method for Preparing Stable Peptide Formulations The present invention relates to the field of medicine. More particularly, the present invention provides an improved method for preparing a powder formulation 5 containing a peptide. The present invention further provides an improved method for preparing a powder formulation containing glucagon or a glucagon analog, wherein said powder formulation is suitable for nasal administration. Peptides are prone to physical instability such as aggregation during and after the manufacturing process. Aggregation is a complex process that originates by several 10 different mechanisms. Aggregation can be typically induced by nucleation of a few peptides or proteins, which form small and soluble aggregates; these then serve as nucleation foci for the subsequent growth oflarger insoluble aggregates. The nucleationgrowth process can increase with time, temperature, protein concentration, and other parameters. During manufacturing, proteins are purified and concentrated using a variety 15 of means such as ul trafiltrati on, affinity chromatography, selective absorption chromatography, ion-exchange chromatography, lyophilization, dialysis, and precipitation or "salting out". Such concentration processes can lead to aggregation (Maggio, BioProcess International 2008; 6(10): 58-65). Removing or solubilizing these aggregates requires extra process steps which can be costly and can compromise the overall product 20 yield. Effects of aggregation can include loss of material, reduced efficacy, altered pharmacokinetics, reduced stability and product shelflife, and induction of unwanted immunogenicity. Aggregation has become a major issue for biopharmaceutical manufacturers particularly because the current trend toward high-concentration solutions increases the 25 likelihood of protein-protein interactions, which in tum favors aggregation. (Maggio, BioProcess International 2008; 6(10): 58-65). Various approaches to limiting aggregation of a peptide have been studied, including, but not limited to, adjusting: pH, buffer conditions, ionic strength, and/or adding other excipients such as cyclodextrins. Glucagon is known for its tendency to aggregate in aqueous solutions (Pedersen 30 JS., J Diabetes Sci Technol. 2010; 4(6): 1357-1367; Beaven et al., lhe European J. Biochem. 1969; 11(1): 37-42; Matilainen et al., European J of Pharmaceutical Sciences 2009; (36): 412-420), which can cause issues during the manufacture of glucagon powder I formulations. Previous methods of preparing glucagon powder formulations suitable for nasal administration are disclosed in WO2016/133863. There exists a need for alternative methods for preparing peptide powder formulations, in particular glucagon or glucagon analog powder formulations. In 5 particular, there is a need for methods which reduce or eliminate aggregation of the peptide in aqueous solution. By reducing, or, preferably, eliminating aggregation, the final powder formulation will retain a very high percentage of active peptide, which is highly advantageous. Preferably, the method results in an aqueous solution prior to drying which is physically and chemically stable for an extended period of time, for 10 example up to 24 hours. This extended stability makes the process much more amenable to large scale manufacture. There is furthermore a need for a method which results in a final powder formulation having a long shelf-life, preferably up to about 24 months. Accordingly, the present invention provides an improved and cost effective method of reducing aggregation of a peptide during the manufacture of a powder 15 formulation. This method incorporates a double filtration step. One such peptide used in the present invention is glucagon or a glucagon analog. The powder formulations prepared according to the present method are particularly suitable for nasal administration. In accordance with one aspect of the invention, a method for preparing a peptide 20 powder formulation is provided. This method comprises the steps of: a. forming a first mixture of an acid, a phospholipid surfactant, and a cyclodextrin in an aqueous earner; b. subjecting the first mixture to a first filtration step wherein the filter comprises a membrane with a pore size of about 0.4 μm to about 0.5 μm; 25 c. adding a peptide to the first filtration product to form a second mixture, and subjecting the second mixture to a second filtration step wherein the filter comprises a membrane with a pore size of about 0.4 μm to about 0.5 μm; and d. drying the second filtration product to form a solid formulation and processing the solid formulation to produce a final powder formulation. 30 In an embodiment, the peptide is glucagon or a glucagon analog. In particular, it is glucagon. 2 In an embodiment, the acid is citric acid or acetic acid. In particular, it is acetic acid. More particularly, the acetic acid is at a concentration of IM. In an embodiment, the surfactant, the cyclodextrin