КОМПОЗИЦИЯ НА ОСНОВЕ ПЕПТИДА GLP-1 И ЕЕ ПРИМЕНЕНИЕ
Patent Information
- Authority / Receiving Office
- EA · EA
- Patent Type
- Applications
- Current Assignee / Owner
- GAN & LEE PHARM CO LTD
- Filing Date
- 2024-10-09
- Publication Date
- 2026-07-13
Abstract
Description
GLP-1 peptide compositions and uses thereof Technical Field
[0001] The present invention belongs to the field of biomedicine and relates to a stable pharmaceutical composition containing GLP-1 peptide. Background Art
[0002] Glucagon-like peptide 1 (GLP-1) and its analogs and derivatives are highly effective in treating type 1 and type 2 diabetes. However, GLP-1 peptides are susceptible to instability, such as chemical instability, in liquid solutions. Therefore, there is a need to optimize formulation systems and provide liquid pharmaceutical compositions of GLP-1 peptides with excellent stability.
[0003] Summary of the Invention
[0004] The present invention provides a liquid pharmaceutical composition of a GLP-1 peptide having excellent chemical stability. In some embodiments, the present invention relates to a kit comprising a pharmaceutical composition as defined herein. In some embodiments, the present invention relates to a pharmaceutical composition as defined herein for medical use.
[0005] In some embodiments of the present invention, GLP-1 peptide formulations are provided that are stable during storage and delivery. A stable formulation is one in which the GLP-1 peptide substantially retains its physical and chemical stability during storage, produces less impurities and generates less high molecular weight proteins under the desired storage conditions, and meets the storage and delivery requirements of injectable formulations. The primary factor determining shelf life is typically the formation of byproducts and degradation products. The formulations of the present invention achieve these desired levels of stability.
[0006] In addition to adequate physical and chemical stability, the formulation should also possess an acceptable pH and osmotic pressure for application. In particular, high concentrations of the active drug can increase aggregation in the formulation. In some embodiments of the present invention, the pharmaceutical formulation can reduce GLP peptide aggregation within a certain concentration range, providing a pharmaceutical formulation that is stable across a range of GLP-1 peptide concentrations from low to high.
[0007] In addition, in some embodiments of the invention, the concentration of the formulation components is optimized from the perspective of patient injection comfort, providing a solution to reduce patient pain.
[0008] In its broadest aspect, the present invention provides a pharmaceutical formulation comprising a GLP-1 peptide as an active ingredient and a buffer system, an osmotic pressure regulator, and an antioxidant (the formulation of the present invention). The formulation of the present invention is a liquid (e.g., an aqueous solution). In some embodiments of the present invention, a stable GLP-1 peptide formulation suitable for subcutaneous administration is provided.
[0009] Specifically, the first aspect of the present invention provides a liquid pharmaceutical composition comprising:
[0010] a. GLP-1 peptide or a pharmaceutically acceptable salt thereof;
[0011] b. Osmotic pressure regulator;
[0012] c. antioxidants; and
[0013] d. buffer;
[0014] Wherein, the GLP-1 peptide is selected from:
[0015] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(19-carboxynonadecanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0016] N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0017] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0018] N-ε 26 -[2-(2-[2-(4-[21-carboxyheneicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0019] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(23-carboxytricosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0020] N-ε 26-[2-(2-[2-(4-[23-carboxytricosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0021] N-ε 26 -(23-carboxytricosanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide,
[0022] N-ε 26 -(19-carboxynonadecanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide,
[0023] N-ε 26 -(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoyl-[Gly8,Arg34]GLP-1-(7-37) peptide,
[0024] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(19-carboxynonadecanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0025] N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0026] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0027] N-ε 26 -[2-(2-[2-(4-[21-carboxyheneicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0028] N-ε 26-[2-(2-[2-(2-[2-(2-[4-(23-carboxytricosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0029] N-ε 26 -[2-(2-[2-(4-[23-carboxytricosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0030] N-ε 26 -(23-carboxytricosanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide,
[0031] N-ε 26 -(19-carboxynonadecanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide,
[0032] N-ε 26 -(21-carboxyheneicosanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide,
[0033] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(20-carboxyeicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0034] N-ε 26 -[2-(2-[2-(4-[20-carboxyeicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0035] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(22-carboxydocosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0036] N-ε 26-[2-(2-[2-(4-[22-carboxydocosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0037] N-ε 26 -(20-carboxyeicosanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide,
[0038] N-ε 26 -(22-carboxydocosanoylamino)-4(S)-carboxybutanoyl-[Gly8,Arg34]GLP-1-(7-37) peptide,
[0039] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(20-carboxyeicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0040] N-ε 26 -[2-(2-[2-(4-[20-carboxyeicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0041] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(22-carboxydocosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0042] N-ε 26 -[2-(2-[2-(4-[22-carboxydocosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide,
[0043] N-ε 26 -(20-carboxyeicosanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide, and
[0044] N-ε 26 -(22-carboxydocosanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide;
[0045] Preferably, the GLP-1 peptide is selected from the following compounds:
[0046] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(19-carboxynonadecanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0047] N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide,
[0048] N-ε 26 -(19-carboxynonadecanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide,
[0049] N-ε 26 -(19-carboxynonadecanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide,
[0050] N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, and
[0051] N-ε 26 -[2-(2-[2-(4-[21-carboxyheneicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide.
[0052] Preferably, the GLP-1 peptide is the following compound:
[0053] In some embodiments, the pharmaceutical composition contains or does not contain a preservative. Preferably, the pharmaceutical composition does not contain a preservative. Preferably, the preservative is phenol or m-cresol. Preferably, the pharmaceutical composition does not contain phenol or m-cresol.
[0054] In some embodiments, the buffer is selected from at least one of acetic acid-sodium acetate buffer, acetic acid-potassium acetate, acetic acid-ammonium acetate, and phosphate buffer, preferably phosphate buffer; preferably, the phosphate buffer is Na2HPO4.
[0055] In some embodiments, the osmotic pressure regulator is NaCl or propylene glycol.
[0056] In some embodiments, the antioxidant is selected from at least one of ascorbic acid, methionine, citric acid and tartaric acid. Preferably, the antioxidant is selected from citric acid and methionine; preferably, the citric acid is citric acid monohydrate.
[0057] In some embodiments, the pharmaceutical composition comprises more than about 0.5 mg / ml Na2HPO4, preferably about 0.5-20 mg / ml Na2HPO4, preferably about 1-20 mg / ml Na2HPO4, preferably about 1-10 mg / ml Na2HPO4, more preferably about 1-5 mg / ml Na2HPO4, and further preferably about 1.40 mg / ml, about 1.41 mg / ml, about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml Na2HPO4.
[0058] In some embodiments, the concentration of NaCl is about 0.5 mg / ml or more, preferably about 1 mg / ml or more, preferably about 1-30 mg / ml, preferably about 3-25 mg / ml, preferably about 5-15 mg / ml, preferably about 5-10 mg / ml, preferably about 5 mg / ml, about 5.5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 8.10 mg / ml, about 8.15 mg / ml, about 8.20 mg / ml, about 8.25 mg / ml, about 8.30 mg / ml, about 9 mg / ml or about 10 mg / ml.
[0059] In some embodiments, the concentration of propylene glycol is about 1 mg / ml or more, preferably about 2 mg / ml or more, preferably about 5-25 mg / ml, preferably about 10-20 mg / ml, preferably about 12-18 mg / ml, preferably about 13 mg / ml, about 13.5 mg / ml, about 14 mg / ml, about 14.5 mg / ml, about 15 mg / ml, about 16 mg / ml or about 17 mg / ml.
[0060] In some embodiments, the concentration of the antioxidant is about 0.5 mM or more, preferably about 1 mM or more, preferably about 1-25 mM, preferably about 1-20 mM, preferably about 1-15 mM, preferably about 1-10 mM, preferably about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM or about 9 mM.
[0061] In some embodiments, the antioxidant is selected from citric acid at a concentration of about 1-25 mM, preferably at a concentration of about 1-20 mM, preferably at a concentration of about 1-15 mM, preferably at a concentration of about 1-10 mM, preferably at a concentration of about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM or about 9 mM; preferably, the citric acid is citric acid monohydrate.
[0062] In some embodiments, the concentration of the GLP-1 peptide is at least about 1 mg / ml, preferably at least about 2 mg / ml, preferably about 1-80 mg / ml, preferably about 1-70 mg / ml, preferably about 1-65 mg / ml, preferably about 1-60 mg / ml, preferably about 2-60 mg / ml, preferably about 2-50 mg / ml, more preferably about 2 mg / ml, about 3 mg / ml, about 4 mg / ml, about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 9 mg / ml, about 10 mg / ml, about 11 mg / ml, about 12 mg / ml, about 13 mg / ml, about 14 mg / ml, about 15 mg / ml, about 16 mg / ml, about 17 mg / ml, about 18 mg / ml, about 19 mg / ml, about 20 mg / ml, about 21 mg / ml, about 22 mg / ml, about 23 mg / ml, about 24 mg / ml, about 25 mg / ml, about 26 mg / ml, about 27 mg / ml, about 28 mg / ml, about 29 mg / ml, about 30 mg / ml, about 31 mg / ml, about 32 mg / ml, about 33 mg / ml, about 34 mg / ml, about 35 mg / ml, about 36 mg / ml, about 37 mg / ml, about 38 mg / ml, about 39 mg / ml, about 40 mg / ml, about 41 mg / ml, about 42 mg / ml, about 43 mg / ml, about 44 mg / ml, about 45 mg / ml, about 46 mg / ml, about 47 mg / ml, about 48 mg / ml, about 49 mg / ml, or about 60 mg / ml. ml, more preferably about 2 mg / ml, about 3 mg / ml, about 5 mg / ml, about 6 mg / ml, about 12 mg / ml, about 15 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, or about 48 mg / ml; further preferably about 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, or about 48 mg / ml.
[0063] In some embodiments, the pH of the pharmaceutical composition is about 6.5 to about 8.5, more preferably about 7.0 to about 8.5, further preferably about 7.1 to about 8.3, further preferably about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, or about 8.2.
[0064] Another embodiment of the present invention also provides a liquid pharmaceutical composition comprising:
[0065] About 1-60 mg / ml, preferably about 1-70 mg / ml, preferably about 1-65 mg / ml, preferably about 1-60 mg / ml, preferably about 2-60 mg / ml, preferably about 2-50 mg / ml, more preferably about 2 mg / ml, about 3 mg / ml, about 4 mg / ml, about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 9 mg / ml, about 10 mg / ml, about 11 mg / ml, about 12 mg / ml, about 13 mg / ml, about 14 mg / ml, about 15 mg / ml, about 16 mg / ml, about 17 mg / ml, about 18 mg / ml, about 19 mg / ml, about 20 mg / ml, about 21 mg / ml, about 22 mg / ml, about 23 mg / ml, about 24 mg / ml, about 25 mg / ml, about 26 mg / ml, about 27 mg / ml, about 28 mg / ml, about 29 mg / ml, about 30 mg / ml, about 31 mg / ml, about 32 mg / ml, about 33 mg / ml, about 34 mg / ml, about 35 mg / ml, about 36 mg / ml, about 37 mg / ml, about 38 mg / ml, about 39 mg / ml, about 40 mg / ml, about 41 mg / ml, about 42 mg / ml, about 43 mg / ml, about 44 mg / ml, about 45 mg / ml, about 46 mg / ml, about 47 mg / ml, about 48 mg / ml, about 49 mg / ml, and further The N-ε of the present invention is preferably about 2 mg / ml, about 3 mg / ml, about 5 mg / ml, about 6 mg / ml, about 12 mg / ml, about 15 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, or about 48 mg / ml; more preferably about 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, or about 48 mg / ml. 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide;
[0066] about 1-30 mg / ml, preferably about 3-25 mg / ml, preferably about 5-15 mg / ml, preferably about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 8.10 mg / ml, about 8.15 mg / ml, about 8.20 mg / ml, about 8.25 mg / ml, about 8.30 mg / ml, about 8.50 mg / ml, about 9 mg / ml or about 10 mg / ml of NaCl;
[0067] About 1-25 mM, preferably about 1-20 mM, more preferably about 1-15 mM, further preferably about 1-10 mM, further preferably about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM or about 9 mM citric acid;
[0068] about 1-20 mg / ml, preferably about 1-10 mg / ml, more preferably about 1-5 mg / ml, and even more preferably about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml of Na2HPO4; and
[0069] The pH of the pharmaceutical composition is about 6.5 to about 8.5, more preferably about 7.0 to about 8.5, further preferably about 7.1 to about 8.3, further preferably about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, or about 8.2.
[0070] Another embodiment of the present invention also provides a liquid pharmaceutical composition comprising:
[0071] About 1-60 mg / ml, preferably about 1-70 mg / ml, preferably about 1-65 mg / ml, preferably about 1-60 mg / ml, Preferably, it is about 2-60mg / ml, preferably about 2-50mg / ml, more preferably about 2mg / ml, about 3mg / ml, about 4mg / ml, about 5mg / ml, about 6mg / ml, about 7mg / ml, about 8mg / ml, about 9mg / ml, about 10mg / ml, about 11mg / ml, about 12mg / ml, about 13mg / ml, about 14mg / ml, about 15mg / ml, about 16mg / ml, about 17mg / ml, about 18mg / ml, about 19mg / ml, about 20mg / ml, about 21mg / ml, about 22mg / ml, about 23mg / ml, about 24mg / ml, about 25mg / ml, about 26mg / ml, about 27mg / ml, about 28mg / ml, about 29mg / ml, about 30mg / ml, about 31mg / ml, about 32mg / ml, about 33mg / ml, about 34mg / ml Preferably, the N-ε is about 2 mg / ml, about 3 mg / ml, about 5 mg / ml, about 6 mg / ml, about 12 mg / ml, about 15 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, or about 48 mg / ml; more preferably, N-ε is about 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, or about 48 mg / ml; more preferably, N-ε is about 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, or about 48 mg / ml. 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide;
[0072] about 5-25 mg / ml, preferably about 10-20 mg / ml, preferably about 12-18 mg / ml, preferably about 13 mg / ml, about 13.5 mg / ml, about 14 mg / ml, about 14.5 mg / ml, about 15 mg / ml, 16 mg / ml or about 17 mg / ml of propylene glycol;
[0073] About 1-25 mM, preferably about 1-20 mM, more preferably about 1-15 mM, further preferably about 1-10 mM, further preferably about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM or about 9 mM citric acid;
[0074] about 1-20 mg / ml, preferably about 1-10 mg / ml, more preferably about 1-5 mg / ml, and even more preferably about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml of Na2HPO4; and
[0075] The pH of the pharmaceutical composition is about 6.5 to about 8.5, more preferably about 7.0 to about 8.5, further preferably about 7.1 to about 8.3, further preferably about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, or about 8.2.
[0076] Another embodiment of the present invention also provides a liquid pharmaceutical composition comprising:
[0077] About 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, about 48 mg / ml, or about 60 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide;
[0078] about 7 mg / ml, about 8 mg / ml, about 8.10 mg / ml, about 8.15 mg / ml, about 8.20 mg / ml, about 8.25 mg / ml, about 8.30 mg / ml, about 8.50 mg / ml, about 9 mg / ml, or about 10 mg / ml of NaCl;
[0079] about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, 9 mM, or about 10 mM citric acid;
[0080] About 1.42 mg / ml of Na2HPO4; and
[0081] The pH of the pharmaceutical composition is about 7.3.
[0082] Another embodiment of the present invention also provides a liquid pharmaceutical composition comprising:
[0083] About 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, or about 48 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide;
[0084] About 14 mg / ml of propylene glycol;
[0085] about 1 mM, 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, 9 mM, or about 10 mM citric acid;
[0086] About 1.42 mg / ml of Na2HPO4; and
[0087] The pH of the pharmaceutical composition is about 7.3.
[0088] The second aspect of the present invention provides a pharmaceutical product comprising a container and the liquid pharmaceutical composition of the first aspect of the present invention placed in the container; preferably, the container is selected from a pen injection device, an automatic injection device, a syringe, and a vial.
[0089] In another embodiment of the present invention, a pharmaceutical product is provided, comprising a container and a liquid pharmaceutical composition placed in the container, wherein the container is deoxygenated and filled with nitrogen before and / or during and / or after the liquid pharmaceutical composition is placed in the container; the liquid pharmaceutical composition comprises:
[0090] About 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, about 48 mg / ml, or about 60 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide;
[0091] about 3 mg / ml, about 4 mg / ml, about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 8.10 mg / ml, about 8.15 mg / ml, about 8.20 mg / ml, about 8.25 mg / ml, about 8.30 mg / ml, about 8.50 mg / ml, about 9 mg / ml, or about 10 mg / ml of NaCl;
[0092] about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml of Na2HPO4; and
[0093] The pH of the pharmaceutical composition is about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.6, or about 7.7.
[0094] In another embodiment of the present invention, a pharmaceutical product is provided, comprising a container and a liquid pharmaceutical composition placed in the container, wherein the container is deoxygenated and filled with nitrogen before and / or during and / or after the liquid pharmaceutical composition is placed in the container; the liquid pharmaceutical composition comprises:
[0095] About 2-60 mg / ml, preferably about 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, or about 48 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide;
[0096] about 13 mg / ml, about 14 mg / ml, about 15 mg / ml, 16 mg / ml, or about 17 mg / ml of propylene glycol;
[0097] about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml of Na2HPO4; and
[0098] The pH of the pharmaceutical composition is about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.6, or about 7.7.
[0099] In some embodiments, the liquid pharmaceutical composition further comprises about 0.005 mM-25 mM, preferably about 0.01 mM-10 mM, and more preferably about 0.1 mM-2.0 mM citric acid or methionine.
[0100] In some embodiments, deoxygenation of the container is achieved by filling the container with an inert gas; preferably, the inert gas is nitrogen.
[0101] In some embodiments, the container is a pen injection device, an autoinjector device, a syringe, or a vial.
[0102] The pharmaceutical composition according to the first aspect of the present invention, or the pharmaceutical product according to the second aspect of the present invention, wherein the pharmaceutical composition or the pharmaceutical product is for parenteral administration; preferably, the parenteral administration is subcutaneous administration.
[0103] A third aspect of the present invention provides a kit comprising a liquid pharmaceutical composition and instructions for use, wherein the liquid pharmaceutical composition is as defined in the first aspect of the present invention.
[0104] In another embodiment of the present invention, a kit is provided comprising a liquid pharmaceutical composition as defined in the first aspect of the present invention and an injection device for administering the composition to a subject, wherein the injection device is selected from a durable pen and a prefilled pen injection device.
[0105] Use of the pharmaceutical composition of the first aspect of the present invention or the pharmaceutical product of the second aspect of the present invention in the preparation of a medicament for treating diabetes, obesity, non-alcoholic fatty liver disease, Alzheimer's disease or Parkinson's disease; preferably, the pharmaceutical composition of the first aspect of the present invention is administered to a subject in need once a week, once every two weeks or less frequently.
[0106] The pharmaceutical composition of the first aspect of the present invention, or the pharmaceutical product of the second aspect of the present invention, is used to treat diabetes, obesity, non-alcoholic fatty liver disease, Alzheimer's disease or Parkinson's disease; preferably, the pharmaceutical composition of the first aspect of the present invention is administered to a subject in need once a week, once every two weeks or less frequently.
[0107] The fourth aspect of the present invention provides a method for treating diabetes, obesity, non-alcoholic fatty liver disease, Alzheimer's disease or Parkinson's disease, the method comprising administering the pharmaceutical composition according to the first aspect of the present invention, or the pharmaceutical product according to the second aspect, to a subject in need thereof; preferably, the pharmaceutical composition according to the first aspect of the present invention is administered to a subject in need thereof once a week, once every two weeks or less frequently.
[0108] A fifth aspect of the present invention provides a method for preparing the pharmaceutical composition of the first aspect, the method comprising:
[0109] (1) dissolving the prescribed amount of buffer, osmotic pressure regulator, and / or antioxidant in water;
[0110] (2) dissolving the prescribed amount of GLP-1 peptide in the solution obtained in step (1), and adjusting the pH using sodium hydroxide and / or dilute hydrochloric acid;
[0111] (3) The solution obtained in step (2) was finally sterilized by filtering through a 0.22 μm sterile filter.
[0112] Another embodiment of the present invention provides a method for preparing the pharmaceutical composition according to the first aspect of the present invention, wherein:
[0113] The method comprises:
[0114] mixing the GLP-1 peptide, buffer, osmotic pressure regulator, and antioxidant according to a prescription;
[0115] Preferably, the method further comprises:
[0116] The mixture obtained in step (2) is sterilized; preferably, the mixture obtained in step (2) is sterilized by filtering through a 0.22 μm sterile filter.
[0117] definition
[0118] For a better understanding of the present invention, definitions and explanations of relevant terms are provided below.
[0119] GLP-1 peptide
[0120] As used herein, the term "GLP-1 peptide" refers to a "GLP-1 analog" or "GLP-1 analog," and refers to a peptide or compound that is a variant of human glucagon-like peptide-1 (GLP-1(7-37)), wherein one or more amino acid residues of GLP-1(7-37) are replaced, and / or one or more amino acid residues are deleted, and / or one or more amino acid residues are added. Specifically, the sequence of GLP-1(7-37) is shown in SEQ ID NO: 1 in the sequence listing. The peptide having the sequence shown in SEQ ID NO: 1 may also be referred to as "native" GLP-1 or "native" GLP-1(7-37).
[0121] In the sequence listing, the first amino acid residue (histidine) of SEQ ID NO: 1 is numbered 1. However, hereinafter, in accordance with established conventions in the art, the histidine residue is numbered 7, and subsequent amino acid residues are numbered accordingly, ending with glycine at position 37. Therefore, generally, the amino acid residue numbering or position numbering of the GLP-1 (7-37) sequence referred to herein is a sequence starting with His at position 7 and ending with Gly at position 37.
[0122] The [Gly8, Arg34]GLP-1-(7-37) peptide is a GLP-1 analog having Gly and Arg at positions corresponding to position 8 and position 34 of GLP-1(7-37) (SEQ ID NO: 1), respectively. The [Arg34]GLP-1-(7-37) peptide is a GLP-1 analog having Arg at a position corresponding to position 34 of GLP-1(7-37) (SEQ ID NO: 1). Specifically, the amino acid sequence of the [Gly8, Arg34]GLP-1-(7-37) peptide is shown in SEQ ID NO: 2 in the sequence listing.
[0123] The GLP-1 peptide described herein can be prepared by referring to the method described in WO2021136303.
[0124] In the context of a GLP-1 peptide or analog thereof, the term "derivative" as used herein refers to a chemically modified GLP-1 peptide or analog wherein one or more substituents have been covalently attached to the peptide. Substituents may also be referred to as side chains.
[0125] Unless otherwise stated, when reference is made to acylation with a lysine residue, this is understood to be to be carried out with the e-amino group thereof.
[0126] The term "peptide" when applied to, for example, the GLP-1 analogs of the present invention, refers to a compound comprising a series of amino acids interconnected by amide (or peptide) bonds.
[0127] In a specific embodiment, the peptide is largely or predominantly composed of amino acids linked to each other by amide bonds (e.g., at least 50%, 60%, 70%, 80%, or at least 90% by molar mass). In another specific embodiment, the peptide is composed of amino acids linked to each other by peptide bonds.
[0128] Amino acids are molecules containing an amino group and a carboxylic acid group, optionally with one or more additional groups, often called side chains.
[0129] The term "amino acid" includes proteinogenic amino acids (encoded by the genetic code, including natural amino acids and standard amino acids), as well as non-proteinogenic (not found in proteins and / or not encoded in the standard genetic code), and synthetic amino acids. Non-proteinogenic amino acids are moieties that can be incorporated into peptides via peptide bonds, but are not proteinogenic amino acids. Synthetic non-proteinogenic amino acids include amino acids produced by chemical synthesis, i.e., D-isomers of amino acids encoded by the genetic code, such as D-alanine and D-leucine, Aib (α-aminoisobutyric acid), Abu (α-aminobutyric acid), 3-aminomethylbenzoic acid, anthranilic acid, desamino-histidine, β-analogs of amino acids such as β-alanine, D-histidine, desamino-histidine, 2-amino-histidine, β-hydroxy-histidine, and homohistidine, etc.
[0130] Non-limiting examples of amino acids not encoded by the genetic code are γ-carboxyglutamate, ornithine, D-alanine, D-glutamine and phosphoserine. Non-limiting examples of synthetic amino acids are the D-isomers of amino acids, such as D-alanine and D-leucine, Aib (α-aminoisobutyric acid), β-alanine and des-amino-histidine (desH, alternative name imidazole propionic acid, abbreviated Imp).
[0131] In the following, all amino acids not indicating the optical isomer are understood to refer to the L-isomer (unless otherwise specified).
[0132] The term "stable" means that all proteins therein substantially retain their physical, chemical, and biological activities after storage at the intended storage temperature, e.g., 0-40°C. A formulation may be considered stable even if the antibodies therein do not retain 100% of their physical, chemical, or biological activities after storage for a specified period of time. A formulation may be considered "stable" if it retains approximately 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or greater of the antibody structure and function after storage for a specified period of time.
[0133] The term "stable pharmaceutical composition" refers to a pharmaceutical composition comprising a GLP-1 peptide, such as a solution or suspension, which composition comprises at least 80% (w / v) of the GLP-1 peptide after storage (e.g., after static storage for 3 months at 25°C). The storage conditions for the stability test may be 2-8°C, such as 5°C, or at least 2.5 years at 5°C. Alternatively, the storage conditions for the stability test may be at least 4 weeks, such as 6 weeks or 3 months, optionally at 30°C. The storage conditions for the stable pharmaceutical composition may be 1 year or 2 years at 5°C. The storage conditions for the stable pharmaceutical composition may be 3 years at 5°C. Alternatively, the storage conditions may be 24 hours or 1 week at 25°C. In yet another alternative, the storage conditions may be two months at room temperature, such as up to two months.
[0134] As used herein, the term "about" when referring to a specifically recited value or range of values generally means within 20%, preferably within 10%, and more preferably within 5% of the given value or range.
[0135] The term "kit" may include a pharmaceutical formulation as described herein and a device for administration, for example, a pharmaceutical formulation may be packaged together with a device for administration, such as a syringe, inhaler, measuring cup, dropper, or applicator. The pharmaceutical formulation may be filled in a container as defined above. The kit may optionally include instructions for use, including instructions for dosage, dosing regimen, and mode of administration.
[0136] The term "treatment" includes therapeutic treatment, prophylactic treatment, and use in reducing the risk of a subject developing a disease or other risk factors. Treatment includes, but is not limited to, complete cure of the disease, as well as alleviation of symptoms or mitigation of potential risks.
[0137] The term "subject" is used interchangeably with "patient" herein and refers to mammals, including but not limited to: humans and non-human primates, including apes and humans; mammalian sport animals (e.g., horses); mammalian farm animals (e.g., sheep, goats, etc.); mammalian pets (dogs, cats, etc.); and rodents (e.g., mice, rats, etc.).
[0138] The term "prescribed amount" includes the amount of the prescribed active substance or excipient in any of the examples herein and the amount of the active substance or excipient in all formulations disclosed.
[0139] The term "related substances" refers to impurities such as starting materials, intermediates, by-products, and degradation products that may be introduced during drug production and storage. The presence of these impurities may affect the quality, safety, and efficacy of the drug. Related substances are an important indicator in drug stability studies.
[0140] The term "high molecular weight protein (HMWP)" or "high molecular weight protein" primarily refers to protein impurities with relatively large molecular weights. During pharmaceutical production and storage, HMWPs can form due to various factors, such as aggregation and deformation. Detection and analysis of HMWPs is crucial for ensuring pharmaceutical quality and safety.
[0141] The "inert gas" mentioned herein refers to rare gases or some physiologically inert gases. Rare gases are elements in Group 18 of the periodic table. Physiologically inert gases include nitrogen and methane. DETAILED DESCRIPTION
[0142] The embodiments of the present invention will be described in detail below with reference to the examples. However, it will be understood by those skilled in the art that the following examples are intended to illustrate the present invention only and should not be construed as limiting the scope of the present invention. Where specific conditions are not specified in the examples, the experiments were carried out under conventional conditions or the conditions recommended by the manufacturer. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be obtained by commercial purchase.
[0143] The pharmaceutically active ingredient GLP-1 peptide is formulated with various excipients using formulation methods known in the art.
[0144] The various formulations were evaluated to determine the overall physicochemical stability of the GLP-1 peptide, the pharmaceutically active ingredient, in the formulations. The formulation stability was analyzed using the following methods:
[0145] (1) Determination of the amount of related substances
[0146] The content of GLP-1 derivative-related impurities was determined by high performance liquid chromatography (HPLC) on a Waters Kromasil 100-3.5-C8 (4.6*250 mm) column at a column temperature of 35°C and a sample cell temperature of 5°C, using an elution phase at a flow rate of 1.0 ml / min. Elution was performed using a mobile phase consisting of the following composition:
[0147] Phase A contained 90 mM potassium dihydrogen phosphate and 10% acetonitrile (v / v), pH 2.4
[0148] Phase B was 75% (v / v) acetonitrile.
[0149] Gradient: linear from 75% / 25% A / B to 55% / 45% A / B from 0-5 min, linear from 5-12 min to 50% / 50% A / B, linear from 12-42 min to 40% / 60% A / B, linear from 42-60 min to 10% / 90% A / B, linear from 60-61 min to 75% / 25% A / B, isocratic from 61-70 min to 85% / 15% A / B.
[0150] The detection wavelength was 214 nm, the flow rate was 1.0 ml / min, and the injection volume was 15 μl. After storage at 4°C, 25°C, and 37°C for several days, the increase in the amount of related substances relative to day 0 was detected.
[0151] (2) Determination of high molecular weight protein (HMWP)
[0152] High molecular weight protein (HMWP) content was determined by high performance liquid chromatography (HPLC) on a Waters TSKgel G2000SWXL (7.8 x 300 mm), 5 μm column at a column temperature of 30°C and a sample cell temperature of 5°C, using a mobile phase containing 300 ml of isopropanol, 400 ml of glacial acetic acid, and 300 ml of purified water at a flow rate of 0.5 ml / min. The detection wavelength was 276 nm, and the injection volume was 25 μl. The increase in HMWP relative to day 0 was measured after storage for several days at 4°C, 25°C, and 37°C.
[0153] Abbreviations
[0154] Na2HPO4 is disodium hydrogen phosphate;
[0155] NaOH is sodium hydroxide;
[0156] OEG is the amino acid residue -NH(CH2)2O(CH2)2OCH2C(O)-;
[0157] OSu is succinimidyl-1-yloxy-2,5-dioxo-pyrrolidin-1-yloxy;
[0158] OtBu is tert-butyloxy;
[0159] HCl is hydrogen chloride;
[0160] γGlu or gGlu is γL-glutamyl;
[0161] HPLC is High Performance Liquid Chromatography (HPLC);
[0162] DCC is dicyclohexylcarbodiimide;
[0163] AEEA is 2-(2-(2-aminoethoxy)ethoxy)acetic acid.
[0164] Example 1
[0165] Preparation of the title compound: N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide (Compound 1)
[0166] 1. N-ε 26 Preparation of -[2-(2-[2-(2-[2-(2-[4-(21-carboxyhenicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide
[0167] [Gly8, Arg34]GLP-1-(7-37) peptide was prepared by protein recombinant expression methods commonly used in the art (for specific methods, see Molecular Cloning: A Laboratory Manual (Fourth Edition), Michael R. Green, Cold Spring Harbor Press, 2012). [Gly8, Arg34]GLP-1-(7-37) peptide (5 g, 1.48 mmol) was dissolved in 100 mM Na2HPO4 aqueous solution (150 mL), acetonitrile (100 mL) was added, and the pH was adjusted to pH 10-12.5 with 1N NaOH. tert-Butyleicosandioyl-γGlu(2xOEG-OSu)-OtBu (1.59 g, 1.63 mmol) was dissolved in acetonitrile (50 mL) and slowly added to the [Gly8, Arg34]GLP-1-(7-37) peptide solution. The pH was maintained at 10-12.5. After 120 minutes, the reaction mixture was added to water (150 mL) and the pH was adjusted to 5.0 with 1N HCl aqueous solution. The precipitate was separated by centrifugation and lyophilized. The crude product was added to a mixed solution of trifluoroacetic acid (60 mL) and dichloromethane (60 mL) and stirred at room temperature for 30 minutes. The mixture was concentrated to about 30 mL and poured into ice-cold n-heptane (300 mL). The precipitated product was separated by filtration and washed twice with n-heptane. After vacuum drying, the product was purified by ion exchange chromatography (Ressource Q, 0.25%-1.25% ammonium acetate gradient in 42.5% ethanol, pH 7.5) and reverse phase chromatography (acetonitrile, water, TFA). The purified fractions were combined, the pH was adjusted to 5.2 with 1N HCl, the precipitate was separated, and lyophilized to obtain the title compound.
[0168] LC-MS (electrospray): m / z = 1035.8 [M+4H] 4+
[0169] 2. Preparation of the intermediate tert-butyl docosanoyl-γGlu-(2xOEG-OSu)-OtBu
[0170] 2.1 tert-Butyldocosanedioyl-OSu
[0171] Under nitrogen, mono-tert-butyl docosandioate (20 g, 50.17 mmol) and NHS (5.77 g, 50.17 mmol) were mixed in dichloromethane (400 mL). Triethylamine (13.95 mL) was added, and the resulting turbid mixture was stirred at room temperature. DCC (11.39 g, 55.19 mmol) was then added and stirred overnight. The mixture was filtered, and the filtrate was concentrated to near dryness. The residue was mixed with cold water and ethyl acetate, stirred for 20 minutes, and separated. The upper organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated to near dryness under reduced pressure and dried overnight in vacuo to obtain 24.12 g (97% yield) of tert-butyl docosandioyl-OSu.
[0172] 2.2 tert-Butyldocosanedioyl-γGlu-OtBu
[0173] tert-Butyldocosanedioyl-OSu (24.12 g, 48.66 mmol) was dissolved in dichloromethane (250 mL) and stirred. H-Glu-OtBu (10.88 g, 53.53 mmol), triethylamine (12.49 mL), and water (25 mL) were added sequentially. The mixture was heated to obtain a clear solution, which was stirred at room temperature for 4 hours. A 10% aqueous citric acid solution (200 mL) was then added, and the layers were separated. The lower organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to near dryness and dried under vacuum overnight. This yielded 27.27 g (96% yield) of tert-Butyldocosanedioyl-γGlu-OtBu.
[0174] 2.3 tert-Butyldocosandioyl-γGlu(OSu)-OtBu.
[0175] Under nitrogen, tert-butyldocosandioyl-γGlu-OtBu (27.27 g, 46.71 mmol) was dissolved in dichloromethane (300 mL). Triethylamine (11.99 mL) was added and stirred for 10 minutes. NHS (5.38 g, 50.17 mmol) was then added, followed by DCC (10.60 g, 51.38 mmol). The mixture was stirred at room temperature overnight. Filtered, the resulting filtrate was concentrated to near dryness, and the residue was mixed with cold water and ethyl acetate, stirred for 20 minutes. The layers were separated, and the upper organic phase was washed with saturated brine. After separation, the upper organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to near dryness under reduced pressure. Methyl tert-butyl ether was added, stirred for 30 minutes, filtered, and the filter cake was dried under vacuum overnight to obtain 25.76 g (81% yield) of tert-butyldocosandioyl-γGlu-(OSu)-OtBu.
[0176] 2.4-tert-Butyldocosanedioyl-γGlu-(2xOEG-OH)-OtBu
[0177] Tert-butyldocosanedioyl-γGlu-(OSu)-OtBu (25.76 g, 37.83 mmol) was dissolved in dichloromethane (250 mL) and stirred. 2xAEEA (11.66 g, 37.83 mmol), triethylamine (9.71 mL), and water (25 mL) were added sequentially. The mixture was heated to obtain a clear solution, which was stirred at room temperature for 4 hours. A 10% aqueous citric acid solution (200 mL) was then added to separate the layers. The lower organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to near dryness and dried under vacuum overnight. This yielded 30.75 g (93% yield) of tert-butyldocosanedioyl-γGlu-(2xOEG-OH)-OtBu.
[0178] 2.5 tert-Butyldocosandioyl-γGlu-(2xOEG-OSu)-OtBu.
[0179] Under nitrogen, tert-butyldocosanedioyl-γGlu-(2xOEG-OH)-OtBu (30.75 g, 35.18 mmol) was dissolved in dichloromethane (300 mL). Triethylamine (9.03 mL) was added and stirred for 10 minutes. NHS (4.05 g, 35.18 mmol) was then added, followed by DCC (7.98 g, 38.70 mmol). The mixture was stirred at room temperature overnight. Filtered, the resulting filtrate was concentrated to near dryness, and the residue was mixed with cold water and ethyl acetate, stirred for 20 minutes, and separated. The upper organic phase was washed with saturated brine. After separation, the upper organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to near dryness under reduced pressure and dried overnight in vacuo to obtain 31.09 g (91% yield) of tert-butyldocosanedioyl-γGlu-(2xOEG-OSu)-OtBu.
[0180] Preparation of the title compound N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide (Compound 2)
[0181] N-ε was prepared in a similar manner to that in Example 1, Part 1 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide
[0182] LC-MS (electrospray): m / z = 992.52 [M+4H] 4+
[0183] The intermediate tert-butyleicosandioyl-γGlu-(OEG-OSu)-OtBu was prepared by a procedure similar to that of Example 1, part 2.
[0184] LC-MS(Scie×100API): m / z=826.54(M+1) +
[0185] Example 2 Effects of the antioxidants citric acid and methionine on the chemical stability of GLP-1 peptide formulations
[0186] 2.1 Preparation of formulations
[0187] Compound 1 was dissolved in sodium hydrogen phosphate solution to the final concentration shown in Table 1. According to the amount of each component in the table below, each auxiliary liquid was added in sequence, and the pH was adjusted to the value in the table below to produce the final formulation shown in Table 1.
[0188] Table 1 Preparation formula
[0189] 2.2 Detection of related substances
[0190] The above-mentioned formulations 2-1 to 2-14 were filled into assembled cartridges and stability studies were conducted at 4°C, 25°C, and 37°C. The increase in the amount of the related substances in each formulation of Example 2 (formulations 2-1 to 2-14) at different days relative to day 0 was measured. The test results are shown in Tables 2, 3, and 4, respectively.
[0191] Table 2 Detection results of related substances of compound 1 preparation at 4°C
[0192] As can be seen from Table 2, the trends in related substances in the formulations containing antioxidants and preservatives during long-term storage at 4°C were essentially the same. The overall increase in related substances in the high-concentration formulation (60 mg / mL) of Compound 1 was lower compared to day 0.
[0193] Table 3 Related substance detection results of compound 1 preparation under accelerated conditions at 25°C
[0194] Table 3 shows that the trends in related substances in the antioxidant-containing Compound 1 formulation and the preservative-containing formulation under accelerated conditions at 25°C were essentially the same. A more pronounced difference was observed at 91 days: in both the 10 mg / mL and 60 mg / mL Compound 1 concentration groups, the trends in related substances following the addition of citric acid were essentially the same as those in the preservative-containing formulation. The increases in related substances relative to day 0 were significantly higher in the citric acid-free group compared to day 0, while the increases in related substances relative to day 0 were significantly lower in the 20 mg / mL and 40 mg / mL groups. Furthermore, the increase in related substances relative to day 0 was even lower in the high-concentration Compound 1 group (60 mg / mL). The addition of antioxidants can slow the increase in related substance levels, with citric acid monohydrate being more effective than methionine in slowing the increase, potentially contributing to formulation stability.
[0195] Table 4 Related substance detection results of compound 1 preparation under accelerated conditions at 37°C
[0196] As can be seen from Table 4, the trends of changes in related substances in the preparations containing antioxidants and those containing preservatives during long-term accelerated temperature at 37°C are basically the same, and the differences are more obvious at 63 days: in the 10 mg / mL and 60 mg / mL compound 1 concentration groups, it can be seen that the increase in related substances without antioxidants relative to day 0 is higher, and in the 20 mg / mL and 40 mg / mL groups, the increase in related substances in the group with citric acid relative to day 0 is significantly lower. The increase in related substances in each group relative to day 0 decreases with the increase in compound 1 concentration, that is, the related substances in high concentrations of compound 1 (for example: 60 mg / mL) are lower; and the addition of antioxidants to the preparation containing NaCl can slow down the increase in related substances compared with the preparation containing propylene glycol, and monohydrate citric acid is more significant than methionine.
[0197] In summary, antioxidants, especially citric acid monohydrate, can slow down the rise of related substances in the formulation. Among them, citric acid monohydrate has a more significant slowing effect than methionine and is more conducive to the stability of the formulation.
[0198] 2.3 Detection of high molecular weight proteins (HMWP)
[0199] As in 2.2, the increase in high molecular weight protein in each formulation of Example 2 relative to day 0 was measured at 4°C, 25°C, and 37°C. The test results are shown in Tables 5, 6, and 7, respectively.
[0200] Table 5 High molecular weight protein detection results of compound 1 preparation at 4°C
[0201] It can be seen from Table 5 that the change trends of high molecular weight proteins in the preparations containing antioxidants and the preparations containing preservatives after long-term storage at 4°C are basically the same.
[0202] Table 6 High molecular weight protein detection results of compound 1 preparation under accelerated conditions at 25°C
[0203] Table 6 shows that at 25°C, all groups showed a significant trend in accelerating the growth of high-molecular-weight protein. For the sodium chloride-added formulation, except for the 10 mg / mL Compound 1 concentration group, the growth of high-molecular-weight protein decreased with increasing Compound 1 concentration. The difference was particularly pronounced at 63 days: in both the 10 mg / mL and 60 mg / mL Compound 1 concentration groups, the increase in high-molecular-weight protein after the addition of citric acid was lower than that at day 0. In the 20 mg / mL and 40 mg / mL groups, the increase in high-molecular-weight protein after the addition of citric acid was significantly lower than that at day 0. The addition of antioxidants can slow the growth of high-molecular-weight protein content, with citric acid monohydrate being more effective than methionine.
[0204] Table 7 Results of high molecular weight protein in compound 1 preparation under accelerated conditions at 37°C
[0205] Table 7 shows that at 37°C, each group showed a clear trend of accelerated growth of high-molecular-weight protein. However, as the concentration of the formulation increased, high-molecular-weight protein decreased, with a significant difference at day 63. In both the 10 mg / mL and 60 mg / mL Compound 1 concentration groups, the increase in high-molecular-weight protein after the addition of citric acid was lower than that at day 0. In the 20 mg / mL and 40 mg / mL groups, the increase in high-molecular-weight protein after the addition of citric acid was significantly lower than that at day 0. The addition of antioxidants can slow the growth of high-molecular-weight protein, with citric acid monohydrate being more effective than methionine.
[0206] In summary, the antioxidant citric acid monohydrate can slow down the increase in the content of high molecular weight protein in the formulation, and the slowing effect of citric acid monohydrate is more significant than that of methionine, which is more conducive to the stability of the formulation.
[0207] Removing the preservative (phenol) and replacing the osmotic pressure regulator (propylene glycol → NaCl) will significantly increase the content of related substances and high molecular weight proteins in the formulation. Adding antioxidants can significantly reduce the increase of related substances and high molecular weight proteins. Monohydrate citric acid is more effective than methionine. Ultimately, monohydrate citric acid was selected as the antioxidant in the formulation of compound 1.
[0208] Example 3 Determination of Chemical Stability of Preparations
[0209] 3.1 Preparation of the drug
[0210] The stability of formulations containing sodium chloride and propylene glycol as osmotic pressure regulators was tested. The formulated formulations were filled into assembled cartridges. The formulation design for the osmotic pressure regulator formulation is shown in Table 8:
[0211] Table 8 Compound 1 formulation
[0212] 3.2 Detection of related substances
[0213] The above-mentioned formulations 3-1 to 3-10 were filled into assembled cartridges and stability studies were conducted at 4°C, 25°C, and 37°C. The increase in the amount of the related substances in each formulation of Example 3 (formulations 3-1 to 3-10) at different days relative to day 0 was measured. The test results are shown in Tables 9, 10, and 11, respectively.
[0214] Table 9 Results of related substances in the preparation of compound 1 at 4°C
[0215] As can be seen from Table 9, the trends of related substances in each group during long-term storage at 4°C were basically the same. The two different osmotic pressure regulators had little effect on the stability of the Compound 1 preparation (related substance content) during long-term storage at 4°C for 56 days.
[0216] Table 10 Contents of related substances in the preparation of compound 1 under accelerated conditions at 25°C
[0217] Table 10 shows that the trends in the changes in related substances during accelerated storage at 25°C were essentially the same across all groups. It can be seen that the growth of related substances was slower when sodium chloride was used as an osmotic pressure regulator than when propylene glycol was used, and that nitrogen replacement reduced the growth rate of related substances. Comparisons between Groups 3-5 and 3-3, and between Groups 3-9 and 3-7, revealed that the addition of citric acid significantly reduced the growth rate of related substances in both groups. Nitrogen replacement, in addition to antioxidants, had no significant effect on reducing the growth rate of related substances.
[0218] Table 11 Results of related substances in the preparation of compound 1 under accelerated conditions at 37°C
[0219] Table 11 shows that the trends in the changes in related substances during accelerated storage at 37°C were essentially consistent across all groups. Comparisons between Groups 3-1, 3-3, and 3-7, and between Groups 3-2, 3-4, and 3-8, show that the removal of phenol resulted in increases in all related substances. The sodium chloride osmotic pressure regulator showed a similar growth rate to propylene glycol, and nitrogen replacement reduced the rate of increase. Comparisons between Groups 3-5 and 3-3, and between Groups 3-9 and 3-7, reveal that the addition of citric acid to the different osmotic pressure regulators reduced the rate of increase in both groups. Comparisons between Groups 3-5 and 3-6, and between Groups 3-9 and 3-10, show that nitrogen replacement, in addition to antioxidants, had no significant effect on reducing the rate of increase in related substances.
[0220] 3.3 High molecular weight protein detection
[0221] As in 2.3, the increase in high molecular weight protein in each formulation of Example 3 relative to day 0 was measured at 4°C, 25°C, and 37°C. The test results are shown in Tables 12, 13, and 14, respectively.
[0222] Table 12 Results of high molecular weight protein in compound 1 preparation under long-term conditions at 4°C
[0223] As can be seen from Table 12, the trends of changes in high molecular weight protein in each group were basically the same during long-term storage at 4°C. The two different osmotic pressure regulators had little effect on the stability (high molecular weight protein content) of the Compound 1 formulation after long-term storage at 4°C for 56 days.
[0224] Table 13 High molecular weight protein content of compound 1 preparation under accelerated conditions at 25°C
[0225] Table 13 shows that the trends in high-molecular-weight protein changes during accelerated storage at 25°C were essentially the same across all groups. Comparisons between Groups 1, 3, and 7, and between Groups 2, 4, and 8, show that high-molecular-weight protein increased after phenol removal. The sodium chloride osmotic pressure regulator group experienced slower growth than the propylene glycol osmotic pressure regulator group, and nitrogen replacement reduced the growth rate. Comparisons between Groups 5 and 9 revealed that the addition of citric acid to the different osmotic pressure regulators reduced the growth rate of high-molecular-weight protein in both groups. Nitrogen replacement, in addition to antioxidants, had no significant effect on reducing the growth of high-molecular-weight protein.
[0226] Table 14 Results of high molecular weight protein in compound 1 preparation under accelerated conditions at 37°C
[0227] The sodium chloride osmotic pressure regulator showed a slightly slower growth rate than the propylene glycol osmotic pressure regulator before day 28. Comparison between groups 3-5 and 3-9 revealed that the addition of citric acid to the different osmotic pressure regulator groups slowed the growth rate of high molecular weight protein in both groups.
[0228] In summary, formulations containing the osmotic pressure regulator sodium chloride showed better stability at 25°C than formulations containing the osmotic pressure regulator propylene glycol. The addition of the antioxidant citric acid monohydrate significantly reduced the growth of related substances and high-molecular-weight proteins, resulting in better stability than formulations containing the preservative phenol at accelerated stability.
[0229] Example 4 Effect of Devices on Preparation Stability
[0230] 4.1 Preparation
[0231] Compound 1 was dissolved in a disodium hydrogen phosphate solution to the final concentration shown in Table 15. The auxiliary liquids were added sequentially according to the amounts of the components in the table below. The pH was adjusted to the values in the table below to produce the final formulation shown in Table 15. The formulation was then filled into the corresponding devices, namely, assembled cartridges, prefilled cartridges, and prefilled syringes.
[0232] Table 15 Compound 1 formulation
[0233] 4.2 Detection of related substances
[0234] The above-mentioned preparations 4-1 to 4-6 were filled into corresponding instruments, and stability studies were conducted at 4°C, 25°C, and 37°C. The increase in the relevant substances in the formulations of Table 15 at different days relative to day 0 was measured. The test results are shown in Tables 16, 17, and 18, respectively.
[0235] Table 16 Results of related substances in compound 1 preparation under long-term conditions at 4°C
[0236] It can be seen from Table 16 that the changing trends of related substances in each group during long-term storage at 4°C were basically consistent, there was basically no difference in related substances between the groups, and different devices had basically no effect on the stability of the Compound 1 preparation.
[0237] Table 17 Comparison of related substance results of compound 1 preparation under accelerated conditions at 25°C
[0238] It can be seen from Table 17 that different devices have little effect on the stability of the compound 1 preparation.
[0239] Table 18 Results of related substances in the preparation of compound 1 under accelerated conditions at 37°C
[0240] It can be seen from Table 18 that different devices have little effect on the stability of the compound 1 preparation.
[0241] 4.3 Detection of high molecular weight proteins
[0242] As in 4.2, the increase in high molecular weight protein in each formulation in Table 18 relative to day 0 was measured at 4°C, 25°C, and 37°C. The test results are shown in Tables 19, 20, and 21, respectively.
[0243] Table 19 High molecular weight protein content of compound 1 preparation under 4℃ long-term conditions in comparative experiment
[0244] As can be seen from Table 19, different devices have little effect on the stability of the Compound 1 preparation (high molecular weight protein content) during long-term storage at 4°C for 28 days.
[0245] Table 20 Changes in high molecular weight protein of compound 1 formulation under accelerated conditions at 25°C
[0246] It can be seen from Table 20 that different devices have little effect on the stability of the compound 1 preparation.
[0247] Table 21 Comparison of compound 1 injection device comparison experiment 37℃ accelerated high molecular weight protein results
[0248] As can be seen from Table 21, the change trends of high molecular weight proteins in each group during accelerated storage at 37°C were basically consistent. Comparing Group 4-3 with Group 4-4, and Group 4-5 with Group 4-6, there was basically no difference in high molecular weight proteins between prefilled cartridges and prefilled syringes. The changes in related substances in Groups 4-3 and 4-4 were slightly better than those in Group 4-1. There was no significant difference between Groups 4-5, 4-6 and 4-2.
[0249] Example 5 Effect of GLP-1 peptide concentration on formulation stability
[0250] 5.1 Preparation of the drug
[0251] The concentration of Compound 1 in the Compound 1 formulation was divided into four concentrations (6 mg / mL, 12 mg / mL, 24 mg / mL, and 48 mg / mL), and the sodium chloride concentration was 6 mg / mL. Stability experiments were conducted on the four concentrations of Compound 1 formulation. Nitrogen replacement was not performed during the experiments. The formulation concentration confirmation experiment design is shown in Table 22 below:
[0252] Table 22 Compound 1 formulation formula
[0253] 5.2 Detection of related substances
[0254] The above-mentioned formulations 5-1 to 5-6 were filled into corresponding devices, and stability studies were conducted at 4°C, 25°C, and 37°C. The increase in the amount of the relevant substances in each formulation in Table 22 at different days relative to day 0 was measured. The test results are shown in Tables 23, 24, and 25, respectively.
[0255] Table 23 Results of related substances in compound 1 preparation under long-term conditions at 4°C
[0256] As can be seen from Table 23, the trends of related substances in each group during long-term storage at 4°C were basically the same. Different formulations had little effect on the stability of Compound 1 preparations (related substance content) during long-term storage at 4°C for 28 days.
[0257] Table 24 Results of related substances of compound 1 preparation under accelerated conditions at 25°C
[0258] As can be seen from Table 24, the trends of changes in related substances in each group during accelerated storage at 25°C were basically the same. There was no significant difference in related substances between Group 5-3 and Group 5-6.
[0259] Table 25 Results of related substances of compound 1 preparation under accelerated conditions at 37°C
[0260] As can be seen from Table 25, the levels of related substances in each group showed an increasing trend during accelerated storage at 37°C. There were no significant differences in the levels of related substances among the groups.
[0261] 5.3 Detection of high molecular weight proteins
[0262] As in 5.2, the increase in high molecular weight protein in each formulation in Table 22 relative to day 0 was measured at 4°C, 25°C, and 37°C. The test results are shown in Tables 26, 27, and 28, respectively.
[0263] Table 26 Results of high molecular weight protein of compound 1 preparation at 4°C
[0264] As can be seen from Table 26, the trends of high molecular weight protein changes in each group were basically the same during long-term storage at 4°C. Different formulations had little effect on the stability (high molecular weight protein content) of Compound 1 formulations during long-term storage at 4°C for 28 days.
[0265] Table 27 Compound 1 formulation concentration confirmation experiment 25°C accelerated high molecular weight protein results
[0266] It can be seen from Table 27 that the high molecular weight proteins in each group showed an increasing trend during accelerated storage at 25°C, and there was no significant difference in the high molecular weight proteins.
[0267] Table 28 Compound 1 formulation concentration confirmation experiment 37°C accelerated high molecular weight protein results
[0268] It can be seen from Table 28 that the relevant substances in each group showed an increasing trend during accelerated storage at 37°C, and there was no significant difference in the relevant substances.
[0269] Example 6 Effect of Osmotic Pressure Regulator Concentration on the Formulation
[0270] The osmotic pressure regulator sodium chloride in the compound 1 preparation is mainly used to adjust the osmotic pressure in the preparation. The different concentrations of compound 1 preparations (the concentrations of the other excipients are the same) lead to different osmotic pressures. The osmotic pressure of the maximum preparation concentration of 48 mg / mL is close to the maximum value of 310 mOsmol / kg of the normal human osmotic pressure of 280-310 mOsmol / kg, and the osmotic pressure of the minimum preparation concentration of 6 mg / mL is close to the minimum value of 280 mOsmol / kg of the normal human osmotic pressure as the standard. Nitrogen replacement was not performed in the experiments and no packaging was performed. The osmotic pressure regulator concentration design and results are shown in Table 29:
[0271] Table 29 Compound 1 formulation and osmotic pressure results
[0272] From the osmotic pressure results in the above table, it can be seen that when the sodium chloride concentration is 7 mg / mL, the osmotic pressure of the minimum preparation concentration of 6 mg / mL is 273 mOsmol / kg, which is close to the minimum osmotic pressure of 280 mOsmol / kg for normal human body; the osmotic pressure of the maximum preparation concentration of 48 mg / mL is 320 mOsmol / kg, which is close to the maximum osmotic pressure of 310 mOsmol / kg for normal human body.
[0273] Example 7 Effect of citric acid concentration on formulation stability
[0274] Citric acid in the formulation can cause pain at the injection site. Lowering the citric acid concentration can improve the pain caused by citric acid (Ya Lan Yang1 and Ted Weita Lai1,2,3. Citric Acid in Drug Formulations Causes Pain by Potentiating Acid-Sensing Ion Channel1[J]. The Journal of Neuroscience, May26, 2021·41(21):4596–4606). Therefore, it is necessary to find a suitable citric acid concentration that can minimize pain while providing good stability.
[0275] 7.1 Study on the Effect of 0.005mM-10mM Citric Acid Concentration on the Stability of the Formulation
[0276] The effect of 0.005 mM-10 mM citric acid concentration on the stability of the formulation was studied, and a 2 mg / mL formulation of compound 1 (the contents of other ingredients were the same as those of formulation 5-1 group) was selected to test its stability under accelerated conditions at 37°C.
[0277] Table 30: Effect of 0.005mM-0.1mM citric acid on stability (high molecular weight protein content)
[0278] Table 31: Effect of 0.1 mM-1 mM citric acid on stability (high molecular weight protein content)
[0279] Table 32: Effect of 1 mM-10 mM citric acid on stability (high molecular weight protein content)
[0280] Table 33: Effect of 1mM-10mM citric acid on stability (content of related substances)
[0281] The three experiments above show that at 0.005mM to 1mM citric acid concentrations, the growth trend of high molecular weight proteins decreases as the citric acid concentration increases, while having no effect on the changes in related substances. At 1mM to 10mM citric acid concentrations, the addition of citric acid decreases the growth trend of related substances and high molecular weight proteins, but this does not change with increasing citric acid concentration.
[0282] 7.2 Study on the Effect of Citric Acid on Preparation Stability
[0283] A 2 mg / mL formulation of Compound 1 was selected to test its stability under accelerated conditions at 37° C. The results of the 37° C. accelerated stability (high molecular weight protein content) study for each formulation are shown in Tables 34 and 35.
[0284] Table 34: Effect of 5 mM citric acid on the stability of each formulation (high molecular weight protein content)
[0285] Table 35: Effect of 5 mM citric acid on the stability of each formulation (related substance content)
[0286] It can be seen from Tables 34 and 35 above that 1 mM-10 mM citric acid concentration helps to improve the stability of the compound 1 preparation, and there is no significant difference in the effect of 1 mM-10 mM citric acid concentration on its stability.
[0287] 7.3 Effect of citric acid concentration on the stability of formulations of compound 1 at different concentrations
[0288] 7.3.1 Preparation of drug formulations
[0289] Low and high concentrations of Compound 1 (Compound 1 concentrations: 3 mg / mL and 48 mg / mL) were selected for citric acid concentration studies. Formulations 1-10 were prepared as shown in Table 36 below.
[0290] Table 36: Compound 1 Formulation
[0291] 7.3.2 Detection of related substances
[0292] The above-described formulations 7.3-1 to 7.3-10 were filled into assembled cartridge bottles and stability studies were conducted at 4°C, 25°C, and 37°C. The increase in the amount of the related substances in each formulation of Example 7.3.1 (formulations 7.3-1 to 7.3-10) at different days relative to day 0 was measured. The test results are shown in Tables 37, 38, and 39, respectively.
[0293] Table 37: Related substance detection results of compound 1 preparation under 4°C long-term conditions
[0294] Table 37 shows that the trends of related substances changes in each group during long-term storage at 4°C were basically the same. Different concentrations of citric acid monohydrate had little effect on the stability trend of Compound 1 preparation (changes in related substances content) during long-term storage at 4°C for 55 days.
[0295] Table 38: Related substance detection results of compound 1 preparation under accelerated conditions of 25°C
[0296] As can be seen from Table 38: in the formulation containing 48 mg / mL compound 1, as the concentration of citric acid monohydrate increases, the growth rate of related substances decreases; in the formulation containing 3 mg / mL compound 1, compared with the formulation without citric acid monohydrate, the concentration of 0.5 mM to 5 mM citric acid monohydrate helps to slow down the growth of related substances, but the effect of 0.5 mM citric acid monohydrate is weaker than that of 1 mM to 5 mM citric acid monohydrate.
[0297] Table 39: Related substance test results of compound 1 preparation under accelerated conditions at 37°C
[0298] As can be seen from Table 39: In the formulation containing 3 mg / mL of compound 1, the stability of related substances in the formulation containing citric acid monohydrate is significantly better than that in the formulation without citric acid. The growth trends of related substances in the concentrations of 2 mM to 5 mM citric acid monohydrate are similar, and both are more stable than the formulation with 0.5 mM citric acid monohydrate. In the formulation containing 48 mg / mL of compound 1, the growth rate of related substances slows down as the concentration of citric acid monohydrate increases.
[0299] 7.3.3 High Molecular Weight Protein Detection
[0300] As in 7.3.2, the increase in high molecular weight protein in each formulation of Example 7.3.1 relative to day 0 was measured at 4°C, 25°C, and 37°C. The test results are shown in Tables 40, 41, and 42, respectively.
[0301] Table 40: Results of high molecular weight protein content test of compound 1 preparation at 4°C for a long time
[0302] Table 40 shows that the trends in high-molecular-weight protein changes were essentially the same across all formulations after long-term storage at 4°C. Different concentrations of citric acid monohydrate had little effect on the growth rate of high-molecular-weight protein in the Compound 1 formulation after 55 days of long-term storage at 4°C.
[0303] Table 41: High molecular weight protein detection results of compound 1 preparation under accelerated conditions at 25°C
[0304] As shown in Table 41, the growth rate of high molecular weight protein in formulations containing 3 mg / mL Compound 1 and varying concentrations of citric acid monohydrate (0.5 mM-5 mM) was essentially consistent, and was superior to that of the 7.3-1 formulation without citric acid monohydrate. In formulations containing 48 mg / mL Compound 1, the growth rate of high molecular weight protein decreased with increasing citric acid monohydrate concentration.
[0305] Table 42: High molecular weight protein detection results of compound 1 preparation under accelerated conditions at 37°C
[0306] Table 42 shows that over 55 days of accelerated growth at 37°C, the growth rates of high molecular weight protein in formulations containing 3 mg / mL Compound 1 at varying citric acid monohydrate concentrations (1 mM to 5 mM) were essentially consistent, all exceeding those in the 7.3-2 formulation containing 0.5 mM citric acid monohydrate. The growth rate of high molecular weight protein in the 7.3-2 formulation containing 0.5 mM citric acid monohydrate was significantly slower than that in the 7.3-1 formulation without citric acid monohydrate. In formulations containing 48 mg / mL Compound 1, the growth rate of high molecular weight protein decreased with increasing citric acid monohydrate concentration.
[0307] Example 8 Stability Study of Compound 1 Formulation
[0308] 8.1 Preparation and filling of drug formulations
[0309] Compound 1 formulations 8-1 to 8-9 were prepared and filled according to Table 43 below.
[0310] Table 43: Compound 1 Formulation
[0311] 8.2 Related Substance Testing
[0312] After the above-mentioned formulations 8-1 to 8-4 were filled into the corresponding devices, stability studies were conducted at 4°C, 25°C, and 37°C. The increase in the amount of the relevant substances in each formulation of Example 8.1 (formulations 8-1 to 8-4) at different days relative to day 0 was measured. The test results are shown in Tables 44, 45, and 46, respectively.
[0313] Table 44: Related substance detection results of compound 1 preparation under 4°C long-term conditions
[0314] As can be seen from Table 44, the trends of changes in related substances in each group of preparations during long-term storage at 4°C were basically the same. There was basically no difference in the stability of Compound 1 preparations with different formulations during long-term storage at 4°C for 53 days.
[0315] Table 45: Related substance test results of compound 1 preparation under accelerated conditions at 25°C
[0316] It can be seen from Table 45 that the stability of formulations 8-3 and 8-4 is significantly better than that of formulations 8-1 and 8-2.
[0317] Table 46: Related substance test results of compound 1 preparation under accelerated conditions at 37°C
[0318] It can be seen from Table 46 that the stability of the formulation 8-3 is significantly better than that of the formulation 8-1, and the stability of the formulation 8-4 is better than that of the formulation 8-2 or is equivalent to that of the formulation 8-2.
[0319] 8.3 High Molecular Weight Protein Detection
[0320] As in 8.2, the increase in high molecular weight protein in each formulation of Example 3 relative to day 0 was measured at 4°C, 25°C, and 37°C. The test results are shown in Tables 47, 48, and 49, respectively.
[0321] Table 47: High molecular weight protein detection results of compound 1 preparation under long-term experimental conditions at 4°C
[0322] As can be seen from Table 47, the trends of changes in high molecular weight protein in each group during long-term storage at 4°C were basically the same. There was basically no difference in the stability (change in high molecular weight protein content) trends of Compound 1 formulations with different formulations during long-term storage at 4°C for 53 days.
[0323] Table 48: High molecular weight protein detection results of compound 1 preparation under accelerated conditions at 25°C
[0324] It can be seen from Table 48 that the stability of formulations 8-3 and 8-4 is significantly better than that of formulations 8-1 and 8-2.
[0325] Table 49: Comparative experiment on compound 1 formulations to accelerate high molecular weight protein at 37°C
[0326] It can be seen from Table 49 that the stability of formulations 8-3 and 8-4 is significantly better than that of formulations 8-1 and 8-2.
[0327] sequence:
[0328] SEQ ID NO.1:
[0329] GLP-1-(7-37) peptide
[0330] SEQ ID NO.2:
[0331] [Gly8, Arg34]GLP-1-(7-37) peptide
[0332] The present invention has been described through the above-described embodiments. However, it should be understood that the above-described embodiments are for illustrative and illustrative purposes only and are not intended to limit the present invention to the described embodiments. Furthermore, it will be understood by those skilled in the art that the present invention is not limited to the above-described embodiments and that further variations and modifications may be made based on the teachings of the present invention, all of which fall within the scope of the present invention. The scope of protection of the present invention is defined by the appended claims and their equivalents.
Claims
1. A liquid pharmaceutical composition comprising: a. GLP-1 peptide or a pharmaceutically acceptable salt thereof; b. Osmotic pressure regulator; c. Antioxidants; and d. buffer; in, The GLP-1 peptide is selected from: N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(19-carboxynonadecanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[21-carboxyheneicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(23-carboxytricosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[23-carboxytricosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -(23-carboxytricosanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -(19-carboxynonadecanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -(21-carboxyheneicosanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(19-carboxynonadecanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy] [(ethoxy)acetyl] [Arg34] GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[21-carboxyheneicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(23-carboxytricosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[23-carboxytricosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -(23-carboxytricosanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide, N-ε 26 -(19-carboxynonadecanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide, N-ε 26 -(21-carboxyheneicosanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(20-carboxyeicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[20-carboxyeicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(22-carboxydocosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[22-carboxydocosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -(20-carboxyeicosanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -(22-carboxydocosanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(20-carboxyeicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[20-carboxyeicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(22-carboxydocosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[22-carboxydocosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37) peptide, N-ε 26 -(20-carboxyeicosanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide, and N-ε 26 -(22-carboxydocosanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide; Preferably, the GLP-1 peptide is selected from the following compounds: N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(19-carboxynonadecanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -(19-carboxynonadecanoylamino)-4(S)-carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide, N-ε 26 -(19-carboxynonadecanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide, N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide, and N-ε 26 -[2-(2-[2-(4-[21-carboxyheneicosanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide; Preferably, the GLP-1 peptide is the following compound:
2. The pharmaceutical composition according to claim 1, wherein The pharmaceutical composition may or may not contain a preservative, preferably the pharmaceutical composition does not contain a preservative, preferably the preservative is phenol or m-cresol, preferably the pharmaceutical composition does not contain phenol or m-cresol; and / or The buffer is selected from at least one of acetic acid-sodium acetate buffer, acetic acid-potassium acetate, acetic acid-ammonium acetate, and phosphate buffer, preferably phosphate buffer; preferably the phosphate buffer is Na2HPO4; and / or The osmotic pressure regulator is NaCl or propylene glycol; and / or The antioxidant is selected from at least one of ascorbic acid, methionine, citric acid and tartaric acid. Preferably, the antioxidant is selected from citric acid and methionine; preferably, the citric acid is citric acid monohydrate.
3. The pharmaceutical composition according to claim 2, characterized in that The pharmaceutical composition comprises about 0.5 mg / ml or more Na2HPO4, preferably about 0.5-20 mg / ml Na2HPO4, preferably about 1-20 mg / ml Na2HPO4, preferably about 1-10 mg / ml Na2HPO4, more preferably about 1-5 mg / ml Na2HPO4, and further preferably about 1.40 mg / ml, about 1.41 mg / ml, about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml Na2HPO4.
4. The pharmaceutical composition according to claim 2 or 3, wherein the concentration of NaCl is about 0.5 mg / ml or more, preferably about 1 mg / ml or more, preferably about 1-30 mg / ml, preferably about 3-25 mg / ml, preferably about 5-15 mg / ml, preferably about 5-10 mg / ml, preferably about 5 mg / ml, about 5.5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 8.10 mg / ml, about 8.15 mg / ml, about 8.20 mg / ml, about 8.25 mg / ml, about 8.30 mg / ml, about 9 mg / ml or about 10 mg / ml; and / or The concentration of propylene glycol is about 1 mg / ml or more, preferably about 2 mg / ml or more, preferably about 5-25 mg / ml, preferably about 10-20 mg / ml, preferably about 12-18 mg / ml, preferably about 13 mg / ml, about 13.5 mg / ml, about 14 mg / ml, about 14.5 mg / ml, about 15 mg / ml, about 16 mg / ml or about 17 mg / ml; and / or The concentration of the antioxidant is above about 0.5 mM, preferably above about 1 mM, preferably about 1-25 mM, preferably about 1-20 mM, preferably about 1-15 mM, preferably about 1-10 mM, preferably about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM or about 9 mM.
5. The pharmaceutical composition according to any one of claims 2 to 4, wherein the antioxidant is selected from citric acid at a concentration of about 1-25 mM, preferably at a concentration of about 1-20 mM, preferably at a concentration of about 1-15 mM, preferably at a concentration of about 1-10 mM, preferably at a concentration of about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM or about 9 mM; preferably the citric acid is citric acid monohydrate; and / or The concentration of the GLP-1 peptide is at least about 1 mg / ml, preferably at least about 2 mg / ml, preferably about 1-80 mg / ml, preferably about 1-70 mg / ml, preferably about 1-65 mg / ml, preferably about 1-60 mg / ml, preferably about 2-60 mg / ml, preferably about 2-50 mg / ml, more preferably about 2 mg / ml, about 3 mg / ml, about 4 mg / ml, about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 9 mg / ml, about 10 mg / ml, about 11 mg / ml, about 12 mg / ml, about 13 mg / ml, about 14 mg / ml, about 15 mg / ml, about 16 mg / ml, about 17 mg / ml, about 18 mg / ml, about 19 mg / ml, about 20 mg / ml, about 21 mg / ml, about 22 mg / ml, about 23 mg / ml, about 24 mg / ml, about 25 mg / ml, about 26 mg / ml, about 27 mg / ml, about 28 mg / ml, about 29 mg / ml, about 30 mg / ml, about 31 mg / ml, about 32 mg / ml, about 33 mg / ml, about 34 mg / ml, about 36 mg / ml, about 37 mg / ml, about 38 mg / ml, about 39 mg / ml, about 40 mg / ml, about 41 mg / ml, about 42 mg / ml, about 43 mg / ml, about 44 mg / ml, about 45 mg / ml, about 46 mg / ml, about 47 mg / ml, about 48 mg / ml, about 49 mg / ml, about 50 mg / ml, about 51 mg / ml, about 52 mg / ml, about 53 mg / ml, about 54 mg / ml, about 56 mg / mg / ml, about 10mg / ml, about 11mg / ml, about 12mg / ml, about 13mg / ml, about 14mg / ml, about 15mg / ml, about 16mg / ml, about 17mg / ml, about 18mg / ml, about 19mg / ml, about 20mg / ml, about 21mg / ml, about 22mg / ml, about 23mg / ml, about 24mg / ml, about 25mg / ml, about 26mg / ml, about 27mg / ml, about 28mg / ml, about 29mg / ml, about 30mg / ml, about 31mg / ml, about 32mg / ml, about 33mg / ml, about 34mg / ml, about 35mg / ml, about 36mg / ml, about 37mg / ml, about 38mg / ml, about 39mg / ml, about 40mg / ml, about 41mg / ml, about 42mg / ml, about 43mg / ml, about 44mg / ml, about 45mg / ml, about 46mg / ml, about 47mg / ml, about 48mg / ml, about 49mg / ml, Or about 60mg / ml, more preferably about 2mg / ml, about 3mg / ml, about 5mg / ml, about 6mg / ml, about 12mg / ml, about 15mg / ml, about 18mg / ml, about 24mg / ml, about 30mg / ml, about 36mg / ml, or about 48mg / ml; further preferably about 3mg / ml, about 6mg / ml, about 12mg / ml, about 18mg / ml, about 24mg / ml, about 36mg / ml, or about 48mg / ml; and / or The pH of the pharmaceutical composition is about 6.5 to about 8.5, more preferably about 7.0 to about 8.5, further preferably about 7.1 to about 8.3, further preferably about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, or about 8.
2.
6. A liquid pharmaceutical composition comprising: About 1-60 mg / ml, preferably about 1-70 mg / ml, preferably about 1-65 mg / ml, preferably about The invention relates to a method for preparing the present invention in an amount of 1-60 mg / ml, preferably about 2-60 mg / ml, preferably about 2-50 mg / ml, more preferably about 2 mg / ml, about 3 mg / ml, about 4 mg / ml, about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 9 mg / ml, about 10 mg / ml, about 11 mg / ml, about 12 mg / ml, about 13 mg / ml, about 14 mg / ml, about 15 mg / ml, about 16 mg / ml, about 17 mg / ml, about 18 mg / ml, about 19 mg / ml, about 20 mg / ml, about 21 mg / ml, about 22 mg / ml, about 23 mg / ml, about 24 mg / ml, about 25 mg / ml, about 26 mg / ml, about 27 mg / ml, about 28 mg / ml, about 29 mg / ml, about 30 mg / ml, about 31 mg / ml, about 32 mg / ml, about 33 mg / ml ml, about 34 mg / ml, about 35 mg / ml, about 36 mg / ml, about 37 mg / ml, about 38 mg / ml, about 39 mg / ml, about 40 mg / ml, about 41 mg / ml, about 42 mg / ml, about 43 mg / ml, about 44 mg / ml, about 45 mg / ml, about 46 mg / ml, about 47 mg / ml, about 48 mg / ml, about 49 mg / ml, more preferably about 2 mg / ml, about 3 mg / ml, about 5 mg / ml, about 6 mg / ml, about 12 mg / ml, about 15 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, or about 48 mg / ml; more preferably about 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, or about 48 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide; about 1-30 mg / ml, preferably about 3-25 mg / ml, preferably about 5-15 mg / ml, preferably about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 8.10 mg / ml, about 8.15 mg / ml, about 8.20 mg / ml, about 8.25 mg / ml, about 8.30 mg / ml, about 8.50 mg / ml, about 9 mg / ml or about 10 mg / ml of NaCl; About 1-25 mM, preferably about 1-20 mM, more preferably about 1-15 mM, further preferably about 1-10 mM, further preferably about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM or about 9 mM citric acid; About 1-20 mg / ml, preferably about 1-10 mg / ml, more preferably about 1-5 mg / ml, and even more preferably about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml of Na2HPO4; and The pH of the pharmaceutical composition is about 6.5 to about 8.5, more preferably about 7.0 to about 8.5, further preferably about 7.1 to about 8.3, further preferably about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, or about 8.
2.
7. A liquid pharmaceutical composition comprising: About 1-60 mg / ml, preferably about 1-70 mg / ml, preferably about 1-65 mg / ml, preferably about 1-60 mg / ml, preferably about 2-60 mg / ml, preferably about 2-50 mg / ml, more preferably about 2 mg / ml, about 3 mg / ml, about 4 mg / ml, about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 9 mg / ml, about 10 mg / ml, about 11 mg / ml, about 12 mg / ml, about 13mg / ml, about 14mg / ml, about 15mg / ml, about 16mg / ml, about 17mg / ml, about 18mg / ml, about 19mg / ml, about 20mg / ml, about 21mg / ml, about 22mg / ml, about 23mg / ml, about 24mg / ml, about 25mg / ml, about 26mg / ml, about 27mg / ml, about 28mg / ml, about 29mg / ml, about 30mg / ml, about 31 48mg / ml, or about 49mg / ml, further The N-ε of about 2 mg / ml, about 3 mg / ml, about 5 mg / ml, about 6 mg / ml, about 12 mg / ml, about 15 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, or about 48 mg / ml is preferred; and the N-ε of about 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, or about 48 mg / ml is further preferred. 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide; About 5-25 mg / ml, preferably about 10-20 mg / ml, preferably about 12-18 mg / ml, preferably about 13 mg / ml, about 13.5 mg / ml, about 14 mg / ml, about 14.5 mg / ml, about 15 mg / ml, 16 mg / ml or about 17 mg / ml of propylene glycol; About 1-25 mM, preferably about 1-20 mM, more preferably about 1-15 mM, further preferably about 1-10 mM, further preferably about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM or about 9 mM citric acid; About 1-20 mg / ml, preferably about 1-10 mg / ml, more preferably about 1-5 mg / ml, and even more preferably about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml of Na2HPO4; and The pH of the pharmaceutical composition is about 6.5 to about 8.5, more preferably about 7.0 to about 8.5, further preferably about 7.1 to about 8.3, further preferably about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, or about 8.
2.
8. A liquid pharmaceutical composition comprising: About 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, about 48 mg / ml or about 60 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide; about 7 mg / ml, about 8 mg / ml, about 8.10 mg / ml, about 8.15 mg / ml, about 8.20 mg / ml, about 8.25 mg / ml, about 8.30 mg / ml, about 8.50 mg / ml, about 9 mg / ml, or about 10 mg / ml of NaCl; about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, 9 mM or about 10 mM citric acid; About 1.42 mg / ml of Na2HPO4; and The pH of the pharmaceutical composition is about 7.
3.
9. A liquid pharmaceutical composition comprising: About 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 30 mg / ml, about 36 mg / ml, or about 48 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide; About 14 mg / ml of propylene glycol; About 1 mM, 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, 9 mM or about 10 mM citric acid; About 1.42 mg / ml of Na2HPO4; and The pH of the pharmaceutical composition is about 7.
3.
10. A pharmaceutical product comprising a container and the liquid pharmaceutical composition of any one of claims 1 to 9 placed in the container; preferably, the container is selected from a pen injection device, an automatic injection device, a syringe, and a vial.
11. A pharmaceutical product comprising a container and a liquid pharmaceutical composition placed in the container, characterized in that: Before the liquid pharmaceutical composition is placed in the container and / or during the process of the liquid pharmaceutical composition being placed in the container and / or after the liquid pharmaceutical composition is placed in the container, the container is deoxygenated and filled with nitrogen; the liquid pharmaceutical composition comprises: About 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, about 48 mg / ml, or about 60 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide; about 3 mg / ml, about 4 mg / ml, about 5 mg / ml, about 6 mg / ml, about 7 mg / ml, about 8 mg / ml, about 8.10 mg / ml, about 8.15 mg / ml, about 8.20 mg / ml, about 8.25 mg / ml, about 8.30 mg / ml, about 8.50 mg / ml, about 9 mg / ml, or about 10 mg / ml of NaCl; about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml of Na2HPO4; and The pH of the pharmaceutical composition is about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.6, or about 7.
7.
12. A pharmaceutical product comprising a container and a liquid pharmaceutical composition placed in the container, characterized in that: Before the liquid pharmaceutical composition is placed in the container and / or during the process of the liquid pharmaceutical composition being placed in the container and / or after the liquid pharmaceutical composition is placed in the container, the container is deoxygenated and filled with nitrogen; the liquid pharmaceutical composition comprises: About 2-60 mg / ml, preferably about 3 mg / ml, about 6 mg / ml, about 12 mg / ml, about 18 mg / ml, about 24 mg / ml, about 36 mg / ml, or about 48 mg / ml of N-ε 26 -[2-(2-[2-(2-[2-(2-[4-(21-carboxyheneicosanoylamino)-4(S)-carboxybutanoylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide or N-ε 26 -[2-(2-[2-(4-[19-carboxynonadecanoylamino]-4(S)-carboxybutanoylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37) peptide; about 13 mg / ml, about 14 mg / ml, about 15 mg / ml, 16 mg / ml, or about 17 mg / ml of propylene glycol; about 1.42 mg / ml, about 1.5 mg / ml, about 2 mg / ml, about 2.5 mg / ml, about 3 mg / ml, about 3.5 mg / ml, about 4 mg / ml, or about 4.5 mg / ml of Na2HPO4; and The pH of the pharmaceutical composition is about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.6, or about 7.
7.
13. The pharmaceutical product according to claim 11 or 12, wherein the liquid pharmaceutical composition further comprises about 0.005 mM-25 mM, preferably about 0.01 mM-10 mM, more preferably about 0.1 mM-2.0 mM citric acid or methionine; and / or Deoxygenating the container is achieved by filling the container with an inert gas; preferably, the inert gas is nitrogen; and / or The container is a pen injection device, an autoinjector device, a syringe, or a vial.
14. The pharmaceutical composition according to any one of claims 1 to 9, or the pharmaceutical product according to any one of claims 10 to 13, wherein the pharmaceutical composition is for parenteral administration; preferably, the parenteral administration is subcutaneous administration.
15. A kit comprising a liquid pharmaceutical composition as defined in any one of claims 1 to 14 and instructions for use.
16. A kit comprising a liquid pharmaceutical composition as defined in any one of claims 1 to 14 and an injection device for administering the composition to a subject, wherein the injection device is selected from a durable pen and a prefilled pen injection device.
17. Use of the pharmaceutical composition according to any one of claims 1 to 9 or the pharmaceutical product according to any one of claims 10 to 14 in the preparation of a medicament for treating diabetes, obesity, non-alcoholic fatty liver disease, Alzheimer's disease or Parkinson's disease; preferably administering the pharmaceutical composition according to any one of claims 1 to 9 to a subject in need once a week, once every two weeks or less frequently.
18. A pharmaceutical composition according to any one of claims 1 to 9, or a pharmaceutical product according to any one of claims 10 to 14, for treating diabetes, obesity, non-alcoholic fatty liver disease, Alzheimer's disease or Parkinson's disease; preferably, the pharmaceutical composition according to any one of claims 1 to 9 is administered to a subject in need once a week, once every two weeks or less frequently.
19. A method for treating diabetes, obesity, non-alcoholic fatty liver disease, Alzheimer's disease or Parkinson's disease, the method comprising administering the pharmaceutical composition of any one of claims 1 to 19, or the pharmaceutical product of any one of claims 10 to 14 to a subject in need thereof; preferably, administering the pharmaceutical composition of any one of claims 1 to 9 to a subject in need thereof once a week, once every two weeks or less frequently.
20. A method for preparing the pharmaceutical composition according to any one of claims 1 to 9, characterized in that: The method comprises: (1) dissolving a prescribed amount of a buffer, an osmotic pressure regulator, and / or an antioxidant in water; (2) dissolving a prescribed amount of GLP-1 peptide in the solution obtained in step (1), and adjusting the pH using sodium hydroxide and / or dilute hydrochloric acid; (3) Finally, the solution obtained in step (2) was sterilized by filtering through a 0.22 μm sterile filter.
21. A method for preparing the pharmaceutical composition according to any one of claims 1 to 9, wherein: The method comprises: mixing the GLP-1 peptide, buffer, osmotic pressure regulator and antioxidant according to the prescription; Preferably, the method further comprises: The mixture obtained in step (2) is sterilized; preferably, the mixture obtained in step (2) is sterilized by filtering through a 0.22 μm sterile filter.