High concentration dosage form of hgh fusion protein

Abstract

The present invention relates to a high-concentration administration dosage form of a human growth hormone (hGH) fusion protein containing Poloxamer 188 and Polysorbate 80, and in the case of containing Poloxamer 188 and Polysorbate 80 according to the present invention, a high-concentration hGH fusion protein dosage form can be prepared, in which agglomeration is significantly reduced and storage stability is improved.

Description

[0001] Case Analysis This application is a divisional application of Chinese patent application No. 202280078653.4, entitled "High-concentration dosage form of HGH fusion protein", which entered the Chinese national phase of PCT international patent application PCT / KR2022 / 095143, filed on October 19, 2022. Technical Field

[0002] This invention relates to a high-concentration dosage form of a human growth hormone (hGH) fusion protein, and more specifically, to a high-concentration dosage form of an hGH fusion protein containing poloxamer 188 and polysorbate 80, thereby significantly reducing aggregation reactions in the high-concentration dosage form of hGH fused with immunoglobulin Fc polypeptide (hereinafter referred to as "hyFc"). Background Technology

[0003] Growth hormone is a hormone secreted by the anterior pituitary gland and is a polypeptide composed of 191 amino acids. Insulin-like growth factor-1 (IGF-1) is expressed in combination with the growth hormone receptor to participate in cell growth and regeneration. It is well known that growth hormone is produced by the pituitary gland in normal individuals, with its production gradually increasing during puberty and decreasing with age.

[0004] The most common growth hormone deficiencies are adult growth hormone deficiency (AGHD) and pediatric growth hormone deficiency (PGHD). Adult AGHD occurs when the pituitary gland is damaged during treatment for brain tumors, brain hemorrhages, or other conditions due to radiation or surgery, or it may occur idiopathically. Abnormal growth hormone secretion can lead to symptoms including weight loss, decreased bone mineral density, increased fat, decreased HDL, increased LDL, and decreased muscle strength, thus reducing quality of life. In adults with AGHD, serum IGF-1 concentrations below -2 (≤-2 SDS) or at the 2.5th percentile (≤2.5th percentile) are considered normal compared to healthy individuals of the same age. The blood growth hormone response can be measured through stimulation tests, such as the insulin tolerance test (ITT), the growth hormone-releasing hormone + arginine (GHRH+ARG) test, the glucagon test, the L-DOPA test, and the clonidine test. Growth hormone deficiency is defined as a peak GH level below 11.0 μg / L in patients with a body mass index (BMI) less than 25 kg / m², below 8.0 μg / L in patients with a BMI between 25 kg / m² and 30 kg / m², and below 4.0 μg / L in patients with a BMI greater than 30 kg / m² (Guidelines for Use of Growth Hormone in Clinical Practice, Endocr. Pract. 2009;15 (Suppl 2)). Growth hormone deficiency in children occurs in the presence of pituitary gland damage or developmental disorders. Growth hormone secretion disorders manifest as short stature, with growth height less than 3% or 5 cm per year in the same age growth curve, and may also include symptoms such as hypoglycemia, decreased endurance, depression, and mental immaturity.Children with growth hormone deficiency can be diagnosed if their height is more than 3 standard deviations (Sd) below the average for their age group, more than 1.5 SD below the average height of their parents, more than 2 SD below the average height and more than 1 SD below the average height growth for their age group for more than one year, more than 0.5 SD below the average for more than two years, or if they do not show signs of low growth but their height remains more than 2 SD below the average for more than one year or more than 1.5 Sd below the average for more than two years. (Consensusguideline for the diagnosis and treatment of GH deficiency in childhood and adolescence: summary statement of the GH Research Society. GH ResearchSociety, J. Clin. Endocrinol. Metab., 2000 Nov; 85(11): 3990-3).

[0005] Growth hormone deficiency is primarily treated with growth hormone. When growth hormone therapy was first introduced in the 1950s, it was extracted from the pituitary glands of cadavers. Due to the extremely limited amount of growth hormone extracted from a single person, the supply was very limited and the cost was high. With the development of recombinant gene technology, growth hormone synthesized in E. coli was introduced (Somatropin, 1981, Genentech, USA). Currently, commercially available recombinant growth hormone (RG) treatments in the United States include Pfizer's Genotropin, Eli Lilly's Humatrope, Genentech's Nutropin, and Novo Nordisk's Norditropin.

[0006] Currently, growth hormone preparations are administered in daily doses, which, especially in the case of pediatric patients, presents the inconvenience of daily injections over a long treatment course of 3 to 4 years, and the psychological stress caused by injections is known to reduce patients' quality of life. Furthermore, unintentional failure to inject frequently occurs, and this is considered a major factor hindering treatment efficacy. In addition, it is known that the number of administration failures increases significantly with the duration of treatment (Endocrine practice, 2008 Mar; 14(2): 143-54). Adherence is reduced in approximately two-thirds of patients due to non-compliance, and in fact, it is known to reduce the rate of height growth (PloS one, 2011 Jan; 6(1): e16223).

[0007] Due to these issues, efforts are ongoing to develop long-acting growth hormone using various technologies. Ascendis Pharma utilizes the Transcon PEG platform, OPKO / Pfizer combines hGH with CTP, and Novo Nordisk combines hGH with albumin to increase the half-life of hGH. This technology allows patients who inject growth hormone daily to receive weekly injections.

[0008] In response, the applicant filed a patent application for and was granted for the substance GX-H9, which, by fusing hGH with hyFc (which combines IgD and IgG4), resulted in lower complement-dependent and antibody-dependent toxicity and improved hGH persistence (US Patent No. 8,529,899). Subsequently, the applicant filed patent applications for dosage methods of GX-H9 that allow for once-weekly or once-bi-weekly administration (US Patent Publication Nos. 2021-0177945 and 2019-0224281).

[0009] In addition, during the initial clinical trials of GX-H9, it was supplied in a 30 mg / ml concentration formulation. Subsequently, as GX-H9 was administered once every two weeks, the dosage (volume) was increased, thus raising concerns about the necessity of higher concentration formulations.

[0010] Therefore, the inventors predicted the structural features of GX-H9 through sequence analysis, and hGH has many hydrophobic patches. Thus, focusing on the fact that proteins themselves can form aggregates due to hydrophobic interactions, and that proteins adsorb onto gas / oil / solid surfaces in drug product (DP) containers to form a protein film, the regions formed by their detachment can form aggregates as insoluble particles, the inventors worked to develop a dosage form that can prevent the formation of aggregates as described above, and confirmed that the liquid-phase dosage form with the simultaneous addition of polysorbate 80 and poloxamer 188 as surfactants significantly improved aggregate formation, thus completing the present invention. Summary of the Invention

[0011] Technical issues The purpose of this invention is to provide a high-concentration formulation of hGH-hyFc fusion protein, wherein when preparing the high-concentration formulation of hGH fusion protein, protein aggregation reaction is significantly reduced and storage stability is maintained.

[0012] Solution to the problem To achieve the above objectives, the present invention provides a liquid-phase drug dosage form comprising human growth hormone (hGH) fusion protein, poloxamer 188, and polysorbate 80.

[0013] The present invention is characterized in that the above-mentioned hGH fusion protein comprises hGH and immunoglobulin Fc polypeptide.

[0014] The present invention is characterized in that the above-mentioned hGH has the amino acid sequence of SEQ ID NO. 2.

[0015] The present invention is characterized in that the above-mentioned immunoglobulin Fc polypeptide has the amino acid sequence of SEQ ID NO. 3.

[0016] The present invention is characterized in that the above-mentioned hGH fusion protein has the amino acid sequence of SEQ ID NO. 1.

[0017] The invention is characterized in that the dosage form contains 30 mg / mL to 150 mg / mL of hGH fusion protein.

[0018] The invention is characterized in that the dosage form contains 0.01% to 0.2% (w / v) of poloxamer 188.

[0019] The invention is characterized in that the above dosage form contains 0.01% to 0.2% (w / v) of polysorbate 80.

[0020] The invention is characterized in that the above dosage form further comprises one or more excipients selected from the group consisting of histidine, arginine, glutamic acid and sodium chloride (NaCl).

[0021] The present invention is characterized in that the above dosage form further comprises a preservative.

[0022] The present invention is characterized in that the above-mentioned preservative is selected from one or more of the group consisting of m-cresol, phenol and benzyl alcohol.

[0023] The present invention is characterized in that the above-mentioned preservative is phenol at a concentration of 1 mg / mL to 2 mg / mL.

[0024] The invention is characterized in that the dosage form contains 1 mM to 20 mM of histidine.

[0025] The invention is characterized in that the above dosage form contains 40 mM to 70 mM of arginine.

[0026] The invention is characterized in that the above dosage form contains 40 mM to 70 mM of glutamic acid.

[0027] The invention is characterized in that the dosage form contains 80 mM to 100 mM of sodium chloride.

[0028] The invention is characterized in that the dosage form is pH 5.5 to pH 7.0.

[0029] The invention is characterized in that the dosage form is preferably pH 6.0 to pH 6.9, more preferably pH 6.0 to pH 6.2.

[0030] The present invention is characterized in that when the above dosage form is refrigerated and stored for 6 months, less than 1.5% of agglomerates are formed.

[0031] The invention is characterized in that the above dosage form is used for subcutaneous or intramuscular administration.

[0032] The invention is characterized in that the dosage form is stored in a container selected from the group consisting of a bottle, microtube, bag, vial, cartridge, injector, and syringe.

[0033] The effects of the invention According to the present invention, when poloxamer 188 and polysorbate 80 are included, a high concentration of hGH fusion protein formulation can be prepared with significantly reduced aggregation reaction and improved storage stability. Attached Figure Description

[0034] Figure 1 The results of the appearance analysis (visual inspection of insoluble foreign matter) were conducted to evaluate the formulation stability of GX-H9 in various dosage forms based on whether it contains polysorbate 80.

[0035] Figure 2 The results are from particulate (insoluble particulate) analysis conducted to evaluate the formulation stability of GX-H9 in various dosage forms based on whether it contains polysorbate 80.

[0036] Figure 3 The results of SE-UPLC analysis were performed to evaluate the formulation stability of GX-H9 in various dosage forms based on whether it contains polysorbate 80.

[0037] Figure 4 The results of RP-HPLC analysis were performed to evaluate the formulation stability of GX-H9 in various dosage forms based on whether it contains polysorbate 80.

[0038] Figure 5 The results of the appearance analysis (visual inspection of insoluble foreign matter) were conducted to evaluate the formulation stability of GX-H9 based on a polysorbate concentration of 80.

[0039] Figure 6 The results are from particulate analysis conducted to assess the formulation stability of GX-H9 based on a polysorbate concentration of 80.

[0040] Figure 7 The results are from purity analysis (SE-UPLC, RP-HPLC) performed to assess the formulation stability of GX-H9 based on polysorbate 80 concentration.

[0041] Figure 8 The results of the appearance analysis (visual inspection of insoluble foreign matter) were conducted to evaluate the formulation stability under various GX-H9 concentrations, polysorbate 80 concentrations, and preservative conditions.

[0042] Figure 9a The results are from purity analyses (SE-UPLC, RP-HPLC) performed to evaluate the stability of various GX-H9 formulations at different concentrations.

[0043] Figure 9b The results are from purity analysis (SE-UPLC, RP-HPLC) performed to evaluate the stability of various polysorbate esters at concentration 80 in the GX-H9 formulation.

[0044] Figure 9cThe results were obtained from purity analysis (SE-UPLC, RP-HPLC) to evaluate the stability of GX-H9 formulations treated with various preservatives. Detailed Implementation

[0045] Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Generally, the nomenclature used in this specification is that which is well-known and commonly used in the art.

[0046] In this invention, when preparing a high-concentration formulation of GX-H9, a significant reduction in aggregate formation was confirmed when polysorbate 80 and poloxamer 188 were simultaneously present as surfactants.

[0047] Therefore, one aspect of the present invention relates to a liquid-phase drug formulation comprising human growth hormone (hGH) fusion protein, poloxamer 188, and polysorbate 80.

[0048] The present invention is characterized in that the above-mentioned hGH fusion protein comprises hGH and immunoglobulin Fc polypeptide.

[0049] The hGH fusion protein "GX-H9" used in this invention refers to the human growth hormone fusion protein hGH-hyFc prepared by fusing human growth hormone (hGH) with Fc. GX-H9, as the hGH fusion protein, can be prepared according to the method disclosed in US Patent No. 8,529,899.

[0050] The present invention is characterized in that the above-mentioned hGH has the amino acid sequence of SEQ ID NO. 2, but is not limited thereto.

[0051] The present invention is characterized in that the above-mentioned immunoglobulin Fc polypeptide has the amino acid sequence of SEQ ID NO. 3, but is not limited thereto.

[0052] The present invention is characterized in that the above-mentioned hGH fusion protein has the amino acid sequence of SEQ ID NO. 1, but is not limited thereto.

[0053] The invention is characterized in that the dosage form comprises 30 mg / mL to 150 mg / mL of hGH fusion protein, preferably 60 mg / mL to 120 mg / mL of GX-H9, but is not limited thereto. The dosage form may comprise GX-H9 at concentrations such as 30 mg / mL, 40 mg / mL, 50 mg / mL, 60 mg / mL, 70 mg / mL, 80 mg / mL, 90 mg / mL, 100 mg / mL, 110 mg / mL, 120 mg / mL, 130 mg / mL, 140 mg / mL, or 150 mg / mL.

[0054] The invention is characterized in that the above dosage form contains 0.01% (w / v) to 0.2% (w / v) of poloxamer 188, preferably 0.05% (w / v) to 0.15% (w / v) or 0.08% (w / v) to 0.12% (w / v) of poloxamer 188, more preferably 0.10% (w / v) of poloxamer 188, but is not limited thereto.

[0055] The invention is characterized in that the above-mentioned dosage form contains 0.01% (w / v) to 0.2% (w / v) of polysorbate 80, preferably 0.05% (w / v) to 0.15% (w / v) or 0.05% (w / v) to 0.10% (w / v) of polysorbate 80, more preferably 0.07% (w / v) of polysorbate 80, but is not limited thereto.

[0056] The invention is characterized in that the above dosage form further comprises one or more excipients selected from the group consisting of histidine, arginine, glutamic acid and sodium chloride (NaCl).

[0057] The invention is characterized in that the above dosage form further includes a preservative.

[0058] The present invention is characterized in that the above-mentioned preservative is selected from one or more of the group consisting of m-cresol, phenol and benzyl alcohol, but is not limited thereto.

[0059] The present invention is characterized in that the above-mentioned preservative is phenol at a concentration of 1 mg / mL to 2 mg / mL, preferably phenol at a concentration of 1 mg / mL, but not limited thereto.

[0060] The invention is characterized in that the above dosage form contains 1 mM to 20 mM of histidine, preferably 5 mM to 15 mM of histidine, more preferably 10 mM of histidine, but is not limited thereto.

[0061] The invention is characterized in that the above dosage form contains 40mM to 70mM of arginine, preferably 50mM to 60mM of arginine, more preferably 55mM of arginine, but is not limited thereto.

[0062] The invention is characterized in that the above dosage form contains 40 mM to 70 mM of glutamic acid, preferably 50 mM to 60 mM of glutamic acid, more preferably 55 mM of glutamic acid, but is not limited thereto.

[0063] The invention is characterized in that the dosage form contains 80 mM to 100 mM of sodium chloride, preferably 85 mM to 95 mM of sodium chloride, more preferably 90 mM of sodium chloride, but is not limited thereto.

[0064] The invention is characterized in that the dosage form is pH 5.5 to pH 7.0, preferably pH 5.9 to pH 6.9, more preferably pH 6.0 to pH 6.5, even more preferably pH 6.0 to pH 6.2, and most preferably pH 6.1, but is not limited thereto.

[0065] The invention is characterized in that when the above-mentioned dosage form is refrigerated for about 6 months, for example, at a temperature of 2°C to 8°C for about 6 months, preferably at a temperature of 4°C to 6°C for about 6 months, less than 3.0% of agglomerates are formed, more preferably less than 2.0% of agglomerates are formed, more preferably less than 1.5% of agglomerates are formed, for example, 0% to 1.5% of agglomerates are formed, but it is not limited thereto.

[0066] The invention is characterized in that the above dosage form is used for subcutaneous or intramuscular administration.

[0067] The present invention is characterized in that the above dosage form is an injectable dosage form.

[0068] The invention is characterized in that the dosage form is stored in a container selected from the group consisting of bottles, microtubes, bags, vials, cartridges, injectors, and syringes, but is not limited thereto.

[0069] For example, the dosage form may be stored in a container selected from, but not limited to, glass vials, glass boxes, plastic boxes, prefilled syringes, pen injectors, and autoinjectors.

[0070] The high-concentration GX-H9 drug formulation of the present invention can be administered to adults or children with growth hormone deficiency. The high-concentration GX-H9 drug formulation of the present invention can be administered to the target body via various methods.

[0071] In this invention, the object can be a mammal, but is not limited thereto; preferably, the object can be a human.

[0072] For example, the above dosage forms can be administered parenterally, such as subcutaneously or intramuscularly.

[0073] In this invention, the term "pharmaceutical dosage form" refers to a formulation containing GX-H9 in a form that enables the bioactivity of GX-H9 to be effective, and which does not contain an amount exceeding an amount of any ingredient that would be toxic to a subject administering the dosage form.

[0074] A “stable” dosage form effectively possesses the physiological and / or chemical stability and / or biochemical activity of GX-H9 during storage, preservation, and distribution. In one approach, the dosage form retains not only its physiological and chemical stability but also its biological activity when stored. Shelf life is typically selected based on the expected shelf life of the dosage form. Stability can be measured at a selected temperature within the selected pre-selected period. For example, in one approach, the liquid dosage form is stable for approximately 2 to 4 weeks, at least approximately 3 months, at least approximately 6 months, at least approximately 9 months, at least approximately 12 months, or at least approximately 18 months at a temperature of approximately 25°C. As another example, the liquid dosage form is stable for approximately 2 to 4 weeks, at least approximately 3 months, at least approximately 6 months, at least approximately 9 months, at least approximately 12 months, at least approximately 18 months, at least approximately 24 months, at least approximately 30 months, or at least approximately 36 months at a temperature of approximately 5°C.

[0075] There are various analytical techniques for measuring protein stability, and references have been made to existing literature [Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10: 29-90 (1993)].

[0076] The stability of liquid formulations can be qualitatively and / or quantitatively assessed using a variety of different methods, including assessments of dimer, polymorphism, and / or aggregate formation (e.g., size exclusion ultra-high performance liquid chromatography (SE-UPLC), size exclusion high performance liquid chromatography (SE-HPLC), reversed-phase ultra-high performance liquid chromatography (RP-UPLC), reversed-phase high performance liquid chromatography (RP-HPLC), matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS), analytical ultracentrifugation, light scattering (photon correlation spectroscopy, dynamic light scattering (DLS), static light scattering, multi-angle laser light scattering). Evaluation of turbidity and / or visual detection by means of scattering (MALLS), flow-based microscopy imaging, resistance impedance (coulter) counters, application of light shielding or other liquid particle measurement systems; cation exchange chromatography (CEX), isoelectric focusing (IEF), such as by applying capillary technique (cIEF), or capillary zone electrophoresis; nucleotide-terminal or carboxyl-terminal sequence analysis; mass spectrometry peptide mapping analysis (e.g., using trypsin / LYS-C); and assessment of antibody biological activity or antigen-binding function.

[0077] In this invention, the aforementioned drug dosage form can be an isotonic formulation, which is essentially an isotonic formulation with the same osmotic pressure as human blood. Isotonic formulations typically have an osmotic pressure of approximately 250 to 350 mOsm / kg. Isotonicity can be measured using a water vapor pressure meter or an ice-cooled osmoremeter.

[0078] In this invention, "buffer" refers to a buffer that resists pH changes through the action of an acid-base binding component. In some embodiments, the buffer of this invention adjusts the pH value of the dosage form to about 5.0 to about 7.5, about 5.8 to about 7.0, about 6.0 to about 6.5, or about 6.0 to about 6.3. In one approach, examples of buffers used alone or in combination to control pH may include acetate, succinate, gluconate, histidine, citrate, phosphate, maleate, cacodylate, 2-[N-morpholino]ethanesulfonic acid (MES), bis(2-hydroxyethyl)iminotris[Hydroxymethyl]methane (bis-tris)methane, N-[2-acetamido]-2-iminodiacetic acid (ADA), glycylglycine, or other organic acid buffers.

[0079] In addition, in this invention, the buffer can be a biological buffer, that is, a buffer known in the art for use in biological systems or in the context of biological systems. For example, the buffer used in this invention can be a mixed buffer containing inorganic and organic salts.

[0080] Furthermore, the preferred mixed buffer solution that can be used in this invention is a biological buffer solution that may contain amino acids. The preferred amino acids that can be used in this invention may be one or more selected from the group consisting of histidine, arginine, and glutamic acid, and preferably, contain all three: histidine, arginine, and glutamic acid.

[0081] In this invention, "surfactant" refers to an agent that reduces the surface tension of a liquid. In one embodiment, the surfactant may be a nonionic surfactant. Preferably, the invention may include both polysorbate 80 and poloxamer 188 as surfactants, but is not limited thereto. Examples of surfactants that may also be included in this invention include those selected from polysorbate (polyoxyethylene sorbitan monolaurate, for example, polysorbate 20), TRITON (polyethylene glycol monooctylphenyl ether, a nonionic detergent, a subsidiary of Dow Chemical's Union Carbide, Mitland, Michigan, USA); sodium dodecyl sulfate. sodium dodecyl sulfate (SDS); sodium octyl glycoside; lauric acid-, myristic acid-, linoleic acid-, or stearoyl-sulfobetaine; lauric acid-, myristic acid-, linoleic acid-, or stearoyl-sarcosine; linoleic acid-, myristic acid-, or cetyl-betaine; dodecylaminopropyl-, cocamidopropyl-, linoleamide-, myristamidopropyl-, palmitamidopropyl-, or isostearamidopropyl-betaine (e.g., dodecylaminopropyl); myristamidopropyl-, palmitamidopropyl-, or isostearamidopropyl-dimethylamine; sodium cocoyl methyl taurate or sodium oleoyl methyl taurate; sorbitol monophosphate; and MONAQUAT series (Mona Industries, Inc., Paterson, New Jersey, USA); polyethylene glycol (polyethyl Surfactants in the group consisting of glycol (PEG), polypropylene glycol (PPG), and copolymers of polyethylene oxide and polypropylene oxide (e.g., Pluronics / Poloxamer, PF68, etc.).

[0082] In one embodiment, the dosage form of the present invention is sterilized and does not contain any preservative. In another embodiment, the dosage form of the present invention may contain any preservative, which may be a preservative free of parabens. Parabens are a series of parabens or esters of parabens, known to cause cytokine release and stimulation and associated with various types of cancer. Examples of parabens include methyl parabens, ethyl parabens, propyl parabens, butyl parabens, heptayl parabens, isobutyl parabens, isopropyl parabens, benzyl parabens, and their sodium salts.

[0083] Exemplary paraben-free preservatives include cresols, such as 3-cresol (m-cresol), phenol, phenethyl alcohol, octyl glycol, phenoxyethanol, sorbitol ester, potassium sorbate, sodium sorbate, sorbic acid, sodium benzoate, benzoic acid, acetylmorphan, oleuropein, carawayol, blueberry extract, gluconolactone, green tea extract, sunflower seed oil (Helianthus annuus), lactic acid bacteria ferment, Usnea barbata extract, polyaminopropyl biguanide, polyglycerol-3 palmitate, polyglycerol-6 caprylate, pomegranate extract, Populus tremuloides bark extract, resveratrol, Rosmarinus officinalis leaf extract, benzyl alcohol, or any combination thereof.

[0084] The dosage forms of the present invention may also include antioxidants. "Antioxidant" refers to an agent that inhibits the oxidation of other molecules. Examples of antioxidants in the present invention include citrate, lipoic acid, uric acid, glutathione, tocopherol, carotene, lycopene, cysteine, and phosphonate compounds, such as hydroxyethyl phosphate, ferric sulfate, and malate.

[0085] The GX-H9 used in this invention is substantially pure (i.e., free from contamination of proteins, etc.) and can be substantially the same. “Substantially pure” means that it contains at least about 90% by weight of GX-H9 based on the total weight of proteins in the pharmaceutical dosage form. Alternatively, it may contain at least about 95% or 97% by weight of GX-H9.

[0086] GX-H9 may include "substantially homologous" GX-H9, which refers to a variant having equivalent activity to the GX-H9 used in this invention. Alternatively, "substantially homologous" may mean substantially identical to the GX-H9 amino acid sequence. A substantially identical sequence means aligning the GX-H9 amino acid sequence of this invention with any other sequence in a manner that maximizes correspondence, representing a sequence with at least 90% homology when analyzed using algorithms commonly used in the art, and most preferably, a sequence representing at least 95%, 96%, 97%, 98%, or 99% homology.

[0087] The present invention will now be described in more detail through embodiments. These embodiments are merely illustrative of the invention, and the scope of the invention is not limited to these embodiments, as will be apparent to those skilled in the art.

[0088] Example 1. Stability assessment of high-concentration hGH-hyFc formulation over 12 weeks. 1-1. Summary of Preliminary Experimental Results In preliminary experiments with the high-volume formulation of hGH-hyFc (hereinafter referred to together with "GX-H9"), anticoagulation under temperature stress was greater in the pH range of 6.0 to 6.5 than in other pH ranges. Furthermore, a higher level of stability was confirmed when using a histidine buffer compared to phosphate buffers. Additionally, high stability was achieved when using poloxamer 188 and polysorbate 80 under freeze / thaw and stirring conditions.

[0089] Overall, SE-HPLC results were confirmed to show greater stability at pH 6.1, while RP-UPLC results showed greater stability in formulations containing sodium chloride at pH 6.5 (data not shown).

[0090] 1-2. Experimental Design The following also includes a phosphate buffer solution at pH 6.9. Thus, for a total of five different formulations, the stability was assessed under temperature (-70°C, 2 to 8°C, 25°C, 45°C) stress, freeze / thaw stress, and mild shaking stress. Table 1 shows the formulation conditions for GX-H9 and the concentrations and types of excipients.

[0091] [Table 1]

[0092] For solutions containing GX-H9, ultrafiltration (UF) / diafiltration (DF) was performed using the various buffer solutions mentioned in Table 1 (except for Poloxamer 188). After confirming that the buffer solution was sufficiently equilibrated by measuring pH and conductivity, it was concentrated to the desired concentration, and then Poloxamer 188 was added to achieve the target concentration.

[0093] Under aseptic conditions, GX-H9 samples prepared from various formulations of oil were filtered through a 0.2 μm polyvinylidene difluoride (PVDF) membrane and then dispensed into pre-sterilized depyrogenated borosilicate vials. The vials were then sealed with aluminum crimping. Each vial was subsequently placed under stress conditions for stability testing, and samples were extracted at the desired time to assess stability (Table 2). Samples were diluted as needed for appropriate analysis, and remaining samples were kept at -70°C for backup testing. Table 2 shows the experimental conditions for the stress conditions and analytical items.

[0094] [Table 2]

[0095] 1-3. Changes in concentration (A280), turbidity (A330), appearance, and pH. Under stress conditions, the concentration, turbidity, visual appearance, and pH changes of each dosage form were confirmed.

[0096] The sample concentration was determined using an absorbance coefficient of 1.0 mg / mL = 1.04 AU and by measuring absorbance at 280 nm. The confirmed final protein concentration was modified relative to scattering ((A280 - A330) / ε). 280 =Modified protein concentration).

[0097] The results of confirming the protein concentration, turbidity, appearance, and pH of the samples after various stress conditions are shown in Tables 3 and 4. No clear differences were observed in protein concentration, turbidity, and pH under various stress conditions. No visible particles were observed in any dosage form under various stress conditions. No identifiable analytical changes were confirmed in any dosage form after 3 days of shaking stress at 5× freeze / thaw or 2–8°C.

[0098] Table 3 shows the changes in concentration, turbidity, appearance, and pH value after temperature stress.

[0099] [Table 3]

[0100] *C = Clear; T = Turbid; O = Opalescent (translucent); LY = Light Yellow; NC = No Color; NP = No Particles Table 4 shows the changes in concentration, turbidity, appearance, and pH after freezing / thawing and shaking stress.

[0101] [Table 4]

[0102] *C = Clear; T = Turbid; O = Opalescent (translucent); LY = Light Yellow; NC = No Color; NP = No Particles The target concentration of GX-H9 was achieved in all tested dosage forms, which was in contrast to the <1% scattering levels observed in most dosage forms after preparation. Following the stress reaction, the results for protein concentration, turbidity, appearance, and pH of the samples were confirmed. No significant differences were observed in protein concentration, turbidity, and pH across all dosage forms under various stress conditions.

[0103] 1-4. SE-UPLC SE-UPLC (also known as gel filtration chromatography) separates proteins into analytical speciations based on size. In this method, high molecular weight proteins (e.g., aggregates, IgG dimers, and oligomers) are dissolved first as pre-peaks compared to the desired (monomer) IgG species. Low molecular weight proteins (e.g., degradation products and fragments) are then dissolved as post-peaks.

[0104] SE-UPLC was performed using Waters Acquity UPLC BEH SEC, 200A (4.6 × 300 mm, 1.7 μm) with 100 mM sodium phosphate, 200 mM arginine-HCl, pH 7.0, and a mobile phase.

[0105] Table 5 shows the SE-UPLC results under temperature stress.

[0106] [Table 5]

[0107] Table 6 shows the SE-UPLC results based on freeze / thaw and shock stress.

[0108] [Table 6]

[0109] SE-UPLC results for samples stored for 12 weeks at -70°C, 2-8°C, 25°C, and 45°C are shown in Tables 5 and 6, with both high molecular weight (HMW) and low molecular weight (LMW) increasing. Endpoint stability comparisons show that GX-H9 exhibits higher stability at pH 6.1 compared to higher pH values. Furthermore, within the same pH 6.1 dosage form, the 100 mg / mL formulation is more stable than the 150 mg / mL formulation. No identifiable trend was observed among all tested dosage forms when exposed to freeze / thaw and 2-8°C shaking stress, likely due to similar results at t=0 and within validation variability (Tables 5 and 6).

[0110] 1-5.RP-UPLC RP-UPLC separates molecules based on differences in hydrophobicity. Separation depends on the solute binding of a mobile phase to a stationary phase containing a fixed hydrophobic ligand. Elution is typically performed in order of increasing molecular hydrophobicity, through changes in the hydrophobicity of the mobile phase in the organic solvent. Solvent gradient separation was performed using a Waters Acquity UPLC Protein BEH300 C4 column (2.1 × 150 mm) with a UV detector (A220) monitored at 220 nm and an ultra-high performance liquid chromatography system (Waters Acquity H-Class UPLC) with integrated software (Chromeleon 7.2).

[0111] Table 7 shows the RP-UPLC results under temperature stress.

[0112] [Table 7]

[0113] Table 8 shows the RP-UPLC results based on freeze / thaw and shock stress.

[0114] [Table 8]

[0115] RP-UPLC results for samples stored at -70°C, 2-8°C, and 25°C for 12 weeks, or at 45°C for 3 weeks, showed that the pH 6.1 formulation containing arginine and glutamic acid exhibited superior stability compared to other high-pH formulations. No identifiable bias was observed among all tested formulations when exposed to freeze / thaw and 2-8°C shaking stress, likely due to similar results at t=0 and within validation variability (Tables 7 and 8).

[0116] Example 2. Stability assessment of hGH-hyFc formulations based on whether they contain polysorbate 80 Under the conditions of 10 mM histidine, 0.1% (w / v) poloxamer 188, 55 mM arginine, 55 mM glutamic acid, 90 mM NaCl, and pH 6.1 (selected as the optimal formulation in Example 1), a tendency for insoluble foreign matter formation and micro-increase was also observed. Thus, the hGH of GX-H9 has many hydrophobic patches. Considering that proteins themselves can form aggregates due to hydrophobic interactions, and that proteins adsorbed on gas / oil / solid surfaces in DP containers can form a protein film, the regions formed by their detachment can form aggregates as insoluble particles. In order to develop a formulation that can prevent the formation of aggregates as described above, it is necessary to confirm a significant improvement in aggregate formation in liquid-phase formulations with the addition of poloxamer 188 and polysorbate 80 as surfactants.

[0117] Compared to using poloxamer 188 alone, using both poloxamer 188 and polysorbate 80 resulted in better suppression of insoluble foreign matter and particulate formation. In four dosage form compositions containing 60 mg / mL hGH-hyFc, the concentration of poloxamer 188 was fixed at 0.10% (w / v), and 0.03% (w / v) of polysorbate 80 was added. Appearance, particulate analysis, SE-UPLC, and RP-HPLC (reversed-phase high performance liquid chromatography) were performed on a total of eight dosage forms.

[0118] 2-1. Appearance analysis (visual inspection of insoluble foreign matter) Various dosage form compositions were prepared using hGH-hyFc stock solution (Drug Substance, DS). After filtration through a 0.22 μm PES filter in a biosafety cabinet, approximately 0.8 mL of the solution was inoculated into glass vials to prevent the inclusion of external dust or other contaminants. Sample containers stored at refrigeration (5°C) and room temperature (25°C) were removed and allowed to thaw at room temperature for approximately 30 minutes. Then, in a foreign matter inspection station, the external surfaces of the containers were cleaned with alcohol or similar substances, and the contents were directly observed with the naked eye to confirm the presence of insoluble foreign matter within 6 months.

[0119] As a result, Figure 1 As shown, when stored refrigerated (5°C), no insoluble foreign matter formed within 6 months under conditions containing polysorbate 80. However, under conditions without polysorbate 80, a significant number of the 10 samples showed the formation of insoluble foreign matter within 6 months of refrigeration. Furthermore, when stored at room temperature (25°C), the formation of insoluble foreign matter was generally suppressed under conditions containing polysorbate 80 compared to conditions without polysorbate 80. In particular, no insoluble foreign matter formed within 3 months in Candidates #1 and #4. However, under dosage forms without polysorbate 80, the formation of insoluble foreign matter was confirmed in all vials after 6 months of storage at room temperature.

[0120] 2-2. Sub-visible particle analysis Take 1 mL of each conditional sample in a glass vial and prepare it. Then, entrust an external company (KBIO, Dosage Form Technical Support Group) to perform particulate analysis using the Microflow Imaging device (MFI 5200, Proteinsimple) and MFI View System Software (MVSS) Version 2-R4.1.0.40.4816 and MFI View Analysis Suite (MVAS) Version 1.4.0.

[0121] As a result, Figure 2 As shown, after 6 months of refrigeration (5°C), candidates #1, #2, #4, and #6 containing polysorbate 80 were observed to effectively inhibit microparticle formation. Regarding microparticle formation, it is known that significantly higher levels of microparticles were formed in formulations that did not contain polysorbate 80, both under refrigeration and at room temperature.

[0122] 2-3. SE-UPLC Analysis Using an ACQUITY UPLC Protein BEH200 (4.6*300mm) (Waters, 186005226) column, 100mM sodium phosphate (pH 7.0), 200mM arginine-HCl, and mobile phase solution were injected at a rate of 0.25mL / min. 10μg of samples under various conditions were then injected, and the peak of GX-H9 protein was confirmed at 280nm absorbance.

[0123] As a result, Figure 3 As shown, candidates #2, #3, and #5 exhibited excellent purity after 6 months of refrigerated (5°C) and room temperature (25°C) storage. In particular, candidate #2 demonstrated exceptionally high purity in dosage forms containing polysorbate 80. It was confirmed that, when candidates #3 and #5 were used as dosage forms without polysorbate 80, their purity was similar to that of candidate #2 after 6 months of refrigerated storage, but relatively lower after 6 months of room temperature storage.

[0124] 2-4. RP-HPLC Using Proteonavi C4 300 A 5 μm (Shiseido, 80205) column was used to inject a mobile phase solution of trifluoroacetic acid (TFA) in 0.05% water (MPA) and trifluoroacetic acid (TFA) in 0.05% acetonitrile (MPB) at a rate of 0.5 mL / min under the gradient conditions shown in Table 9. 20 μg of sample was injected, and the peak of the GX-H9 protein was confirmed at an absorbance of 220 nm.

[0125] [Table 9]

[0126] As a result, Figure 4 As shown, when stored under refrigeration (5°C) for 6 months, the purity was similar among all dosage forms, but when stored at room temperature (25°C) for 6 months, Candidate #4 showed the best relative purity.

[0127] Example 3. Evaluation of the stability of hGH-hyFc formulation based on polysorbate 80 content. Based on Example 2, in Candidate #2, which was determined to be the most suitable dosage form, the concentration of polysorbate 80 was set at different levels, and the dosage form stability analysis of hGH-hyFc was performed using the same experimental methods as in Example 2 (appearance analysis, particulate analysis, SE-UPLC analysis, RP-HPLC analysis).

[0128] As a result, in appearance analysis, it was confirmed that a formulation containing more than 0.07% (w / v) of polysorbate 80 was suitable for storage at room temperature (25°C) for more than 3 months. Figure 5 Furthermore, in particulate analysis, substantially similar particles were identified at concentrations of polysorbate 80 ranging from 0.03% (w / v) to 0.15% (w / v). Figure 6 In the SE-UPLC and RP-HPLC results, there was no significant difference in purity based on the concentration of polysorbate 80 when stored under refrigeration (5°C). In the SE-UPLC results, when stored at room temperature, the purity decreased when the concentration of polysorbate 80 was above 0.10% (w / v). In the RP-HPLC results, the purity difference based on the concentration of polysorbate 80 was not significant when stored at room temperature. Figure 7 ).

[0129] Example 4. Stability assessment of high-capacity hGH-hyFc dosage form In summary, regarding the above embodiments, for (i) the case where polysorbate 80 is fixed at 0.07% (w / v), and 60, 80, 100, and 120 mg / ml of hGH-hyFc are treated respectively and stored in a glass box; (ii) the case where hGH-hyFc is fixed at 120 mg / ml, and 0.03, 0.05, 0.07, 0.10, and 0.15% (w / v) of polysorbate 80 are treated respectively and stored in a glass box; and (iii) the case where hGH-hyFc is fixed at 60 mg / ml and polysorbate 80 is fixed at 0.07% (w / v), and m-cresol, phenol, or benzyl alcohol are treated respectively. The preservative (alcohol) was stored in a glass vial. By analyzing the appearance of the sample under refrigeration (5°C) and room temperature (25°C), the presence of insoluble foreign matter was confirmed, and changes in purity were identified, thereby assessing the stability of the dosage form.

[0130] 4-1. Appearance analysis (visual inspection of insoluble foreign matter) Various dosage form compositions were prepared using hGH-hyFc stock solution (DS). After filtration through a 0.22 μm PES filter in a biosafety cabinet, approximately 1.0 mL of the solution was inoculated into glass vials to prevent the inclusion of external dust or other contaminants. Sample containers stored at refrigeration (5°C) and room temperature (25°C) were removed and allowed to thaw at room temperature for approximately 30 minutes. Then, in a foreign matter inspection station, the external surfaces of the containers were cleaned with alcohol or similar substances, and the contents were directly observed with the naked eye to confirm the presence of insoluble foreign matter within 6 months.

[0131] As a result, (i) when polysorbate 80 was fixed at 0.07% (w / v), treated with 60-100 mg / ml hGH-hyFc, and stored in glass boxes, it was confirmed that no insoluble foreign matter was generated within 6 months of refrigeration and room temperature storage; (ii) when hGH-hyFc was fixed at 120 mg / ml, treated with 0.05-0.15% (w / v) polysorbate 80, and stored in glass boxes, it was confirmed that no insoluble foreign matter was generated within 6 months of refrigeration and room temperature storage; (iii) when hGH-hyFc was fixed at 60 mg / ml and polysorbate 80 was fixed at 0.07% (w / v), treated with 1 mg / mL phenol as a preservative, and stored in glass vials, it was confirmed that no insoluble foreign matter was generated within 6 months of refrigeration and room temperature storage. Figure 8 ).

[0132] 4-2. SE-UPLC and RP-HPLC analysis Take the sample from the glass box or vial according to the conditions, dilute it to 2 mg / mL with mobile phase buffer and formulation buffer, filter it through a 0.22 μm tubular filter for cellulose acetate centrifugation, put it into an LC vial, and perform SE-UPLC or RP-HPLC analysis.

[0133] For SE-UPLC, using an ACQUITY UPLC Protein BEH200 (4.6*300mm) (Waters, 186005226) column, 100mM sodium phosphate (pH 7.0), 200mM arginine-HCl, and mobile phase solution were injected at a rate of 0.25mL / min, and 10μg of sample was injected. The peak of GX-H9 protein was confirmed at an absorbance of 280nm.

[0134] For RP-HPLC, use Proteonavi C4 300. A 5 μm (Shiseido, 80205) column was used to inject a mobile phase solution of trifluoroacetic acid (TFA) in 0.05% water (MPA) and trifluoroacetic acid (TFA) in 0.05% acetonitrile (MPB) at a rate of 0.5 mL / min under the gradient conditions shown in Table 9. 20 μg of sample was injected, and the peak of the GX-H9 protein was confirmed at an absorbance of 220 nm.

[0135] As a result, (i) when polysorbate 80 was fixed at 0.07% (w / v), and hGH-hyFc at concentrations of 60–120 mg / mL was treated and stored in a glass container, in SE-UPLC, the purity difference between hGH-hyFc concentrations of 60 mg / mL and 120 mg / mL was 0.5% when stored refrigerated (5°C), which was negligible. However, when stored at room temperature (25°C), the purity difference between hGH-hyFc concentrations of 60 mg / mL and 120 mg / mL was confirmed to be 3.6%. Furthermore, when stored at room temperature, the purity difference between hGH-hyFc concentrations of 60 mg / mL and 120 mg / mL, as measured by RP-HPLC, was 0.8%, which was also negligible. Figure 9a ).

[0136] (ii) When hGH-hyFc was fixed at 120 mg / ml, and 0.03-0.15% (w / v) of polysorbate 80 were treated and stored in glass containers, there was no purity difference between the concentrations of 0.03% (w / v) and 0.15% (w / v) of polysorbate 80 when refrigerated, and the difference between the concentrations of 0.03% (w / v) and 0.15% (w / v) of polysorbate 80 when stored at room temperature was also negligible. Figure 9b ).

[0137] (iii) When m-cresol, phenol, or benzyl alcohol preservatives were treated with hGH-hyFc fixed at 60 mg / ml and polysorbate 80 fixed at 0.07% (w / v) and stored in glass vials, SE-UPLC results showed that the purity reduction was minimal when phenol was present at 1 mg / ml. RP-HPLC results showed no significant difference in purity among all preservatives. Figure 9c ).

[0138] The foregoing has described specific parts of the present invention in detail. It will be clear to those skilled in the art that this specific description is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the essential scope of the present invention is defined by the appended claims and their equivalents.

[0139] Sequence List 1. GX-H9 (hGH-hyFc5) Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe Arg Asn Thr Gly Arg Gly Gly Glu Glu Lys Lys Lys Glu Lys Glu Lys Glu Glu Gln Glu Glu Arg Glu Thr Lys Thr Pro Glu Cys Pro Ser His Thr Gln Pro Leu Gly Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 2. hGH (GenBank: AAA98618.1) (excluding the signal sequence) Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 3.hyFc5 Arg Asn Thr Gly Arg Gly Gly Glu Glu Lys Lys Lys Glu Lys Glu Lys Glu Glu Gln Glu Glu Arg Glu Thr Lys Thr Pro Glu Cys Pro Ser His Thr Gln Pro Leu Gly Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

Claims

1. A liquid-phase drug dosage form comprising, as shown in SEQ ID NO: 1, a human growth hormone fusion protein, 1 mM to 20 mM of histidine, 40 mM to 70 mM of arginine, 40 mM to 70 mM of glutamate, 80 mM to 100 mM of sodium chloride, and 0.01% (w / v) to 0.2% (w / v) of poloxamer 188, and a pH of 5.5-7.

0.

2. The dosage form according to claim 1, characterized in that, The dosage form contains 5 mM to 15 mM of histidine.

3. The dosage form according to claim 1, characterized in that, The dosage form contains 10 mM of histidine.

4. The dosage form according to claim 1, characterized in that, The dosage form contains 50 mM to 60 mM of arginine.

5. The dosage form according to claim 1, characterized in that, The dosage form contains 55 mM of arginine.

6. The dosage form according to claim 1, characterized in that, The dosage form contains 50 mM to 60 mM of glutamic acid.

7. The dosage form according to claim 1, characterized in that, The dosage form contains 55 mM of glutamic acid.

8. The dosage form according to claim 1, characterized in that, The dosage form contains 85 mM to 95 mM sodium chloride.

9. The dosage form according to claim 1, characterized in that, The dosage form contains 90 mM sodium chloride.

10. The dosage form according to claim 1, characterized in that, The dosage form contains 0.05% (w / v) to 0.15% (w / v) of poloxamer 188.

11. The dosage form according to claim 1, characterized in that, The dosage form contains 0.08% (w / v) to 0.12% (w / v) of poloxamer 188.

12. The dosage form according to claim 1, characterized in that, The dosage form contains 0.10% (w / v) of poloxamer 188.

13. The dosage form according to claim 1, characterized in that, The dosage form is pH 6.0 to pH 6.

5.

14. The dosage form according to claim 1, characterized in that, The dosage form is pH 6.0 to pH 6.

2.

15. The dosage form according to claim 1, characterized in that, The dosage form is pH 6.

1.

16. The dosage form according to claim 1, characterized in that, The dosage form comprises human growth hormone fusion protein as shown in SEQ ID NO: 1, 5 mM to 15 mM histidine, 50 mM to 60 mM arginine, 50 mM to 60 mM glutamic acid, 85 mM to 95 mM sodium chloride, and 0.05% (w / v) to 0.15% (w / v) poloxamer 188, and has a pH of 6.0-6.

5.

17. The dosage form according to claim 1, characterized in that, The dosage form comprises human growth hormone fusion protein as shown in SEQ ID NO: 1, 10 mM histidine, 55 mM arginine, 55 mM glutamic acid, 90 mM sodium chloride, and 0.10% (w / v) poloxamer 188, and has a pH of 6.

1.

18. The dosage form according to claim 1, characterized in that, The dosage form contains 30 mg / mL to 150 mg / mL of the human growth hormone fusion protein as shown in SEQ ID NO:

1.

19. The dosage form according to claim 1, characterized in that, The dosage form contains 60 mg / mL to 120 mg / mL of the human growth hormone fusion protein as shown in SEQ ID NO:

1.

20. The dosage form according to claim 1, characterized in that, The dosage form is intended for subcutaneous or intramuscular administration.

21. The dosage form according to claim 1, characterized in that, The dosage form is stored in a container selected from the group consisting of vials, microtubes, packs, vials, boxes, injectors, and syringes.

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