Stable pharmaceutical composition of Anti-rankl antibody

Abstract

A stable pharmaceutical composition containing an anti-RANKL antibody, and more specifically, a pharmaceutical composition of denosumab. The composition has ensured stability and reduced insoluble particles in the preparation, which effectively solves the problems in high-concentration antibody preparations with respect to drug manufacturing, storage, transportation, and delivery.

Description

Stable anti-RANKL antibody pharmaceutical compositions

[0001] This application claims priority to Chinese Patent Application No. 2024117184294, filed on November 27, 2024, entitled "Pharmaceutical Composition of Stable Anti-RANKL Antibody", and Chinese Patent Application No. 2025117346363, filed on November 21, 2025, entitled "Pharmaceutical Composition of Stable Anti-RANKL Antibody", the entire contents of which are incorporated herein by reference. Technical Field

[0002] This disclosure relates to a stable anti-RANKL antibody pharmaceutical composition, and more specifically, to an anti-RANKL antibody pharmaceutical composition and its therapeutic use. Background Technology

[0003] Denosumab is a RANK ligand (RANKL) inhibitor. Osteoporosis progenitor (OPG), acting as a decoy receptor, binds to RANKL on osteoblasts, blocking the RANKL-RANK interaction between osteoblasts and osteoclast precursors, thereby inhibiting the differentiation of osteoclast precursors into mature osteoclasts. In June 2010, the FDA approved denosumab for the treatment of osteoporosis in postmenopausal women, and subsequently approved it for other indications such as osteoporosis in men, bone loss due to androgen deprivation therapy for prostate cancer, and bone loss due to aromatase inhibitor therapy for breast cancer.

[0004] Current technologies still have many shortcomings and areas for improvement. For example, the concentration of therapeutic antibodies in drug formulations varies widely depending on the route of administration. When limited by injection volume, high-concentration antibody formulations are often required. However, increasing the concentration of protein formulations can cause stability problems, such as the formation of high molecular weight substances (HMWS) aggregates, increased viscosity, and a series of other issues related to drug stability. Therefore, it is necessary to find a stable protein drug that is resistant to degradation, aggregation, and misfolding caused by protein hydrolysis and denaturation. Improving the stability of drug compositions is challenging. Considering the properties of the excipients themselves, it is essential to continue optimizing and screening suitable formulation excipients to develop a stable, high-concentration liquid formulation of denosumab suitable for all patients.

[0005] Invention Overview

[0006] In pursuit of a superior formulation composition suitable for subcutaneous injection, this disclosure discloses a denosumab formulation containing an amino acid and sugar / alcohol system through formulation screening. By using a combination of amino acids and sugar / alcohol as a stabilizer, the amino acid-containing formulation of this disclosure exhibits superior SEC and IEC purity compared to conventional formulations. This improves product stability while effectively reducing formulation viscosity, facilitating drug delivery.

[0007] This disclosure provides an aqueous pharmaceutical composition for an anti-RANKL antibody comprising about 80-200 mg / ml of an anti-RANKL antibody and a stabilizer, said stabilizer comprising an amino acid and a combination of sorbitol, sucrose or trehalose;

[0008] The amino acid is selected from one or more of glycine, proline, lysine, histidine, glutamic acid, aspartic acid, and arginine, and the content of the amino acid is about 5mM-100mM.

[0009] In some embodiments, the anti-RANKL antibody in the pharmaceutical composition of this disclosure is in a high concentration, approximately 80-200 mg / ml. For example, the pharmaceutical composition contains anti-RANKL antibodies at concentrations of approximately 80 mg / ml, 85 mg / ml, 90 mg / ml, 95 mg / ml, 100 mg / ml, 105 mg / ml, 110 mg / ml, 115 mg / ml, 120 mg / ml, 125 mg / ml, 130 mg / ml, 135 mg / ml, 140 mg / ml, 145 mg / ml, 150 mg / ml, 155 mg / ml, 160 mg / ml, 165 mg / ml, 170 mg / ml, 175 mg / ml, 180 mg / ml, 185 mg / ml, 190 mg / ml, 195 mg / ml, and 200 mg / ml.

[0010] In some embodiments, the amino acids in the pharmaceutical compositions of this disclosure are selected from one or more of glycine, proline, lysine, histidine, glutamic acid, aspartic acid, and arginine, preferably histidine and glutamic acid, histidine and aspartic acid, or arginine and glutamic acid; the content of the amino acids is about 5 mM-100 mM, for example, about 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, etc. M, 20mM, 21mM, 22mM, 23mM, 24mM, 25mM, 26mM, 27mM, 28mM, 29mM, 30mM, 35mM, 40mM, 45mM, 50mM, 55mM, 60mM, 65mM, 70mM, 75mM, 80mM, 85mM, 90mM, 95mM, 100mM, preferably about 10mM, 15mM, 20mM, 25mM, 28mM, 30mM, 37mM, 38mM, 46mM, 55mM.

[0011] In some embodiments, the stabilizer in the pharmaceutical composition of this disclosure contains sorbitol, sucrose, or trehalose, wherein the sorbitol content is about 3%-5%, for example, about 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, or 5%, preferably, the sorbitol content is about 3.5%, 3.7%, or 4%. 0%; or the sucrose content is about 5%-10%, for example, about 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, preferably, the sucrose content is about 7%, 7.6%, 8%; or the trehalose content is 5%-10%, for example, about 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, preferably, the trehalose content is about 7%, 7.6%, 8%.

[0012] In some embodiments, the pharmaceutical composition of this disclosure further includes a buffer selected from acetate, succinate, and citrate; the content of the buffer is about 20-50 mM, for example, about 20 mM, 22 mM, 25 mM, 27 mM, 30 mM, 32 mM, 35 mM, 37 mM, 40 mM, 42 mM, 45 mM, 47 mM, or 50 mM, preferably about 25 mM, 30 mM, 32 mM, or 35 mM.

[0013] In some embodiments, the pharmaceutical compositions of this disclosure further include surfactants, such as polysorbate 80, polysorbate 20, and poloxamer 188; the surfactant is present in an amount of about 0.005% to 0.03%, for example, about 0.005%, about 0.01%, about 0.015%, about 0.02%, about 0.025%, or about 0.03%, preferably about 0.01% or 0.02%.

[0014] In some embodiments, the anti-RANKL antibody disclosed herein is denosumab, the heavy chain sequence of which is shown in SEQ ID NO:1 and the light chain sequence of which is shown in SEQ ID NO:2.

[0015] In some embodiments, the pH of the pharmaceutical composition disclosed herein is about 4.5-5.5, for example, pH values ​​of about 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, and 5.5, preferably, pH values ​​of about 5.1, 5.2, and 5.3.

[0016] In some preferred embodiments, this disclosure provides an aqueous pharmaceutical composition for an anti-RANKL antibody comprising about 80-200 mg / ml of denosumab, about 20-50 mM of acetate, about 5-100 mM of a combination of histidine and glutamic acid, or a combination of histidine and aspartic acid, or a combination of arginine and glutamic acid, about 0.005%-0.03% of polysorbate 20, about 3%-5% of sorbitol, or about 5%-10% of sucrose, or about 5%-10% of trehalose, with a pH of about 4.5-5.5.

[0017] Further preferably, this disclosure provides an aqueous pharmaceutical composition for an anti-RANKL antibody comprising about 100-150 mg / ml of denosumab, about 30-40 mM of acetate, about 10-50 mM of a combination of histidine and glutamic acid, or a combination of histidine and aspartic acid, or a combination of arginine and glutamic acid, about 0.005%-0.02% of polysorbate 20, about 3.5%-4% of sorbitol, or about 5%-8% of sucrose, or about 5%-8% of trehalose, with a pH of about 5.0-5.5.

[0018] In addition, this disclosure also provides a lyophilized formulation, which is obtained by lyophilizing the pharmaceutical composition of this disclosure, or by reconstituted the lyophilized formulation to obtain the pharmaceutical composition of this disclosure.

[0019] Furthermore, this disclosure also provides a method for treating bone diseases, the method comprising administering a subject the pharmaceutical composition or lyophilized formulation of this disclosure. The bone disease is selected from osteoporosis, rheumatoid arthritis, giant cell tumor of bone, multiple myeloma, and bone metastases of solid tumors, etc.

[0020] This disclosure also provides the use of the pharmaceutical composition or lyophilized formulation in the preparation of a medicament for treating bone diseases. The bone diseases are selected from osteoporosis, rheumatoid arthritis, giant cell tumor of bone, multiple myeloma, and bone metastases of solid tumors, etc. Detailed Implementation

[0021] All publications, patents and patent applications mentioned in this specification are incorporated herein by reference as if specifically and individually indicated to be incorporated by reference.

[0022] Before this disclosure is described in detail below, it should be understood that this disclosure is not limited to the specific methodologies, procedures, and reagents described herein, as these can vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of this disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0023] Some embodiments disclosed herein include numerical ranges, and certain aspects of this disclosure may be described using ranges. Unless otherwise stated, it should be understood that numerical ranges or descriptions using ranges are for purposes of brevity and convenience only and should not be considered as a strict limitation of the scope of this disclosure. Therefore, descriptions using ranges should be considered as specifically disclosing all possible subranges and all possible specific numerical points within those ranges, as these subranges and numerical points have been explicitly stated herein. The above principles apply equally regardless of the breadth of the numerical values ​​described. When a range description is used, the range includes the endpoints of the range.

[0024] The term “about” includes and describes the value or parameter itself. For example, “about x” includes and describes “x” itself. As used herein, when used in conjunction with a measured value or to modify a value, unit, constant, or series of values, the term “about” refers to a variation of ±1% to 10% in addition to including the value or parameter itself. In some embodiments, when used in conjunction with a measured value or to modify a value, unit, constant, or series of values, the term “about” refers to a variation of ±1%, ±2%, ±3%, ±4%, ±5%, ±6%, ±7%, ±8%, ±9%, or ±10%.

[0025] The term "Denosumab" is a RANK ligand (RANKL) inhibitor. "Denosumab" is the generic name for the antibody, but can also refer solely to the active ingredient itself, which comprises two homologous heavy chains and two homologous light chains, wherein the sequence of the heavy chain is shown in SEQ ID NO:1 and the sequence of the light chain is shown in SEQ ID NO:2.

[0026] The terms “formulation” or “pharmaceutical formulation” or “prescription” or “drug prescription” are used interchangeably herein and refer to a form in which the active pharmaceutical ingredient is present in a valid manner and which does not contain any other components that would be toxic to the subject to whom the formulation is administered.

[0027] The term "pharmaceutical composition" refers to a formulation or combination of formulations containing one, two, or more active ingredients, wherein the active ingredients contained therein are present in a biologically effective form and do not contain any additional ingredients that would have unacceptable toxicity to a subject administering the formulation. In some narrower contexts, such as when containing only one active pharmaceutical ingredient, "pharmaceutical composition" may be used interchangeably with "pharmaceutical formulation" or "pharmaceutical prescription." When a "pharmaceutical composition" exists as a combination of individual formulations containing two or more different active ingredients, it can be administered simultaneously, sequentially, separately, or at intervals, with the aim of exerting the biological activity of multiple active ingredients together for the treatment of a disease.

[0028] The term "aqueous" pharmaceutical composition refers to a pharmaceutical composition that includes water.

[0029] The term "high concentration" refers to a formulation containing an active pharmaceutical ingredient (API) at a concentration of approximately 80-200 mg / ml.

[0030] The terms "pharmaceuticalally acceptable excipient" or "excipient" refer to substances / reagents that can be reasonably administered to a subject to provide an effective amount of a stable formulation of an active pharmaceutical ingredient. Suitable pharmaceutically acceptable excipients are well known in the art and include, but are not limited to, buffers, stabilizers, surfactants, etc.

[0031] The term "buffer" refers to a reagent that maintains the pH of a formulation solution within an acceptable range.

[0032] The term "stabilizer" is a pharmaceutically acceptable excipient used to protect an active pharmaceutical ingredient and / or formulation from chemical and / or physical degradation during production, storage, and application.

[0033] The term "surfactant" refers to a pharmaceutically acceptable excipient used to protect protein formulations from mechanical stresses such as agitation and shearing.

[0034] The term "lyophilized formulation" refers to a composition obtained or available through a freeze-drying process of a liquid formulation. Preferably, it is a solid composition with a water content of less than 5%, more preferably less than 3%.

[0035] The term "insoluble microparticles" includes, but is not limited to, microparticles introduced during product formulation, fatty acid particles generated from the degradation of polysorbate, silicone oil, large molecular particles caused by protein aggregation, and other unknown particulate matter, such as possible packaging material debris and excipient crystallization.

[0036] The term "treatment" refers to a clinical intervention in an attempt to alter an individual's or treat a disease caused by cells, which can be preventative or intervention in the clinicopathological process. Therapeutic effects include, but are not limited to, preventing the onset or recurrence of disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, improving or alleviating the condition, and mitigating or improving prognosis.

[0037] The terms “individual,” “subject,” or “patient” refer to any animal, such as a mammal or marsupial. Individuals in this disclosure include, but are not limited to, humans, non-human primates (such as cynomolgus monkeys or rhesus monkeys or other types of macaques), mice, pigs, horses, donkeys, cattle, sheep, rats, and any kind of poultry.

[0038] The terms “disease,” “symptom,” or “disorder” refer to any alteration or dysregulation that impairs or interferes with the normal function of cells, tissues, or organs. For example, “disease” includes, but is not limited to: tumors, pathogen infections, autoimmune diseases, T-cell dysfunction, or deficiencies in immune tolerance (such as transplant rejection).

[0039] The term "tumor" refers to a disease characterized by the pathological proliferation of cells or tissues, and their subsequent migration or invasion into other tissues or organs. Tumor growth is typically uncontrolled and progressive, neither inducing nor inhibiting the proliferation of normal cells. Tumors include "cancer," which broadly refers to all malignant tumors.

[0040] Protein content determination: Protein content was determined using a Lunatic micro-spectroscopy analyzer. A blank was calculated using purified water, and the 280nm ultraviolet absorption was corrected for 330nm light scattering. Protein concentration was calculated according to Formula 1, where the extinction coefficient ε was 1.5 (L / g·cm⁻¹). -1 ).

[0041] Size exclusion chromatography (SE-HPLC): This method uses size exclusion chromatography to quantify polymers, monomers, and fragments. This assay was performed using a TSK gel G3000 SWXL 7.8 mm × 30 cm 5 μm column on a Waters e2695-2489 HPLC system. The mobile phase was sodium phosphate buffer. The sample was diluted to 1 mg / mL with the mobile phase, and the injection volume was 25 μL. Proteins were eluted isocratically at a flow rate of 0.5 mL / min for 30 min, and the absorbance of the eluent was measured at 215 nm. Integration was performed using Empower3 software.

[0042] Non-reducing capillary electrophoresis (nrCE-SDS): The purity of the main peak was determined by non-reducing capillary electrophoresis (nrCE-SDS). This determination was performed on a SCIEX PA800 plus capillary electrophoresis system using a 50μm ID uncoated quartz capillary with an effective separation length of 20cm (total length 30.2cm); detection was performed using a PDA at 220nm and a bandwidth of 10nm.

[0043] Insoluble microparticles are studied using microfluidic imaging analysis (MFI). The MFI microfluidic imaging particle analysis system is a precision instrument that integrates digital microscopy, microfluidics, and image processing techniques into a fully automated analysis of particulate matter. As the sample flows through the flow cell (detection zone), the MFI captures the particulate matter in the sample and counts the particles using software.

[0044] The freezing point depression method for determining osmolality indirectly measures the osmolality of a solution by measuring its freezing point depression. In an ideal dilute solution, the freezing point depression follows the relationship ΔTf = Kf·m, where ΔTf is the freezing point depression, Kf is the freezing point depression constant (1.86 when water is the solvent), and m is the molality. Osmotic pressure follows the relationship Po = ko·m, where Po is the osmotic pressure, ko is the osmotic pressure constant, and m is the molality. Since the concentrations in both equations are equivalent, the freezing point depression method can be used to determine the osmolality of a solution.

[0045] Viscosity was measured using an m-VROC type viscometer: via Rheosense e-VROC built with technology TM It is an excellent tool for simultaneously measuring tensile and shear viscosity. The viscometer uses a MEMS pressure sensor to measure the pressure upstream and downstream of the contraction and records changes in flow rate. The fluid undergoes a nearly constant stretching through contraction / expansion, allowing the apparent tensile viscosity to be calculated.

[0046] Example 1: Effect of different amino acids on formulation stability

[0047] The change in the percentage of high molecular weight surfactants (HMWS) in six formulations containing high concentrations (approximately 120 mg / ml) of denosumab with different amino acid groups over time was tested. The component contents of the six formulations are shown in Table 1. The preparation method is as follows: buffer exchange was performed using the expected excipient solution, and the antibody concentration was concentrated to the required level by ultrafiltration. The corresponding polysorbate 20 stock solution was added, and the prepared formulation was aseptically filtered through a 0.22 μm low protein binding filter and dispensed. The presence (%) of high molecular weight surfactants (HMWS, LMWS) and the main peak (Monomer / MAIN) was determined by SE-HPLC. The change in the percentage of HMWS in the formulations over time was detected by SE-HPLC at 40℃±2℃ to assess the stability of the antibody during storage at high temperature (40℃). The test results are shown in Table 2. At the same time, the viscosity, osmotic pressure, and insoluble particles of the formulations were measured. The test results are shown in Tables 3 and 4.

[0048] Table 1. Composition of formulations containing denosumab

[0049] Table 2. Stability results of the formulation under high temperature conditions

[0050] Table 3. Viscosity and osmotic pressure of the formulation under high temperature conditions (40℃, 4W)

[0051] Table 4. Insoluble microparticles of the formulation under high temperature conditions (40℃, 4W)

[0052] The above experimental results show that the aggregates in all formulations did not increase significantly during storage. The increase was smallest in test case A5, with only a 0.7% increase after 4 weeks, indicating that histidine and glutamate components effectively inhibited aggregate formation and demonstrated excellent stabilizing effects on denosumab. All formulations had viscosities less than 5 mPa·s and osmotic pressures between 280-330 mOsmol / kg. Notably, formulations A4 and A5 had low viscosity and low levels of insoluble particles, meeting the requirements for subcutaneous injection. Furthermore, adding L-phenylalanine to the formulations is unsuitable for patients with phenylketonuria. Replacing L-phenylalanine with amino acids such as histidine and glutamate not only expands the target population but also reduces the risk of medication errors.

[0053] Example 2: Stability comparison of formulations containing different sugars / alcohols

[0054] To further investigate the effects of sorbitol, sucrose, and trehalose on the stability of the pharmaceutical composition, experimental formulations B1-B4 containing different sugar / alcohol ratios were prepared. The component contents of each formulation are shown in Table 5, where the concentration of denosumab in formulation B1-B4 is approximately 120 mg / mL. In this example, the percentages of Monomer / MAIN, HMWS (polymer), and LMWS (fragment) of formulations B1-B4 were measured by SE-HPLC and nr-SDS at 0 h and 40℃ ± 2℃. The results are shown in Table 6.

[0055] Table 5. Composition of formulations containing denosumab

[0056] Table 6 Stability data of the formulation

[0057] The results above show that high-purity, high-concentration formulations of sorbitol, sucrose, and trehalose components B1-B4 can be obtained. Furthermore, after 4 weeks of storage at 40℃±2℃, the SE-HPLC main peak purity of formulations B1-B4 was between 97.8% and 98.0%, and the nr-SDS main peak purity was between 95.8% and 96.3%, indicating that each formulation has excellent purity levels. Therefore, formulations containing different sugars and alcohols all exhibit excellent stability, meaning that sorbitol, sucrose, and trehalose have a significant protective effect against denosumab.

[0058] Example 3: Stability comparison of formulations containing different amino acid combinations

[0059] To further investigate the effect of different amino acid combinations on the stability of the drug composition, formulations C1-C6 containing different amino acid combinations were prepared. The contents of each component are shown in Table 7, where the concentration of denosumab in C1-C6 is approximately 120 mg / mL. In this example, the percentages of Monomer / MAIN, HMWS (polymer), and LMWS (fragment) of formulations C1-C6 were measured by SE-HPLC and nrCE-SDS at 0 h and 40℃±2℃. The results are shown in Table 8.

[0060] Table 7. Composition of formulations containing denosumab

[0061] Table 8 Stability data of the formulation

[0062] The results above show that combinations of sorbitol with histidine and glutamic acid, sorbitol with histidine and aspartic acid, and sorbitol with arginine and glutamic acid can all prepare high-concentration formulations with high purity. Furthermore, after incubation at 40℃±2℃ for 4 weeks, the SE-HPLC peak purity trends of formulations C1-C6 were generally consistent, ranging from 97.6% to 97.9%. The nr-SDS peak purity ranged from 95.2% to 96.4%, indicating excellent purity levels for all formulations. Therefore, combinations of sorbitol with histidine and glutamic acid, sorbitol with histidine and aspartic acid, and sorbitol with arginine and glutamic acid can all provide significant protection against denosumab.

[0063] The embodiments described above are merely exemplary, and any person skilled in the art will recognize or be able to identify numerous equivalents of specific compounds, materials, and operations without the need for extraordinary experimentation. All such equivalents are within the scope of this disclosure and are encompassed by the claims.

Claims

1. An aqueous pharmaceutical composition of an anti-RANKL antibody, comprising about 80-200 mg / ml of an anti-RANKL antibody and a stabilizer comprising an amino acid in combination with sorbitol, sucrose or trehalose, wherein the amino acid is selected from one or more of glycine, proline, lysine, histidine, glutamic acid, aspartic acid, arginine, and the content of the amino acid is about 5 mM-100 mM.

2. The aqueous pharmaceutical composition of claim 1, wherein the content of the amino acid is about 10 mM, 15 mM, 20 mM, 25 mM, 28 mM, 30 mM, 37 mM, 38 mM, 46 mM, 55 mM.

3. The aqueous pharmaceutical composition of claim 1 or 2, wherein the aqueous pharmaceutical composition comprises a buffer.

4. The aqueous pharmaceutical composition of claim 3, wherein the buffer is selected from acetate, succinate, citrate, and the content of the buffer is about 20-50 mM, preferably, the content is about 25 mM, 30 mM, 32 mM, 35 mM.

5. The aqueous pharmaceutical composition of any one of claims 1-4, wherein the aqueous pharmaceutical composition comprises a surfactant.

6. The aqueous pharmaceutical composition of claim 5, wherein the surfactant is preferably polysorbate 80, polysorbate 20, poloxamer 188; and the content of the surfactant is about 0.005%-0.03%, preferably, the content is about 0.01%, 0.02%.

7. The aqueous pharmaceutical composition of any one of claims 1-6, wherein the content of the sorbitol is about 3%-5%, the content of the sucrose is about 5%-10%, or the content of the trehalose is about 5%-10%; preferably, the content of the sorbitol is about 3.7%, the content of the sucrose is about 7%, or the content of the trehalose is about 7.6%.

8. The aqueous pharmaceutical composition of any one of claims 1-7, wherein the anti-RANKL antibody is denosumab, and the heavy chain sequence of the denosumab is set forth in SEQ ID NO: 1, and the light chain sequence of the denosumab is set forth in SEQ ID NO:

2.

9. The aqueous pharmaceutical composition of any one of claims 1-8, wherein the pH of the aqueous pharmaceutical composition is about 4.5-5.

5.

10. An aqueous pharmaceutical composition of an anti-RANKL antibody, comprising about 80-200 mg / ml of denosumab, about 20-50 mM of acetate, about 5-100 mM of a combination of histidine and glutamic acid, or a combination of histidine and aspartic acid, or a combination of arginine and glutamic acid, about 0.005%-0.03% of polysorbate 20, about 3%-5% of sorbitol, or about 5%-10% of sucrose, or about 5%-10% of trehalose, and the pH is about 4.5-5.

5.

11. A lyophilized formulation obtained by lyophilizing an aqueous pharmaceutical composition according to any one of claims 1-10, or by reconstituted the lyophilized formulation to obtain an aqueous pharmaceutical composition according to any one of claims 1-10.

12. A method for treating a bone disease, the method comprising administering to a subject the aqueous pharmaceutical composition of any one of claims 1-10 or the lyophilized formulation of claim 11.

13. The method of claim 12, wherein the bone disease is osteoporosis, rheumatoid arthritis, giant cell tumor of bone, multiple myeloma, or bone metastasis of solid tumors.

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