Injectable preparation containing isoxazoline derivative and its manufacturing method

Abstract

The present invention relates to an injectable formulation comprising, as an active pharmacological ingredient (API), an isoxazoline derivative useful as a caspase inhibitor, or a pharma- ceutically acceptable salt thereof. The injectable formulation according to the present invention comprises a first agent comprising a high-dose API and a second agent comprising a reconstitution solution, and can be prepared by mixing the first agent and the second agent just before administration to a patient. Since the injectable formulation stably contains the active API, it has the advantage that it can be expected to have an effective medicinal effect when administered to a patient.

Description

[Technical field]

[0001] This application claims the benefit of priority based on Korean Patent Application No. 10-2022-0023373 filed on February 23, 2022, and all contents disclosed in the documents of that Korean patent application are incorporated as part of this specification.

[0002] The present invention relates to an injectable formulation containing an isoxazoline derivative or a pharma- ceutically acceptable salt thereof, which is useful as a caspase inhibitor, as an active pharmacological ingredient (API), and a method for producing the same. [Background technology]

[0003] Caspase is a cysteine ​​protease, and caspase inhibitors are compounds that can regulate inflammation and apoptosis caused by the action of caspases by inhibiting the action of caspases. Diseases that can be treated with these compounds to eliminate or alleviate symptoms include osteoarthritis, rheumatoid arthritis, degenerative arthritis, destructive bone disease, liver disease caused by viral hepatitis, acute hepatitis, liver sclerosis, brain damage caused by viral hepatitis, human sudden liver failure, sepsis, organ transplant rejection, ischemic heart disease, dementia, stroke, brain damage caused by AIDS, diabetes, and gastric ulcers.

[0004] As caspase inhibitors, various kinds of isoxazoline derivatives and prodrugs of isoxazoline derivatives are known. Among the isoxazoline derivatives, (R)-N-((2S,3S)-2-(fluoromethyl)-2-hydroxy-5-oxotetrahydrofuran-3-yl)-5-isopropyl-3-(isoquinolin-1-yl)-4,5-dihydroisoxazole-5-carboxamide (hereinafter referred to as "compound of formula 1") has already been proven to be effective in liver diseases caused by viral hepatitis, liver fibrosis, and nonalcoholic steatohepatitis (NASH). [ka]

[0005] Recently, there have been attempts to apply the compound of Formula 1 to diseases related to bones and joints, such as osteoarthritis (OA), rheumatoid arthritis, degenerative arthritis, and destructive bone disorders.

[0006] In the treatment of diseases related to bones and joints, the general method of drug administration is to administer the drug directly into the joint cavity, which is filled with synovial fluid, using a syringe. When administered orally, the drug has a systemic effect, but systemic side effects cannot be avoided. On the other hand, when administered directly into the joint cavity, local administration of the drug has the advantage of not only achieving the effect of local treatment where the drug is directly delivered, but also of being able to administer a high concentration of the drug and minimizing systemic side effects.

[0007] The compound of Chemical Formula 1 has been confirmed to have therapeutic effects for existing liver diseases as an oral preparation, but since the compound of Chemical Formula 1 is poorly soluble in water, it is difficult to formulate it into an injectable preparation.

[0008] Therefore, there is a need to develop a high-concentration injectable formulation that can use the compound of Chemical Formula 1 as an injection. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] Korean Patent Publication No. 0774999 (November 2, 2007) Isoxazoline derivatives and their manufacturing method [Patent Document 2] Korean Patent Application No. 2021-0102476 (August 4, 2021) Use of caspase inhibitors in alleviating or treating osteoarthritis [Non-patent literature]

[0010] [Non-Patent Document 1] Ratziu et al.,'A Phase 2, Randomized, Double‐Blind, Placebo‐Controlled Study of GS‐9450 in Subjects with Nonalcoholic Steatohepatitis' HEPATOLOGY, Vol.55,No.2,2012 Summary of the Invention [Problem to be solved by the invention]

[0011] As a result of extensive research conducted by the present inventors to solve the above problems, the present invention aims to provide an injectable formulation containing a high content of the compound of Chemical Formula 1 as an active pharmacological ingredient (API).

[0012] In addition, the present invention aims to provide a freeze-dried injectable formulation that can be stored for a long period of time by increasing the stability of the active pharmacological ingredient (API) in an injectable formulation containing a high content of the compound of Chemical Formula 1. [Means for solving the problem]

[0013] The present invention provides an injectable formulation comprising a first agent in powder form comprising a compound of Formula 1 below, a phosphate buffer, a solubilizing agent, and a pH adjusting agent; and a second agent comprising a reconstitution solution having a pH of 5 to 12: [ka]

[0014] The present invention also provides an injectable formulation wherein the phosphate buffer is a mixture of sodium phosphate dibasic heptahydrate and sodium phosphate monobasic monohydrate.

[0015] The present invention also provides an injectable preparation, wherein the content of the phosphate buffer is 1 to 15 mM based on the first agent.

[0016] The present invention also provides an injectable formulation, wherein the solubilizing agent is beta-cyclodextrin.

[0017] The present invention also provides an injectable preparation, wherein the beta-cyclodextrin is any one selected from the group consisting of hydroxymethyl-beta-cyclodextrin, hydroxyethyl-beta-cyclodextrin, and hydroxypropyl-beta-cyclodextrin.

[0018] The present invention also provides an injectable preparation, wherein the content of the beta-cyclodextrin is 10 to 30% by weight based on the first agent.

[0019] The present invention also provides an injectable preparation, wherein the pH of a reconstituted solution obtained by mixing the first agent and the second agent is 5.0 to 7.0.

[0020] The present invention also provides an injectable preparation, wherein the osmotic pressure of a reconstituted solution obtained by mixing the first agent and the second agent is 300 mOsm / kg to 400 mOsm / kg.

[0021] The present invention also provides an injectable formulation having a content of the compound of Formula 1 of 1 to 20 mg / ml based on a reconstituted solution obtained by mixing the first agent and the second agent.

[0022] The present invention also provides a method for preparing a first solution having a pH of 6 to 8 by dissolving a compound represented by the following Formula 1 in the prepared solvent: (S1) preparing a solvent having a pH of 10 to 12, the solvent comprising a phosphate buffer, a solubilizing agent, and a basic pH adjusting agent; (S2) preparing a first solution having a pH of 6 to 8 by dissolving a compound represented by the following Formula 1 in the prepared solvent; [ka]

[0023] The present invention provides a method for producing an injectable formulation, comprising the steps of: (S3) adding an acidic pH adjuster to the first solution to produce a second solution having a pH of 1.0 to 3.0; (S4) lyophilizing the second solution to produce a first agent in powder form; and (S5) producing a second agent comprising a reconstituted solution having a pH of 5 to 12.

[0024] The present invention also provides a method for preparing an injectable formulation, the content of the compound of Formula 1 being 1 to 20 mg / ml based on the reconstituted solution.

[0025] The present invention also provides a method for preparing an injectable formulation, wherein the phosphate buffer is a mixture of sodium phosphate dibasic heptahydrate and sodium phosphate monobasic monohydrate.

[0026] The present invention also provides a method for preparing an injectable formulation, wherein the content of the phosphate buffer is 1 to 15 mM based on the solvent.

[0027] The present invention also provides a method for preparing an injectable formulation, wherein the solubilizing agent is beta-cyclodextrin.

[0028] The present invention also provides a method for preparing an injectable formulation, wherein the beta-cyclodextrin is any one selected from the group consisting of hydroxymethyl-beta-cyclodextrin, hydroxyethyl-beta-cyclodextrin, and hydroxypropyl-beta-cyclodextrin.

[0029] The present invention also provides a method for preparing an injectable formulation, wherein the beta-cyclodextrin content is 10 to 30% by weight based on the solvent.

[0030] The present invention also provides an injectable preparation for treating or preventing a disease selected from the group consisting of osteoarthritis, rheumatoid arthritis, degenerative arthritis, destructive bone disorders, liver disease due to viral hepatitis, acute hepatitis, liver sclerosis, brain damage due to viral hepatitis, sudden human liver failure, sepsis, organ transplant rejection, ischemic heart disease, dementia, stroke, brain damage due to AIDS, diabetes, and gastric ulcer.

[0031] The present invention also provides an injectable preparation, wherein the disease is selected from the group consisting of osteoarthritis, rheumatoid arthritis, degenerative arthritis and destructive bone disorders.

[0032] The present invention also provides an injectable preparation in which the first and second agents are mixed in a single dose and used in a solution state.

[0033] The present invention also provides an injectable preparation which is prepared by mixing the first and second agents and then administering the mixture into the articular cavity.

[0034] The present invention also provides an injectable preparation which is administered into the articular cavity within 30 minutes after mixing the first and second agents.

[0035] The present invention also provides an injectable formulation which is administered once after the first and second agents are mixed.

[0036] The present invention also provides an injectable formulation comprising: a first agent in powder form, the first agent comprising a compound of the following formula 1, a phosphate buffer, a solubilizing agent, and a pH adjusting agent; and a second agent comprising a reconstitution solution having a pH of 5 to 12; [ka] The injectable formulation provides an injectable formulation in which, when the contents of the compound of Chemical Formula 2 and the compound of Chemical Formula 3 are measured within 30 minutes after mixing the first and second agents, the content of the compound of Chemical Formula 2 is 94% or more of the total content of the compound of Chemical Formula 2 and the compound of Chemical Formula 3: [ka] [ka] Effect of the Invention

[0037] The injectable formulation according to the present invention comprises a first agent in a lyophilized dosage form and a second agent containing a reconstitution solution, and the first and second agents are mixed together just before administration to a patient to prepare a drug, which is then administered to the patient within 30 minutes, thereby providing an injectable formulation with a high content of active API.

[0038] It is also possible to provide a freeze-dried injectable preparation that enhances the stability of the active pharmacological ingredient (API) and can be stored for a long period of time. [Brief description of the drawings]

[0039] [Figure 1] 1 is a graph showing the results of measuring API purity in an API mixed solution versus stirring time. [Diagram 2] FIG. 5(a) shows the TGA measurement result of the lyophilized final dosage form of API prepared in Example 52, and FIG. 5(b) shows the DSC measurement result of the lyophilized final dosage form of API prepared in Example 52. [Diagram 3] 1 shows the XRD pattern measurement results of the API lyophilized final dosage form prepared in Example 52. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0040] The present invention will now be described in more detail.

[0041] All technical terms used in the present invention are used in the same sense as commonly understood by those having ordinary skill in the art unless otherwise defined. In addition, although preferred methods and samples are described in this specification, similar or equivalent methods and samples are also included in the scope of the present invention. The contents of all publications cited as references in this specification are incorporated herein by reference in their entirety.

[0042] Since the compound of the present invention represented by Chemical Formula 1 is almost insoluble in water, the solubility in solvents of various compositions was analyzed in order to find a solvent capable of dissolving the compound of Chemical Formula 1 at a high content.

[0043] As a result, it was confirmed that the compound of Chemical Formula 1 is hardly soluble in an acidic solvent of pH 4 or less, among solvents containing a phosphate buffer, a solubilizing agent, and a pH adjuster, but is highly soluble in a weakly acidic solvent of pH 6 or more and a basic solvent of pH 7.5 or more. It was also confirmed that the compound of Chemical Formula 1 has the property of decreasing the pH of the solution while dissolving in the solvent.

[0044] In addition, it was found that when the compound of Chemical Formula 1 is dissolved in an aqueous solvent, the lactone ring is cleaved and the compound is converted to the compound of Chemical Formula 2 below. At this time, the compound of Chemical Formula 2 is in an active form, but the compound of Chemical Formula 2 is quickly and irreversibly converted to the structural isomer, the compound of Chemical Formula 3, in an aqueous solution, and the compound of Chemical Formula 3 is in an inactive form that does not exhibit caspase inhibitory activity, unlike the compound of Chemical Formula 2.

[0045] [ka]

[0046] [ka]

[0047] [ka]

[0048] In addition, the conversion rate of the compound of Chemical Formula 2 to the compound of Chemical Formula 3 was examined depending on the pH of the solvent. As a result, it was confirmed that the compound of Chemical Formula 3 was hardly produced in an acidic solvent, but was rapidly produced in a neutral or more basic solvent.

[0049] Therefore, in order to dissolve the compound of Chemical Formula 1, a basic solvent with a high pH must be used, but it was found that not only does the compound of Chemical Formula 1 dissolve, lowering the pH of the solution to reduce the solubility of undissolved compounds, but also the active form of the dissolved compound is rapidly converted to an inactive form in the basic solvent. This mechanism not only reduces the purity of the active ingredient in an injectable dosage form in which the compound of Chemical Formula 1 is the API, thereby affecting the efficacy, but also causes a problem in that it is difficult to adjust the pH when preparing a final drug for administration to an individual, since the compound of Chemical Formula 1 dissolves and lowers the pH of the solution.

[0050] Therefore, the present inventors have studied various injectable formulations and have completed a freeze-dried injectable formulation having improved stability during long-term storage by preparing an acidic solution containing the compound of Chemical Formula 1 and then freeze-drying it.

[0051] Manufacturing method of the injectable preparation According to one embodiment of the present invention, a method for preparing an injectable formulation comprises the following steps: (S1) preparing a solvent having a pH of 10 to 12, comprising a phosphate buffer, a solubilizing agent, and a basic pH adjusting agent; (S2) preparing a first solution having a pH of 6 to 8 by dissolving a compound of Formula 1 in the prepared solvent: [ka] (S3) adding an acidic pH regulator to the first solution to prepare a second solution having a pH of 1.0 to 3.0; (S4) freeze-drying the second solution to prepare a first agent in powder form; and (S5) preparing a second agent comprising a reconstituted solution having a pH of 5 to 12.

[0052] In one embodiment of the present invention, a method for preparing an injectable formulation may include preparing (S1) a solvent having a pH of 10 to 12, comprising a phosphate buffer, a solubilizing agent, and a basic pH adjusting agent.

[0053] The solvent containing the buffer, the solubilizing agent, and the basic pH adjusting agent may be a solvent composition capable of dissolving the compound of Chemical Formula 1.

[0054] The solvent may contain a buffer. The buffer may be a common buffer that can be used in injection preparations, and specific examples include, but are not limited to, phosphate buffer, Tris buffer, MES buffer, etc. When a phosphate buffer is used, it may be selected from the group consisting of sodium phosphate dibasic, sodium phosphate monobasic, potassium phosphate dibasic, potassium phosphate monobasic, and mixtures thereof. When a Tris buffer is used, TRIZMA® buffer may be used.

[0055] The buffer may have a concentration of 1 to 15 mM based on the solvent, specifically, 1 to 15 mM, 2 to 15 mM, 3 to 15 mM, 4 to 15 mM, 5 to 15 mM, 1 to 14 mM, 2 to 14 mM, 3 to 14 mM, 4 to 14 mM, 5 to 14 mM, 1 to 13 mM, 2 to 13 mM, 3 to 13 mM, 4 to 13 mM, 5 to 13 mM, 1 to 12 mM, 2 to 12 mM, 3 to 12 mM, 4 to 12 mM, 5 to 12 mM, 1 to 11 mM, 2 to 11 mM, 3 to 11 mM, 4 to 11 mM, 5 to 11 mM, 1 to 10 mM, 2 to 10 mM, 3 to 10 mM, 4 to 10 mM, or 5 to 10 mM.

[0056] The solvent may include a solubilizing agent. The solubilizing agent may include cyclodextrin or a derivative thereof. The cyclodextrin may improve the dissolution stability of the compound of Formula 1 and may affect the formation of a stable and uniform cake during lyophilization.

[0057] The cyclodextrin is a cyclic oligosaccharide in which 6 to 12 glucose molecules are linked by alpha-1,4-glycosidic bonds, and includes alpha-cyclodextrin with 6 glucose molecules linked, beta-cyclodextrin with 7 glucose molecules linked, and gamma-cyclodextrin with 8 glucose molecules linked, etc. Cyclodextrin that can be used in the present invention can include all cyclodextrin derivatives in addition to the above three types.

[0058] The cyclodextrin derivatives include alpha-cyclodextrin, 2-hydroxypropyl-beta-cyclodextrin, sulfobutylether-beta-cyclodextrin, etc., and may be specifically 2-hydroxypropyl-beta-cyclodextrin.

[0059] The solubilizer may be included in an amount of 10 to 30% by weight based on the solvent. Specifically, it may contain 10 to 30% by weight, 10.5 to 30% by weight, 11 to 30% by weight, 11.5 to 30% by weight, 12 to 30% by weight, 10 to 29.5% by weight, 10.5 to 29.5% by weight, 11 to 29.5% by weight, 11.5 to 29.5% by weight, 12 to 29.5% by weight, 10 to 29% by weight, 10.5 to 29% by weight, 11 to 29% by weight, 11.5 to 29% by weight, 12 to 29% by weight, 10 to 28.5% by weight, 10.5 to 28.5% by weight, 11 to 28.5% by weight, 11.5 to 28.5% by weight, 12 to 28.5% by weight, 10 to 28% by weight, 10.5 to 28% by weight, 11 to 28% by weight, 11.5 to 28% by weight, or 12 to 28% by weight.

[0060] The solvent may include a basic pH adjuster, which may be used to prepare a solvent having a pH of 10 to 12, such as, but not limited to, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, ammonia, sodium bicarbonate, etc.

[0061] The basic pH adjusting agent has a concentration of 20 to 40 mM based on the solvent.

[0062] The solvent can be adjusted to a pH of 10 to 12 using the buffer and basic pH adjusting agent.

[0063] In one embodiment of the present invention, the method for preparing an injectable formulation may include the step of (S2) dissolving a compound of Formula 1 in the prepared solvent to prepare a first solution having a pH of 6 to 8: [ka]

[0064] The compound of Formula 1 may be dissolved in a solvent having a pH of 10 to 12. As the compound of Formula 1 is dissolved in the solvent, the pH of the first solution is lowered, so the pH of the first solution obtained by mixing the compound of Formula 1 and the solvent may be 6 to 8. If the pH of the mixed solution of the compound of Formula 1 is outside the range of 6 to 8, it may not be suitable for dissolving API.

[0065] In one embodiment of the present invention, the method for preparing an injectable formulation may include the step of (S3) adding an acidic pH regulator to the first solution to prepare a second solution having a pH of 1.0 to 3.0.

[0066] The compound of Formula 1, which is an API, is dissolved in a solvent with a pH of 10 to 12, and the active form of the dissolved compound of Formula 1 is rapidly converted to an inactive form in a basic solution. Therefore, the second solution is prepared to prevent the active form of the API from being rapidly hydrolyzed in an aqueous solution and converted to an inactive form as the pH of the first solution containing the API increases.

[0067] The preparation of the second solution should not cause precipitation of the API, and the stability (purity) of the API should be maintained after preparation of the second solution. To meet these conditions, the pH of the second solution may be in the range of 1.0 to 3.0. Specifically, the pH of the second solution may be 1.0 to 3.0, 1.0 to 2.9, 1.0 to 2.8, 1.0 to 2.7, 1.0 to 2.6, or 1.0 to 2.5.

[0068] The acidic pH adjuster added for preparing the second solution may be hydrochloric acid, but is not limited thereto. Perchloric acid, nitric acid, sulfuric acid, etc., which can satisfy the pH of the second solution between 1.0 and 3.0, may be used.

[0069] In one embodiment of the present invention, the method for preparing an injectable formulation may include (S4) freeze-drying the second solution to prepare a first agent in powder form.

[0070] As used herein, the term "lyophilization" refers to a process in which the material to be dried is first frozen and then the ice or frozen solvent is removed by sublimation in a vacuum environment.

[0071] The freeze-drying can be carried out by freezing at -80°C for 3 to 5 hours and then drying at 10 to 20 Pa for 10 to 50 hours.

[0072] The first agent may be in the form of a powder, a dry powder, or a lyophile. The first agent can be stored for a long period of time and is easily formulated as an injection. In addition, since there are no additives that may cause side effects, it is completely harmless to the human body.

[0073] In one embodiment of the present invention, the method for preparing an injectable formulation may include preparing a second agent comprising a reconstitution solution having a pH of 5 to 12 (S5).

[0074] The term "reconstitution" as used herein means returning a solid state to a liquid state by adding water or a solution to a dry powder such as a lyophilized powder, and a reconstituted solution according to the present invention means a solution to be added to the first agent to produce a liquid state.

[0075] The second agent may include a reconstitution solution that includes a reconstitution medium such as sterile purified water, water for injection, saline, or a combination thereof, and further includes sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, or a combination thereof.

[0076] The reconstituted solution may have a pH of 5 to 12. The second agent containing the reconstituted solution having a pH of 5 to 12 is prepared to reconstitute the first agent prepared in a powder form by freeze-drying the second solution having a pH of 1.0 to 3.0, and is for adjusting the pH of the injectable formulation to a suitable value in the body to prevent the patient from inducing pain before the injectable formulation is administered to the body.

[0077] The second agent may further include other additives that do not affect pH in addition to the reconstituted solution. The additives may further include preservatives, surfactants, etc., but are not limited to these examples, and may selectively include additives commonly used in the art. The preservative is for preventing secondary contamination, and a pharma- ceutically acceptable preservative may be used. The surfactant may be, but is not limited to, a non-ionic surfactant containing a polyoxyethylene group, and a pharma-ceutically acceptable surfactant may be used.

[0078] Injectable preparations According to one embodiment of the present invention, there may be provided an injectable formulation comprising: a first agent in powder form, comprising a compound of Formula 1 below, a phosphate buffer, a solubilizing agent, and a pH adjusting agent; and a second agent comprising a reconstitution solution having a pH of 5 to 12: [ka]

[0079] The first agent may include a buffer, a solubilizing agent, and a pH adjusting agent.

[0080] The buffer may be a common buffer that can be used in injection preparations, and specific examples include, but are not limited to, phosphate buffer, Tris buffer, MES buffer, etc. When a phosphate buffer is used, it may be selected from the group consisting of sodium phosphate dibasic, sodium phosphate monobasic, potassium phosphate dibasic, potassium phosphate monobasic, and mixtures thereof. When a Tris buffer is used, TRIZMA® buffer may be used.

[0081] The buffer has a concentration of 1 to 15 mM based on the first agent, specifically, 1 to 15 mM, 2 to 15 mM, 3 to 15 mM, 4 to 15 mM, 5 to 15 mM, 1 to 14 mM, 2 to 14 mM, 3 to 14 mM, 4 to 14 mM, 5 to 14 mM, 1 to 13 mM, 2 to 13 mM, 3 to 13 mM, 4 to 13 mM, 5 to 13 mM, 1 to 12 mM, 2 to 12 mM, 3 to 12 mM, 4 to 12 mM, 5 to 12 mM, 1 to 11 mM, 2 to 11 mM, 3 to 11 mM, 4 to 11 mM, 5 to 11 mM, 1 to 10 mM, 2 to 10 mM, 3 to 10 mM, 4 to 10 mM, or 5 to 10 mM.

[0082] The solubilizing agent may include cyclodextrin or a derivative thereof. The cyclodextrin may improve the dissolution stability of the compound of Formula 1 and may affect the formation of a stable and uniform cake during lyophilization.

[0083] The cyclodextrin is a cyclic oligosaccharide in which 6 to 12 glucose molecules are linked by alpha-1,4-glycosidic bonds, and includes alpha-cyclodextrin with 6 glucose molecules linked, beta-cyclodextrin with 7 glucose molecules linked, and gamma-cyclodextrin with 8 glucose molecules linked, etc. Cyclodextrins that may be used in the present invention may include all cyclodextrin derivatives in addition to the above three types.

[0084] The cyclodextrin derivatives include alpha-cyclodextrin, 2-hydroxypropyl-beta-cyclodextrin, sulfobutylether-beta-cyclodextrin, etc., and may be specifically 2-hydroxypropyl-beta-cyclodextrin.

[0085] The solubilizer may be included in an amount of 10 to 30% by weight based on the first agent. Specifically, it may contain 10 to 30% by weight, 10.5 to 30% by weight, 11 to 30% by weight, 11.5 to 30% by weight, 12 to 30% by weight, 10 to 29.5% by weight, 10.5 to 29.5% by weight, 11 to 29.5% by weight, 11.5 to 29.5% by weight, 12 to 29.5% by weight, 10 to 29% by weight, 10.5 to 29% by weight, 11 to 29% by weight, 11.5 to 29% by weight, 12 to 29% by weight, 10 to 28.5% by weight, 10.5 to 28.5% by weight, 11 to 28.5% by weight, 11.5 to 28.5% by weight, 12 to 28.5% by weight, 10 to 28% by weight, 10.5 to 28% by weight, 11 to 28% by weight, 11.5 to 28% by weight, or 12 to 28% by weight.

[0086] The pH adjusting agent can include a basic pH adjusting agent and an acidic pH adjusting agent, as described above.

[0087] The first agent may be in the form of a powder, a dry powder, or a lyophile. The first agent can be stored for a long period of time and can be easily formulated into an injection. In addition, since the first agent does not contain any additives that may cause side effects, it is completely harmless to the human body.

[0088] The second agent may comprise a reconstitution solution having a pH of 5 to 12. The reconstitution solution is as described above.

[0089] In the injectable formulation according to an embodiment of the present invention, the pH of the reconstituted solution obtained by mixing the first and second agents may be 5.0 to 7.0. If the pH of the reconstituted solution is out of the range of 5.0 to 7.0, the patient may feel pain when administering the injectable formulation, making the injectable formulation unsuitable for use as an injectable formulation.

[0090] In the injectable formulation according to an embodiment of the present invention, the osmotic pressure of the reconstituted solution obtained by mixing the first agent and the second agent may be 300 mOsm / kg to 400 mOsm / kg. If the osmotic pressure of the reconstituted solution is out of the range of 300 mOsm / kg to 400 mOsm / kg, the patient may feel pain when administering the injectable formulation, and therefore the injectable formulation is not suitable for use as an injectable formulation.

[0091] The content of the compound of Formula 1 in the injectable preparation according to one embodiment of the present invention may be varied depending on the condition of the patient to be administered and the degree of the desired treatment.

[0092] The injectable formulation may contain the compound of Chemical Formula 1 in an amount of 1 to 20 mg / ml based on the reconstituted solution obtained by mixing the first agent and the second agent. Specifically, the compound of Chemical Formula 1 may have an amount of 1 to 20 mg / ml, 5 to 20 mg / ml, 10 to 20 mg / ml, 1 to 15 mg / ml, 5 to 15 mg / ml, 10 to 15 mg / ml, 5 mg / ml, 10 mg / ml, 15 mg / ml, or 20 mg / ml based on the reconstituted solution, which is preferred for one dose.

[0093] When the content of the compound of Formula 1 is a low concentration of less than 1 mg / ml, a large amount of injection solution is required to exhibit a sufficient therapeutic effect, which may cause difficulty in administering the compound to the affected area of ​​the patient. When the content of the compound of Formula 1 is a high concentration of more than 20 mg / ml, the API may precipitate when dissolved in a solvent, or may cause pain when administered to the affected area of ​​the patient.

[0094] The injectable formulation according to one embodiment of the present invention may be in a transparent state in which the first agent containing the compound of Chemical Formula 1 is mixed with the second agent and the compound of Chemical Formula 1 is completely dissolved.

[0095] The injectable formulation according to one embodiment of the present invention can be used by reconstituting a first agent containing the compound of Chemical Formula 1 into a second agent containing the reconstituted solution just before administration to a patient, and administering the reconstituted solution to the affected area of ​​the patient.

[0096] In an embodiment of the injectable formulation of the present invention, the first and second agents are mixed together immediately before administration to a patient, and therefore may not need to contain additional excipients for preservation such as antioxidants and preservatives. However, in the case of the second agent containing a reconstitution solution, a preservative may be included to prevent secondary contamination.

[0097] The present invention provides a method for treating or preventing a caspase-related disease using the injectable formulation of the present invention.

[0098] The term "treatment" means stopping or delaying the progression of a disease when used on a subject who shows symptoms of the disease, and the term "prevention" means stopping or delaying the onset of the disease when used on a subject who does not show symptoms of the disease but is at high risk of the disease.

[0099] The "caspase-related disease" of the present invention refers to a disease that can be treated or prevented by inhibiting caspase, and examples thereof include osteoarthritis, rheumatoid arthritis, degenerative arthritis, destructive bone disorders, liver disease due to viral hepatitis, acute hepatitis, hepatosclerosis, brain damage due to viral hepatitis, sudden human liver failure, sepsis, organ transplant rejection, ischemic heart disease, dementia, stroke, brain damage due to AIDS, diabetes, and gastric ulcer, but are not limited to the above diseases.

[0100] The injectable preparation of the present invention can be administered to an individual in need of treatment or prevention to treat or prevent a caspase-related disease in the individual.

[0101] The term "administration" or "administering" as used herein refers to a method of administering a dose of the injectable formulation of the present invention to a vertebrate or invertebrate animal, including a mammal, a bird, a fish, or an amphibian, by injection. A preferred administration method can be selected from common injection methods such as intravascular injection, subcutaneous injection, transdermal injection, intramuscular injection, spinal injection, and intradermal injection depending on the type of disease to be treated, the site of the disease, and the severity of the disease, and in particular, in the case of osteoarthritis, rheumatoid arthritis, degenerative arthritis, or destructive bone disorders, which are diseases related to bone and joints, intra-articular injection can be selected.

[0102] When the injectable preparation of the present invention is injected into a joint cavity, it can be guided by using an imaging method such as ultrasound, or a local anesthetic can be administered before or in combination with the injection.

[0103] The term "individual" as used herein refers to a human or a non-human mammal, such as a dog, cat, mouse, rat, cow, sheep, pig, goat, non-human ape, or bird. In some embodiments, the individual is a human.

[0104] The injectable formulation of the present invention is administered to an individual within 30 minutes after the first agent and the second agent are mixed to prepare a mixed solution. Thus, the injectable formulation of the present invention is for single administration for injection or administration to a single patient.

[0105] In addition, the injectable preparation of the present invention can be administered in combination with drugs such as painkillers, anti-inflammatory drugs, steroids, etc. Specifically, it can be administered in combination with drugs such as ibuprofen, naproxen, aspirin, acetaminophen, indomethacin, diclofenac, meloxicam, celecoxib, piroxicam, etodolac, nabumetone, lumiracoxib, valdecoxib, etoricoxib, parecoxib, fenoprofen, oxaprozin, mefenamic acid, diflunisal, flurbiprofen, ketoprofen, and other nonsteroidal anti-inflammatory drugs (NSAIDs); corticoid drugs such as prednisone and methylprednisolone, narcotic painkillers such as codeine, pentanyl, morphine, and meperidine, or injections of hyaluronic acid derivatives.

[0106] The present invention provides a kit comprising the injectable formulation of the present invention. In one embodiment of the present invention, the kit may include an injectable formulation comprising a first agent comprising a compound of Formula 1 of the present invention and a second agent comprising a reconstitution solution; an administration system, such as one or more syringes, for administering the formulation; a method for mixing the first and second agents; and instructions for using the kit, including instructions for treating a patient.

[0107] In one embodiment of the present invention, the kit may include a label indicating that the formulation and contents described herein are administered to a patient suffering from osteoarthritis, rheumatoid arthritis, degenerative arthritis, or destructive bone disorders, which are diseases associated with bone and joints, wherein selecting an appropriate dosage for a patient is within the knowledge of a person of ordinary skill in the art, taking into account the symptoms, age, etc., of the patient.

[0108] Unless otherwise indicated, all numbers used in the specification and claims, whether stated or not, should be understood to be modified in all cases by the term "about". Also, the precise numerical values ​​used in the specification and claims should be understood to form additional embodiments of the present disclosure. Efforts have been made to ensure the accuracy of the numerical values ​​disclosed in the examples. However, all measured numerical values ​​inherently contain certain error values ​​generated from the standard deviations observed with the respective measuring techniques. EXAMPLES

[0109] Various evaluations in the examples and comparative examples were carried out as follows.

[0110] Manufacturing Example In the present invention, the compound of formula 1, (R)-N-((2S,3S)-2-(fluoromethyl)-2-hydroxy-5-oxotetrahydrofuran-3-yl)-5-isopropyl-3-(isoquinoline-1-yl)-4,5-dihydroisooxazol-5-carboxamide) (hereinafter referred to as "API") used as an active pharmacological ingredient (API) was prepared according to KR10-0774999.

[0111] Test Example 1: Confirmation of API dissolution depending on the solvent composition, pH of the solvent, and stability of API in API mixed solution (Examples 1 to 16) Solvents for preparing API mixed solutions with a concentration of 10 mg / ml were prepared by dissolving APIs according to the composition shown in Table 1 below. Solvents containing 0 to 100 mM sodium phosphate buffer (PB), 24 to 36 mM sodium hydroxide (NaOH), and 10 to 30% (w / v) hydroxypropyl beta cyclodextrin (HPβCD) were prepared as shown in Table 1 below.

[0112] Sodium phosphate buffer was prepared at pH 7.4 by mixing sodium phosphate dibasic heptahydrate and sodium phosphate monobasic monohydrate. HPβCD was a parenteral grade reagent with a molar substitution range of 0.81–0.99 and a molecular weight of approximately 1500 g / mol.

[0113] As shown in Table 1 below, 10 mg of API was added to 1 ml of the solvent prepared according to Examples 1 to 16, and stirred at about 800 rpm for 30 minutes to prepare an API mixed solution.

[0114] [Table 1]

[0115] Table 2 below shows the results of confirming the pH of the API mixed solution obtained by mixing the API in the solvent having the composition shown in Table 1, and the API stability (purity) in the API mixed solution 30 minutes after mixing.

[0116] Since the active form of API of Chemical Formula 1 is quickly hydrolyzed in an aqueous solution and turns into an inactive form, the peak area (%) was measured using HPLC to confirm the stability of API (purity or content of active form in API mixture solution) 30 minutes after preparation of API mixture solution.

[0117] [Table 2]

[0118] It was confirmed that the solvents in which the API is not soluble (Examples 3, 5, and 6), the solvents in which the API mixed solution has a pH outside the range of 6 to 8 (Examples 1, 7, and 12 to 16), or the API purity (peak area%) is less than 94% 30 minutes after preparation of the API mixed solution (Examples 1 and 10 to 16) are not suitable for dissolving the API of the present invention. Therefore, it was confirmed that the solvents prepared in Examples 2, 4, 8, and 9 are suitable for dissolving the API.

[0119] In Examples 1 to 4, it was confirmed that the solvent containing the phosphate buffer (Examples 2 and 4) is a suitable solvent for easily controlling the pH and dissolving the API and maintaining the purity of the API at 94% or more 30 minutes after preparing the API mixed solution.

[0120] When the phosphate buffer concentration is 10 mM or less and the NaOH concentration is 30 to 36 mM (Examples 6, 7, and 11 to 16), the pH of the API mixture solution is 7.2 or more, so it is desirable to reduce the NaOH concentration to an appropriate level. This is because the higher the pH of the API mixture solution, the faster the active form of API of Formula 1 is hydrolyzed in aqueous solution and converted to an inactive form, resulting in a problem of lowering the purity of the API.

[0121] On the other hand, when the concentration of phosphate buffer was 10 mM or less, NaOH was 24 mM, and the content of HPβCD was 12.5 wt% to 15 wt% (Examples 8 and 9), the API mixture solution had a pH range of 6 to 7 and the purity of API was maintained at 94% or more, confirming that the solvents prepared in Examples 8 and 9 are suitable for preparing API mixture solutions.

[0122] Test Example 2: Preparation of Acidified Mixture and Confirmation of API Stability in Acidified Mixture (Examples 17 to 22) An acid (6N HCl) was added to the mixed solutions of API containing the solvents of Examples 2, 4, 8 and 9 prepared above to prepare acidified mixed solutions.

[0123] The acidified mixed solution is prepared to prevent the active form of API of Formula 1 from being rapidly hydrolyzed in an aqueous solution and converted to an inactive form as the pH of the API mixed solution becomes higher.

[0124] As shown in Table 3 below, 60 to 150 mM HCl was added to the mixed solutions of API containing the solvents prepared in Examples 2, 4, 8, and 9 using 6N HCl as an acidifying solution to prepare acidified mixed solutions.

[0125] [Table 3]

[0126] Table 4 below shows the pH of the acidified mixed solution prepared by adding an acidifying solution to the mixed solution of the APIs shown in Table 3, and the results of confirming the presence or absence of API precipitation and API stability (purity) in the acidified mixed solution.

[0127] [Table 4]

[0128] From the results in Table 4, it was confirmed that the APIs prepared in Examples 17 to 22 were dissolved in the acidified mixed solutions without being precipitated.

[0129] In Examples 17 and 18, the pH of the acidified mixed solution containing the API was about 6.0, and the purity of the acidified mixed solution dropped rapidly after preparation, decreasing to about 60% one day after preparation of the acidified mixed solution. This confirmed that the API is unlikely to exist stably when the pH of the acidified mixed solution containing the API is about 6.0.

[0130] In Examples 19 and 20, the pH of the acidified mixed solution containing API was about 2.0 to 2.7, and one day after preparation of the acidified solution, the purity remained stable with little change. However, it was confirmed that the lower the pH of the acidified mixed solution, the less the purity changed and the more stable the API.

[0131] In Examples 21 and 22, the pH of the acidified mixed solution containing API was about 1.7, and the purity remained stable with little change one day after preparation of the acidified mixed solution.

[0132] Therefore, it was confirmed that the API exists stably when the pH of the acidified solution containing the API is between 1.0 and 2.0.

[0133] Test Example 3: Preparation of freeze-dried dosage forms (Examples 23 to 25) The solutions of Examples 20 to 22 prepared above were freeze-dried under the conditions shown in Table 5 below. That is, the freeze-drying conditions were freezing at -80°C for 3 hours, and then drying at 10 to 20 Pa for 16 or 40 hours.

[0134] [Table 5]

[0135] The freeze-dried dosage forms of API prepared in Examples 23 to 25 had a uniform white cake, indicating that freeze-drying was successful.

[0136] Test Example 4: Preparation of reconstituted solution, reconstitution time, pH and osmolality after reconstitution, and confirmation of stability after reconstitution (Examples 26 to 29) To reconstitute the previously prepared lyophilized dosage forms of Examples 23 to 25, a reconstitution solution was prepared as shown in Table 6 below. The administration volume of the reconstitution solvent was calculated by calculating the volume of water removed after lyophilization in Examples 23 to 25. The reconstitution solvent was added to the lyophilized dosage form and then quickly shaken to prepare a reconstitution solution.

[0137] [Table 6]

[0138] Table 7 below shows the results of measuring the time required for the reconstituted solutions prepared in Examples 26 to 29 to become transparent upon reconstitution, the pH and osmotic pressure of the reconstituted solutions, and the purity of the reconstituted solutions. The purity of the reconstituted solutions was confirmed by UPLC.

[0139] [Table 7]

[0140] From the results in Table 7, it was confirmed that the pH of the reconstituted solutions prepared in Examples 26 to 29 was 6.4 to 6.9, which is a suitable pH for use as an injection.

[0141] However, the preferred osmolality range of a dosage form for use as an injection is 300-400 mOsm / kg, and if it is below or above this range, the patient may feel pain. Therefore, when applying this standard, it was confirmed that Examples 28 and 29, in which the osmolality of the reconstituted solution is 308 mOsm / kg and 365 mOsm / kg, respectively, are preferred as injection dosage forms containing 10 mg / ml of API.

[0142] Test Example 5: Confirmation of API dissolution depending on the solvent composition, pH of the solvent, and stability of API in API mixed solution (Examples 30 to 34) Solvents were prepared to prepare API mixed solutions with a concentration of 15 mg / ml by dissolving the APIs according to the composition shown in Table 8 below. Solvents containing 5 to 10 mM sodium phosphate buffer (PB), 35 to 40 mM sodium hydroxide (NaOH), and 17.5 to 20.0% (w / v) hydroxypropyl beta cyclodextrin (HPβCD) were prepared as shown in Table 8 below.

[0143] Sodium phosphate buffer was prepared by mixing sodium phosphate dibasic heptahydrate and sodium phosphate monobasic monohydrate at pH 7.4. HPβCD was a parenteral grade reagent with a molar substitution range of 0.81–0.99 and a molecular weight of approximately 1500 g / mol.

[0144] As shown in Table 8 below, 15 mg of API was added to 1 ml of the solvent prepared in Examples 30 to 34, and stirred at about 800 rpm for 30 minutes to prepare an API mixed solution.

[0145] [Table 8]

[0146] Table 9 below shows the results of confirming the pH of the API mixed solution obtained by mixing the API in the solvent having the composition shown in Table 8, and the API stability (purity) in the API mixed solution 30 minutes after mixing.

[0147] Since the active form of API of Chemical Formula 1 is quickly hydrolyzed in an aqueous solution and turns into an inactive form, the peak area (%) was measured using HPLC to confirm the stability of API (purity or content of active form in API mixture solution) 30 minutes after preparation of API mixture solution.

[0148] [Table 9]

[0149] From the results in Table 9, it was confirmed that the solvents prepared in Examples 30 to 34 were suitable for dissolving APIs.

[0150] It was confirmed that when the concentration of phosphate buffer is 10 mM or less, the higher the NaOH concentration (Examples 31 and 34), the higher the pH of the API mixture solution. It is preferable to lower the NaOH concentration to an appropriate level. This is because the higher the pH of the API mixture solution, the faster the active form of API of Formula 1 is hydrolyzed in aqueous solution and converted to an inactive form, resulting in a problem of lowering the purity of the API.

[0151] Test Example 6: Preparation of Acidified Mixture and Confirmation of API Stability in Acidified Mixture (Examples 35 to 39) An acid (6N HCl) was added to the mixed solution of API containing the solvent of Examples 30 to 34 prepared above to prepare an acidified mixed solution.

[0152] The acidified mixed solution is prepared to prevent the active form of API of Formula 1 from being rapidly hydrolyzed in an aqueous solution and converted to an inactive form as the pH of the API mixed solution becomes higher.

[0153] As shown in Table 10 below, 50 to 60 mM HCl was added to the mixed solutions of API and solvent prepared in Examples 30 to 34 using 6N HCl as an acidifying solution to prepare acidified mixed solutions.

[0154] [Table 10]

[0155] Table 11 below shows the results of confirming the pH of the acidified mixed solution prepared by adding an acidifying solution to the mixed solution of the APIs shown in Table 10, the presence or absence of API precipitation in the acidified mixed solution, and the API stability (purity).

[0156] [Table 11]

[0157] From the results in Table 11, it was confirmed that the API was not precipitated but dissolved in the acidified mixed solution prepared in Examples 35 to 39. In Examples 35 and 36, the pH of the acidified mixed solution containing the API was about 1.8 to 2.0, and the purity of the acidified mixed solution remained stable one day after preparation with little change.

[0158] In Examples 37 to 39, the pH of the acidified solutions containing API was about 2.0 to 2.2, which was on the lower pH side, but there was a small change in purity one day after preparation of the acidified solutions.

[0159] Therefore, it was confirmed that the API exists stably when the pH of the acidified solution containing the API is 2.0 or less.

[0160] Test Example 7: Preparation of freeze-dried dosage forms (Examples 40 to 44) The solutions of Examples 35 to 39 prepared above were freeze-dried under the conditions shown in Table 12 below. That is, the freeze-drying conditions were freezing at -80°C for 3 hours, followed by drying at 10 to 20 Pa for 40 hours.

[0161] [Table 12]

[0162] The properties of the freeze-dried API dosage forms prepared in Examples 40 to 44 were that an opaque white cake was uniformly formed, confirming that freeze-drying was successful.

[0163] Test Example 8: Preparation of reconstituted solution, reconstitution time, pH and osmotic pressure of the solution after reconstitution, and confirmation of stability after reconstitution (Examples 45 to 49) To reconstitute the previously prepared lyophilized dosage forms of Examples 40 to 44, a reconstitution solution was prepared as shown in Table 13 below. The administration volume of the reconstitution solvent was calculated from the volume of water removed after lyophilization in Examples 40 to 44. The reconstitution solvent was added to the lyophilized dosage form and then quickly shaken to prepare a reconstitution solution.

[0164] [Table 13]

[0165] Table 14 below shows the results of measuring the time required for the reconstituted solutions prepared in Examples 45 to 49 to become transparent upon reconstitution, the pH and osmotic pressure of the reconstituted solutions, and the purity of the reconstituted solutions, which were confirmed by UPLC.

[0166] [Table 14]

[0167] From the results in Table 14, it was confirmed that the pH of the reconstituted solutions prepared in Examples 45 to 49 was about 5.8 to 7.0, all of which were suitable for use as injections.

[0168] However, the preferred osmolality range of a dosage form for use as an injection is 300-400 mOsm / kg, and if it is below or above this range, the patient may feel pain. Therefore, when applying this standard, it was confirmed that Examples 47 and 48, in which the osmolality of the reconstituted solution is 372 mOsm / kg and 367 mOsm / kg, respectively, are preferred as injection dosage forms containing 15 mg / ml of API.

[0169] In the case of Example 49, the purity immediately after reconstitution was about 93%, and the purity 30 minutes after reconstitution was about 92%, confirming that the purity of the API itself was too low to be suitable.

[0170] Test Example 9: Preparation of API mixed solution using mixing equipment In the production of API mixed solution, the change in purity of API in API mixed solution due to mixing equipment was confirmed.

[0171] As shown in Table 15 below, API mixed solutions were prepared according to API concentration and solvent composition using different stirring equipment, a homogenizer and a magnetic stirrer.

[0172] [Table 15]

[0173] In Example 50, where a homogenizer was used as the stirring equipment, the API did not have good wettability, but it was confirmed that the API and the solvent were mixed well and uniformly without floating on the solvent. On the other hand, in Example 51, where a magnetic stirrer was used as the stirring equipment, the API did not dissolve immediately on the solvent, but floated, and it took more time to dissolve.

[0174] Figure 1 shows the results of measuring the API purity in the API mixed solution versus the mixing time. As in Figure 1, the API purity was 96.4% in Example 50 using a homogenizer, and 96.1% in Example 51 using a magnetic stirrer, so it is unlikely that the mixing equipment has any effect on the purity change. However, it was confirmed that the purity is greatly dependent on the dissolution time.

[0175] Therefore, it was confirmed that a homogenizer capable of producing a mixed solution quickly and uniformly is suitable as the stirring equipment.

[0176] Test Example 10: Changes in API purity during the manufacture of lyophilized dosage forms The API lyophilized final dosage forms were manufactured as described in Table 16 below.

[0177] [Table 16]

[0178] In order to confirm the change in purity of the API during the preparation of the lyophilized final dosage form, the purity of the API powder before dissolving in the solvent composition, the purity of the API mixed solution, and the purity after lyophilization were measured using UPLC.

[0179] [Table 17]

[0180] As shown in Table 17, when preparing the final lyophilized dosage form of API, the purity mainly decreased during the process of preparing the API mixture solution, and there was almost no change in purity during the process of lyophilization from the API mixture solution.

[0181] Test Example 11: Thermal analysis and XRD pattern analysis of freeze-dried dosage form The lyophilized final dosage form of API prepared in Example 52 was subjected to TGA and DSC measurements at a temperature range of 30 to 350° C. and a heating rate of 10° C. / min.

[0182] FIG. 2(a) shows the TGA measurement result of the lyophilized final dosage form of API prepared in Example 52, and FIG. 2(b) shows the DSC measurement result of the lyophilized final dosage form of API prepared in Example 52.

[0183] In FIG. 2, the initial weight loss was estimated to be due to residual moisture, the amount of which was about 1.8%, and since no endothermic peak of the API was found, it was confirmed that the API was in an amorphous state.

[0184] Therefore, it was confirmed that the residual moisture was 1.8% (<2%) and the API was in an amorphous state, so that the freeze-drying was successful.

[0185] The XRD pattern of the lyophilized final dosage form of API prepared in Example 52 was measured under the conditions of a voltage of 45 kV, a current of 40 mA, and a scan range of 4° to 40°.

[0186] FIG. 3 shows the XRD pattern of the lyophilized final dosage form of the API prepared in Example 52.

[0187] In Figure 3, Example 52, which is the final lyophilized dosage form of the API, showed the same pattern as HPβCD, and no powder peak of the API was found. Therefore, it was confirmed that the API was well dissolved in the solution and was well lyophilized, turning into an amorphous API.

[0188] Test Example 12: Solid-state stability analysis of lyophilized dosage forms In order to predict the shelf life of the lyophilized formulation of API in solid state in Example 52, Example 52 was stored under the conditions shown in Table 18 below, and then a qualitative analysis was performed using UPLC.

[0189] [Table 18]

[0190] Table 18 shows the % area value of IMP that increased the most in the stability test of Example 52. It was confirmed that IMP increased more significantly as the storage temperature and time increased.

[0191] Therefore, the lyophilized dosage form of the API of Example 52 was confirmed to be a stable dosage form having a shelf life of about 3 years or less at 5°C.

Claims

1. A first agent in powder form comprising a compound of the following formula 1, a phosphate buffer, a solubilizing agent, and a pH adjusting agent; and a second agent comprising a reconstitution solution having a pH of 5 to 12; 【Chemistry 1】

2. 2. The injectable formulation of claim 1, wherein the phosphate buffer is a mixture of sodium phosphate dibasic heptahydrate and sodium phosphate monobasic monohydrate.

3. The injectable preparation according to claim 1, wherein the content of the phosphate buffer is 1 to 15 mM based on the first agent.

4. The injectable formulation of claim 1 , wherein the solubilizing agent is a cyclodextrin.

5. The injectable formulation according to claim 4, wherein the cyclodextrin is any one selected from the group consisting of alpha-cyclodextrin, 2-hydroxypropyl-beta-cyclodextrin, and sulfobutylether-beta-cyclodextrin.

6. The injectable preparation according to claim 4, wherein the content of the cyclodextrin is 10 to 30% by weight based on the first agent.

7. The injectable formulation according to claim 1 , wherein the pH of the solution obtained by mixing the first agent and the second agent and reconstituting the solution is 5.0 to 7.

0.

8. The injectable preparation according to claim 1 , wherein the osmotic pressure of a solution obtained by mixing the first agent and the second agent and reconstituting the solution is 300 mOsm / kg to 400 mOsm / kg.

9. The injectable formulation according to claim 1, wherein the content of the compound of Chemical Formula 1 is 1 to 20 mg / ml based on a reconstituted solution obtained by mixing the first agent and the second agent.

10. (S1) preparing a solvent having a pH of 10 to 12, comprising a phosphate buffer, a solubilizing agent, and a basic pH adjusting agent; (S2) preparing a first solution having a pH of 6 to 8 by dissolving a compound of Formula 1 in the prepared solvent: 【Chemistry 2】 (S3) adding an acidic pH adjuster to the first solution to prepare a second solution having a pH of 1.0 to 3.0; (S4) freeze-drying the second solution to prepare a first agent in powder form; and (S5) preparing a second agent containing a reconstituted solution having a pH of 5 to 12.

11. The method for preparing an injectable formulation according to claim 10, wherein the content of the compound of Formula 1 is 1 to 20 mg / ml based on the solution after reconstitution.

12. The method for producing an injectable formulation according to claim 10, wherein the phosphate buffer is a mixture of sodium phosphate dibasic heptahydrate and sodium phosphate monobasic monohydrate.

13. The method for preparing an injectable formulation according to claim 10, wherein the content of the phosphate buffer is 1 to 15 mM based on the solvent.

14. The method for producing an injectable formulation according to claim 10, wherein the solubilizing agent is cyclodextrin.

15. The method for producing an injectable formulation according to claim 14, wherein the cyclodextrin is any one selected from the group consisting of alpha-cyclodextrin, 2-hydroxypropyl-beta-cyclodextrin, and sulfobutylether-beta-cyclodextrin.

16. The method for preparing an injectable formulation according to claim 14, wherein the content of the cyclodextrin is 10 to 30% by weight based on the solvent.

17. The injectable formulation according to claim 1, which is for treating or preventing a disease selected from the group consisting of osteoarthritis, rheumatoid arthritis, degenerative arthritis, destructive bone disorders, liver disease due to viral hepatitis, acute hepatitis, liver sclerosis, brain damage due to viral hepatitis, sudden human liver failure, sepsis, organ transplant rejection, ischemic heart disease, dementia, stroke, brain damage due to AIDS, diabetes and gastric ulcer.

18. The injectable formulation according to claim 17, wherein the disease is selected from the group consisting of osteoarthritis, rheumatoid arthritis, degenerative arthritis and destructive bone disorders.

19. The injectable preparation according to claim 1 , wherein the injectable preparation is used in a single dose in the form of a mixed solution of the first agent and the second agent.

20. The injectable preparation according to claim 19 , wherein the injectable preparation is administered into an articular cavity after the first agent and the second agent are mixed.

21. The injectable preparation according to claim 19, wherein the injectable preparation is administered into the articular cavity within 30 minutes after the first agent and the second agent are mixed.

22. The injectable formulation according to claim 19, wherein the injectable formulation is administered once after the first agent and the second agent are mixed.

23. A first agent in powder form comprising a compound of the following formula 1, a phosphate buffer, a solubilizing agent, and a pH adjusting agent; and a second agent comprising a reconstitution solution having a pH of 5 to 12; 【Chemistry 3】 The injectable preparation is characterized in that, when the contents of the compound of Chemical Formula 2 and the compound of Chemical Formula 3 are measured within 30 minutes after mixing the first and second agents, the content of the compound of Chemical Formula 2 is 94% or more of the total content of the compound of Chemical Formula 2 and the compound of Chemical Formula 3: 【Chemistry 4】 【Chemistry 5】

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