Injectable depot composition comprising enavogliflozin and low-dose liraglutide

Abstract

The present invention provides an injectable depot composition comprising enavogliflozin and a low-dose liraglutide. A combination regimen of enavogliflozin and a low-dose liraglutide provides a synergistic effect in the treatment of obesity compared to the sole administration of either of the drugs. In addition, compared to the sole administration of liraglutide, the administered dose of liraglutide can be reduced, and thus direct causes of gastrointestinal and cardiovascular side effects of liraglutide can be mitigated. In addition, the present invention has the advantage of additionally providing the effect of alleviating metabolic diseases and cardiovascular diseases related to blood glucose, blood lipids, blood pressure, fatty liver, and the like through combined administration with enavogliflozin. Furthermore, the depot composition of enavogliflozin and liraglutide according to the present invention can significantly improve medication convenience and medication adherence for a patient compared to existing liraglutide formulations, which are subcutaneously injected once a day, by unifying the route of administration and improving the administration cycle.

Description

Injectable depot composition containing inavogliflozin and low-dose liraglutide

[0001] It relates to an injectable depot composition comprising inabogliflozin and low-dose liraglutide.

[0002] GLP-1 receptor agonists (GLP-1RAs) are drugs that activate the GLP-1 receptor, reducing blood sugar and energy intake. GLP-1RAs mimic the action of GLP-1, an endogenous incretin hormone secreted by the intestines after meals. GLP-1RAs suppress glucagon secretion and stimulate insulin production, reducing hyperglycemia in patients with type 2 diabetes and reducing food intake, making them effective in treating obesity through weight loss. GLP-1RAs suppress glucagon secretion in proportion to glucose concentration. This means that glucagon secretion is suppressed at high blood sugar levels and not suppressed at low blood sugar levels, which has the advantage of not causing serious problems in hypoglycemic situations. Liraglutide, a GLP-1RA drug better known under the brand name Saxenda, was initially developed as a diabetes treatment. However, it has been shown to be effective in obesity when administered in high doses, and is now widely used in the treatment of obesity. However, the obesity prescription dose of liraglutide is 3 to 5 times higher than the diabetes prescription dose, and unlike when administering the diabetes prescription dose, when administering the obesity prescription dose, it is known to have many side effects, including gastrointestinal side effects such as nausea and diarrhea due to high dose prescription, and cardiovascular side effects such as increased heart rate.

[0003] Meanwhile, SGLT-2 inhibitors are drugs with mechanisms of action that promote weight loss and lower blood pressure through osmotic diuresis and caloric loss due to glucose excretion. Enavogliflozin, an SGLT-2 inhibitor developed by the applicant, recently received marketing approval as an oral diabetes treatment. According to the applicant's preliminary research, enavogliflozin has been confirmed to have an excellent anti-obesity effect based on its ability to inhibit glucose reabsorption.

[0004] Accordingly, the inventors of the present invention hypothesized that the combined administration of inavogliflozin and low-dose liraglutide would have a synergistic effect in the treatment of obesity, thereby improving the anti-obesity effect while reducing side effects compared to existing treatments, and conducted research. As a result, it was confirmed that the combined administration of inavogliflozin and low-dose liraglutide showed a synergistic effect in weight loss compared to when the two drugs were administered alone while reducing the side effects of liraglutide. Furthermore, it was confirmed that these drugs could be manufactured into an injectable depot composition to provide a long-acting combination preparation, thereby completing the present invention.

[0005] The present invention aims to provide a novel anti-obesity pharmaceutical composition that can improve the anti-obesity effect while reducing the side effects of high-dose administration of liraglutide, and at the same time improve the duration of the drug effect, convenience of taking the medication, and compliance of the patient.

[0006] In the course of conducting research to reduce the side effects of high-dose liraglutide administration, the present inventors were able to confirm that co-administration of inavogliflozin (administered orally once daily) and low-dose liraglutide (administered subcutaneously once daily) reduces the side effects of liraglutide while showing a synergistic effect on weight loss compared to when the two drugs are administered alone.

[0007] Although the difference in the administration route of each drug does not affect the achievement of the enhanced anti-obesity effect, unifying the administration route can contribute to improving the patient's convenience and compliance with medication, so it is desirable to administer both drugs through a single administration route.

[0008] Furthermore, once-daily administration, especially for injections, not only causes patients to reject injections but is also extremely uncomfortable. Anti-obesity treatment typically lasts for two to four weeks or longer, and once-daily subcutaneous injections can significantly reduce patient convenience and compliance. Therefore, improving this once-daily subcutaneous injection regimen was also a problem the inventors sought to address.

[0009] Accordingly, the inventors of the present invention have conducted ongoing research to develop a combination formulation that unifies the route of administration and dosage regimen for low-dose co-administration of inavogliflozin and liraglutide. To avoid the need for a single daily subcutaneous injection, the inventors conceived of a depot composition that could be developed into a sustained-release injection. As a result, they confirmed that manufacturing an injectable depot composition containing inavogliflozin and low-dose liraglutide as active ingredients could improve the duration of efficacy, patient convenience, and medication compliance.

[0010] Accordingly, the present invention provides an injectable depot composition comprising microspheres comprising inabogliflozin and liraglutide as active ingredients and a biodegradable polymer.

[0011] A depot composition comprising microspheres containing a drug and a biodegradable polymer can be used as a sustained-release injection that exhibits long-term efficacy as the drug is slowly released during the decomposition process of the microspheres.

[0012] In the depot composition, it is possible to formulate multiple drugs into a single dosage form by encapsulating multiple drugs in a single microparticle or encapsulating two or more drugs in two or more types of microparticles.

[0013] However, since inavogliflozin is a compound drug and liraglutide is a peptide drug, it is not easy to formulate drugs with different properties into a single dosage form. Inavogliflozin, as a compound drug, has low water solubility (less than 1 mg / ml), so the use of organic solvents is inevitable for the manufacture of microspheres. On the other hand, liraglutide is a peptide drug with excellent water solubility (approximately 50 mg / ml) and is not dissolved at all in volatile organic solvents (e.g., methylene chloride) that are essential for the manufacture of microspheres. These significant differences in the physical and pharmacological properties of the two drugs pose significant challenges throughout the manufacturing process of microspheres, the drug encapsulation process, and the control of drug release. In order to solve these problems, the inventors of the present invention attempted to control the amount of organic solvent used to dissolve inabogliflozin and various manufacturing processes that affect the encapsulation of liraglutide (control of primary emulsification conditions, control of secondary emulsification conditions, control of volatilization process, solubility control through additives, and change of the salt of liraglutide), and as a result, they showed that it was possible to manufacture a complex depot composition capable of releasing two drugs at a constant effective concentration over a target period of time.

[0014] In the pharmaceutical composition of the present invention, inabogliflozin or liraglutide may be present in the form of a pharmaceutically acceptable salt, a pharmaceutically acceptable amide, or a pharmaceutically acceptable ester. For example, in one embodiment of the present invention, liraglutide may be present as liraglutide acetate.

[0015] Techniques for manufacturing microspheres containing drugs and biodegradable polymers are well known in the art.

[0016] For example, the biodegradable polymer may be selected from the group consisting of polylactide (PLA), polyglycolide (PGA), poly(lactide-co-glycolide) (PLGA), and mixtures thereof. The ratio of polylactide to polyglycolide in the poly(lactide-co-glycolide) copolymer may be from 50:50 to 95:5, for example, 50:50, 60:40, 65:35, 70:30, 75:25, 80:20, or 85:15.

[0017] Although not limited thereto, the biodegradable polymer may have a weight average molecular weight of 4,000 to 50,000. For example, the weight average molecular weight of the biodegradable polymer includes all lower numerical ranges within the above range, such as a weight average molecular weight of 4,000 to 15,000, a weight average molecular weight of 7,000 to 17,000, a weight average molecular weight of 5,000 to 20,000, a weight average molecular weight of 8,000 to 30,000, a weight average molecular weight of 8,000 to 20,000, a weight average molecular weight of 10,000 to 18,000, a weight average molecular weight of 18,000 to 28,000.

[0018] For example, RESOMER from Evonik Rohm GmbH is a biodegradable polymer used in the present invention. TM Polylactide, polyglycolide, poly(lactide-co-glycolide) under the trade name can be used or a blend thereof can be used. For example, R202H, R202S, R203H, R203S, RG502H, RG503H, RG653H, RG752H, RG752S, RG753H, RG753S can be used alone or in a blend.

[0019] The appropriate molecular weight or blending ratio of the biodegradable polymer can be appropriately selected by those skilled in the art, taking into account the biodegradable polymer's decomposition rate and the resulting drug release rate. In one specific example, but not limited thereto, the biodegradable polymer may be polylactide having a weight average molecular weight of 8,000 to 30,000.

[0020] In a specific example of the present invention, the microspheres may be manufactured by a solvent evaporation method or a spray drying method.

[0021] The solvent evaporation method refers to a method of forming microspheres by preparing an O / W or W / O / W emulsion and then evaporating the solvent therefrom. An emulsion is obtained by mixing a mixture of a biodegradable polymer and a drug dissolved in a solvent with an aqueous medium, and the solvent is evaporated from the emulsion to form microspheres containing the drug.

[0022] Spray drying is a method that sprays a liquid containing an active ingredient through a nozzle at a specific rate and simultaneously dries it with hot air to obtain a powder. A mixture of a biodegradable polymer and a drug in an organic solvent is sprayed as a liquid through a nozzle and then dried simultaneously to recover microparticles containing the drug of a specific size.

[0023] For example, the above microspheres may be W / O / W type microspheres manufactured by preparing a W / O / W emulsion and then evaporating the solvent therefrom.

[0024] The encapsulation of drugs within microspheres may vary depending on the manufacturing method. As explained above, liraglutide is water-soluble, whereas inavogliflozin is poorly water-soluble. Therefore, it is difficult to utilize the O / W method, which mixes a solvent capable of dissolving multiple drugs simultaneously with an organic solvent containing a biodegradable polymer and prepares the microspheres. This is because the solubilities of inavogliflozin and liraglutide are different, and a co-solvent capable of dissolving both drugs simultaneously in an organic solvent and an aqueous phase could not be found. Therefore, in one embodiment of the present invention, liraglutide is dissolved in an aqueous phase, and various organic solvents capable of dissolving inavogliflozin are mixed with an organic solvent containing a biodegradable polymer to create a W / O emulsion, and liraglutide in the aqueous phase is remixed, thereby manufacturing microspheres using the W / O / W manufacturing method.

[0025] In the case of W / O / W type microspheres, the oil phase solution and the first phase solution are mixed with a homomixer to form a W / O emulsion, which is then mixed with the second phase solution to form W / O / W type microspheres. At this time, the first phase solution may be water, for example, first distilled water. The first phase solution may also include a PBS buffer. Although not limited thereto, the oil phase solvent may be methylene chloride, acetic acid, or the like. The oil phase solution may include a biodegradable polymer such as polylactide, polyglycolide, poly(lactide-co-glycolide), or a blend thereof. The organic solvent capable of dissolving inavogliflozin may include methanol, dimethyl sulfoxide, acetonitrile n-methyl-2-pyrrolidone, or the like. The second phase solution may be a surfactant aqueous solution. Polyvinyl alcohol can be used as a surfactant in the second water solution.

[0026] A W / O emulsion can be prepared by mixing an oil phase solution and a first phase solution and homogenizing them by stirring using a homogenizer. Specifically, a W / O emulsion can be prepared by mixing an oil phase solution (O) containing a first phase solution (W) containing liraglutide, a solution of inavogliflozin dissolved in an organic solvent, and a solution of a polymer dissolved in an organic solvent.

[0027] The prepared W / O emulsion can be added again to a second aqueous solution containing a surfactant to prepare a W / O / W type emulsion. In this case, the emulsion can be prepared by homogenization through stirring using a homogenizer. The stirring speed and time for homogenization are not limited thereto, but generally, stirring can be performed at 500 to 20,000 rpm for 1 to 5 minutes. The solvent is evaporated from the homogenized emulsion, and the evaporation of the solvent can be performed at a temperature of 5 to 40°C for 1 to 4 hours while stirring at 50 to 600 RPM.

[0028] In the present invention, the drug encapsulation efficiency of inavogliflozin was examined by considering the solubility of the organic solvent, and among them, methanol was mainly used because it can be easily removed compared to other organic solvents during the volatilization and freeze-drying processes, and thus leaves less solvent residue in the microspheres. In addition, in order to increase the drug encapsulation efficiency of inavogliflozin, an optimal ratio was searched where inavogliflozin was not precipitated by methylene chloride, which is used as an oil-phase cosolvent, and conversely, a biodegradable polymer dissolved in methylene chloride was not precipitated by the organic solvent used to dissolve inavogliflozin, thereby establishing the optimal concentration range of inavogliflozin and the amount of organic solvent. Although not limited thereto, the concentration of inavogliflozin in the organic solvent may be 10 to 20 w / w%. Additionally, the weight ratio of solvent (e.g., methanol) to inabogliflozin relative to oil solvent (e.g., methylene chloride) may preferably be 4 to 6:1.

[0029] In one specific example of the present invention, when manufacturing W / O / W type microspheres, sodium acetate or magnesium hydroxide may be included as an additive during the preparation of a W / O emulsion. Using the W / O / W method, in order to increase the drug encapsulation efficiency of liraglutide, a manufacturing process was secured in which sodium acetate was added to the first aqueous solution to adjust the pH, and the stability and solubility of the peptide drug liraglutide in the aqueous solution were controlled, thereby inducing an increase in the drug encapsulation efficiency of liraglutide in the microspheres. In another specific example, magnesium hydroxide was added as an additive to enable physical and chemical bonding with liraglutide by evenly mixing it with a biodegradable polymer during the preparation of the emulsion, thereby increasing the drug encapsulation efficiency and suppressing the initial drug release (initial burst), and inducing an effect of sustained drug release for a long period of time compared to a control liraglutide drug administered once a day.

[0030] In another specific embodiment of the present invention, the microspheres may be prepared by evaporating the solvent from an S / O / W emulsion. For example, liraglutide may exist in a solid state of liraglutide, such as liraglutide acetate, within the microspheres. In the case of microspheres containing liraglutide, initial drug burst may be a problem, and in this case, side effects may occur in patients. In order to control the initial drug burst of microspheres encapsulating liraglutide, in a specific embodiment of the present invention, liraglutide was prepared as liraglutide acetate and used by utilizing hydrophobic ion pairing. Hydrophobic ion pairing is a method in which a charged hydrophilic molecule electrostatically binds to an oppositely charged hydrophobic counterion to form a complex having greater hydrophobicity than the original molecule. Liraglutide acetate exhibits greater hydrophobicity at a specific pH in the aqueous phase, causing it to precipitate as a solid. This property can be utilized to recover liraglutide acetate in a solid phase and utilize it for microsphere formulation using the S / O / W method. Furthermore, this solid liraglutide acetate exhibits lower initial drug release than in vivo, and it is more stably encapsulated in microspheres, potentially making it more suitable for achieving a certain level of drug release in vivo.

[0031] In the case of S / O / W type microspheres, a solid dispersion (S / O) is prepared by mixing an oil solution and a solid liraglutide acetate using a homogenizer for several minutes, and then mixing it with a second aqueous solution containing a surfactant using a homogenizer, to ultimately form S / O / W type microspheres. An S / O emulsion can be prepared by homogenizing the oil solution and the solid liraglutide acetate through stirring and mixing using a homogenizer. The stirring speed and time for homogenization are not limited thereto, but generally, homogenization can be performed at 500 to 25,000 rpm for 30 seconds to 3 minutes. At this time, the particle size of the solid liraglutide acetate can be 0.1 to 10 μm, for example, 0.1 to 5 μm. Although not limited thereto, the oil solvent may be methylene chloride. The oil solution may include a biodegradable polymer such as polylactide, polyglycolide, or poly(lactide-co-glycolide), or a blend thereof. In addition, the oil solvent may include acetic acid, and may include an organic solvent capable of dissolving inavogliflozin, such as methanol, dimethyl sulfoxide, or acetonitrile n-methyl-2-pyrrolidone. The second aqueous solution may be an aqueous surfactant solution. The second aqueous solution may also include polyvinyl alcohol. The prepared S / O emulsion may be added to the second aqueous solution to prepare an S / O / W type emulsion. In this case, the emulsion may be prepared by homogenization through stirring using a homogenizer. The stirring speed and time for homogenization are not limited thereto, but generally, stirring can be performed at 500 to 20,000 rpm for 1 to 5 minutes. The solvent is evaporated from the homogenized emulsion, and the evaporation of the solvent can be performed at a temperature of 5 to 40°C for 1 to 4 hours while stirring at 50 to 600 RPM.

[0032] In one embodiment of the present invention, a method for preparing microspheres is exemplified by mixing solid-state liraglutide, for example, liraglutide acetate, with an O / W emulsion containing inavogliflozin.

[0033] S / O / W type microspheres containing liraglutide in a solid state may be advantageous in that they can suppress excessive initial release of liraglutide while increasing drug encapsulation efficiency.

[0034] In another specific embodiment of the present invention, a depot composition is provided comprising microspheres comprising inavogliflozin and microspheres comprising liraglutide. In this case, inavogliflozin may be manufactured as O / W type microspheres, and liraglutide may be manufactured as W / O / W or S / O / W type microspheres, and the two types of microspheres may be prepared by mixing them. In addition, inavogliflozin and liraglutide may each be manufactured by a spray-drying method, and then the content of the active ingredient contained in each microsphere may be measured, and then the amount of microsphere powder may be quantified in consideration of the administration dose, and then the composition may be mixed for several minutes using a powder Lab Mixer for use.

[0035] In the present invention, the microspheres may have an average particle size of 10 to 100 μm. Although not limited thereto, microspheres having an average particle size of 10 to 50 μm are suitable for use as an injection.

[0036] The desired drug content ratio within the entire microspheres is not limited thereto, but may be from 10 to 40 wt%, including, but not limited to, all lower numerical ranges within the above range, such as 10 to 20 wt%, 15 to 35 wt%, 15 to 30 wt%, 20 to 30 wt%, 20 to 27 wt%, and 20 to 24 wt%.

[0037] In one specific example of the present invention, in the case of inavogliflozin, the drug content ratio of inavogliflozin within the microparticles may be 10 to 20 wt%, but is not limited thereto. In the case of liraglutide, the drug content ratio of liraglutide within the microparticles may be 10 to 20 wt%, but is not limited thereto.

[0038] In the present invention, the depot composition may be for the prevention or treatment of obesity.

[0039] Although not limited thereto, the depot composition may be injected subcutaneously once every two to four weeks. The administration cycle of the depot composition can be controlled by controlling the amount of drug loaded in the microspheres, the drug release rate, and the amount of the depot composition injected at a time. Although not limited thereto, the depot composition may be injected subcutaneously once every two weeks.

[0040] Inavogliflozin is currently approved as a diabetes treatment drug and is sold under the product name Enblo Tablet. The dosage of inavogliflozin as a diabetes treatment drug is approved as 0.3 mg per day, but the dosage of inavogliflozin as an obesity treatment drug has not been determined. The present invention relates to the use of inavogliflozin for the treatment of obesity in combination with low-dose liraglutide. Based on the examples below, the dosage of inavogliflozin for combination with low-dose liraglutide may be 0.3 to 1.5 mg / day.

[0041] Here, low-dose liraglutide refers to a lower dosage compared to the dosage of liraglutide currently used to treat obesity. Liraglutide is currently approved for two uses: diabetes and obesity. The dosage for diabetes treatment is 0.6 to 1.8 mg per day, whereas the dosage for obesity treatment is 3.0 mg per day, which means that the approved dosage for obesity treatment is much higher than the approved dosage for diabetes treatment. The low-dose liraglutide intended in the present invention means a dosage less than 3.0 mg per day, which is the current approved dosage for obesity treatment, and is not limited thereto, but the dosage of liraglutide may be 1.2 to 2.4 mg / day. That is, when inavogliflozin and low-dose liraglutide are co-administered, in one specific example, the dosage of inavogliflozin may be 0.3 to 1.5 mg / day, and the dosage of liraglutide may be 1.2 to 2.4 mg / day. In another specific example, the dosage of inavogliflozin may be 0.6 to 1.2 mg / day, and the dosage of liraglutide may be 1.5 to 2 mg / day.

[0042] In one specific example, the depot composition may be formulated as an injection for parenteral administration. When formulated as an injection, the sustained-release drug microspheres may be formulated as an aqueous or oily suspension by adding appropriate excipients. For example, when the drug microspheres are formulated as a suspension, those skilled in the art can select a dispersion medium that exhibits excellent dispersibility for the microspheres. Furthermore, preservatives, isotonic agents, and the like commonly used in suspensions may be added.

[0043] In one embodiment, when the depot composition is formulated as an injection, the depot composition may be present in a separate vial from the dispersion medium and may be prepared into a suspension immediately prior to administration to the patient. In one embodiment, the present invention provides a kit comprising the depot composition, the dispersion medium, and a syringe. Alternatively, the depot composition and the suspension may be filled into a syringe, but may exist independently of each other in separate compartments within the syringe.

[0044] Combination therapy with inavogliflozin and low-dose liraglutide offers a synergistic effect in the treatment of obesity compared to either drug alone. Furthermore, compared to liraglutide alone, the dose of liraglutide can be reduced, thereby reducing the direct cause of liraglutide's gastrointestinal and cardiovascular side effects. Furthermore, combination therapy with inavogliflozin offers the additional benefit of improving metabolic and cardiovascular conditions, including blood sugar, blood lipids, blood pressure, and fatty liver.

[0045] Furthermore, the depot composition of inabogliflozin and liraglutide according to the present invention can significantly improve patient convenience and compliance with medication compared to the conventional once-daily subcutaneous injection form of liraglutide through unification of the administration route and improvement of the administration cycle.

[0046] Figure 1 is a graph showing the body weight reduction rate in an obesity-induced rat model following administration of a depot composition according to the present invention.

[0047] Figure 2 is a photograph showing a decrease in the percentage of fat within the tissue as a result of body composition analysis of an obesity-induced rat model following administration of a depot composition according to the present invention.

[0048] Figure 3 shows the PK analysis results of inabogliflozin according to administration of each test substance in test groups G1 to G9.

[0049] Figure 4 shows the PK analysis results of liraglutide according to administration of each test substance in test groups G1 to G9.

[0050] The advantages and features of the present invention, and the methods for achieving them, will become clearer with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various different forms. These embodiments are provided solely to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the invention. The present invention is defined solely by the scope of the claims.

[0051] The present inventors have conceived the idea of ​​producing a combination formulation that can reduce the dosage of liraglutide and co-administer it with inavogliflozin, while improving the unification of the administration route and the dosage regimen of the existing two drugs, which are administered once a day. As follows, a depot composition containing inavogliflozin and liraglutide was produced and its anti-obesity efficacy was evaluated.

[0052] For convenience, in the tables and drawings of the examples below, liraglutide is also expressed by the abbreviation LIRA, and inavogliflozin is also expressed by the abbreviation ENAVO.

[0053] [Manufacturing Examples 1 and 2]

[0054] Depot compositions containing inabogliflozin and liraglutide were prepared according to the composition and manufacturing method of Table 1, and the depot compositions of Manufacturing Examples 1 and 2 were obtained.

[0055] Manufacturing Example Manufacturing Method Polymer Concentration / Type Additives and Features ENAVO Concentration (w / w%) LIRA Concentration (w / w%) Continuous Phase (CP) Conditions 1st Homogenization Condition (RPM) 2nd Homogenization Condition (RPM) CP Injection Speed / Injection Temperature (°C) Discontinuous Phase (DP) Injection Speed ​​Volatilization Conditions Temperature / Time / RPM Size D10 / D50 / D90 / D[4,3] 1 W / O / W 35% PLGARG 75 2 HNaAc 1% 16.67 27.17 0.25% PVA 3L+1% mannitol 4,000 RPM 3 min 4,000 RPM 3 min 1.5 L / min 20℃ 5ml / min 25℃ / 3 hours / 150 3.4 / 26.8 / 65.1 / 30.92 W / O / W 30% PLAR202HMg(OH)210%16.6727.170.50% PVA 5L+1% mannitol4,000 RPM3min4,000 RPM3min1.5 L / min20℃5ml / min30℃ / 3hrs / 1503.8 / 23.1 / 63.7 / 29.1

[0056] Liraglutide was sufficiently dissolved in distilled water for more than 30 minutes to prepare a first phase solution (W). Inavogliflozin was sufficiently dissolved in methanol for more than 30 minutes before use. The biodegradable polymer was dissolved in methylene chloride, an oil solvent. An oil phase solution (O) was prepared by mixing a solution of inavogliflozin dissolved in methanol and a solution of the biodegradable polymer dissolved in methylene chloride. The first phase solution (W) and the oil phase solution (O) were mixed and homogenized using a homogenizer to prepare a water-on-oil (W / O) emulsion. In addition, an aqueous solution containing polyvinyl alcohol (PVA) as a surfactant was prepared as a second phase solution. When the W / O emulsion was injected through a rubber pipe using a micro-injector, it was exposed to the surfactant aqueous solution and simultaneously mixed by the homo-mixer to form microspheres. The organic solvent present in the microparticles was removed by rotating the mixture of the water-soluble emulsion and the surfactant aqueous solution at a constant speed. To remove the organic solvent, the temperature of the preparation vessel was adjusted to 15-35℃ and maintained for 3 hours to remove the organic solvent. After the organic solvent was removed, the suspension was repeatedly washed with distilled water to remove the residual polyvinyl alcohol, and the recovered microparticles were freeze-dried.

[0057]

[0058] [Experimental Example 1] Evaluation of the anti-obesity efficacy of Manufacturing Examples 1 and 2

[0059] To evaluate the anti-obesity efficacy of the depot compositions of Manufacturing Examples 1 and 2, obesity was induced in 5-week-old male SD rats and used as an animal model.

[0060] Obesity was induced in rats by feeding them a high-fat diet for 13 weeks. To minimize stress on the experimental animals due to subcutaneous administration of the test substance, all animals were administered saline subcutaneously for 2 weeks prior to administration of the test substance to provide prior adaptation training for subcutaneous administration.

[0061] The test groups were set up as follows: vehicle group (G1), inavogliflozin monotherapy group (G2), Saxenda™ (high-dose liraglutide) monotherapy group (G3), and groups administered a depot formulation containing inavogliflozin and low-dose liraglutide every 2 or 4 weeks (G4 to G6) (n=8 per group). All test groups were administered the test substance via subcutaneous (SC) injection of 2 mg / kg containing the test substance and vehicle for 4 weeks.

[0062] Group Group name Test substance and single dose Dosage G1 Vehicle Vehicle (PBS) Once a day G2 Inavogliflozin Inavogliflozin (0.30 mg / kg) Once a day G3 Saxenda™ Liraglutide (0.31 mg / kg) Once a day G4 Manufacturing example 1 (2W) ENAVO (5.32 mg / kg), LIRA (2.17 mg / kg) Once every 2 weeks G5 Manufacturing example 2 (2W) ENAVO (4.20 mg / kg), LIRA (2.17 mg / kg) Once every 2 weeks G6 Manufacturing example 2 (4W) ENAVO (8.40 mg / kg), LIRA (4.34 mg / kg) Once every 4 weeks

[0063] As a result of analyzing the weight loss rate by group, as can be seen in Figure 1, the weight loss effect of G2 was minimal compared to G1 (-3.39%), but the combination depot groups G4 (-8.85%) and G5 (-9.26%) administered at 2-week intervals in combination with low-dose liraglutide showed a better weight loss effect than the high-dose liraglutide alone group G3 (-7.09%). In addition, the combination depot group G6 (-6.75%) administered at 4-week intervals in combination with low-dose liraglutide showed a similar weight loss effect to the high-dose liraglutide alone group G3 (-7.09%). In the case of G6, it is a formulation with significantly improved patient convenience in that it achieves a weight loss effect with just one injection compared to G3, which requires 28 injections over 4 weeks. In addition, as a result of analyzing the fat weight and the ratio of fat in the tissue through body composition analysis, as can be seen in Table 3 and Figure 2, it was confirmed that the weight loss in the depot injection group G5 containing inavogliflozin and low-dose liraglutide was similar to that in the high-dose liraglutide alone group G3 compared to the placebo group (the red part in Figure 2 represents fat).

[0064] Body fat mass and muscle mass group of SD rats fed 60% HFD administered test substances during the experimental period. Value (g) BMD (g / cm 2 )Bone Area(cm 2 )Bone Volume(cm 3)FAT In Tissue (%)Total MassBMCFATLEANG1비히클NMean856.1619.4574358.8684477.83790.244679.569411.768642.68938SD65.281.006658.857028.74930.00953.46830.60884.3588G2이나보글리플로진NMean820.0318.8277311.1294490.07050.242377.726311.387738.62818SD73.051.077752.643730.36520.00594.45650.65183.6568G3Saxenda ™ NMean793.2419.1914293.3549480.69090.242579.075411.607737.70668SD65.201.623847.818826.41590.00924.80240.98213.5796G4제조예1(2W)NMean778.9118.5477304.1375456.22130.235078.946811.218439.76807SD79.421.430854.677332.01080.00745.73220.86543.4135G5제조예2(2W)NMean775.8118.6810289.3078467.82310.239178.114411.299037.93768SD72.930.911154.330829.17600.00463.04890.55104.2511G6제조예2(4W)NMean794.1218.4799297.5711478.07340.2340*79.058811.177438.19648SD55.280.580246.407426.70880.00953.44530.35094.0630

[0065] BMC: bone mineral content; BMD: bone mineral density. Significant difference from G1 HFD vehicle control group by One-way ANOVA followed by the Dunnett's test: * p<0.05, ** p<0.01.

[0066] In addition, as shown in Table 4, in the serological analysis, the LDL-C reduction results were significantly reduced in the combination depot group G4 containing inavogliflozin and low-dose liraglutide compared to the high-dose liraglutide monotherapy group G3. In addition, the TG levels in the combination depot groups G5 and G6 containing inavogliflozin and low-dose liraglutide also significantly decreased compared to the G1 group.

[0067] Blood chemistry test results of SD rats fed 60% HFD administered test substances during the experimental period Group Blood chemistry test AST (U / L) ALT (U / L) TG (mg / dL) T-Chol (mg / dL) LDL-C (mg / dL) HDL-C (mg / dL) G1 Vehicle NMean 339.63 198.1379.25 96.75 6.88 22.638 SD30 6.04 180.52 21.83 21.17 1.55 4.84 G2 Inavogliflozin NMean 226.50 123.38 61.1386.385.1324.008 SD15 1.1996.09 18.70 18.57 0.64 3.25 G3 Saxenda ® NMean184.25105.5040.88**80.506.1322.888SD84.1299.4710.2615.720.994.64G4 Manufacturing Example 1 (2W) NMean167.2972.8652.7178.864.71*22.577SD57.8338.3712.2024.410.956.05G5 Joye2(2W)NMean157.7573.6349.63*74.385.0022.258SD79.2843.2310.9316.071.413.96G 6 Manufacturing example 2 (4W)NMean135.8855.7551.50*90.756.7521.888SD42.8327.143.8912.161.832.17

[0068] Significant difference from G1 vehicle control group by One-way ANOVA followed by the Dunnett's test: * p<0.05, ** p<0.01.AST: aspartate aminotransferase, ALT: alanine aminotransferase, TG: triglyceride T-Chol: total cholesterol, LDL-C: low density lipoprotein-cholesterol, HDL-C: high density lipoprotein-cholesterol.

[0069]

[0070] In addition, as shown in Table 5, which compares the internal organ weights of SD rats through autopsy after the end of the test, the absolute organ weight of the liver was significantly reduced in the combination depot injection groups G4, G5, and G6 containing inavogliflozin and low-dose liraglutide compared to the high-dose liraglutide monotherapy group G3. In addition, the relative organ weight in the combination depot injection group G6 containing inavogliflozin and low-dose liraglutide was also significantly reduced compared to the G1 group.

[0071] Absolute organ weight (g) and relative organ weight (organ weight / body weight ratio) at the time of sacrifice Group Body weight (g) Absolute organ weight (g) Relative organ weight Hepatorenal fat Hepatorenal fat G1 Vehicle NMean 811.13 19.50 1526.14 350.02 400.03 218 SD6 0.05 3.09 7 24.44 36 0.00 300.00 44 G2 Inavogliflozin NMean 777.17 18.68 1723.29 77 0.02 410.02 95 8 SD71.98 2.82 277.09 75 0.00 320.00 65 G3 Saxenda ®NMean744.0917.163221.52370.02310.02878SD62.642.37004.58150.00270.0041G4 Manufacturing Example 1 (2W )NMean729.4215.7746*21.62600.02180.02967SD75.521.47513.90170.00250.0042G5 Manufacturing Example 2 ( 2W)NMean735.4615.2485*20.64180.02060.02818SD69.092.75443.21070.00240.0041G6 Manufacturing example 2(4W)NMean751.3414.5738**23.48280.0194**0.03118SD52.721.11086.10030.00130.0071

[0072] Significant difference from G1 HFD vehicle control group by One-way ANOVA followed by the Dunnett's test: * p<0.05, ** p<0.01.

[0073] In conclusion, in a rat model in which obesity was induced by feeding a high-fat diet for 13 weeks, we confirmed that the combination depot injections G4 and G5 containing inavogliflozin and low-dose liraglutide administered once every two weeks had a superior weight loss effect than the group (G3) administered Saxenda™ (high-dose liraglutide), an existing obesity treatment. In addition, the combination depot injection G6 administered once every four weeks showed a similar weight loss effect to G3. In addition, the results of fat weight, serological analysis, and absolute and relative liver tissue weights were combined to show that the combination depot injection containing inavogliflozin and low-dose liraglutide has additional benefits related to improving obesity metabolism that cannot be confirmed with high-dose liraglutide administration, while at the same time reducing side effects, showing a similar weight loss effect, and further improving patient medication convenience. This was confirmed through an anti-obesity efficacy evaluation in SD rats.

[0074] Accordingly, the following study was conducted to conduct further research on the sustained drug release of the depot composition.

[0075]

[0076] [Manufacturing Examples 3 to 10]

[0077] In order to ensure sustained drug release of inavogliflozin and liraglutide, a depot composition containing inavogliflozin and liraglutide was manufactured through various attempts (S / O / W formulation method, spray-drying formulation method, and freeze-drying recovery and post-mixing formulation method after separating inavogliflozin and liraglutide and encapsulating them into microspheres). The specific manufacturing method is as follows.

[0078] In Manufacturing Examples 3 to 10, inabogliflozin and liraglutide acetate were used as active ingredients. The formulations were manufactured using the S / O / W manufacturing method, the spray drying manufacturing method, or the method of separating each active ingredient, encapsulating it in microspheres, and then post-mixing them.

[0079] First, to prepare liraglutide acetate using hydrophobic ion pairing, 10 g of liraglutide and 4.2 g of sodium acetate were dissolved in 400 ml of distilled water and mixed for 30 minutes. 10 ml of acetic acid (Glacial, Sigma) or 10 ml of 1 N HCl was added to the mixture to lower the pH, thereby precipitating liraglutide acetate, which was then further reacted for 4 hours. After repeated washing with distilled water and methanol, and filtering through a 635 mesh sieve, the recovered liraglutide acetate was lyophilized and recovered.

[0080] In Manufacturing Example 3, the PLA polymer of R202H was dissolved in methylene chloride at a weight ratio of 30 w / w% and used, and inabogliflozin was completely dissolved in methanol at a weight ratio of 16.7 w / w%. Then, the polymer solution and solid liraglutide acetate were mixed using a homomixer at 10,000 RPM for 3 minutes to prepare a first mixture (S / O emulsion). After that, the first mixture (S / O emulsion) was put into a 5 L water tank containing a surfactant (PVA) at a weight ratio of 0.5%, and at the same time, a homomixer was applied at 4500 RPM for 3 minutes to prepare a second mixture (S / O / W microspheres). For the remaining volatilization process, the water tank temperature was adjusted to 30℃ and stirring was performed at 150 RPM using an overhead stirrer for 3 hours. Afterwards, the microspheres were recovered through centrifugation at 1500~3000 RPM for 3 minutes, and washed three times with distilled water to wash away the unencapsulated active ingredient and surfactant. Afterwards, the microsphere powder was recovered by freeze-drying at -40℃~5℃ for 48 hours. The recovered microspheres were finally secured by passing them through a 200 mesh sieve.

[0081] In Manufacturing Example 4, the PLA polymer of R203H was dissolved in methylene chloride at a weight ratio of 23 w / w% and used, and inabogliflozin was completely dissolved in methanol at a weight ratio of 16.7 w / w%. Then, the polymer solution and solid liraglutide acetate were mixed using a homomixer at 10,000 RPM for 3 minutes to prepare a first mixture (S / O emulsion). After that, the first mixture (S / O emulsion) was put into a 1 L water tank containing a surfactant (PVA, Gohsenol EG-40PW) at a weight ratio of 1.0 w / w%, and at the same time, a homomixer was applied at 3500 RPM for 3 minutes to prepare the second mixture (S / O / W microspheres). The remaining volatilization process was performed by adjusting the water tank temperature to 30℃ and stirring was performed at 150 RPM using an overhead stirrer for 3 hours. After that, the microspheres were recovered through centrifugation at 1500 to 3000 RPM for 3 minutes, and the washing was repeated 3 times using primary distilled water to wash away the unencapsulated active ingredient and surfactant. After that, the microsphere powder was recovered by freeze-drying at -40℃ to 5℃ for 48 hours. The recovered microspheres were finally obtained by passing through a 200 Mesh sieve.

[0082] In Manufacturing Example 5, the PLA polymer of R203H was dissolved in methylene chloride at a weight ratio of 15 w / w% and used, and inabogliflozin was completely dissolved in methanol at a weight ratio of 16.7 w / w%. Then, the polymer solution and solid liraglutide acetate were mixed using a homomixer at 15,000 RPM for 3 minutes to create a mixture (S / O emulsion), and then microspheres were prepared using a spray dryer (B”U”CHI, Mini B-290). Here, the mixture was injected at a speed of 1.0 ml / min using an injection pump, the inlet temperature was set to 55 to 75°C, the outlet temperature was set to 36 to 42°C, and the mixture was sprayed at a pressure of 20 to 50 psi to create microspheres and then recovered.

[0083] In Manufacturing Example 6, the PLA polymer of R203H was dissolved in methylene chloride at a weight ratio of 23 w / w% and used, and inabogliflozin was completely dissolved in methanol at a weight ratio of 16.7 w / w%. Then, the polymer solution and solid liraglutide acetate were mixed using a homomixer at 10,000 RPM for 3 minutes to prepare a first mixture (S / O emulsion). After that, the first mixture (S / O emulsion) was put into a 1 L water tank containing a surfactant (PVA, Merck 05P) at a weight ratio of 1.0%, and at the same time, a homomixer was applied at 3500 RPM for 3 minutes to prepare the second mixture (S / O / W microspheres). The remaining volatilization process was performed at 150 RPM for 3 hours using an overhead stirrer while adjusting the water tank temperature to 35℃. After that, the microspheres were recovered through centrifugation at 1500-3000 RPM for 3 minutes, and the washing was repeated 3 times using primary distilled water to wash away the unencapsulated active ingredient and surfactant. After that, the microsphere powder was recovered by freeze-drying at -40℃ to 5℃ for 48 hours. The recovered microspheres were finally secured by passing through a 200 Mesh sieve.

[0084] In the case of Manufacturing Example 7, inabogliflozin and liraglutide acetate were prepared separately without mixing. In the case of biodegradable polymers, PLA polymer of R203H was dissolved in methylene chloride at a weight ratio of 23% and used, and after inabogliflozin was completely dissolved in methanol at a weight ratio of 16.7%, the polymer solution was mixed using a homomixer at 10,000 RPM for 3 minutes to create a first mixture (S / O emulsion), and the subsequent process was to put it into a 1 L water tank containing a surfactant (PVA, Merck 05P) at a weight ratio of 1.0% and simultaneously apply a homomixer at 3500 RPM for 3 minutes to create a second mixture (S / O / W microspheres). The remaining volatilization process was performed using an overhead stirrer at 150 RPM for 3 hours by adjusting the water tank temperature to 30℃. Afterwards, the microspheres were recovered through centrifugation at 1500~3000 RPM for 3 minutes, and washed three times using distilled water to wash away the unencapsulated active ingredient and surfactant. Afterwards, the microsphere powder was recovered by freeze-drying at -40℃~5℃ for 48 hours. The recovered microspheres were finally secured by passing through a 200 mesh sieve. In addition, liraglutide acetate was used by dissolving the PLA polymer of R203H in methylene chloride at a weight ratio of 23%, and the other manufacturing processes were manufactured in the same conditions as the preparation of microspheres in which inavogliflozin was separated and encapsulated.

[0085] In the case of Manufacturing Example 8, the PLA polymer of R202H was dissolved in methylene chloride at a weight ratio of 30% and used, and after inabogliflozin was completely dissolved in methanol at a weight ratio of 16.7%, the polymer solution and solid liraglutide acetate were mixed using a homomixer at 15,000 RPM for 3 minutes to create a first mixture (S / O emulsion), and then the subsequent process was to put it into a 1 L water bath containing a surfactant (PVA, Merck 05P) at a weight ratio of 1.0% and simultaneously apply a homomixer at 3500 RPM for 3 minutes to create a second mixture (S / O / W microspheres). The remaining volatilization process was performed using an overhead stirrer at 150 RPM for 3 hours while adjusting the water bath temperature to 30℃. Afterwards, the microspheres were recovered through centrifugation at 1500~3000 RPM for 3 minutes, and washed once with PBST (Tween 80, 0.05%) to wash away the unencapsulated active ingredient and surfactant, and then washed three times with distilled water. Afterwards, the microsphere powder was recovered by freeze-drying at -40℃~5℃ for 48 hours. The recovered microspheres were finally secured by passing through a 200 mesh sieve.

[0086] The PLA polymer of R203H of Manufacturing Example 9 was used by dissolving it in methylene chloride at a weight ratio of 23%, and after completely dissolving inavogliflozin in methanol at a weight ratio of 16.7%, 1N HCl was used instead of acetic acid to precipitate the polymer solution and solid liraglutide acetate, and using a homomixer, mixing at 15,000 RPM for 2.5 minutes to make the first mixture (S / O emulsion), and the subsequent process was to put it into a 1 L water bath containing a surfactant (PVA, Merck 05P) at a weight ratio of 1.0% and apply the homomixer at 3500 RPM for 3 minutes to make the second mixture (S / O / W microspheres), and for the other volatilization process, the water bath temperature was adjusted to 30℃ and an overhead stirrer was used at 150 RPM for 3 hours. The microspheres were recovered by centrifugation at 1500-3000 RPM for 3 minutes, and then washed once with PBST (Tween 80, 0.05%) to wash away the unencapsulated active ingredient and surfactant, and then washed three times with distilled water. Afterwards, the microsphere powder was recovered by freeze-drying at -40℃ to 5℃ for 48 hours. The recovered microspheres were finally secured by passing through a 200 mesh sieve.

[0087] In the case of Manufacturing Example 10, the PLA polymer of R203H was dissolved in methylene chloride at a weight ratio of 23% and used, and inabogliflozin was completely dissolved in methanol at a weight ratio of 16.7%, and then the polymer solution and solid liraglutide acetate were precipitated and recovered, and then washed repeatedly with distilled water for the first time and then washed with methanol for the final time to secure the precipitation, and then mixed for 3 minutes at 10,000 RPM using a homomixer to make the first mixed solution (S / O emulsion), and then the subsequent process was to put it into a 1 L water bath containing a surfactant (PVA, Merck 05P) at a weight ratio of 1.0% and apply the homomixer at 3500 RPM for 3 minutes to make the second mixed solution (S / O / W microspheres), and for the other volatilization processes, the water bath temperature was adjusted to 30℃ and an overhead stirrer was used for 3 hours. The mixture was centrifuged at 150 RPM. Afterwards, the microspheres were recovered through centrifugation at 1500-3000 RPM for 3 minutes, and washed once with PBST (Tween 80, 0.05%) to wash away the unencapsulated active ingredient and surfactant, and then washed three times with distilled water. Afterwards, the microsphere powder was recovered by freeze-drying at -40℃ to 5℃ for 48 hours. The recovered microspheres were finally secured by passing through a 200 mesh sieve.

[0088] To confirm the diameter and particle size distribution of the composite microspheres containing inabogliflozin and liraglutide acetate, analysis was conducted using a Malvern particle size analyzer.

[0089] Specifically, the depot compositions of Manufacturing Examples 3 to 10 were obtained according to the composition and manufacturing method of Table 6.

[0090] Manufacturing Example Manufacturing Method Polymer Concentration and Type Additives and Features ENAVO Concentration w / w % Continuous Phase Conditions 1st Homogenization Conditions (RPM) 2nd Homogenization Conditions (RPM) Volatilization Conditions Temperature / Time / RPM Size D10 / D50 / D90 / D[4,3]3 S / O / W 30% PLAR20 2 HLIRA-Acetate 16.7 0.50% PVA 5L Gohsenol EG-40 PW 10,000 3 min 4,500 3 min 30°C / 3 / 150 2.3 / 13.0 / 37.4 / 16.94 S / O / W 23% PLAR20 3 HLIRA-Acetate 16.7 1.0%, PVA 1L Gohsenol EG-40 PW 10,000 3 min 3,500 3 min 30°C / 3 / 1501.9 / 13.0 / 34.2 / 15.75SprayDrying15% PLAR203HLIRA-Acetate16.71.0%, PVA 1LMerckPVA05P15,0003minN.AN.A2.7 / 17.6 / 44.2 / 21.66S / O / W23% PLAR203HLIRA-Acetate16.71.0%, PVA 1LMerckPVA05P10,0003min3,5003min35°C / 3 / 1502.0 / 15.6 / 36.4 / 17.57O / W23% PLAR202HENAVOSeparating agent16.71.0%, PVA 1LMerckPVA05P10,0003min3,5003min30°C / 3 / 1502.0 / 14.6 / 33.3 / 18.1S / O / W23% PLAR203HLIRA-AcetateN.A1.0%, PVA 1LMerckPVA05P10,0003 minutes3,5003 minutes30°C / 3 / 1502.0 / 16.6 / 37.6 / 22.28S / O / W30% PLAR202HLIRA-Acetate / PBST0.05%washing16.71.0%, PVA 1LMerckPVA05P15,0003 minutes3,5003 minutes30°C / 3 / 1508.0 / 31.2 / 64.0 / 34.09S / O / W23% PLAR203HLIRA-Acetate(HCl)16.71.0%, PVA 1LMerckPVA05P15,0002.5min3,5003min30°C / 3 / 1505.2 / 24.4 / 51.2 / 26.910S / O / W30% PLAR202HLIRA-Acetate / MeOH Washing16.71.0%, PVA 1LMerckPVA05P10,0003 minutes3,5003 minutes30°C / 3 / 1507.4 / 28.8 / 65.1 / 32.1.

[0091] [Experimental Example 2] PK analysis according to administration of the depot compositions of Manufacturing Examples 3 to 10

[0092] For PK analysis according to administration of the depot compositions of Manufacturing Examples 3 to 10, 5-week-old male SD rats were used as an animal model.

[0093] The test groups were set as the inabogliflozin + liraglutide combination administration group (G1) and the depot formulation administration group of Manufacturing Examples 3 to 10 containing inabogliflozin and liraglutide (G2 to G9), as shown in Table 7 below (G1: n=3, G2 to G9: n=2 per group).

[0094] Test substance Dosage Dose (mg / kg) Volume (mL / kg) G1 Inavogliflozin + liraglutide SC Once a day Inavogliflozin 0.093 + liraglutide 0.186 Inavogliflozin 0.93 + liraglutide 1.86 G2 Manufacturing example 3 SC Single Inavogliflozin 1.87 + liraglutide 2.60 G3 Manufacturing example 4 Inavogliflozin 2.86 + liraglutide 2.60 G4 Manufacturing example 5 Inavogliflozin 2.77 + liraglutide 2.60 G5 Manufacturing example 6 Inavogliflozin 3.00 + liraglutide 2.60 G6 Manufacturing example 7 Inavogliflozin 2.60 + liraglutide 2.60 G7 Manufacturing example 8 Inavogliflozin 2.30 + Liraglutide 2.60G8 Manufacturing Example 9 Nabogliflozin 1.68 + Liraglutide 2.60G9 Manufacturing Example 10 Nabogliflozin 1.86 + Liraglutide 2.60

[0095] Plasma samples were collected at specified times and stored at -70°C until analysis using an LC-MS / MS system. Protein precipitation was performed on 20 μL of plasma samples using 180 μL of acetonitrile containing internal standards. After mixing and centrifugation, the supernatant (150 μL) was analyzed using an LC-MS / MS system with MRM mode. LC / MS / MS data were analyzed using Analyst 1.6.3. PK parameters were calculated using noncompartmental analysis using WinNonlin software. Figure 3 shows the PK analysis results of inavogliflozin according to the administration of each test substance.

[0096] As a result of evaluating the level of sustained drug release through blood drug concentration measured 4 weeks (total 672 h) after administration of the test substance, the drug was detected at 2 weeks (336 h point) in G2 / G4 / G7 / G9 compared to the ENAVO control drug administration group (G1), and the drug was detected up to 4 weeks (672 h point) in G7 and G9 compared to the ENAVO control drug administration group (G1), confirming that the drug was continuously released.

[0097] Figure 4 shows the PK analysis results of liraglutide according to administration of each test substance.

[0098] The sustained drug release level was evaluated through the blood drug concentration measured 4 weeks (total 672 h) after administration of the test substance. As a result, the drug was detected at 2 weeks (336 h point) in G2 / G7 / G9 compared to the LIRA control drug administration group (G1), confirming that the drug was continuously released.

[0099] In summary, it was confirmed that in the case of the depot compositions of Manufacturing Examples 3, 8, and 10, both inagliflozin and liraglutide continued to release the drugs for more than two weeks.

[0100] More preferably, it was confirmed that the liraglutide drug release of the depot composition of Manufacturing Example 10 was maintained at a relatively higher level than other Manufacturing Examples at 2 weeks (336 h point).

Claims

1. A depot composition comprising microspheres comprising inabogliflozin and liraglutide as active ingredients; and a biodegradable polymer.

2. A depot composition in the first paragraph, wherein the biodegradable polymer is selected from the group consisting of polylactide (PLA), polyglycolide (PGA), poly(lactide-co-glycolide) (PLGA), and mixtures thereof.

3. A depot composition according to claim 1, wherein the biodegradable polymer has a weight average molecular weight of 4,000 to 50,000.

4. A depot composition according to claim 1, wherein the microparticles are prepared by a solvent evaporation method or a spray drying method.

5. A depot composition in the first paragraph, wherein the microparticles are W / O / W type microparticles.

6. A depot composition in claim 5, wherein the W / O / W type microparticles contain sodium acetate or magnesium hydroxide as an additive during the production of a W / O emulsion.

7. A depot composition in the first paragraph, wherein the microparticles are S / O / W type microparticles.

8. A depot composition in claim 7, wherein the S / O / W type microparticles are liraglutide and contain liraglutide acetate in a solid state.

9. A depot composition according to claim 1, wherein the microparticles have an average particle size of 10 to 100 um.

10. A depot composition in claim 1, wherein the drug content ratios of inabogliflozin and liraglutide in the microparticles are each 10 to 20 wt%.

11. In the first paragraph, the depot composition is a depot composition for preventing or treating obesity.

12. A depot composition according to claim 1, wherein the depot composition is injected subcutaneously once every 2 to 4 weeks.

13. A depot composition in claim 1, wherein the dosage of inabogliflozin is 0.3 to 1.5 mg / day, and the dosage of liraglutide is 1.2 to 2.4 mg / day.

14. A depot composition in claim 1, wherein the dosage of inabogliflozin is 0.6 to 1.2 mg / day, and the dosage of liraglutide is 1.5 to 2 mg / day.

15. A kit comprising a depot composition, a dispersion medium and a syringe according to any one of claims 1 to 14.

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