Pharmaceutical composition in form of eye drops comprising enagliflozin

Abstract

The present invention provides a pharmaceutical composition in the form of eye drops, the pharmaceutical composition comprising: enagliflozin or a pharmaceutically acceptable salt thereof as an active ingredient; and a combination of polysorbate and polyoxyethylene 40 stearate as a solubilizing agent and a stabilizing agent. According to the pharmaceutical composition disclosed by the invention, through the combination containing the specific surfactants, the stability can be obviously improved, and the exposure of the enagliflozin in eyeballs can be obviously increased.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical composition in the form of eye drops containing enagliflozin. More specifically, the present invention relates to a pharmaceutical composition in the form of eye drops, comprising: enagliflozin or a pharmaceutically acceptable salt thereof as an active ingredient; and a combination of specific surfactants, wherein the pharmaceutical composition provides excellent stability and increased intraocular exposure. Background Art

[0002] Enagagliflozin has the chemical structure of the following Chemical Formula 1. It has inhibitory activity against sodium-dependent glucose cotransporter 2 (SGLT2) present in the intestine and kidney and can be useful for treating metabolic disorders, particularly diabetes (WO2012 / 165914, WO2017 / 217792, etc.).

[0003] <Chemical Formula 1>

[0004]

[0005] The present inventors have found that an SGLT2 inhibitor comprising enagliflozin or a pharmaceutically acceptable salt thereof has excellent preventive or therapeutic activity for diabetic eye diseases (e.g., diabetic retinopathy) (Korean Patent Publication No. 10-2022-0079480). In addition, the present inventors have found that an SGLT2 inhibitor comprising enagliflozin or a pharmaceutically acceptable salt thereof has excellent preventive or therapeutic activity for macular degeneration (Korean Patent Publication No. 10-2023-0007963).

[0006] Enagliptin is a drug with low water solubility, so it is difficult to formulate it into an aqueous solution, such as an eye drop in the form of an aqueous solution. In order to solve these problems, Korean Patent Publication Nos. 10-2022-0079480 and 10-2023-0007963 disclose the use of polyoxyethylene 35 castor oil (Kolliphor) as a solubilizer. TM ELP) and polysorbate 80 (Tween TM 80) is used to solubilize enagliflozin eye drops.

[0007] On the other hand, in order to treat posterior segment diseases such as diabetic retinopathy and macular degeneration, drug delivery in the eyeball is required. In particular, in order for the drug to reach the affected area (e.g., retina), it is not only necessary to effectively solubilize the drug, but also to increase the exposure of the drug in the eyeball by allowing the dissolved drug to reach the retina through the cornea or conjunctiva. In addition, patients with posterior segment diseases such as diabetic retinopathy and macular degeneration need long-term repeated administration, so it is necessary to ensure excellent stability and prevent precipitation of the drug or generation of degradation substances from the preparation (e.g., eye drops). Summary of the Invention

[0008] (1) Technical issues to be resolved

[0009] The present inventors have found that existing eye drops containing enagliflozin (for example, the eye drops disclosed in Korean Patent Publication No. 10-2022-0079480 and No. 10-2023-0007963) may have significantly reduced stability during long-term storage, and furthermore cannot provide satisfactory eyeball exposure. In order to solve these problems, the present inventors have conducted various formulation studies. As a result, the present inventors found that when a specific combination of surfactants is used for formulation, the physical stability and chemical stability of the eye drops containing enagliflozin can be significantly increased. That is, the present inventors found that the combination of the specific surfactants not only acts as a solubilizer in the eye drops containing enagliflozin, but also acts as a stabilizer. In addition, the present inventors found that the eye drops obtained using the specific combination of surfactants can provide excellent pharmacological activity by significantly increasing the exposure of enagliflozin in the eyeball.

[0010] Therefore, an object of the present invention is to provide a pharmaceutical composition in the form of eye drops, comprising enagliflozin or a pharmaceutically acceptable salt thereof and a combination of the specific surfactant.

[0011] (2) Technical solution

[0012] According to one embodiment of the present invention, a pharmaceutical composition in the form of eye drops is provided, comprising: enagliflozin or a pharmaceutically acceptable salt thereof as an active ingredient; and a combination of polysorbate and polyoxyethylene 40 stearate as a solubilizer and stabilizer.

[0013] The concentration of enagliflozin or a pharmaceutically acceptable salt thereof may be 0.1-10 w / v% (weight / volume %), preferably 0.3-8 w / v%, and more preferably 0.5-5 w / v%.

[0014] The concentration of the combination of polysorbate and polyoxyethylene 40 stearate may be 1-15 w / v%, preferably 4-10 w / v%. The weight ratio of polysorbate to polyoxyethylene 40 stearate may be 1:1 to 10, preferably 1:2 to 8.

[0015] The pharmaceutical composition of the present invention can form nanomicelles with an average particle size of 1-500 nm, preferably nanomicelles with an average particle size of 5-50 nm.

[0016] The pharmaceutical composition of the present invention may further comprise one or more additives selected from a penetration enhancer, an osmotic pressure regulator, and a pH regulator. In one embodiment, the penetration enhancer may be D-α-tocopheryl polyethylene glycol succinate. In another embodiment, the osmotic pressure regulator may be glycerol. In another embodiment, the pharmaceutical composition of the present invention may have an osmotic pressure of 230-350 mOsmol / kg and / or a pH of pH 6.0 to pH 7.5.

[0017] (3) Beneficial effects

[0018] The pharmaceutical composition in the form of an eye drop according to the present invention can significantly increase physical and chemical stability by including a specific surfactant combination, namely, a combination of polysorbate and polyoxyethylene 40 stearate. Thus, the present invention discloses that the combination of polysorbate and polyoxyethylene 40 stearate functions as a solubilizer and stabilizer. Furthermore, the pharmaceutical composition of the present invention, including the specific surfactant combination, can provide excellent pharmacological activity by significantly increasing the exposure of enagliflozin to the eyeball. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The surfactant mixture (Tween TM 80 and Kolliphor TM The results were obtained by measuring the saturation solubility of enagliflozin (a mixture of EL).

[0020] Figure 2 In the preparation process of Formulation Example-1, the appearances in the dissolution-1 process (40° C.), the dissolution-2 process (80° C.), and the filtration process (25° C.) are shown.

[0021] Figure 3a 、 Figure 3b and Figure 3c Preparation Example-3 ( Figure 3a )、Preparation Example-4( Figure 3b ) and Preparation Example-5( Figure 3c ) of the preparation.

[0022] Figure 4 The drug concentration curve in the rat eyeball obtained by performing a pharmacokinetic study is shown. DETAILED DESCRIPTION

[0023] The present invention provides a pharmaceutical composition in the form of eye drops, comprising: enagliflozin or a pharmaceutically acceptable salt thereof as an active ingredient; and a combination of polysorbate and polyoxyethylene 40 stearate as a solubilizer and stabilizer.

[0024] The pharmaceutical composition of the present invention may contain enagliflozin or a pharmaceutically acceptable salt thereof in an appropriate amount for the treatment of posterior ocular diseases such as diabetic retinopathy and macular degeneration. For example, the concentration of enagliflozin or a pharmaceutically acceptable salt thereof in the pharmaceutical composition may be 0.1-10 w / v%, preferably 0.3-8 w / v%, and more preferably 0.5-5 w / v%.

[0025] The present invention discloses that the use of a specific surfactant combination, namely, a combination of polysorbate and polyoxyethylene 40 stearate, can significantly enhance the physical and chemical stability of eye drops containing enagliflozin. Furthermore, the present invention discloses that pharmaceutical compositions comprising this specific surfactant combination can provide superior pharmacological activity by significantly increasing the intraocular exposure of enagliflozin.

[0026] The polysorbate can be polysorbate 20, polysorbate 40, polysorbate 80, etc., and can preferably be polysorbate 80. The polyoxyethylene 40 stearate (polyoxyl 40 stearate) is also known as polyethylene glycol monostearate (polyethylene glycol monostearate). In the pharmaceutical composition of the present invention, the concentration of the combination of the polysorbate and polyoxyethylene 40 stearate in the pharmaceutical composition can be 1-15w / v%, and can preferably be 4-10w / v%. The weight ratio of the polysorbate and polyoxyethylene 40 stearate can be 1:1 to 10, and can preferably be 1:2 to 8. In a specific embodiment, the concentration of polysorbate in the pharmaceutical composition can be 0.1-10w / v%, and can preferably be 1-4w / v%; within the above-mentioned weight ratio, the concentration of polyoxyethylene 40 stearate in the pharmaceutical composition can be 0.1-10w / v%, and can preferably be 3-7w / v%. The pharmaceutical composition of the present invention forms nanomicelles with an average particle size of 1-500 nm, preferably 5-50 nm, by comprising a combination of polysorbate and polyoxyethylene 40 stearate.

[0027] The pharmaceutical composition of the present invention can contain the additive that is commonly used in eye drop field.For example, the pharmaceutical composition of the present invention can further include one or more additives selected from penetration enhancer, osmotic pressure regulator and pH regulator.In a specific embodiment, the penetration enhancer can be D-α-tocopheryl polyethylene glycol succinate (TPGS), and its concentration can be for example 0.1-0.5w / v%.In another specific embodiment, the osmotic pressure regulator can be glycerol, and its concentration can be for example 0.5-6w / v%.In another specific embodiment, the pharmaceutical composition of the present invention can have the osmotic pressure of 230-350mOsmol / kg and / or the pH of pH6.0 to pH7.5.

[0028] In one embodiment of the present invention, a pharmaceutical composition is provided, comprising: 0.5-5 w / v% enagliflozin or a pharmaceutically acceptable salt thereof; 4-10 w / v% of a combination of polysorbate and polyoxyethylene 40 stearate; 0.1-0.5 w / v% of D-α-tocopheryl polyethylene glycol succinate; 0.5-6 w / v% of glycerol; and a pH adjuster in an aqueous medium, wherein the pharmaceutical composition forms nanomicelles having an average particle size of 5-50 nm. In this embodiment, the weight ratio of the polysorbate to the polyoxyethylene 40 stearate can be 1:2 to 8.

[0029] Hereinafter, the present invention will be described in more detail by way of examples. However, these examples are merely for illustrating the present invention and the present invention is not limited to these examples.

[0030] Example 1: Formulation Study-1

[0031] (1) Evaluation of solubility

[0032] Using the FDA inactive ingredient database (https: / / www.accessdata.fda.gov / scripts / cder / iig / index.cfm), additives that can be used for ophthalmic administration were dissolved in purified water at the maximum concentration recommended by the FDA, and then enagliflozin was dissolved to saturation to measure the saturated solubility.

[0033] (1-1) Preparation of standard solution

[0034] According to Table 1 below, place the corresponding amount of enagliflozin for each standard solution and 70 mL of methanol into a volumetric flask. After complete dissolution using ultrasonic agitation, add methanol to bring the volume to 100 mL. Filter this solution through a 0.45 μm regenerated cellulose (RC) membrane filter, discard the first 2 mL, and use the filtrate as the standard solution.

[0035] [Table 1]

[0036]

[0037] (1-2) Preparation of test solution

[0038] Each additive was dissolved in 5 mL of purified water at the maximum concentration recommended by the FDA (Table 2). Enagagliflozin was then added until no further dissolution occurred and stirred for approximately 12 hours. After confirming that no further dissolution had occurred after 12 hours, each sample was centrifuged at 4000 rpm for 30 minutes. The clear supernatant (1 mL) was collected and diluted with methanol. This solution was filtered through a 0.45 μm RC membrane filter, and the first 2 mL was discarded. The filtrate was used as the test solution.

[0039] (1-3) HPLC analysis

[0040] The peak areas of the standard solution and the test solution were analyzed under the following HPLC conditions.

[0041] -Detector: UV spectrophotometer (measurement wavelength is 225nm)

[0042] -Chromatographic column: Capcellpak C18 (4.6×250mm, 5μm)

[0043] - Column temperature: about 35°C

[0044] - Sample temperature: about 25°C

[0045] -Mobile phase: buffer: acetonitrile = 25:75 (v / v)

[0046] (Buffer solution: Dissolve 100 μL of trifluoroacetic acid in 1000 mL of water)

[0047] -Flow rate: 1.0mL / min

[0048] -Injection volume: 10μL

[0049] -Analysis time: about 20 minutes

[0050] (1-4) Solubility of each additive

[0051] The measurement results of the saturation solubility of enagliflozin in various additives are shown in Table 2 below.

[0052] [Table 2]

[0053]

[0054] As can be seen from the results in Table 2, Tween TM 80 and Kolliphor TM EL has the highest improvement on the solubility of enagliflozin. TPGS, as a penetration enhancer, also has a relatively high improvement on the solubility of enagliflozin. Kolliphor, as a surfactant TM HS 15, Twain TM 20 and Kolliphor TM RH 40 also has a similar solubilizing effect, but its limitation is that its maximum usage as an eye drop additive is limited.

[0055] (2) Evaluation of saturated solubility according to surfactant concentration

[0056] According to the results of (1), the surfactant Tween was selected TM 80 and Kolliphor TM EL is used as an additive to solubilize enagliflozin. The target concentration of enagliflozin in eye drops is 0.5-5.0 w / v%, but saturation solubility does not reach this level. Therefore, the two most effective solubilizing ingredients are mixed, and the saturation solubility of the main ingredient is measured based on the content ratio.

[0057] (2 / 1) Test methods

[0058] Will Twain TM 80 with Kolliphor TM EL was mixed according to the ratio in Table 3 below, heated to 40°C, and a certain amount of enagliflozin was added to dissolve it. The point at which enagliflozin no longer dissolved and became turbid was taken as the saturation solubility.

[0059] [Table 3]

[0060]

[0061] (2 / 2) Test Results

[0062] Saturation solubility was analyzed by summing the fraction of enagliflozin that dissolved to the node that maintained its appearance. Figure 1 The results showed that the solubility of enagliflozin increased proportionally with the increase in the total concentration of surfactants, and in order to dissolve it to the target concentration of 5.0 w / v%, the total concentration of surfactants had to be at least 5.0 w / v%.

[0063] (3) Preparation and evaluation of preparations

[0064] (3-1) Preparation of preparations

[0065] According to the test results, eye drops containing enagliflozin were prepared according to the ingredients and contents in Table 4 below (Formulation Example 1: 5.0 w / v%, Formulation Example 2: 2.0 w / v%). TM 80. Kolliphor TM EL was used as a surfactant, TPGS was used as a penetration enhancer, and glycerol was used as an osmotic pressure regulator. TM 80. Kolliphor TM EL, glycerol, anhydrous sodium dihydrogen phosphate, sodium hydroxide) and water for injection are heated to about 40°C and stirred (dissolution-1 process). The main ingredient (enagliflozin) is added and heated to about 80°C (dissolution-2 process). Thereafter, the solution is cooled to 25°C and then filtered through a 0.2μm RC, PTFE or PVDF membrane filter (filtration process). The obtained individual eye drops are filled in polyethylene tubes.

[0066] [Table 4]

[0067]

[0068] (3-2) Evaluation of preparations

[0069] Filters made of RC, PTFE, or PVDF were used as membrane filters. The main component content before and after filtration was analyzed by HPLC using the same method as described in (1) to confirm whether the main component was adsorbed on the filter membrane. The results are shown in Table 5 below.

[0070] [Table 5]

[0071]

[0072] From the results in Table 5, it can be seen that no matter what the material of the filter used in the filtration process is, the main component does not show obvious adsorption, and the most commonly used RC material is selected as the filter for the filtration process.

[0073] In addition, the appearance, content, pH, and osmotic pressure of Preparation Example 1 and Preparation Example 2 were evaluated. The appearance was visually confirmed, and the content was analyzed by HPLC analysis using the same method as described in (1). The pH was measured using a Metrohm 913 product, and the osmotic pressure was measured using an OSMOMAT 3000D product. The results of the evaluation of the preparations as described above are shown in Table 6 below.

[0074] [Table 6]

[0075]

[0076] During the dissolution process of the main component, a phenomenon was observed during heating where the main component was not fully dissolved and was suspended. This is presumably due to the precipitation of the main component at a temperature above the cloud point of the nonionic surfactant. It is presumed that during the subsequent cooling process, when the temperature is cooled below the cloud point, the surfactant will reassemble and form nanomicelles, thereby dissolving the main component. (Refer to Figure 2 )

[0077] The osmotic pressure of Formulation Example 1 slightly exceeded the target standard, suggesting that the dosage of the osmotic pressure regulator may need to be adjusted. Although the osmotic pressure of Formulation Example 1 did not meet the target standard, Formulation Example 1 was used as a sample in Formulation Study 2 of Example 2 to observe the trends of degradation substances.

[0078] Example 2: Formulation Study-2

[0079] (1) Preparation of preparations

[0080] Based on the test results of Example 1 and the components and contents in Table 7, the concentrations of the main ingredient were set at 1.0 w / v%, 3.0 w / v%, and 5.0 w / v%, respectively. Eye drops were prepared using the same method as in Example 1 (3-1) (filtration using an RC membrane filter). Since the eye drops containing 1.0 w / v% of the main ingredient were considered sufficient in terms of saturation solubility, the amount of surfactant was reduced by 30%. In addition, the amounts of pH adjusters and osmotic pressure adjusters were adjusted to meet the pH and osmotic pressure standards (Table 7).

[0081] [Table 7]

[0082]

[0083] (2) Evaluation of appearance, content, pH, and osmotic pressure

[0084] Formulation Example 3, Formulation Example 4, and Formulation Example 5 were evaluated for appearance, content, pH, and osmotic pressure using the same method as in (3-2) of Example 1. The results are shown in Table 8 below.

[0085] [Table 8]

[0086]

[0087] The results in Table 8 confirm that the formulations of Formulation Example 3, Formulation Example 4, and Formulation Example 5 meet the target standards. Considering that the pH values measured at all concentrations were nearly identical, it is believed that the pH adjuster fully exerted its buffering effect. Furthermore, the osmotic pressure did not increase proportionally with the amount of the main ingredient or surfactant.

[0088] (3) Evaluation of particle size distribution and zeta potential

[0089] Tween, a solubilizing agent used to achieve the target concentration of enagliflozin as the main ingredient TM 80 and Kolliphor TM EL forms micelles at a concentration above the critical micelle concentration. Since the formation of micelles or nanomicelles also affects the drug delivery efficiency, it is confirmed whether the preparations of Preparation Example-3, Preparation Example-4 and Preparation Example-5 form nanomicelles. Dynamic light scattering (DLS) is used to analyze the distribution of particles formed in the solution, and the zeta potential (zeta-potential) is measured simultaneously to further confirm the charge (charge) formed on the particle surface. DLS measurement equipment uses Otsuka's ELSZ-2000 for measurement, and the measurement conditions are shown in Table 9 below.

[0090] [Table 9]

[0091]

[0092] The analysis results of the particle size distribution of the preparations of Preparation Example-3, Preparation Example-4 and Preparation Example-5 are shown in Table 10 and Figure 3 (i.e., Figure 3a 、 Figure 3b 、 Figure 3c In addition, the measurement results of Zeta potential are shown in Table 10 below.

[0093] [Table 10]

[0094]

[0095] The results in Table 10 show that the average particle size (Z-average) ranged from 11.9 to 29.4 nm, showing an increasing trend with increasing proportion of the main component. However, all formulations exhibited particle sizes below 10 nm using the number of distributed particles (distribution number) criterion. Zeta potential was confirmed to be weakly negative.

[0096] Example 3: Formulation Study-3

[0097] (1) Evaluation of stability

[0098] Contains Kolliphor TM EL and Twain TM The results of evaluating the stability of Formulation Example 2 of the combination of 80% and 1% PEG-100 was shown in Table 11 below while being stored under accelerated conditions for 2 months and at room temperature for 4 months.

[0099] [Table 11]

[0100]

[0101] As shown in the results of Table 11, Formulation Example 2 exhibited precipitation or exhibited a content below the standard value during stability testing at room temperature for 4 months, accelerated for 2 months, and accelerated for 4 months, making it unsuitable. While a decrease in the content of the main component due to decomposition can significantly increase the amount of degradation substances, the increase in the amount of degradation substances in enagliflozin was not significant.

[0102] (2) Improvement of formulation

[0103] The use of surfactants improves the solubility of poorly soluble enagliflozin, leading to speculation that the interaction between the main ingredient and the surfactant may affect stability during storage. Therefore, an indirect assessment can be made by measuring the cloud point, where solubility plummets when the water-soluble groups of the nonionic surfactant are dehydrated, during heating. This led to the identification of the cause of the formulation instability, and further formulation studies were conducted by modifying the solubilizing agent to further stabilize the formulation.

[0104] (2-1) Evaluation of Solubilization of Polyoxyethylene 40 Stearate

[0105] Polyoxyl 40 stearate (Myrj TM S40) Dissolve the mixture in 5 mL of purified water at a concentration of 7.0 w / v%, add enagliflozin until it no longer dissolves, and then stir for about 12 hours. Thereafter, the degree of solubilization was evaluated using the same method as in Example 1.

[0106] (2-2) Measurement of cloud point

[0107] The cloud point is measured by visually observing the point at which the test formulation becomes opaque when slowly heated. 5 mL of the test formulation (i.e., the formulations of Formulation Examples 6, 7, and 8, and the formulation of Formulation Example 1, prepared in (2-3) below) is placed in a clear glass bottle and stirred while heating in a water bath by increasing the temperature by 0.5°C. The cloud point is the temperature at which the formulation changes from a clear, transparent liquid to a turbid suspension.

[0108] (2-3) Preparation of preparations

[0109] According to the components and contents in Table 12, the concentrations of the main components were set to 1.0 w / v%, 3.0 w / v%, and 5.0 w / v%, respectively, and eye drops were prepared by the same method as in Example 1 (3-1).

[0110] [Table 12]

[0111]

[0112] (2-4-1) Evaluation of Solubilization of Polyoxyethylene 40 Stearate

[0113] According to the FDA's use case for additives, Kolliphor TM EL can be used for eye drops at a concentration of 5.0 w / v%, and polyoxyethylene 40 stearate can be used at a concentration of up to 7.0 w / v%. Based on this, the comparison results of the solubilization degree of the two additives are shown in Table 13 below.

[0114] [Table 13]

[0115]

[0116] From the results in Table 13, it can be seen that when the maximum usage is assumed, polyoxyethylene 40 stearate is more effective than Kolliphor TM EL can solubilize enagliflozin approximately twice.

[0117] (2-4-2) Evaluation of appearance, content, pH, osmotic pressure, and cloud point

[0118] Based on the above results, Kolliphor TM Formulations of Formulation Examples 6, 7, and 8 were prepared by replacing EL with polyoxyethylene 40 stearate, and their appearance, content, pH, osmotic pressure, and cloud point were evaluated. For comparison, the cloud point of the formulation of Formulation Example 1 was measured. The results are shown in Table 14 below.

[0119] [Table 14]

[0120]

[0121] As shown in Table 14, when the concentrations of the main components of Formulation Example 1 and Formulation Example 8 are compared, the cloud point is increased by about 40°C. This indicates that the TM Compared to EL, polyoxyethylene 40 stearate can maintain its solubilization even when the water-soluble portion undergoes some dehydration due to heating. The formulations of Formulation Examples 6, 7, and 8 showed suitable values in all evaluation items.

[0122] (2-4-3) Evaluation of stability

[0123] The accelerated stability of Formulation Example 1, Formulation Example 2, Formulation Example 6, Formulation Example 7, and Formulation Example 8 was evaluated by measuring the content of the main component while storing the formulations under accelerated conditions for 6 months. The results are shown in Table 15 below.

[0124] [Table 15]

[0125]

[0126] As shown in Table 15, the content of Formulation Example 1 decreased by more than 90% after 6 months of storage under accelerated conditions. However, the formulation of Formulation Example 8, containing the same concentration, exhibited significantly superior stability. Formulation Examples 6 and 7, which are low-concentration formulations, also exhibited significantly superior stability. Therefore, it was confirmed that the use of polyoxyethylene 40 stearate can significantly improve the stability of eye drops containing enagliflozin.

[0127] Example 4: Formulation Study-4

[0128] (1) Preparation of preparations

[0129] Based on the test results, the concentration of the main component was set to 2.0 w / v% according to the components and contents in Table 16, and eye drops were prepared in the same manner as in Example 1 (3-1).

[0130] [Table 16]

[0131]

[0132] (2) Evaluation of stability

[0133] The preparations of Preparation Example 9 and Preparation Example 10 were stored under accelerated conditions for 6 months, and the accelerated stability was evaluated by measuring the content of the main component and the content of the total degradation substances. The results are shown in Table 17 below.

[0134] [Table 17]

[0135]

[0136] From the results in Table 17, it can be confirmed that the use of polyoxyethylene 40 stearate according to the present invention exhibits more excellent stability.

[0137] (3) Pharmacokinetic study in rat eyes

[0138] In order to compare the exposure of each preparation in the rat eyes, eye drops were administered. Each preparation was administered to 10 rats. After the rats were fixed outside the cage, the upper and lower eyelids were opened, and 5 μL of the test substance was dropped into the right eyeball and the left eyeball respectively using a pipette. After the instillation, it was kept fixed for a period of time to prevent outflow. At the sampling times of 1 hour, 2 hours, 4 hours, 6 hours, and 12 hours after the instillation, both eyes were anesthetized by inhalation of isoflurane, and then the left and right eyes were removed respectively with surgical scissors and forceps. The retinal tissue separated from the left / right eye was collected into a 1.5 mL Eppendorf tube immediately after collection. The retinal tissue of each individual collected at each sampling point was placed in a 1.5 mL Eppendorf tube and collected (pooling). The retinal tissue was immediately immersed in liquid nitrogen for quick freezing after weighing, and the concentration of enagliflozin in the retina was analyzed by LC-MS / MS.

[0139] The drug concentration curve in the eyeball obtained by conducting the pharmacokinetic test in the rat eyeball as described above is as follows Figure 4 As shown. Figure 4 The results show that the eye drops obtained by using polyoxyethylene 40 stearate according to the present invention maintain a significantly higher concentration in the eyeball. Therefore, the eye drops obtained according to the present invention can provide excellent pharmacological activity by increasing the exposure of enagliflozin in the eyeball.

Claims

1. A pharmaceutical composition in the form of eye drops, comprising: enagliflozin or a pharmaceutically acceptable salt thereof as an active ingredient; and a combination of polysorbate and polyoxyethylene 40 stearate as a solubilizer and stabilizer.

2. The pharmaceutical composition according to claim 1, characterized in that The concentration of enagliflozin or a pharmaceutically acceptable salt thereof is 0.1-10 w / v%.

3. The pharmaceutical composition according to claim 1, characterized in that The concentration of enagliflozin or a pharmaceutically acceptable salt thereof is 0.3-8 w / v%.

4. The pharmaceutical composition according to claim 1, characterized in that The concentration of enagliflozin or a pharmaceutically acceptable salt thereof is 0.5-5 w / v%.

5. The pharmaceutical composition according to claim 1, characterized in that The concentration of the combination of polysorbate and polyoxyethylene 40 stearate is 1-15 w / v %.

6. The pharmaceutical composition according to claim 1, characterized in that The concentration of the combination of polysorbate and polyoxyethylene 40 stearate is 4-10 w / v %.

7. The pharmaceutical composition according to claim 1, characterized in that The weight ratio of the polysorbate to polyoxyethylene 40 stearate is 1:1 to 10.

8. The pharmaceutical composition according to claim 1, characterized in that The weight ratio of the polysorbate to polyoxyethylene 40 stearate is 1:2 to 8.

9. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition forms nanomicelles with an average particle size of 1-500 nm.

10. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition forms nanomicelles with an average particle size of 5-50 nm.

11. The pharmaceutical composition according to any one of claims 1 to 10, wherein The pharmaceutical composition further comprises one or more additives selected from the group consisting of a penetration enhancer, an osmotic pressure regulator, and a pH regulator.

12. The pharmaceutical composition according to claim 11, characterized in that The penetration enhancer is D-α-tocopheryl polyethylene glycol succinate.

13. The pharmaceutical composition according to claim 11, characterized in that The osmotic pressure regulator is glycerol.

14. The pharmaceutical composition according to claim 13, characterized in that The pharmaceutical composition has an osmotic pressure of 230-350 mOsmol / kg.

15. The pharmaceutical composition according to claim 11, characterized in that The pharmaceutical composition has a pH of pH 6.0 to pH 7.

5.

16. The pharmaceutical composition according to claim 1, wherein In an aqueous medium, the pharmaceutical composition comprises: 0.5-5 w / v% of enagliflozin or a pharmaceutically acceptable salt thereof; 4-10 w / v% of a combination of polysorbate and polyoxyethylene 40 stearate; 0.1-0.5 w / v% of D-α-tocopheryl polyethylene glycol succinate; 0.5-6 w / v% of glycerol; and a pH adjuster. The pharmaceutical composition forms nanomicelles with an average particle size of 5-50 nm.

17. The pharmaceutical composition according to claim 16, characterized in that The weight ratio of the polysorbate to polyoxyethylene 40 stearate is 1:2 to 8.

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