A pharmaceutical composition comprising a metabolite of enavogliflozin and use thereof

Abstract

The present invention relates to a pharmaceutical composition comprising a metabolite of inabogliflozin and the use thereof. The inabogliflozin M1 metabolite of Formula 1 is an SGLT1 / SGLT2 dual inhibitor and exhibits a pharmacological mechanism different from that of inabogliflozin, which is an SGLT2 selective inhibitor. A pharmaceutical composition comprising the inabogliflozin M1 metabolite as an SGLT1 / SGLT2 dual inhibitor as an active ingredient can be usefully used for the prevention or treatment of diabetes or heart failure.

Description

Technology Field

[0001] The present invention relates to a pharmaceutical composition comprising a metabolite of inabogliflozin and its use. Background Technology

[0002] SGLT2 (sodium glucose cotransporter 2) is a transporter responsible for glucose reabsorption in the kidneys along with SGLT1 (sodium glucose cotransporter 1), with SGLT2 playing the majority of the role. Therefore, when SGLT2 inhibitors suppress the SGLT2 transporter, the amount of glucose excreted in urine increases, which ultimately lowers blood glucose levels and further excretes the calories contained in the blood glucose, resulting in weight loss.

[0003] One of the drugs developed as an SGLT2 inhibitor that can be usefully used as a treatment for type 2 diabetes due to such effects is Enavogliflozin, represented by the following structural formula (chemical formula A), and has been disclosed in Korean Patent Publication No. 2014-0022086 (Patent Document 1).

[0005] [Chemical Formula A]

[0006]

[0007] Compound Name: (2S,3R,4R,5S,6R)-2-(7-chloro-6(4-cyclopropylbenzyl)-2,3-dihydrobenzofuran-4-yl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol

[0008] Inabogliflozin is a drug that treats type 2 diabetes by selectively inhibiting sodium-glucose cotransporter 2 (SGLT2). It demonstrates efficacy equivalent to or greater than that of existing SGLT2 inhibitors with only 0.3 mg, which is less than one-thirtieth of the dosage. It received marketing approval after demonstrating superior efficacy in lowering glycated hemoglobin (HbA1c) and fasting blood glucose, as well as safety, compared to existing commercially available drugs through Phase 3 clinical trials conducted on patients with type 2 diabetes.

[0009] According to Non-Patent Literature 1, inabogliflozin was identified to produce a total of five metabolites—M1, M2, M3, U1, and U2—in human hepatocytes. The formation of M1 and M2 from inabogliflozin was catalyzed by CYP3A4 and CYP2C19. M3 was produced by the hydroxylation of M1 and was catalyzed by CYP3A4. The formation of U1 was catalyzed by UGT2B7, while the formation of U2 was catalyzed by UGT1A4, UGT1A9, and UGT2B7.

[0010] Non-patent literature 1 is a paper on the drug metabolism of inabogliflozin in the liver and the identification of metabolites and metabolic enzymes of inabogliflozin, but it does not disclose at all what activity the metabolites of inabogliflozin exhibit. Prior art literature

[0012] Republic of Korea Published Patent Application No. 2014-0022086

[0013] Ju-Hyun Kim et al., Pharmaceutics. 2020 Sep 11;12(9):865. doi: 10.3390 / pharmaceutics12090865. The problem to be solved

[0014] The purpose of the present invention is to provide a pharmaceutical use for a metabolite of inabogliflozin. means of solving the problem

[0015] In the process of studying metabolites of inabogliflozin, the inventors confirmed that the inabogliflozin M1 metabolite of Formula 1 exhibits a different pharmacological mechanism from inabogliflozin, which is an SGLT2 selective inhibitor, as an SGLT1 / SGLT2 dual inhibitor, and completed the present invention.

[0016] [Chemical Formula 1]

[0017]

[0018] (2S,3R,4R,5S,6R)-2-(7-chloro-6-(4-cyclopropylbenzyl)-2-hydroxy-2,3-dihydrobenzofuran-4-yl)-6-(hy droxymethyl)tetrahydro-2H-pyran-3,4,5-triol

[0020] SGLT-2 inhibitors target SGLT2, which is responsible for the reabsorption of more than 90% of glucose filtered in the kidneys, while SGLT1 inhibitors target SGLT1, which is responsible for the reabsorption of the remaining 10% of glucose in the small intestinal epithelial cells.

[0021] Most conventionally developed SGLT2 inhibitors were drugs with high selective binding affinity to SGLT2, and Inabogliflozin, developed by the applicant, is also a drug with very high selectivity for SGLT2 compared to SGLT1, and was recently approved as a treatment for type 2 diabetes.

[0022] Lexicon Pharmaceuticals' sotagliflozin, an SGLT1 / SGLT2 dual inhibitor, was approved in the EU for type 1 diabetes and by the US FDA for heart failure (HF).

[0023] The present invention provides a use of the inabogliflozin M1 metabolite as an SGLT1 / SGLT2 dual inhibitor.

[0024] The present invention will be described in more detail below.

[0025] The present invention provides the use of an inabogliflozin M1 metabolite or a pharmaceutically acceptable salt thereof as an SGLT1 / SGLT2 dual inhibitor, a pharmaceutical composition for the prevention or treatment of diabetes or heart failure comprising an inabogliflozin M1 metabolite or a pharmaceutically acceptable salt thereof as an active ingredient, and a method for the prevention or treatment of diabetes or heart failure comprising administering an effective amount of inabogliflozin or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0026] The present invention provides a pharmaceutical composition for the prevention or treatment of diabetes or heart failure comprising an inabogliflozin M1 metabolite of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0027] In addition, the present invention provides an SGLT1 / SGLT2 dual inhibitor comprising an inabogliflozin M1 metabolite of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0028] As can be seen in the following examples, the inabogliflozin M1 metabolite has binding affinity for SGLT2 while simultaneously exhibiting strong binding affinity for SGLT1. Therefore, it is a drug candidate capable of acting on both indications treatable as an SGLT2 inhibitor and indications treatable as an SGLT1 / SGLT2 dual inhibitor.

[0029] In the present invention, the diabetes may be type 1 diabetes or type 2 diabetes.

[0030] A representative indication that can be treated with SGLT2 inhibitors may be type 2 diabetes. In Example 3 below, the excellent antidiabetic efficacy of the inabogliflozin M1 metabolite was confirmed through the oral glucose tolerance test (OGTT) and urinary glucose excretion capacity (UGE) in a normal rat model.

[0031] Representative indications treatable with SGLT1 / SGLT2 dual inhibitors may include type 1 diabetes and heart failure.

[0032] Inabogliflozin M1 metabolite can simultaneously block SGLT2 present in the kidneys and SGLT1 present in the small intestine, so it can effectively act on type 1 diabetes with impaired insulin secretion. In Example 4 below, the antidiabetic efficacy of inabogliflozin M1 metabolite was demonstrated in a rat model of type 1 diabetes. Example 4 suggests that inabogliflozin M1 metabolite may be the most effective drug for type 1 diabetes compared to inabogliflozin or sotagliflozin.

[0033] In one embodiment, the pharmaceutical composition may be administered to a patient with diabetes or a patient with prediabetes.

[0034] A pharmaceutical composition comprising an inabogliflozin M1 metabolite or a pharmaceutically acceptable salt thereof as an active ingredient may be used in the form of single administration or combination administration.

[0035] Inabogliflozin M1 metabolite alone is sufficient to prevent or treat diabetes and / or heart failure. However, since diabetes or heart failure treatments are administered in combination for various purposes and may exhibit complementary or synergistic effects when administered in combination, a pharmaceutical composition containing inabogliflozin M1 metabolite as an active ingredient can be administered in combination with other diabetes or heart failure treatments.

[0036] For example, it may be administered in combination with biguanide drugs such as metformin, a known diabetes treatment, DPP4 inhibitors, sulfonylurea insulin secretagogues and / or insulin.

[0037] In addition, another indication treatable with an SGLT1 / SGLT2 dual inhibitor may be heart failure. The preventive or therapeutic effect of sotagliflozin as an SGLT1 / SGLT2 dual inhibitor on heart failure has been proven through U.S. FDA approval.

[0038] The term “prevention” as used in this specification refers to any act of suppressing or delaying the onset of diabetes or heart failure by administering a pharmaceutical composition according to the present invention.

[0039] In addition, the term “treatment” as used in this specification refers to any act in which diabetes or heart failure is improved or beneficially altered by the administration of a pharmaceutical composition according to the present invention.

[0040] The dose of inabogliflozin M1 metabolite that may be used for the prevention or treatment of diabetes and / or heart failure in patients with or at risk of diabetes and / or heart failure is not specifically limited and may be appropriately adjusted based on the severity of the patient's disease, body weight, age, sex, presence or absence of other complications, etc.

[0041] Inabogliflozin M1 metabolites or pharmaceutically acceptable salts thereof may be administered orally or parenterally. The duration of administration may be appropriately adjusted by a clinician based on the preventive or therapeutic effect of administration on diabetes and / or heart failure in patients with or at risk of diabetes and / or heart failure.

[0042] The pharmaceutical composition according to the present invention may further comprise a pharmaceutically acceptable carrier in addition to the active ingredient, the inabogliflozin M1 metabolite of Formula 1 or a pharmaceutically acceptable salt thereof, and may be formulated together with the carrier.

[0043] In the present invention, the term "pharmaceuticalally acceptable carrier" refers to a carrier or diluent that does not irritate living organisms and does not impair the biological activity and properties of the administered compound. Acceptable pharmaceutical carriers for compositions formulated as liquid solutions include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components; additionally, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed. Furthermore, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into injectable formulations such as aqueous solutions, suspensions, and emulsions, as well as pills, capsules, granules, or tablets.

[0044] In the present invention, the pharmaceutical composition may have a formulation for oral administration, such as a tablet or capsule. In one embodiment of the present invention, the pharmaceutical composition may have a tablet formulation.

[0045] In the case of a formulation for oral administration, the pharmaceutical composition of the present invention may include excipients, disintegrants, and binders as additives.

[0046] Examples of excipients include lactose (including hydrates), dextrin, mannitol, sorbitol, starch, microcrystalline cellulose [e.g., Celphere™], silicified microcrystalline cellulose [e.g., Prosolv™], calcium phosphate hydrate, anhydrous calcium phosphate, calcium carbonate, sugars, or mixtures thereof. In embodiments of the present invention, a preferred excipient is microcrystalline cellulose.

[0047] Examples of disintegrants include crospovidone, sodium croscarmellose, sodium starch glycolate, and low-substituted hydroxypropylcellulose. In a specific embodiment of the present invention, a preferred excipient is sodium croscarmellose.

[0048] Examples of binders include polyvinylpyrrolidone, povidone, gelatin, starch, sucrose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylalkylcellulose (e.g., hydroxypropylmethylcellulose), and mixtures thereof. In a specific embodiment of the present invention, a preferred binder is hydroxypropylcellulose.

[0049] Examples of other additives include lubricants and coloring agents.

[0050] The above lubricant comprises stearic acid, stearate (e.g., magnesium stearate), hard anhydrous silica, talc, corn starch, carnauba wax, magnesium silicate, synthetic aluminum silicate, hardened oil, white wax, titanium oxide, microcrystalline cellulose, macrogol 4000 and 6000, isopropyl myristate, calcium hydrogen phosphate, and mixtures thereof.

[0051] In the present invention, the pharmaceutical composition may have a formulation for parenteral administration. Although not limited thereto, for example, a parenteral administration formulation containing the composition of the present invention as an active ingredient may be formulated in an injectable form such as subcutaneous injection, intravenous injection, or intramuscular injection.

[0052] In order to formulate it into an injectable formulation, the composition of the present invention may be mixed in water with a stabilizer or a buffer to prepare a solution or suspension, and this may be formulated for unit administration in ampoules or vials.

[0053] Alternatively, the composition of the present invention may be formulated into various forms of parenteral administration formulations, such as eye drops, microneedles, patches, and depots.

[0054] The composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease, and the effective dose level may be determined based on factors including the type and severity of the patient's disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as a single or multiple doses. That is, the total effective amount of the composition of the present invention may be administered to the patient as a single dose, or administered via a fractionated treatment protocol involving long-term administration of multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects by considering all the aforementioned factors, and this can be easily determined by a person skilled in the art.

[0055] The titrated once-daily dose of inabogliflozin identified during clinical trials is 0.1 mg to 0.5 mg, and based on the results of the following examples, it is expected that the inabogliflozin M1 metabolite can also be taken at a dose of 0.1 mg to 0.5 mg, similar to that of inabogliflozin.

[0056] When the above pharmaceutical composition is formulated into a unit dosage form, the content of the active ingredient in the pharmaceutical composition may be 0.1 to 0.5 mg.

[0057] The pharmaceutical composition according to the present invention may be administered once to three times a day, for example, once a day, but is not limited thereto. Effects of the invention

[0058] A pharmaceutical composition comprising the inabogliflozin M1 metabolite of the present invention as an active ingredient can be usefully used for the prevention or treatment of diabetes or heart failure. Brief explanation of the drawing

[0059] Figure 1 is a graph showing the inhibitory effects of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin on the SGLT1 activity of CHO-SGLT1 on AMG transport into cells. Figure 2 is a graph showing the inhibitory effects of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin on the SGLT2 activity of CHO-SGLT2 AMG transport into cells. Figure 3 is a graph showing the changes in the inhibitory effects of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin on SGLT1 activity according to the pretreatment time of the drugs. Figure 4 is a graph showing the changes in the inhibitory effects of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin on SGLT2 activity according to the pretreatment time of the drugs. Figure 5 shows the results of comparing the activity recovery ability of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin against SGLT1. Figure 6 shows the results of comparing the activity recovery capabilities of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin against SGLT2. Figure 7 shows the results of comparing the binding affinity and dissociation degree of inabogliflozin, M1, M2, and dapagliflozin to SGLT1. Figure 8 shows the results of comparing the binding affinity and dissociation degree of inabogliflozin, M1, M2, and dapagliflozin to SGLT2. Figure 9 is a graph showing changes in blood glucose over time after administration of inabogliflozin, M1, or M2 in normal rats. Figure 10 is a graph showing the area under the blood glucose concentration-time curve after administration of inabogliflozin, M1, or M2 in normal rats. Figure 11 is a graph showing non-fasting blood glucose levels observed up to 72 hours after administration of the test substance in STZ-induced type 1 diabetic rats. Figure 12 is a graph showing blood glucose levels according to an oral glucose tolerance test in STZ-induced type 1 diabetic rats. Figure 13 is a graph showing the area under the blood glucose concentration-time curve according to an oral glucose tolerance test in STZ-induced type 1 diabetic rats. Figure 14 is a graph showing changes in inflammation-related cytokine levels over time after administration of the test substance in STZ-induced type 1 diabetic rats. Specific details for implementing the invention

[0060] The advantages and features of the present invention, and the methods for achieving them, will become clear by referring to the experimental and manufacturing examples described in detail below. However, the present invention is not limited to the experimental and manufacturing examples disclosed below, but may be implemented in various different forms, and is provided merely 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.

[0062] [Example]

[0063] [Example 1] Evaluation of SGLT1 and SGLT2 Inhibitory Effects of Inabogliflozin Metabolites

[0064] According to the applicant's prior research, it was confirmed that inabogliflozin (also referred to as DWP16001 in the following examples) exhibits higher inhibitory activity on SGLT2 relative to SGLT1 compared to competing drugs, dapagliflozin and ipragliflozin, and that there was no change in inhibitory activity after pretreatment for up to 2 hours. Through Dixon plot and Replot results regarding the evaluation of inhibitory activity according to substrate concentration, the inhibition of SGLT2 exhibited a reversible competitive inhibition mechanism. However, when intracellular drug concentrations were quantified after 24 hours of incubation, it was confirmed that the amount of inabogliflozin remaining in the cells was significantly higher compared to dapagliflozin and ipragliflozin. Furthermore, in the case of inabogliflozin, a time-dependent delay in the recovery of intracellular SGLT2 activity was observed, whereas this was not the case for the competing drugs, dapagliflozin and ipragliflozin. These results indicate that inabogliflozin may have better sustained efficacy compared to dapagliflozin and ipragliflozin.

[0065] The inventors intend to evaluate through this study whether the same results appear for the M1 and M2 metabolites of inabogliflozin (also referred to as M1 and M2 in this specification, respectively). In addition, they intended to evaluate and compare them with dapagliflozin and empagliflozin, which have recently been the subject of active clinical research.

[0067] Example 1-1. Evaluation of the inhibitory activity of inabogliflozin metabolites against SGLT subtypes

[0068] (1) Evaluation of SGLT1 inhibitory activity

[0069] When SGLT1-overexpressing cell lines (CHO-SGLT1 stable cells) and mock cells (CHO-mock cells) are cultured and 80% confluence is reached, 1 x 10⁶ cells are transferred to a 96-well plate. 5 Seeded cells / well.

[0070] After 24 hours, the medium was removed, and pre-incubation buffer [10 mM HEPES, 5 mM Tris, 140 mM choline chloride, 2 mM KCl, 1 mM CaCl2, 1 mM MgCl2 pH 7.4] was added and incubated for 1 hour.

[0071] After 1 hour, remove the pretreatment buffer and 10 μM [the substrate of SGLT1] 14 Incubation buffer [10 mM HEPES, 5 mM Tris, 140 mM NaCl, 2 mM KCl, 1 mM CaCl2, 1 mM MgCl2, pH 7.4] containing [C]AMG and inhibitory evaluation substances at various concentrations was added and reacted for 2 hours in a thermomicromixer at 37℃.

[0072] The concentrations of the inhibitory evaluation substances used in the experiment are as follows.

[0073] Inabogliflozin: 1, 10, 100, 500, 1000, 5000, 20000, 50000 nM

[0074] M1: 1, 10, 100, 500, 1000, 5000, 20000, 50000 nM

[0075] M2: 1, 10, 100, 500, 1000, 5000, 20000, 50000 nM

[0076] Dapagliflozin: 1, 10, 100, 500, 1000, 5000, 20000, 50000 nM

[0077] Empagliflozin: 1, 10, 100, 500, 1000, 5000, 20000, 50000 nM

[0079] For reference, the M2 metabolite has the following compound name and the structure of Chemical Formula 2.

[0080] Compound name: (2S,3R,4R,5S,6R)-2-(7-chloro-6-(4-(1-hydroxycyclopropyl)benzyl)-2,3-dihydrobenzofuran-4-yl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol)

[0081] [Chemical Formula 2]

[0082]

[0084] After a set time, the washing was performed twice with 200 μL of ice-cold washing buffer [10 mM HEPES, 5 mM Tris, 140 mM choline chloride, 2 mM KCl, 1 mM CaCl2, 1 mM MgCl2, 100 mM AMG, pH 7.4].

[0085] After removing the culture medium, 40 μL of 10% SDS solution was added to lyse the cells, then the sample was transferred to a liquid scintillation counter plate, 150 μL of OptiPhase Supermix® isotope cocktail was added, and the mixture was incubated overnight to ensure that the cell solution was homogeneously mixed with the cocktail solution.

[0086] Among the samples [ 14 C]AMG was measured using a liquid scintillation counter.

[0087] In SGLT1-overexpressing cell lines [ 14 Compare the intracellular uptake rate of C]AMG with the uptake rate in CHO-mock cells, and substrate drug [ 14 From the graph of the intracellular uptake rate of [C]AMG (% of control) and the concentration of the applied inhibitory evaluation substance, the IC50 that exhibits a 50% inhibition value (half maximal inhibitory concentration) 50Values ​​were calculated. The inhibitory effect Sigmoid Emax model was used, and the WinNonlin program (ver 2.0) was utilized. All data values ​​were expressed as the mean ± SD of three independent experiments.

[0089] (2) Evaluation of SGLT2 inhibitory activity

[0090] The evaluation was performed in the same manner as the SGLT1 inhibition evaluation method, except that a cell line with overexpression of SGLT2 (CHO-SGLT2 stable cells) was used and the inhibitory evaluation substance was used as follows.

[0091] The concentrations of the inhibitory evaluation substances are as follows.

[0092] Inabogliflozin: 0.01, 0.05, 0.2, 2, 10, 50, 100, 500 nM

[0093] M1: 0.01, 0.05, 0.2, 2, 10, 50, 100, 500 nM

[0094] M2: 0.01, 0.05, 0.2, 2, 10, 50, 100, 500 nM

[0095] Dapagliflozin: 0.01, 0.05, 0.2, 2, 10, 50, 100, 500 nM

[0096] Empagliflozin: 0.01, 0.05, 0.2, 2, 10, 50, 100, 500 nM

[0098] (3) Evaluation of the inhibitory activity of inabogliflozin and M1 and M2 metabolites in SGLT1 and SGLT2 overexpressing cell lines

[0099] For SGLT1 or SGLT2 The inhibitory activities of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin were evaluated and are shown in Table 1 and Figures 1 and 2 below.

[0100] [Table 1] Inhibitory effects of Inabogliflozin, M1, M2, Dapagliflozin, and Empagliflozin on SGLT1 or SGLT2 activity

[0101]

[0102] The inhibition resolution constants calculated from the evaluation results were similar to previous study results, and compared to dapagliflozin and empagliflozin, inabogliflozin was evaluated to have stronger inhibition of SGLT2 and better selectivity as the ratio of inhibition resolution constants of SGLT1 to SGLT2. In addition, both metabolites M1 and M2 of inabogliflozin showed inhibition of SGLT2.

[0103] Notably, unlike inabogliflozin, M2, dapagliflozin, and empagliflozin, which selectively inhibit SGLT2, metabolite M1 also exhibits excellent SGLT1 activity inhibitory effects (IC 50 (value at the 28.4 nM level), SGLT1 / SGLT2 IC 50 The ratio showed a very low value, confirming that it can act as a dual inhibitor for both SGLT1 and SGLT2.

[0105] Example 1-2. Evaluation of the inhibitory mechanisms of Inabogliflozin and M1 and M2 metabolites on SGLT1 and SGLT2

[0106] (1) Comparative evaluation of changes in inhibitory activity according to drug exposure time in SGLT1 overexpressing cell lines

[0107] CHO-SGLT1 cells were pretreated with inabogliflozin, M1 and M2 dapagliflozin, and empagliflozin at various concentrations in sodium-free pretreatment buffer (Na-negative pretreatment buffer) for 1 or 2 hours. Subsequently, the concentration-dependent inhibitory effect of inabogliflozin on AMG transport into CHO-SGLT1 cells was evaluated and compared with the unpretreated group. All data were expressed as the mean ± SD of four independent experiments.

[0108] As shown in Table 2 and Figure 3, the inhibitory activity of inabogliflozin, M2, dapagliflozin, and empagliflozin did not increase or change significantly as the exposure time increased (p>0.05). On the other hand, the inhibition constant of M1 significantly increased as the exposure time increased (p<0.05).

[0109] [Table 2] IC50 of Inabogliflozin, M1, M2, Dapagliflozin, and Empagliflozin for SGLT1 activity according to pretreatment time 50 Change in (nM) value

[0110]

[0111] P value was assessed by one-way ANOVA

[0113] (2) Evaluation of inhibitory activity according to the concentration of substrate AMG in SGLT1 overexpressing cell lines

[0114] To determine the inhibition mechanism and inhibition constant (Ki) for SGLT1, the concentration-dependent inhibition of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin was evaluated by varying the concentrations of the substrate AMG in CHO-SGLT1 cell lines.

[0115] The absorption rate of AMG and the concentrations of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin were plotted using a Dixon plot, and the AMG concentration and Dixon slopes were plotted graphically. Additionally, the results were plotted using an LB plot, and the inhibitor concentrations and LB slopes were plotted graphically (not shown). From this, the mechanism of inhibition was identified and the inhibition constant was calculated. The inhibition constant and the mechanism of inhibition calculated from the slopes of the LB plot are shown in Table 3.

[0116] [Table 3] Ki values ​​of Inabogliflozin, M1, M2, Dapagliflozin, and Empagliflozin for SGLT1 activity

[0117]

[0118] As can be seen in Table 3, the Ki value of the M1 metabolite for SGLT1 activity was 84.8 nM, which was the smallest among the test substances. A smaller Ki value indicates a higher affinity, and this result indicates that M1 has a stronger inhibition of SGLT1 because it has a greater affinity for SGLT1 compared to the test substances.

[0120] (3) Comparative evaluation of changes in inhibitory activity according to drug exposure time in SGLT2 overexpressing cell lines

[0121] CHO-SGLT2 cells were pretreated for 1 or 2 hours with inabogliflozin, M1, M2, dapagliflozin, and empagliflozin at various concentrations in sodium-free pretreatment buffer. The concentration-dependent inhibitory effect of inabogliflozin on AMG transport into CHO-SGLT2 cells was evaluated and compared with the unpretreated group. All data were expressed as the mean ± SD of four independent experiments.

[0122] As shown in Table 4 and Figure 4, the inhibitory activity of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin did not increase or change significantly even with increased exposure time (p>0.05).

[0123] [Table 4] IC50 of Inabogliflozin, M1, M2, Dapagliflozin, and Empagliflozin for SGLT2 activity according to pretreatment time 50 Change in (nM) value

[0124]

[0125] P value was assessed by one-way ANOVA

[0127] (4) Evaluation of inhibitory activity according to the concentration of substrate AMG in SGLT2 overexpressing cell lines

[0128] To determine the inhibition mechanism and inhibition constant (Ki) for SGLT2, the concentration-dependent inhibitory effects of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin were evaluated in CHO-SGLT2 cell lines by varying the concentration of the substrate AMG.

[0129] The absorption rate of AMG and the concentrations of inabogliflozin, M1, M2, dapagliflozin, and empagliflozin were plotted using a Dixon plot, and the AMG concentration and Dixon slopes were plotted graphically. Additionally, the results were plotted using an LB plot, and the inhibitor concentrations and LB slopes were plotted graphically (not shown). From this, the mechanism of inhibition was identified and the inhibition constant was calculated. The inhibition constant and the mechanism of inhibition calculated from the slopes of the LB plot are shown in Table 5.

[0130] [Table 5] Ki values ​​of Inabogliflozin, M1, M2, Dapagliflozin, and Empagliflozin for SGLT2 activity

[0131]

[0133] Examples 1-3. Comparative evaluation of activity recovery ability for SGLT1 and SGLT2 after 24-hour pretreatment with inabogliflozin and metabolites.

[0134] (1) Recovery of activity of inabogliflozin and metabolites after washout in SGLT1 overexpressing cell lines Comparative evaluation

[0135] When SGLT1-overexpressing cell lines (CHO-SGLT1 stable cells) are cultured and reach 80% confluence, 1 x 10⁶ cells are transferred to a 96-well plate. 5 Cells were seeded per well. After 24 hours, the medium was removed and replaced with medium containing the evaluation substance at different concentrations, and cultured for 24 hours.

[0136] The concentrations of the evaluation substances used in the experiment are as follows.

[0137] Inabogliflozin: 50, 500, 5000, 20000 nM

[0138] M1: 50, 500, 5000, 20000 nM

[0139] M2: 50, 500, 5000, 20000 nM

[0140] Dapagliflozin: 50, 500, 5,000, 20,000 nM

[0141] Empagliflozin: 50, 500, 5000, 20000 nM

[0142] Pre-incubation buffer [10 mM HEPES, 5 mM Tris, 140 mM choline chloride, 2 mM KCl, 1 mM CaCl2, 1 mM MgCl2 pH 7.4], which was free of the evaluation substance, was added to SGLT1-overexpressing cell lines treated with the drug for 24 hours, and the cells were incubated for 1, 2, 4, 8, and 24 hours to allow the drug bound to SGLT1 to dissociate.

[0143] After 1 hour, remove the pretreatment buffer and 10 μM [the substrate of SGLT1] 14Incubation buffer containing [10 mM HEPES, 5 mM Tris, 140 mM NaCl, 2 mM KCl, 1 mM CaCl2, 1 mM MgCl2, pH 7.4] was added and reacted in a Thermo Micro Mixer at 37°C for 2 hours. After 2 hours, it was washed twice with 200 μL of ice-cold washing buffer [10 mM HEPES, 5 mM Tris, 140 mM choline chloride, 2 mM KCl, 1 mM CaCl2, 1 mM MgCl2, 100 mM AMG, pH 7.4].

[0144] After removing the culture medium, 40 μL of 10% SDS solution was added to lyse the cells, and the sample was transferred to a liquid scintillation counter plate. 150 μL of OptiPhase Supermix® isotope cocktail was added, and the mixture was incubated overnight to ensure the cell suspension was homogeneously mixed with the cocktail. Among the samples, [ 14 After measuring [C]AMG with a liquid scintillation counter, substrate drugs in SGLT1-overexpressing cell lines [ 14 The intracellular uptake rate of [C]AMG (% of control) is shown in Figure 5. All data values ​​were expressed as the mean ± SD of four independent experiments.

[0145] As can be seen in Figure 5, the reduced SGLT1 activity after treating CHO-SGLT1 cell lines with inabogliflozin, M1, M2, dapagliflozin, or empagliflozin for 24 hours showed differences in the ability to recover SGLT1 activity depending on the time after removing the drug from the medium.

[0146] That is, when treated with inabogliflozin, activity was incompletely recovered up to 2 hours after washout, but recovered after 4 hours (Fig. 5A).

[0147] When treated with M1, activity was partially recovered up to 2 hours after washout at low concentrations (50, 500, 5000 nM), but in the 20000 nM treatment group, the recovery of activity was incomplete even up to 24 hours after washout (Fig. 5B).

[0148] The case treated with M2 showed a trend very similar to that of inabogliflozin. That is, activity was incompletely recovered up to 2 hours after washout, but recovered after 4 hours (Fig. 5C).

[0149] When treated with dapagliflozin or empagliflozin, activity was fully restored 2 hours after washout, and it was confirmed that recovery was faster at the same concentration than in the case of inabogliflozin (Figs. 5D and 5E).

[0150] From the above results, it was confirmed that the dissociation of M1 to SGLT1 is the slowest, while the dissociation of inabogliflozin and M2 is fast, and the dissociation of dapagliflozin and empagliflozin is fast. The above results indicate that among the evaluated substances, M1 binds to SGLT1 for the longest period.

[0152] (2) Recovery of activity of inabogliflozin and metabolites after washout in SGLT2 overexpressing cell lines Comparative evaluation

[0153] The evaluation method is the same as that for SGLT1 overexpressing cells, except that a cell line with SGLT2 overexpression (CHO-SGLT2 stable cells) is used and the concentration of the evaluation substance is used as follows.

[0154] Inabogliflozin: 0.5, 5, 50, 500 nM

[0155] M1: 0.5, 5, 50, 500 nM

[0156] M2: 0.5, 5, 50, 500 nM

[0157] Dapagliflozin: 0.5, 5, 50, 500 nM

[0158] Empagliflozin: 0.5, 5, 50, 500 nM

[0160] As can be seen in Figure 6, the reduced SGLT2 activity after treating CHO-SGLT2 cell lines with inabogliflozin, M1, M2, dapagliflozin, or empagliflozin for 24 hours showed differences in the ability to recover SGLT2 activity depending on the time after removing the drug from the medium.

[0161] When treated with inabogliflozin, the recovery of activity over time after washout varied depending on the concentration of inabogliflozin. At 0.5 nM treatment, activity was recovered after 2 hours, whereas at 5 nM treatment, recovery occurred after 24 hours. At 50 and 500 nM treatments, the recovery of activity remained unstable even after 24 hours, showing a recovery of less than 50%. These results indicate that the recovery of activity was slow because the intracellular concentration of inabogliflozin was maintained or dissociation from SGLT2 was delayed (Fig. 6A).

[0162] When treated with M1, activity was recovered after 2 hours at a low concentration (0.5 nM), but when washed out after treatment with 5 and 50 nM, it recovered over time up to 8 hours and was almost fully recovered by 24 hours. It was confirmed that the recovery of activity was delayed when treated with 500 nM (Fig. 6B).

[0163] In the case of treatment with M2, it was confirmed that there was no significant difference depending on the concentration, and that activity recovered over time. That is, activity recovered over time up to 4 hours after washout, and showed results of recovery after 8 hours (Fig. 6C).

[0164] When treated with dapagliflozin, a trend similar to M2 is observed. That is, the difference according to concentration is not significant, and it was confirmed that activity recovers over time. Activity recovers over time up to 4 hours after washout, and recovery is observed after 8 hours (Fig. 6D).

[0165] When treated with empagliflozin, activity was fully restored after 2 hours of washout, and it was confirmed that recovery was faster at the same concentration than in the case of inabogliflozin, M1, M2, and dapagliflozin (Fig. 6E).

[0166] The above results show that the dissociation of inabogliflozin against SGLT2 is the slowest, and at high concentrations, activity recovery is incomplete even after 24 hours, showing a recovery of less than 50%. In the case of M1, it was confirmed that the recovery of activity is delayed depending on the concentration and washout time. For M2, the recovery of activity of dapagliflozin does not show a significant difference depending on the concentration and shows a delay depending on the washout time, while empagliflozin was confirmed to have the fastest recovery of activity without any delay depending on the pretreatment concentration or time.

[0168] [Example 2] Evaluation of Drug-Target Binding Affinity and Dissociation of Inabogliflozin Metabolite

[0169] Cell lines overexpressing SGLT1 and SGLT2 (CHO-SGLT1, CHO-SGLT2 stable cells) were cultured until 60% confluence was reached, at which point the cells were treated with 0.05% trypsin to detach them from the bottom of the plate, suspended in 200 μL of culture medium, and used at room temperature.

[0170] To evaluate the target binding affinity and dissociation of inabogliflozin, M1, M2, and dapagliflozin on SGLT1 and SGLT2, glucose currents were recorded using a whole-cell mode patch clamp, and drug-target binding affinity and dissociation were confirmed by evaluating how long the drugs remained bound to the target through washout.

[0171] Glucose reabsorption in the kidneys is induced by the Na+ electrochemical potential gradient via SGLT2, and glucose diffuses across the cell membrane into the blood via glucose transporter 2 (GLUT-2), which enables facilitated glucose transport. One Na+ per glucose molecule via SGLT2 + While this is absorbed, SGLT1 has two Na₂ per glucose molecule. + Ions are absorbed. Therefore, by changing the composition of the buffer, Na + Only glucose current was recorded, excluding current.

[0173] The experiment was conducted in the following manner.

[0174] 1) After preparing a glass micropipette in two stages at 62.7°C and 57°C using a Narishige puller, it was filled with an internal solution (145 mM CsCl, 5 mM NaCl, 10 mM HEPES, 11 mM EGTA, pH 7.2) and inserted into the Ag / AgCl2 electrode of the parchment clamp head-stage.

[0175] 2) SGLT1 or SGLT2 cells were placed in a chamber pre-coated with poly-L-lysine, and then an external solution (150 mM NaCl, 10 mM HEPES, 1 mM CaCl2, 1 mM MgCl2, pH 7.4) was flowed to ensure the cells were sufficiently submerged. A temperature control device was used to maintain the chamber temperature at 36.5 ± 0.5℃.

[0176] 3) Using a micromanipulator, a glass micropipette was attached to the cell membrane to create a giga-ohm seal and ruptured to detach the cell membrane (Whole-cell mode, 3~5 MΩ).

[0177] 4) Fix the cell at -60 mV (holding potential) and set the current [I in gap-free data acquisition mode. total The current (Na+ / Glucose current (pA)) was measured.

[0178] 5) Continuously flow the external solution into the chamber containing the cells to supply an electric current (I total After recording for approximately 30 seconds to 1 minute (stabilization), an external solution containing glucose (Na+ / Glucose external) (150 mM NaCl, 10 mM HEPES, 1 mM CaCl2, 1 mM MgCl2, 100 mM Glucose, pH 7.4) was applied to obtain a glucose current (I glucose ... was recorded. Then, cells were exposed to an external solution containing glucose with one concentration of the compound for 2 minutes, and the glucose current inhibited by the compound was recorded.

[0179] 6) After confirming the inhibited current, the external solution was replaced with a fresh glucose solution free of the compound and the change in current was monitored while washing out, and recorded for a minimum of 3 minutes to a maximum of 10 minutes (if the reaction continued longer and the seal could be maintained, it was recorded up to 15 minutes).

[0180] 7) After confirming the recovery of the current, the solution was replaced with a sodium-free external solution (Na+-free external) (150 mM Choline-Cl, 1 mM CaCl2, 1 mM MgCl2, pH 7.4) and the Na+ current was recorded. After the current stabilized, the solution was switched back to the external solution to record the current in the resting state, where the cells are not stimulated, thereby ensuring the reliability of the experimental system.

[0181] pCLAMP10.4 (Molecular Devices, LLC, San Jose, CA, USA) or Origin6.0 (OriginLab, Northampton, MA, USA) was used for the statistical analysis of the data.

[0182] Test results were expressed as mean ± standard deviation (SD) of the mean. Statistical significance was verified using the non-parametric Kruskal-Wallis test with SPSS for Windows (version 26.0, IBM Corp., Armonk, NY, USA). The significance level was set at p<0.05.

[0183] The results of the evaluation of binding affinity and dissociation of inabogliflozin, M1, M2, and dapagliflozin on SGLT1 are as shown in Figure 7 and Table 6.

[0184] Half-time of recovery (T) of inabogliflozin for SGLT1 1 / 2,off )) is 34.9 seconds, and it was confirmed that M2 (23.5 seconds) and dapagliflozin (30.5 seconds) dissociate at similar rates. On the other hand, M1's (T 1 / 2,off It was confirmed that ) dissociated at a rate 1.7 times slower than inabogliflozin and 1.96 times slower than dapagliflozin in 59.8 seconds.

[0185] These results suggest that M1 has an effect on binding to and maintaining SGLT1 protein more effectively than inabogliflozin and its competitor, dapagliflozin, and that efficacy can be maintained.

[0187] [Table 6] Comparison of binding and dissociation degrees of Inabogliflozin, M1, M2, and Dapagliflozin in SGLT1

[0188]

[0189] Each data point represents the mean ± SD (n = 4 - 5); t 1 / 2,on : half-time on rate; t 1 / 2,off: half-time off rate; *: p < 0.05 compared to M2, +: p < 0.05 compared with dapagliflozinby post hoc analysis followed by the significant nonparametric Kruskal-Wallis test.

[0191] In addition, the results of the evaluation of binding affinity and dissociation of inabogliflozin, M1, M2, and dapagliflozin in SGLT2 are as shown in Figure 8 and Table 7.

[0192] Half-time of recovery of inabogliflozin for SGLT2 (T 1 / 2,off )) was 745.5 seconds, showing the slowest dissociation rate among the evaluated substances. It was about 2.8 times slower than the competitor dapagliflozin (265.2 seconds), and M1 (369.9 seconds) and M2 (540.4 seconds) showed dissociation rates that were 2 times and 1.38 times slower than inabogliflozin, respectively.

[0193] These results indicate that inabogliflozin binds to the SGLT2 protein for the longest period, implying that inhibitory activity can be maintained as the binding is sustained. Additionally, M1 and M2 also indicate that efficacy can be maintained because they bind to the SGLT2 protein and dissociate at a slower rate than their competitor, dapagliflozin.

[0194] In conclusion, similar to how M1 exhibits a dual inhibitory effect on SGLT-1 / SGLT-2, the recovery of inhibitory activity upon wash-out from the SGLT-1 target showed a slower dissociation time than M2. Additionally, M2 was found to maintain binding to the SGLT-2 target for a longer period than M1.

[0195] [Table 7] Comparison of binding and dissociation degrees of Inabogliflozin, M1, M2, and Dapagliflozin in SGLT2

[0196]

[0197] Each data point represents the mean ± SD (n = 3 - 6); t 1 / 2,on : half-time on rate; t 1 / 2,off : half-time off rate; *: p < 0.05 compared to enavogliflozin, +: p < 0.05 compared with dapagliflozin, #: p < 0.05 compared with empagliflozin by post hoc analysis followed by the significant nonparametric Kruskal-Wallis test.

[0199] [Example 3] Evaluation of Antidiabetic Efficacy Following Intravenous Administration of Inabogliflozin Metabolite

[0200] We intended to evaluate the antidiabetic efficacy of a single intravenous dose of inabogliflozin, M1, or M2 administered to normal rats.

[0201] To determine the oral glucose tolerance test (OGTT) and urinary glucose excretion capacity (UGE), normal 8-week-old male rats were intravenously administered inabogliflozin, M1, or M2 at a dose of 1 mg / kg, respectively, followed by the administration of a glucose solution (2 g / kg) to load glucose. The oral glucose tolerance was evaluated by measuring blood glucose levels before the administration of the glucose solution and at 5, 10, 15, 20, 30, 40, 60, 90, and 120 minutes after administration. Additionally, urinary glucose excretion capacity (UGE) was evaluated by measuring urinary glucose excretion at 6, 24, 48, and 72 hours after the administration of the glucose solution.

[0202] Figure 9 is a graph showing changes in blood glucose levels over time after intravenous administration of each test substance at a dose of 1 mg / kg to normal rats. Figure 10 is a graph showing the area under the blood glucose concentration-time curve analyzed for 120 minutes after administration of the test substance.

[0203] As can be seen in Figures 9 and 10, inabogliflozin, M1, and M2 were found to improve glucose tolerance when administered intravenously to normal rats at a dose of 1 mg / kg. Metabolite M1 showed superior glucose tolerance compared to inabogliflozin when administered intravenously, while M2 showed a pattern similar to that of inabogliflozin.

[0204] When compared by blood glucose AUC, M1 showed the most significant decrease in blood glucose AUC, while inabogliflozin and M2 showed a lower rate of decrease than M1. The inhibition rate of glucose tolerance improved by 27% for inabogliflozin, 49% for M1, and 23% for M2 compared to the vehicle (Fig. 10).

[0205] Meanwhile, when urinary glucose excretion (UGE) was confirmed, UGE was observed for metabolites M1 and M2 starting from the 6th hour after intravenous administration of 1 mg / kg. Although the amount and pattern of glucose excreted by M1 and inabogliflozin were similar, in the case of M2, less glucose was excreted than that of inabogliflozin and M1 up to 24 hours.

[0206] Based on the above results, it is determined that the intravenous administration of the test drug exerts in vivo activity through the action of SGLT1 and SGLT2 in the kidneys.

[0207] IC of metabolite M1 for SGLT2 50 Since it is higher than inabogliflozin but exhibits a more potent effect on SGLT1, metabolite M1 collectively acts as a dual inhibitor of SGLT1 and SGLT2, and as a result, it is judged that M1 exhibits antidiabetic efficacy similar to inabogliflozin.

[0208] M2 exhibits efficacy for up to 24 hours and shows weaker potency than M1; this is presumed to be due to 1) M2 having a lower pharmacological effect because its IC50 for SGLT1 is higher than M1's, and 2) rapid elimination occurring (t 1 / 2(Confirmed as =0.8h).

[0210] [Example 4] Evaluation of Antidiabetic Efficacy of Inabogliflozin Metabolite in a Type 1 Diabetic Rat Model

[0211] To evaluate the antidiabetic efficacy of inabogliflozin metabolites in a rat model of type 1 diabetes induced by STZ, test groups were established as shown in Table 8 below and experiments were conducted.

[0212] [Table 8]

[0213]

[0215] STZ was administered to 5-week-old SD male rats on Day 0 (G2 to G7, excluding G1), and non-fasting blood glucose was measured on Day 3 to determine if Type 1 diabetes was induced. On Day 7, the experimental substance was administered to each group, and blood and urine samples were collected at 6h, 24h, 48h, and 72h to measure non-fasting blood glucose. On Day 14, the animals were fasted for 16 hours, and on Day 15, a glucose solution at a dose of 2 g / kg was administered orally following the administration of the experimental substance. Subsequently, blood glucose was measured at 15, 30, 60, 90, and 120 minutes after glucose administration. After blood collection, serum was isolated, and the IL-6 inflammatory response was evaluated using an Interleukin-6 (IL-6) ELISA kit.

[0217] (1) Non-fasting blood sugar

[0218] Figure 11 shows the results of measuring non-fasting blood glucose at 6, 24, 48, and 72 hours after the first administration of the test drug in a rat model of type 1 diabetes induced by STZ.

[0219] As shown in Figure 11, low blood glucose levels were observed in G5 (inabogliflozin M1 metabolite) and G6 (inabogliflozin) under the same dose condition of 1 mg / kg, and this phenomenon confirmed that statistically significant hypoglycemic efficacy persisted compared to the vehicle group up to 48 hours after administration. In particular, G5 showed the lowest blood glucose level at 48 hours after administration, allowing for the observation of a sustained hypoglycemic effect.

[0220] Compared to G7, the 1 mg / kg sotagliflozin group approved for type 1 diabetes, G5 and G6, which have the same dosage, were found to have superior blood glucose-lowering effects.

[0222] (2) Oral glucose tolerance

[0223] Blood glucose levels and AUC measured before glucose administration (0 min) and at 15, 30, 60, and 120 minutes after glucose administration were calculated and shown in Figures 12 and 13.

[0224] As shown in Figure 12, blood glucose reduction was observed in G5 administered 1 mg / kg of inabogliflozin M1 metabolite, G6 administered 1 mg / kg of inabogliflozin, and G7 administered 1 mg / kg of sotagliflozin, and dose-dependent improvement in glucose tolerance was confirmed in the case of the M1 metabolite. In particular, the G5 group administered 1 mg / kg of inabogliflozin M1 metabolite showed the fastest and strongest blood glucose-lowering effect, and this effect was much superior to that of the G7 group administered 1 mg / kg of sotagliflozin.

[0225] As can be seen in Figure 13, the G5 group administered 1 mg / kg of inabogliflozin M1 metabolite showed the best improvement in oral glucose load in terms of AUC value, and G5 showed an AUC value similar to that of the control group G1 administered the vehicle, confirming the superior blood glucose control effect of inabogliflozin M1 metabolite in the oral glucose load experiment.

[0227] (3) Evaluation of inflammatory response

[0228] Figure 14 shows the results of measuring cytokines after administering the test drug to a rat model of type 1 diabetes induced by STZ.

[0229] As can be seen in Figure 14, interleukin-6 (IL-6), a cytokine that regulates inflammatory responses in the vehicle group, increased at all times (6, 24, 48, and 72 hours) and was found to decrease with the administration of M1. Compared to the slight decrease with inabogliflozin and sotagliflozin, M1 showed a significant decrease in all dose groups (0.01, 0.1, and 1 mg / kg), and showed a more significant decreasing trend, particularly in the 0.01 mg / kg group at 6 hours and in the 0.1 mg / kg and 1 mg / kg groups at 72 hours.

[0230] These results suggest that M1 can regulate the inflammatory response associated with type 1 diabetes.

Claims

Claim 1 A pharmaceutical composition for the prevention or treatment of diabetes or heart failure as an SGLT1 / SGLT2 dual inhibitor, comprising as an active ingredient an inabogliflozin M1 metabolite of Chemical Formula 1 below or a pharmaceutically acceptable salt thereof. [Chemical Formula 1] Claim 2 A pharmaceutical composition for the prevention or treatment of diabetes or heart failure as an SGLT1 / SGLT2 dual inhibitor, wherein the diabetes is type 1 diabetes. Claim 3 A pharmaceutical composition for the prevention or treatment of diabetes or heart failure as an SGLT1 / SGLT2 dual inhibitor, wherein the diabetes is type 2 diabetes. Claim 4 In claim 1, the pharmaceutical composition is a pharmaceutical composition for the prevention or treatment of diabetes or heart failure as an SGLT1 / SGLT2 dual inhibitor for oral or parenteral administration. Claim 5 A pharmaceutical composition for the prevention or treatment of diabetes or heart failure as an SGLT1 / SGLT2 dual inhibitor, wherein the once-daily dose of the inabogliflozin M1 metabolite of Chemical Formula 1 or a pharmaceutically acceptable salt thereof is 0.1 to 0.5 mg. Claim 6 A pharmaceutical composition for the prevention or treatment of diabetes or heart failure as an SGLT1 / SGLT2 dual inhibitor, wherein the pharmaceutical composition is administered once a day. Claim 7 delete

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