A method for regulating blood glucose levels using a dual AMPK activator and a mitochondrial uncoupling agent.

Abstract

This specification provides a method for regulating blood glucose levels in subjects requiring it, by an action that manifests as an increase in the glucose effect. The subjects are insulin-deficient and are taking or need to take exogenous insulin prior to treatment with the compounds of this specification. In some embodiments, the method involves a compound of formula (I) described herein: [Formula 1] JPEG2026526084000015.jpg24169 Alternatively, administering a pharmaceutically acceptable salt, solvate, or prodrug thereof; monitoring the subject's blood glucose levels; and delaying the need for the use of exogenous insulin, or reducing and / or ultimately discontinuing the need for exogenous insulin in the subject, as long as the subject is being administered a compound of formula (I) as described herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 514,696, filed on 20 July 2023, which is incorporated herein by reference in its entirety. Reference to electronic sequence listings

[0002] The contents of the electronic sequence listing (286502000840SEQLIST.xml, size: 43,302 bytes, created: July 18, 2024) are incorporated herein by reference in their entirety.

[0003] This disclosure generally relates to methods for regulating blood glucose levels using a dual AMPK activator and a mitochondrial uncoupler, and includes methods for treating type 1 diabetes (T1D) and other diseases, disorders, or conditions in subjects who are insulin deficient and who are taking or need to take exogenous insulin (or an external source of insulin) prior to treatment with the compounds herein. [Background technology]

[0004] In humans, glucose processing is mediated by both insulin-dependent and insulin-independent glucose uptake. In humans, the ability of glucose to stimulate its own uptake independently of insulin, known as the "glucose effect," accounts for approximately 50% of glucose processing. In insulin-deficient conditions, such as those seen in patients with type 1 diabetes (T1D), which causes hyperglycemia, an increased glucose effect can be advantageous. Currently available and clinically developing antidiabetic drugs typically do not enhance the glucose effect in diabetic individuals.

[0005] In individuals with T1D, insulin-producing β-cells are destroyed by the immune system. Without insulin, the body cannot regulate glucose levels in the bloodstream, leading to hyperglycemia, also known in this art. Individuals with T1D rely on exogenous insulin to control their blood glucose levels. However, if these individuals take in more insulin than necessary, their blood glucose levels can become too low, leading to hypoglycemia. Therefore, there is a need in this art for more effective methods of controlling blood glucose levels in patients who rely on exogenous insulin, such as therapies for T1D patients that increase glucose processing by enhancing the insulin-independent glucose effect. [Overview of the project]

[0006] In some embodiments, this specification provides a method for regulating blood glucose levels in a subject requiring it by an effect manifesting as an increase in the glucose effect. In some embodiments, this specification provides a method for regulating blood glucose levels in a subject requiring it by a dual antihyperglycemic effect manifesting as an increase in the glucose effect and preservation of β-cells. The subject is insulin-deficient and is taking or needs to take exogenous insulin before treatment with the compounds of this specification. In some embodiments, this method involves administering a compound of formula (I) to the subject: [ka] This includes administering a pharmaceutically acceptable salt, solvate, or prodrug thereof; monitoring the subject's blood glucose levels after administration of the compound; and reducing or eliminating the dose of exogenous insulin administered to the subject to maintain the subject's blood glucose levels within the subject's normal range.

[0007] In some embodiments, the compound is administered repeatedly to the subject over a period of time. In some variations, when the compound is administered over a period of time, the dose of exogenous insulin administered to the subject is gradually reduced until the dose of exogenous insulin is discontinued, as long as the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered to the subject. In one variation, the dose of exogenous insulin is discontinued after the compound has been repeatedly administered to the subject for approximately two weeks.

[0008] In one embodiment, a treatment method for T1D is provided to delay the need for insulin use in a subject requiring insulin therapy before initiation of insulin therapy, comprising administering to the subject a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In another embodiment, a treatment method for T1D is provided to reduce the need for insulin in a subject requiring it, comprising administering to the subject a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In yet another embodiment, a treatment method for unstable diabetes is provided in a subject requiring it, wherein the subject has high and low glucose levels despite using insulin, comprising administering to the subject a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0009] In certain embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in a treatment comprising any of the methods described herein is also provided. For example, in some embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is provided for regulating blood glucose levels in subjects requiring it, by an action manifesting as an increase in the glucose effect. In some variant embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is provided for regulating blood glucose levels in subjects requiring it, by a dual antihyperglycemic effect manifesting as an increase in the glucose effect and preservation of β-cells.

[0010] In certain embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for the manufacture of a drug is also provided. In some embodiments, the drug is for any of the methods described herein. For example, in some embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in the manufacture of a drug that regulates blood glucose levels in a subject having the effect of increasing the glucose effect. In some modified embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in the manufacture of a drug that regulates blood glucose levels in a subject having the effect of increasing the glucose effect and preserving β-cells, is provided.

[0011] In one embodiment, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is provided for the treatment of T1D, delaying the need for insulin use before insulin therapy is initiated. In another embodiment, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is provided for the treatment of T1D, reducing the need for insulin. In yet another embodiment, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is provided for the treatment of unstable diabetes in subjects with high and low glucose levels despite insulin use.

[0012] In some variations of the aforementioned embodiments, the subject is a human being. [Brief explanation of the drawing]

[0013] This application can be understood by referring to the following description, which is considered together with the attached drawings. [Figure 1]This shows the avoidance of hyperglycemia by administering the compound of formula (I) to STZ mice. a is the fasting glucose level and area under the curve (AUC) (n=6-8 / group) in the control group (n=7) and STZ mice that were untreated or treated with 0.25 and 0.5 mg / g of the compound of formula (I), respectively, from day 5. b is the insulin level and area under the curve (AUC) (n=6-8 / group) in the control group (n=7) and STZ mice that were untreated or treated with 0.25 and 0.5 mg / g of the compound of formula (I), respectively, from day 5. c is the total insulin content in the pancreas (n=7-8 / group) in the control group and STZ mice that were untreated or treated with 0.25 and 0.5 mg / g of the compound of formula (I), respectively, from day 5. d is the pancreatic islet cell area in STZ mice in the control group (n=3-5) and in untreated mice (n=3-5) or treated with the compound of formula (I) at 0.25 mg / g (n=3-5) and 0.5 mg / g (n=3-6), respectively. e is the fraction of insulin-positive (Ins+) cells in STZ mice in the control group (n=3-5) and in untreated mice (n=3-5) or treated with the compound of formula (I) at 0.25 mg / g (n=3-5) and 0.5 mg / g (n=3-6), respectively. f is the fraction of glucagon-positive (Glu+) cells in STZ mice in the control group (n=3-5) and in untreated mice (n=3-5) or treated with the compound of formula (I) at 0.25 mg / g (n=3-5) and 0.5 mg / g (n=3-6), respectively. g represents fasting glucose levels and area under the curve (AUC) (n=8-16 / group) in the control group (n=7) and STZ mice that were untreated or treated with 0.25 and 0.5 mg / g of the compound of formula (I), respectively, from day 15. h indicates the avoidance of hyperglycemia by administering the compound of formula (I) to STZ mice. i represents the total insulin content in the pancreas (n=8-16 / group) in the control group (n=7) and STZ mice that were untreated or treated with 0.25 and 0.5 mg / g of the compound of formula (I), respectively, from day 15.j represents the islet cell area in STZ mice in the control group (n=3-5) and in untreated mice (n=6-10) or treated with the compound of formula (I) at doses of 0.25 (n=3-5) and 0.5 mg / g (n=6-10), respectively. k represents the fraction of Ins+ cells in STZ mice in the control group (n=3-5) and in untreated mice (n=6-10) or treated with the compound of formula (I) at doses of 0.25 (n=3-5) and 0.5 mg / g (n=6-10), respectively. l represents the fraction of Glu+ cells in STZ mice in the control group (n=3-5) and in untreated mice (n=6-10) or treated with the compound of formula (I) at doses of 0.25 (n=3-5) and 0.5 mg / g (n=6-10), respectively. Note that the control mice in (a) to (f) are the same as the control mice in the corresponding panels in (g) to (l). Data are shown as mean ± SEM. Statistical significance between untreated STZ mice and STZ mice treated with the compound of formula (I) was determined by Welch's ANOVA followed by Games-Howell post-hoc tests ((a)-(d), (g)-(j)) or one-way ANOVA followed by Turkey's post-hoc tests ((e)-(f), (k)-(l)) (*p<0.05, **p<0.01, *p<0.001). Statistical significance between control mice and STZ mice was determined by Wilcoxon tests ((a)-(d), (g)-(j)) or Student's t-tests ((e)-(f), (k)-(l)) (#p<0.05, ##p<0.01, ###p<0.001). [Figure 2]This shows the stimulation of glucose uptake in skeletal muscle after administration of the compound of formula (I) to STZ mice. a is the timeline of days between STZ treatment and FDG-PET scans. b is representative PET / CT images of FDG uptake in untreated mice (upper panel) and mice treated with the compound of formula (I) (lower panel). c is the estimated maximum muscle and cardiac glucose uptake (MRglu) at baseline (scan 1), 9-10 days after the last STZ injection (scan 2), and after 1 week of treatment with 0.5 mg / g of the compound of formula (I) (scan 3) (n=5) or in the untreated group (n=5). d is the muscle and cardiac glycogen content in the control group (n=6-18) and in STZ mice from day 15 onwards in the untreated (n=14-36) or treated with the compound of formula (I) (n=13-27) groups. e represents the relative mRNA levels of Txnip, Slc2a1, Slc2a4, Hk2, Pkm, Ppargc1a, Pdk4, Pdha1, Shda, and Cox8b in the muscle of control mice (n=5-7), untreated STZ mice (n=5-9), and STZ mice treated with the compound of formula (I) (n=6-8). f represents the relative mRNA levels of Txnip, Slc2a1, Slc2a4, Pdk4, Ucp2, and Ucp3 in the heart of control mice (n=5), untreated STZ mice (n=9), and STZ mice treated with the compound of formula (I) (n=6-7). g represents mitral valve Doppler in STZ-treated mice at baseline (Scan 1), 15 days after the start of STZ (Scan 2), and after 1 week of treatment with 0.5 mg / g of the compound of formula (I) (Scan 3) (n=9) or in the untreated group (n=9). Data are shown as mean ± SEM.Statistical significance between untreated STZ mice and STZ mice treated with the compound of formula (I) was determined by Student's t-test ((c): muscle, (g)) or Wilcoxon test ((c): heart, (d)~(f)) (*p<0.05, **p<0.01, ***p<0.001), statistical significance between control mice and STZ mice was determined by Wilcoxon test ((d)~(f)) (#p<0.05, ##p<0.01, ###p<0.001), and statistical significance between scans ((c), (g)) was determined by repeated one-way ANOVA followed by paired Student's t-test (○p<0.05). [Figure 3]This shows that the compound of formula (I) avoids hyperglycemia in a dose-dependent manner in db / db mice. a is the fasting glucose level and area under the curve (AUC) (n=15-20 / group) in BKS mice and db / db mice that were untreated or treated with 0.5 and 1.0 mg / g of the compound of formula (I), respectively. b is the insulin level and area under the curve (AUC) (n=15-20 / group) in BKS mice and db / db mice that were untreated or treated with 0.5 and 1.0 mg / g of the compound of formula (I), respectively. c is the HOMA-IR (calculated from (a) and (b)) and area under the curve (AUC) in BKS mice and db / db mice that were untreated or treated with 0.5 and 1.0 mg / g of the compound of formula (I). d is the HOMA-β (calculated from (a) and (b)) and area under the curve (AUC) in untreated or treated BKS mice and db / db mice with 0.5 and 1.0 mg / g of the compound of formula (I). e is the glycogen content in the muscle and heart of BKS mice (n=5-9) and untreated (n=9-10) or (n=6-7) db / db mice treated with the compound of formula (I). f is the relative mRNA levels of Txnip, Slc2a1, Slc2a4, Hk2, Pkm, Ppargc1a, Pdk4, Pdha1, Sdha, Cox8b, Ucp2, and Ucp3 in the muscle of BKS mice (n=6-13) and untreated (n=15-22) or (n=8-15) db / db mice treated with 1.0 g / kg of the compound of formula (I). g represents the relative mRNA levels of Txnip, Slc2a1, Slc2a4, Pdk4, Ucp2, and Ucp3 in the hearts of BKS mice (n=4-5), untreated db / db mice (n=8-9), and db / db mice treated with the compound of formula (I) (n=6-7). Data are shown as mean ± SEM.Statistical significance between untreated db / db mice and db / db mice treated with a compound of formula (I) was determined by Welch's ANOVA and subsequent Games-Howell post hoc test ((a)-(d)), Student's t-test ((e)), or Wilcoxon test ((f), (g)) (*p < 0.05, **p < 0.01, ***p < 0.001), and statistical significance between BKS mice and db / db mice was determined by Wilcoxon test (((a)-(d), (f), (g)) (#p < 0.05, ##p < 0.01, p < 0.001), or Student's t-test ((e)). [Figure 4] Induction of mitochondrial uncoupling by the compound of formula (I) is shown. a is a respirometry plot showing oxygen consumption rate (OCR) measured by sequentially injecting oligomycin, FCCP, and a cocktail of rotenone and antimycin into intact differentiated C2C12 myotubes treated with 0.625, 1.25, and 2.5 μM of the compound of formula (I) for ±4 h. b is a respirometry plot showing extracellular acidification rate (ECAR) measured by sequentially injecting oligomycin, FCCP, and a cocktail of rotenone and antimycin into intact differentiated C2C12 myotubes treated with 0.625, 1.25, and 2.5 μM of the compound of formula (I) for ±4 h. c is an OCR vs. ECAR plot obtained from the last baseline measurement (measurement 3 in each panel of (a) and (b)). d is a mitochondrial function parameter calculated from the OCR data in the panel of (a). Data are shown as mean ± SEM. n = 5 / condition. Statistical significance between untreated cells and cells treated with the compound of formula (I) in (d) was determined by one-way ANOVA and subsequent Tukey's post hoc test (○p < 0.05, ○○p < 0.01). [Figure 5]This shows the conservation of β-cell mass and expression of β-cell markers in db / db mice after administration of the compound of formula (I). a is representative immunostaining (n=5 / group and antibody) for insulin (green) and glucagon, Glut2, Ipf1 / Pdx1, Nkx6-1, Mafa, and Raldh3 (all red) from pancreases of 15-week-old BKS and db / db mice, either untreated or treated with 0.5 and 1.0 mg / g of the compound of formula (I), respectively. b is quantification of pancreatic islet cell area. This is from 6-week-old BKS and db / db mice, and 15-week-old BKS and db / db mice, either untreated or treated with 0.5 and 1.0 mg / g of the compound of formula (I), respectively (n=4-6 / group). c is the fraction of insulin (Ins+) and glucagon (Glu+) positive cells in 15-week-old BKS and db / db mice that were untreated or treated with 0.5 and 1.0 mg / g of the compound of formula (I) (n=5 in all groups). d is the total insulin content in the pancreas of 15-week-old BKS mice (n=9) and db / db mice that were untreated (n=7-10) or treated with 0.5 mg / g (n=9) and 1.0 mg / g (n=6-8) of the compound of formula (I). e is the insulin / proinsulin ratio in the pancreas of 15-week-old BKS mice (n=9) and db / db mice that were untreated (n=7-10) or treated with 0.5 mg / g (n=9) and 1.0 mg / g (n=6-8) of the compound of formula (I). f represents the quantification of Glut2, Ipf1 / Pdx1, Nkx6-1, Mafa, and Raldh3 expression in the pancreatic islets of 15-week-old db / db mice treated with formula (I) at 0.5 mg / g (n=5) and 1.0 mg / g (n=4-5). g represents the relative mRNA levels of Ins1 / 2, Slc2a2, Pdx1, Nkx6-1, Mafa, Ucn3, Trpm5, Txnip, Aldh1a3, Hspa5, Erp29, and Edem2 in the pancreatic islets of 15-week-old db / db mice treated with formula (I) at 0.5 mg / g (n=14-15) and 1.0 mg / g (n=9-10). Data are shown as mean ± SEM.Statistical significance between untreated db / db mice and db / db mice treated with a compound of formula (I) was determined by Welch's ANOVA followed by Games-Howell post hoc test ((b)–(f)), or Kruskal-Wallis test followed by Dunn's post hoc test (*p < 0.05, **p < 0.01, ***p < 0.001), and statistical significance between BKS mice and db / db mice was determined by Wilcoxon test, and statistical significance between BKS mice and db / db mice was determined by Wilcoxon test ((b)–(e)) (#p < 0.05, ##p < 0.01, p < 0.001). [Figure 6a] Shows the alleviation of hyperglycemia-induced pancreatic gene expression changes in islets cultured ex vivo by the compound of formula (I). MA plot showing differential expressed genes between mouse islets cultured in 22 mM (G22) vs. 11 mM (G11) glucose, and 22 mM glucose + 5 μM of the compound of formula (I) (G22 + C(I)) vs. 11 mM glucose. Data are shown as mean ± SEM. Statistical significance was determined by Wilcoxon test (##p < 0.01) for 22 mM glucose vs. 11 mM glucose, and by Wilcoxon test (*p < 0.05, **p < 0.01) for 22 mM glucose + 5 μM of the compound of formula (I) vs. 22 mM glucose. [Figure 6b] Shows the alleviation of hyperglycemia-induced pancreatic gene expression changes in islets cultured ex vivo by the compound of formula (I). Overrepresentation analysis (ORA) of the Molecular Signature Hallmark gene set (MSIG) in mouse islets cultured in 22 mM vs. 11 mM glucose, and 22 mM glucose + 5 μM of the compound of formula (I) vs. 22 mM glucose. Data are shown as mean ± SEM. Statistical significance was determined by Wilcoxon test (##p < 0.01) for 22 mM glucose vs. 11 mM glucose, and by Wilcoxon test (*p < 0.05, **p < 0.01) for 22 mM glucose + 5 μM of the compound of formula (I) vs. 22 mM glucose. [Figure 6c]This study demonstrates the mitigation of hyperglycemia-induced changes in pancreatic islet gene expression in ex vivo cultured islets by the compound of formula (I). The log2 multiple change in the normalized read count of Txnip (all groups n=5) is shown for mouse islets cultured with 11 mM glucose, 22 mM glucose, and 22 mM glucose + 5 μM of the compound of formula (I). Data are presented as mean ± SEM. Statistical significance was determined by the Wilcoxon test (##p<0.01) for 22 mM glucose vs. 11 mM glucose, and by the Wilcoxon test (*p<0.05, **p<0.01) for 22 mM glucose + 5 μM of the compound of formula (I) vs. 22 mM glucose. [Figure 6d] This shows the mitigation of hyperglycemia-induced changes in pancreatic islet gene expression in ex vivo cultured pancreatic islets by the compound of formula (I). (b) is a heatmap of normalized read counts of DEGs derived from the enhancement category and KEGG:TCA_CYCLE. The number of genes was limited to the 20 most significant DEGs. [Figure 7]This shows the mitigation of the effects of chronic hyperglycemia on GSIS, mTORC1, and AMPK signaling in INS-1E cells by the compound of formula (I). a is the GSIS (n=4-5 / group) of INS-1E cells cultured for 4 days under 11 mM or 25 mM glucose conditions, both untreated and treated with 5 μM of the compound of formula (I) for 4 days. b is the GSIS (n=4-5 / group) of INS-1E cells cultured for 4 days under 11 mM or 25 mM glucose conditions, both untreated and treated with 5 μM of the compound of formula (I) for 2 hours. c is the protein expression ratio (n=4-5 / group) of phosphorylated (P-) and total ACC, AMPK, RAPTOR, and S6 in INS-1E cells cultured for 4 days under 11 mM or 25 mM glucose conditions, both untreated and treated with 5 μM of the compound of formula (I) for 4 days. d represents the protein expression ratios (n=4-5 / group) of phosphorylated (P-) and total amounts of ACC, AMPK, RAPTOR, and S6 in INS-1E cells cultured for 4 days under 11 mM or 25 mM glucose conditions, for untreated cells and cells treated with 5 μM of compound (I) for 2 hours (n=4-5 / group). Data are shown as mean ± SEM. Statistical significance was determined between untreated cells and cells treated with compound (I) by two-way ANOVA followed by Tukey's post-hoc test (*p<0.05, **p<0.01, ***p<0.001), and between cells cultured at 11 mM and 25 mM glucose by the same test (○p<0.05, ○○p<0.01). [Figure 8]The immunohistochemical and metabolic analyses of control mice and STZ mice are shown. a is a representative double immunostaining (n=5 in total) for insulin (green) and glucagon (red) in the pancreases of control mice and STZ mice that were untreated or treated with 0.25 and 0.5 mg / g of the compound of formula (I) from day 5. b is the total insulin content in the pancreas (n=7-8 / group) in STZ mice that were untreated or treated with 0.25 and 0.5 mg / g of the compound of formula (I), respectively, from day 5. c is the islet cell area in STZ mice that were untreated (n=3-5) or treated with 0.25 mg / g (n=3-5) and 0.5 mg / g (n=3-6), respectively, from day 5. d is the fraction of insulin (Ins+) cells in STZ mice that were untreated from day 5 (n=3-5) or treated with the compound of formula (I) at 0.25 mg / g (n=3-5) and 0.5 mg / g (n=3-6), respectively. e is the non-fasting glucose level and area under the curve (AUC) in control mice (n=6) and STZ mice that were untreated from day 15 (n=9) or treated with the compound of formula (I) at 0.5 mg / g (n=9), respectively. f is representative double immunostaining for insulin and glucagon in the pancreas of STZ mice that were untreated from day 15 or treated with the compound of formula (I) at 0.25 and 0.5 mg / g, respectively (all groups n=5). g is the total insulin content in the pancreas (n=8-16 / group) in STZ mice that were untreated or treated with the compound of formula (I) at 0.25 mg / g and 0.5 mg / g, respectively, from day 15. h is the islet cell area in STZ mice that were untreated (n=6-10) or treated with the compound of formula (I) at 0.25 mg (n=3-5) and 0.5 mg / g (n=6-10), respectively, from day 15. i is the fraction of Ins+ cells in STZ mice that were untreated (n=6-10) or treated with the compound of formula (I) at 0.25 mg (n=3-5) and 0.5 mg / g (n=6-10), respectively, from day 15. Data are shown as mean ± SEM.*P<0.05, **P<0.01, and ***P<0.001 were determined by Welch's ANOVA followed by Games-Howell post-hoc tests ((b), (c), (g), (h)), Tukey's post-hoc tests ((d), (i)) followed by one-way ANOVA, or Student's t-test ((f)). [Figure 9] This shows standardized uptake (SUV) values ​​during FDG-PET scans. The SUV profiles of the gastrocnemius muscle and heart are from 40-minute dynamic FDG-PET scans at baseline (Scan 1), 9-10 days after the last STZ injection (Scan 2), and after 1 week of treatment with 0.5 mg / g of compound (I) (n=5) (Scan 3) or untreated (n=5). Data are shown as mean ± SEM. [Figure 10] This shows the quantitative analysis of protein levels of p-T172 AMPK, p-S79 ACC, and TXNIP in vastus muscle extracts from control mice (n=5) and STZ mice that were either untreated (n=9) or treated with 0.5 mg / g of the compound of formula (I) from day 15 (n=9), as determined by Western blot analysis. [Figure 11]This shows that treatment with the compound of formula (I) does not increase serum lactate levels in STZ mice and db / db mice. a is the fasting glucose level of STZ mice on day 15 (i.e., before treatment) and on day 22 after 1 week of treatment with 0.5 mg / g of the compound of formula (I) (n=9 / group). b is the lactate level of STZ mice on day 15 (i.e., before treatment) and on day 22 after 1 week of treatment with 0.5 mg / g of the compound of formula (I) (n=9 / group). c is the fasting glucose level and area under the curve (AUC) (n=7-9 / group) in untreated BKS mice and db / db mice treated with 0.5 and 1.0 mg / g of the compound of formula (I), respectively. d represents the lactate levels and area under the curve (AUC) (n=7-9 / group) in BKS mice and db / db mice that were untreated or treated with 0.5 mg / g of the compound of formula (I), respectively. Data are shown as mean ± SEM. The statistical significance between time points in (a) was determined by Student's t-test (*P<0.05), the statistical significance between untreated db / db mice and db / db mice treated with the compound of formula (I) was determined by Welch's ANOVA followed by Games-Howell post-hoc test (*P<0.05, **P<0.01, ***P<0.001), and the statistical significance between BKS mice and db / db mice in (c) and (d) was determined by Wilcoxon test (##P<0.01). [Figure 12]The fractions of PP+ and Som+ cells, as well as the total proinsulin content in the pancreas, are shown for BKS mice and db / db mice. 'a' represents the fractions of pancreatic polypeptide (PP+) and somatostatin (Som+) positive cells (n=3 per group) in 15-week-old BKS mice and db / db mice treated with formula (I) at 0.5 and 1.0 mg / g, respectively. 'b' represents the total proinsulin content in the pancreas (n=8-10 / group) in 15-week-old BKS mice (n=8) and db / db mice treated with formula (I) at 0.5 and 1.0 mg / g, respectively. Data are shown as mean ± SEM. *P<0.05 was determined between untreated db / db mice and db / db mice treated with formula (I) by Welch ANOVA followed by Games-Howell post-hoc testing. The value #P<0.05 was determined by Welch's ANOVA between BKS mice and db / db mice. [Figure 13a] This shows the effect of the compound of formula (I) on gene expression signatures in ex vivo cultured mouse and human pancreatic islets. The MA plot shows differential gene expression between mouse pancreatic islets cultured with 22 mM (G22) versus 11 mM (G11) glucose. [Figure 13b] This shows the effect of the compound of formula (I) on gene expression signatures in ex vivo cultured mice and human pancreatic islets. The MA plot shows differential gene expression between 22 mM glucose + 5 μM of the compound of formula (I) (G22 + C(I)) versus 22 mM glucose. [Figure 13c] This shows the effect of the compound of formula (I) on gene expression signatures in ex vivo cultured mice and human pancreatic islets. The MA plot shows differential gene expression between 22 mM glucose + 5 μM of the compound of formula (I) versus 11 mM glucose. [Figure 13d] This shows the effect of the compound of formula (I) on gene expression signatures in ex vivo cultured mouse and human pancreatic islets. The MA plot shows differential gene expression between human pancreatic islets cultured with 25 mM (G25) versus 5.5 mM (G5) glucose. [Figure 13e]This shows the effect of the compound of formula (I) on gene expression signatures in ex vivo cultured mice and human pancreatic islets. The MA plot shows differential gene expression between 25 mM glucose + 5 μM of the compound of formula (I) (G25 + C(I)) versus 25 mM glucose. [Figure 13f] This shows the effect of the compound of formula (I) on gene expression signatures in ex vivo cultured mice and human pancreatic islets. The MA plot shows differential gene expression between 25 mM glucose + 5 μM of the compound of formula (I) versus 5.5 mM glucose. [Figure 13g] This study demonstrates the effect of the compound of formula (I) on gene expression signatures in ex vivo cultured mouse and human pancreatic islets. It includes overexpression analysis (ORA) of the Molecular Signature Hallmark gene set in human pancreatic islets cultured with 22 mM vs. 11 mM glucose, and with 22 mM glucose + 5 μM of the compound of formula (I) vs. 22 mM glucose. [Figure 14] Representative immunoblots are shown for INS-1E cells cultured for 4 days under 11 mM or 25 mM glucose conditions, both untreated and treated with 5 μM of compound (I) for 4 days (4d) or 2 hours (2h). [Figure 14] Representative immunoblots are shown for INS-1E cells cultured for 4 days under 11 mM or 25 mM glucose conditions, both untreated and treated with 5 μM of compound (I) for 4 days (4d) or 2 hours (2h). [Figure 15] A schematic model of the antidiabetic effect of the compound of formula (I) is shown. As a dual AMPK activator and mitochondrial uncoupler, the compound of formula (I) stimulates glucose uptake and utilization in muscles, and in part, through the reduction of mTORC1 signaling and Pdk expression, avoids the glucotoxic effects of hyperglycemia on β-cell function. [Modes for carrying out the invention]

[0014] The following description concerns exemplary compositions, methods, parameters, etc. However, it should be noted that such description is not intended to limit the scope of this disclosure, but rather is provided as a description of exemplary embodiments.

[0015] This specification provides a method for regulating blood glucose levels in subjects requiring it. The method herein regulates blood glucose levels through a dual antihyperglycemic effect, manifesting as increased glucose effect and preservation of β-cells. In some variants, the subject is insulin-deficient and is taking or requires exogenous insulin. In some of the aforementioned variants, the subject is human.

[0016] In some embodiments, this method targets compounds of formula (I): [ka] This includes administering a pharmaceutically acceptable salt, solvate, or prodrug thereof; monitoring the subject's blood glucose levels after administration of the compound; and reducing or eliminating the dose of exogenous insulin administered to the subject to maintain the subject's blood glucose levels within the subject's normal range.

[0017] In some embodiments, normal blood glucose levels are as defined by the American Diabetes Association in the United States or equivalent organizations in other countries. In some variations, the normal range for the subject is as follows: • A1C: Less than 7% A1C may sometimes be reported as eAG: less than 154 mg / dL. • Before meals (pre-meal plasma glucose): 80-130 mg / dL • 1-2 hours after the start of a meal (postprandial plasma glucose): Less than 180 mg / dL It should be understood that blood glucose levels can be measured using any suitable method known in the art, including, for example, a blood glucose meter or a continuous glucose meter.

[0018] In other embodiments, the method further includes monitoring the urine of the subject for ketones, for example, to identify if blood glucose levels are higher than normal.

[0019] As described above, the compound of formula (I), or its pharmaceutically acceptable salts, solvates, or prodrugs, enhances the glucose effect while preserving β-cells. The "glucose effect" refers to the process by which glucose itself, independently of insulin, promotes both its own uptake from the circulatory system into cells and its subsequent metabolism. Administration of the compound of formula (I), or its pharmaceutically acceptable salts, solvates, or prodrugs provided herein enhances the glucose effect while maintaining β-cell function under hyperglycemic conditions, even in the absence of exogenous insulin.

[0020] Surprisingly, the compounds provided herein have been observed to act as exercise mimics, enhancing cardiac function and exercise capacity without promoting glycogen accumulation in cardiac tissue. The compounds provided herein promote mitochondrial uncoupling within myotubes. Mitochondrial uncoupling dissipates the potential-energy gradient across the inner mitochondrial membrane. This potential energy is converted into thermal energy rather than being used for oxidative phosphorylation (the process of converting ADP to ATP). Therefore, administration of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, generates a metabolic demand for glucose that promotes glucose utilization rather than glycogen accumulation within cells.

[0021] Surprisingly, the compounds provided herein have also been observed to act on glucose control without insulin. Specifically, these compounds improve glucose efficacy independently of insulin, showing improvement in subjects with type 1 diabetes (T1D), a loss of β-cells, a genetic condition resulting in loss of insulin secretion, or a genetic defect affecting the ability of tissues to respond to insulin (e.g., congenital hyperinsulinism).

[0022] Therefore, the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, can be used in a variety of ways. In some embodiments, the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered to a subject with T1D before insulin therapy is initiated, in order to delay the need for insulin use. In some embodiments, administration of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, to a subject (e.g., a patient) who needs it lowers the subject's diastolic blood pressure. In one embodiment, the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered to a subject with T1DM accompanied by unstable diabetes, where the subject has high and low glucose levels despite using insulin. In further embodiments, the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered to subjects suffering from conditions in which glucose control is favored independently of insulin action, such as those with genetic secretory deficiencies and genetic deficiencies in tissue insulin signaling.

[0023] Insulin deficiency Beta cells produce insulin, which the body needs to regulate glucose levels in the bloodstream. For example, if a subject's beta cells are destroyed by cell-mediated autoimmune destruction of pancreatic beta cells, the subject will experience high blood glucose levels, also known as hyperglycemia in this field. For example, in individuals with T1D, beta cells are generally destroyed by the immune system, and the beta cells produce little to no insulin. When there is insufficient insulin, glucose accumulates in the bloodstream instead of being taken up by cells. This accumulation of glucose in the blood is called hyperglycemia.

[0024] Insulin deficiency can be absolute insulin deficiency or relative insulin deficiency. In one embodiment, this specification provides a method for regulating blood glucose levels in a subject suffering from absolute insulin deficiency, which occurs when the body is unable to produce any insulin due to autoimmune damage to pancreatic β-cells or complete resection of the pancreas. The pancreas is responsible for producing insulin, a hormone that regulates blood glucose levels in the body. In absolute insulin deficiency, because there is no insulin in the blood to regulate blood glucose levels, exogenous insulin (or an external source of insulin) is required to perform the functions that insulin performs in the body, including controlling blood glucose levels, managing the body's energy levels, and responding to glucose storage and distribution in the body's cells.

[0025] In some embodiments, the individual has T1D, chronic pancreatitis, pancreatectomy, cystic fibrosis, monogenic diabetes, or a post-infectious condition following severe acute respiratory syndrome (SARS) infection or another viral infection. In one embodiment, the individual has T1D. In some embodiments, the subject has juvenile mature diabetes mellitus (MODY) or adult latent autoimmune diabetes mellitus (LADA).

[0026] In another aspect, this specification provides a method for regulating blood glucose levels in an individual suffering from relative insulin deficiency, which typically occurs when the body develops insulin resistance and is unable to produce enough insulin to manage the blood glucose present in the body.

[0027] In one embodiment, the Specified Information provides a method for regulating blood glucose levels in an individual suffering from acute insulin deficiency. In one embodiment, the individual suffers from diabetic ketoacidosis. In another embodiment, the individual suffers from unstable diabetes.

[0028] In some variants, individuals suffer from T1D, a condition characterized by loss of β-cells. In other variants, individuals suffer from a genetic condition resulting in loss of insulin secretion. In yet another variant, individuals suffer from a genetic defect that affects the insulin responsiveness of tissues. In one variant, individuals suffer from congenital hyperinsulinism.

[0029] Exogenous insulin In some modified embodiments, the subject is a human. Subjects described in the manner provided herein are taking or need to take exogenous insulin (or an external source of insulin) prior to the first dose of the compound of formula (I), or its pharmaceutically acceptable salt, solvate, or prodrug. Subjects may be taking, or need to take, any appropriate exogenous insulin therapy, including sustained-release insulin (also known as long-acting insulin) or rapid-acting insulin (also known as fast-acting or short-acting insulin), or a combination thereof. Sustained-release insulin typically supplies the body with the insulin it needs throughout the day and night. Examples include insulin glargine (Lantus), insulin detemir (Levemir), and insulin degludec (Tesiba). Rapid-acting insulin is typically taken before the subject eats or drinks anything containing carbohydrates. Examples of rapid-acting insulins include insulin aspart (Novorapid), insulin aspart (Fiasp), insulin lispro (Humalog), and insulin glulisine (Apidra). Such exogenous insulins may be administered to subjects using any preferred method known in the art, including injection, insulin pumps, and insulin pens.

[0030] Remarkably, after administering the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, over time, and after the subject has adapted to the effects of the compound, the need for exogenous insulin decreases and eventually disappears over time as long as the subject continues to take the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0031] In some embodiments of the methods provided herein, the blood glucose levels of the subject are monitored before and after administration of the compound. Due to the remarkable dual antihyperglycemic effect, which manifests as increased glucose effect and preservation of β-cells, the insulin levels of the subject gradually return to normal levels without the need for exogenous insulin.

[0032] In some embodiments, the administration of exogenous insulin is not required, for example, after the subject has been administered the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof for two weeks, as long as the subject continues to receive the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the administration of exogenous insulin is initially reduced and then later eliminated.

[0033] therapeutic compounds In some of the embodiments described above, the therapeutic compound includes exemplary compounds, which are described in more detail below. Compounds used in the methods provided herein may include salts, solvates, or prodrugs thereof.

[0034] In some embodiments, the compound is 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide, or a salt thereof, solvate, or prodrug.

[0035] In one embodiment, the compound is an alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. "Alkali metals" are metals that, along with hydrogen, belong to Group I of the periodic table. Alkali metals include lithium, sodium, potassium, rubidium, cesium, and francium. Therefore, it can be understood that an "alkali metal salt" is a compound consisting of an aggregate of one or more alkali metal cations and anions associated with them. Therefore, the term "alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide" refers to compounds containing an alkali metal cation (e.g., lithium, rubidium, cesium, sodium, and potassium) and an anion of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. For example, alkali metal salts of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide are as follows: [ka] In the formula, X + This represents an alkali metal cation (e.g., lithium, rubidium, cesium, sodium, or potassium).

[0036] It will be understood that a "sodium salt" is a compound consisting of a sodium cation and an anion associated with it. Therefore, the term "sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide" refers to a compound containing a sodium cation and the anion of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. For example: [ka] In the formula, Na + This represents a sodium cation.

[0037] Those skilled in the art will recognize that, when dissolved in a suitable solvent (e.g., water), the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide can dissociate into its anionic and cationic components.

[0038] Throughout this specification, structures may or may not be presented by their chemical names. In the event of any question regarding nomenclature, the structure takes precedence. Where a compound can exist as a tautomer (e.g., in an alternative resonance form), the structure shown represents one of the possible tautomer forms, and the tautomer form(s) actually observed may vary depending on environmental factors such as solvent, temperature, or pH. All tautomer (and resonance) forms and mixtures thereof are included within the scope of this invention. For example, the following tautomers are included within the scope of this invention: [ka]

[0039] To avoid misunderstanding, the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide is solid under ambient conditions, and therefore the scope of the present invention includes all of its amorphous, crystalline, and partially crystalline forms.

[0040] Alkali metal salts of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide can be prepared according to techniques well known to those skilled in the art. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide may be reacted with a suitable alkali metal hydroxide or an alternative alkali metal base compound. Salt switching techniques can also be used to convert one salt to another.

[0041] The sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide can be prepared according to techniques well known to those skilled in the art. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide may be reacted with sodium hydroxide or an alternative sodium base compound. Salt switching techniques can also be used to convert one salt to another.

[0042] When the salt is prepared from 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide can be prepared according to the art well known to those skilled in the art. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide can be produced according to the art described in international patent application WO2011 / 004162.

[0043] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention pertains.

[0044] In certain embodiments, the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide is the sodium or potassium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. In one embodiment, the salt is the sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0045] In some embodiments, the compounds used in the methods provided herein are compounds of formula (II): [ka] or a salt thereof, where R 1 The compound is selected from the group consisting of -C(O)-C2H4-CO2H and -PO3H2, or their salts or solvates. In some embodiments, the compound can be metabolized in vivo to form 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. In some embodiments, the compound is a salt. For example, salts of the compound include: [ka] In the formula, X + is an alkali metal, alkaline earth metal, or quaternary ammonium (e.g., lithium, magnesium, calcium, ammonium, tetramethylammonium, especially sodium and potassium) cation, and appropriate stoichiometric adjustments are made considering the ion's charge. In one embodiment, X + This represents an alkali metal cation (e.g., lithium, rubidium, cesium, or especially sodium or potassium).

[0046] Pharmaceutical dosage forms The therapeutic compound is administered to human subjects in need in the form of a pharmaceutical formulation, which is also referred to herein as a pharmaceutical dosage form.

[0047] In one embodiment, the therapeutic compound is the sole active pharmaceutical ingredient present in the dosage form. In further embodiments, the therapeutic compound may be present in the dosage form together with one or more other active pharmaceutical ingredients, or administered as part of a combination therapy with one or more other active pharmaceutical ingredients.

[0048] In certain embodiments, the therapeutic compound is provided in the form of particles having a particle size distribution defined by D90 less than about 10 μm (e.g., measured using laser diffraction). In one embodiment, particles containing the therapeutic compound may have a particle size distribution defined by D90 less than about 10 μm (e.g., about 5 μm to about 10 μm) (e.g., measured using laser diffraction). Alternatively, the particle size distribution may be defined by D90 less than about 8 μm (e.g., about 5 μm to about 8 μm). In further embodiments, particles consisting of the therapeutic compound may have a particle size distribution defined by D50 less than about 6 μm (e.g., about 0.5 μm to about 6 μm). In even further embodiments, the particle size distribution of particles consisting of the therapeutic compound is further defined by D10 less than about 2 μm (e.g., about 0.2 μm to about 2 μm). The above particle size distribution parameters may be applied individually or in combination. For example, in certain embodiments, the dosage form comprises particles containing a therapeutic compound, the particles having a particle size distribution defined by D90 less than about 10 μm and D50 less than about 6 μm. Furthermore, the particles may have a particle size distribution defined by D90 less than 9 μm, D50 less than 6 μm or less than 5 μm, and D10 less than 2 μm or less than 1.5 μm. The particle size distribution of the particles containing the therapeutic compound may be measured, for example, by laser diffraction using a commercially available particle size analyzer.

[0049] dose Those skilled in the art will understand that the pharmaceutical dosage forms described herein may act systemically and, therefore, may be administered using appropriate techniques known to those skilled in the art. The pharmaceutical dosage forms described herein are typically administered orally, for example, as oral pharmaceutical dosage forms. Accordingly, several variants provide oral pharmaceutical dosage forms comprising about 200 to about 1000 mg of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide or a pharmaceutically acceptable salt, solvate, or prodrug thereof. One variant provides an oral pharmaceutical dosage form comprising about 200 to about 1000 mg of the sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0050] In certain embodiments, the pharmaceutical dose may contain about 200 mg to about 800 mg, about 200 mg to about 600 mg, or about 200 mg to about 400 mg of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some variant forms, the pharmaceutical dosage form contains about 200 to about 400 mg of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for example, the sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0051] Enteric coating Dosage forms intended for oral administration may further include an enteric coating to prevent or minimize dissolution or disintegration in the gastric environment. Thus, oral formulations coated with an enteric coating (e.g., capsules or tablets) may result in targeted release of the therapeutic compound in the small intestine. For example, the enteric coating may be present on the surface of the formulation (e.g., the surface of the tablet or capsule), or each particle containing the therapeutic compound may be coated with the enteric coating. Therefore, in certain embodiments, the pharmaceutical dosage forms used in the method of the present invention further include an enteric coating.

[0052] In certain embodiments, the enteric coating is present on the pharmaceutical dosage form, and in some variations, the coating may be provided as an outer layer on the pharmaceutical dosage form.

[0053] Alternatively, particles containing the therapeutic compound may be individually coated with an enteric coating, and the coated particles may be prepared into a pharmaceutical dosage form. Thus, in certain embodiments, the pharmaceutical dosage form contains particles containing the therapeutic compound, each particle being coated with an enteric coating.

[0054] The term "enteric coating" refers to a substance (e.g., a polymer) incorporated into an oral medication (e.g., applied to the surface of a tablet, capsule, particle, or pellet) that inhibits the dissolution or breakdown of the medication in the gastric environment. Enteric coatings are typically stable at the highly acidic pH found in the stomach but rapidly decompose at the relatively basic pH of the small intestine. Therefore, the enteric coating prevents the release of the active ingredient in the medication until it reaches the small intestine.

[0055] Any enteric coating known to those skilled in the art can be used in the present invention. Specific enteric coating materials that may be mentioned include beeswax, shellac, alkylcellulose polymer resins (e.g., ethylcellulose polymer, carboxymethylethylcellulose, or hydroxypropylmethylcellulose phthalate), or acrylic polymer resins (e.g., acrylic acid and methacrylic acid copolymer, methyl methacrylate copolymer, ethoxyethyl methacrylate, cyanoethyl methacrylate, methacrylate copolymer, methacrylic acid copolymer, aminoalkyl methacrylate copolymer, poly(acrylic acid), poly(methacrylic acid), alkyl methacrylate amide copolymer, poly(methyl methacrylate), poly(methacrylic acid) (anhydride), polymethacrylate, methyl methacrylate copolymer, poly(methyl methacrylate) copolymer, polyacrylamide, poly(methacrylic anhydride), and glycidyl methacrylate copolymer).

[0056] Dosage form In some variations, the therapeutic compound may be provided in the form of a tablet or, in particular, a capsule. For example, capsules such as soft gelatin capsules may be prepared containing the therapeutic compound alone or together with a suitable vehicle, such as vegetable oil or fat. Similarly, hard gelatin capsules may contain the therapeutic compound alone or in combination with solid powder components such as disaccharides (e.g., lactose or saccharose), alcoholic sugars (e.g., sorbitol or mannitol), vegetable starches (e.g., potato starch or corn starch), polysaccharides (e.g., amylopectin or cellulose derivatives), or gelling agents (e.g., gelatin).

[0057] The pharmaceutical dosage forms described herein may be prepared in accordance with standard and / or known pharmaceutical practice. The pharmaceutical dosage forms are generally provided as mixtures comprising a therapeutic compound and one or more pharmaceutically acceptable excipients. The one or more pharmaceutically acceptable excipients may be selected in accordance with standard pharmaceutical practice, taking into account the intended route of administration. Such pharmaceutically acceptable excipients are preferably chemically inert to the active compound and preferably free from adverse side effects or toxicity under the conditions of use. Suitable pharmaceutical formulations can be found, for example, in Remington, The Science and Practice of Pharmacy, 19th ed., Mack Printing Company, Easton, Pennsylvania (1995). A brief overview of drug delivery methods can also be found, for example, in Langer, Science 249, 1527 (1990).

[0058] Excipients In some modified embodiments, the pharmaceutical composition comprises a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and at least one pharmaceutically acceptable excipient. In particular, the at least one pharmaceutically acceptable excipient may be a lubricant, binder, filler, surfactant, diluent, anti-adhesion agent, coating agent, flavoring agent, coloring agent, fluidizer, preservative, sweetener, disintegrant, adsorbent, buffer, antioxidant, chelating agent, dissolution accelerator, dissolution retarder, or wetting agent.

[0059] Specific pharmaceutically acceptable excipients that may be mentioned include mannitol, PVP (polyvinylpyrrolidone) K30, lactose, saccharose, sorbitol, starch, amylopectin, cellulose derivatives, gelatin, or other suitable ingredients, as well as disintegrants and lubricants such as sodium lauryl sulfate, sodium doxate, magnesium stearate, calcium stearate, sodium stearyl fumarate, and polyethylene glycol wax. In the preparation of pharmaceutical dosage forms of therapeutic compounds for oral administration, particles containing the therapeutic compound (preferably ground) may be mixed with mannitol, PVP (polyvinylpyrrolidone) K30, and sodium lauryl sulfate, either together or separately.

[0060] In the preparation of pharmaceutical dosage forms, the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, may be mixed with one or more pharmaceutical excipients (including basic excipients), either together or separately.

[0061] A mixture of a therapeutic compound and one or more pharmaceutically acceptable excipients can be processed into pellets or granules, or compressed into tablets. Therefore, the pharmaceutical dosage forms of the method of the present invention may be tablets, small tablets, blocks, pellets, particles, granules, or orally administered powders.

[0062] Pharmaceutical formulations that may be mentioned include those in which the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is present in an amount of at least 1% by weight (or at least 10% by weight, at least 30% by weight, or at least 50% by weight) of the total weight of the formulation. That is, the weight ratio of the therapeutic compound to the total weight of the components of the pharmaceutical formulation (i.e., the therapeutic compound and all pharmaceutical excipients, e.g., adjuvants, diluents, and carriers) is at least 1:99 (or at least 10:90, at least 30:70, or at least 50:50).

[0063] As used herein, “therapeutably effective amount,” “effective amount,” or “dosage” refers to an amount of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, sufficient to produce a desired effect, which may be a therapeutic and / or beneficial effect. The effective amount or dosage will vary depending on the age or general condition of the individual or subject (e.g., human), the severity of the condition being treated, the specific drug administered, the duration of treatment, the nature of any concurrent treatments, the pharmaceutically acceptable carrier used, and similar factors within the scope of the knowledge and expertise of those skilled in the art. Where appropriate, the “therapeutably effective amount,” “effective amount,” or “dosage” may be determined by those skilled in the art in each individual case by referring to the relevant literature and / or by conducting routine experiments. Those skilled in the art will understand that the therapeutic effect does not need to be complete or curative, as long as some benefit is provided to the subject.

[0064] Those skilled in the art will understand that a suitable formulation may be administered in various doses (for example, via one or more preparations described herein), and that a suitable dose can be easily determined by those skilled in the art. The total dose of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof administered to a subject requiring it may range from about 0.01 to about 2000 mg / kg / day, about 0.1 to about 500 mg / kg / day, or about 1 to about 100 mg / kg / day.

[0065] When administered orally, treatment with such formulations (including capsules containing such formulations) may involve the administration of a unit dose formulation containing about 0.01 mg to about 3000 mg of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for example, about 0.1 mg to about 2000 mg, or about 1 mg to about 1000 mg (e.g., about 10 mg to about 500 mg) of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, treatment involves a single daily administration of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof (including capsules containing said formulations). Alternatively, the total daily dose of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, may be administered in two, three, or four divided doses per day (for example, 100 mg, 250 mg, 500 mg, or 1000 mg twice daily, as described herein). An experienced physician will recognize that the dosage will vary depending on the patient.

[0066] In certain embodiments, the daily dose of the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug administered to the subject is in the range of approximately 1 mg to approximately 3000 mg. In certain embodiments, the daily dose of the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug administered to the subject is in the range of approximately 1 mg to approximately 1,000 mg.

[0067] As used herein, the term “about” when referring to measurable values ​​such as the amount, dose, time, or temperature of a compound refers to a variation of a specific amount of 20%, 10%, 5%, 1%, 0.5%, or even 0.1%. In each example, it is intended that such a term may be replaced by notation such as “±10%” or similar notation (or by indicating a variation of a specific amount calculated based on the relevant value). It is also intended that such a term may be deleted in each example.

[0068] It should be understood that the dosages and pharmacokinetic parameters described below pertain to the treatment of specific diseases, particularly patients with T1D as described herein.

[0069] In any of the prior embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug can be orally administered once daily to a patient in need at a dose of approximately 100 mg to approximately 1,000 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug can be orally administered once daily at a dose of approximately 200 mg to approximately 1,000 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug can be orally administered once daily at a dose of approximately 400 mg to approximately 800 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug can be orally administered once daily at a dose of approximately 100 mg to approximately 300 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily in doses of about 100 mg, about 200 mg, about 300 mg, about 400 mg, or about 500 mg.

[0070] In any of the prior embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof can be administered orally once daily to a subject requiring it, wherein the steady-state plasma concentration of the compound of formula (I) is approximately 40 μg / mL to approximately 200 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered orally once daily to a subject or patient requiring it, wherein the steady-state plasma concentration of the compound of formula (I) is approximately 40 μg / mL to approximately 80 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered orally once daily to a subject or patient requiring it, wherein the steady-state plasma concentration of the compound of formula (I) is approximately 70 μg / mL to approximately 120 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug is administered orally once daily to a subject or patient in need, resulting in a steady-state plasma concentration of approximately 90 μg / mL to approximately 160 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug is administered orally once daily to a subject or patient in need, resulting in a steady-state plasma concentration of approximately 100 μg / mL to approximately 150 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug is administered daily to a subject (e.g., a human), resulting in a steady-state plasma concentration of approximately 120 μg / mL to approximately 140 μg / mL.

[0071] In any of the prior embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof can be administered orally once daily to a patient in need, wherein the administration provides a steady-state AUC of the compound of formula (I). 0-24is about 1,000 h*μg / mL to about 4,000 h*μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof can be orally administered once daily to a subject or patient in need thereof, wherein, by said administration, the AUC at steady state of the compound of formula (I) 0-24 is about 1,000 h*μg / mL to about 2,000 h*μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof can be orally administered once daily to a subject or patient in need thereof, wherein, by said administration, the AUC at steady state of the compound of formula (I) 0-24 is about 1,500 h*μg / mL to about 4,000 h*μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof can be orally administered once daily to a subject or patient in need thereof, wherein, by said administration, the AUC at steady state of the compound of formula (I) 0-24 is about 1,800 h*μg / mL to about 3,100 h*μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof can be orally administered once daily to a subject or patient in need thereof, wherein, by said administration, the AUC at steady state of the compound of formula (I) 0-24 is about 1,500 h*μg / mL to about 2,000 h*μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof can be orally administered once daily to a subject or patient in need thereof, wherein, by said administration, the AUC at steady state of the compound of formula (I) 0-24 is about 2,500 h*μg / mL to about 4,000 h*μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof can be orally administered once daily to a subject or patient in need thereof, wherein, by said administration, the AUC at steady state of the compound of formula (I) 0-24This ranges from approximately 3,000 μg / mL to approximately 3,500 μg / mL.

[0072] In any of the prior embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a patient in need, wherein the C of the compound of formula (I) max The concentration is approximately 40 μg / mL to approximately 200 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject or patient in need, wherein the administration provides the C of the compound of formula (I). max The concentration is approximately 70 μg / mL to approximately 200 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject or patient in need, wherein the administration provides the C of the compound of formula (I). max The concentration is approximately 80 μg / mL to approximately 140 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject or patient in need, wherein the administration provides the C of the compound of formula (I). max The concentration is approximately 70 μg / mL to approximately 100 μg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject or patient in need, wherein the administration provides the C of the compound of formula (I). max This ranges from approximately 1200 μg / mL to approximately 150 μg / mL.

[0073] In some of the variations described above, the subject or patient requiring it is human. To avoid misunderstanding, in the context of the present invention, the dose administered to the subject (e.g., human) should be sufficient to produce a therapeutic effect on the subject over a reasonable period of time. Those skilled in the art will recognize that the precise dose and composition, as well as the selection of the most appropriate delivery regimen, are also influenced, among other things, by the pharmacological properties of the formulation, the nature and severity of the condition being treated, the recipient's health status and mental sensitivity, the potency of the particular compound, the age, condition, weight, sex and response of the subject being treated, and the stage / severity of the disease.

[0074] In any case, a physician or other person skilled in the art can routinely determine the most appropriate actual dosage for the subject. The above dosages are illustrative for an average case, but naturally, there may be individual cases that deserve higher or lower dosages, and these are within the scope of this disclosure.

[0075] Use of compounds In certain embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in a treatment comprising any of the methods described herein is provided. For example, in some embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is provided for regulating blood glucose levels in a subject requiring it, by an action manifesting as an increase in the glucose effect. In some variant embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is provided for regulating blood glucose levels in a subject requiring it, by a dual antihyperglycemic effect manifesting as an increase in the glucose effect and preservation of β-cells. In one embodiment, the treatment comprises administering the compound to a subject; monitoring the subject's blood glucose levels after administration of the compound; and reducing or discontinuing the dose of exogenous insulin administered to the subject in order to maintain the subject's blood glucose levels within the subject's normal range.

[0076] In certain embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, for the manufacture of a drug is also provided. In some embodiments, the drug is for any of the methods described herein. For example, in some embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in the manufacture of a drug that regulates blood glucose levels in a subject having the effect of increasing the glucose effect. In some modified embodiments, the use of a compound of formula (I), or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in the manufacture of a drug that regulates blood glucose levels in a subject having the effect of increasing the glucose effect and preserving β-cells, is provided.

[0077] In one embodiment, a drug containing a compound is administered to a subject, the subject's blood glucose level is monitored after administration of the compound, and the dose of exogenous insulin administered to the subject is reduced or discontinued in order to maintain the subject's blood glucose level within the subject's normal range. [Examples]

[0078] The subject matter of this disclosure will be better understood by referring to the following embodiments, which are provided as examples of the present invention, rather than as limitations.

[0079] In this specification, unless otherwise specified, “Cmpd-(I)” or “compound of formula (I)” as referred to in the experiments detailed in the following examples is 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0080] Mice. Mice were housed in a certified animal facility under a 12-hour light / dark cycle and given free access to their respective diets. Leptin receptor-deficient male BKS.Cg-Dock7m + / + Leprdsb / J(db / db) mice and C57BLKS / J(BKS) mice were obtained. Male C57BL / 6J mice were mated with female CBA / CaCrl mice to obtain F1 mice. Male F1 mice at 9 weeks of age were treated with multiple low doses of streptozotocin (50 mg / kg* days for 5 consecutive days; freshly prepared in 0.1 mM sodium citrate (pH 4.5)) to induce diabetes. Mice were freely administered either a D10001 diet or D10001 formulated with the compound of formula (I), containing 0.25 mg / g, 0.5 mg / g, and 1 mg / g of the compound of formula (I) (CAS number 1261289-04-6). Cohorts of BKS, db / db, and F1 mice were housed in groups of 4-5 mice / cage. Mice with more than 10% weight loss or obvious health problems such as fighting were excluded to avoid intergroup differences. In different cohorts, mice were randomly assigned to cages based on body weight and fasting blood glucose, and assigned to different treatments per cage or, where possible, per individual, to minimize the effects of starting body weight and glucose homeostasis. Samples were randomly selected from 5-9 mice / diet for in vitro analyses such as Western blotting, qPCR, histological, and immunohistochemical studies. All in vivo analyses were performed between 9 a.m. and 3 p.m.

[0081] PET analysis of in vivo glucose uptake. All in vivo experiments were successfully repeated 2-3 times. In vitro tissue analysis was successfully repeated 2-3 times in different cohorts. Mice were starved for 3 hours before each scan. Before scanning, mice were sedated with less than 2% isoflurane in oxygen (800 mL / min), and a cannula was inserted via the tail vein using a 27G needle and a custom-made catheter. Dynamic PET acquisition for 40 minutes (frames 8x30s, 8x60s, 6x180s, 2x300s) was initiated by injecting 50-70 μL of 13.9+2.3 MBq[18F]-fluorodeoxyglucose (FDG) dissolved in physiological saline. The maximum metabolic rate of glucose, MRglu, was calculated using Patlak analysis of PET data combined with the measured glucose concentration. PET / CT imaging was performed on a 0.375x0.375x0.377 mm scale. 3 The study began with a 50kV, 0.088mAs helical CT acquisition, reconstructed into voxel-sized images. PET images were reconstructed to voxel size (0.4x0.4x0.4) in four iterations and four subsets using Tera-Tomo 3D iterative reconstruction with attenuation, scattering, and random correction. Image analysis and Patlak pharmacokinetic calculations were performed using imlook4d software. During scanning, mice were monitored on temperature-controlled beds, and blood glucose was measured using tail vein blood 10, 20, and 40 minutes after injection. Regions of interest (ROIs) consisting of both left and right gluteus medius / rectus femoris muscles were defined as cylinders with a 10-pixel diameter and 10 slices in the coronal view. Myocardial ROIs were defined by thresholding the search volume, manually depicted in the last PET frame, to 40% of the maximum voxels. Vena cava ROIs were defined as voxels exceeding 60% of the maximum voxels in the first frame showing uptake. The image derivation input function was approximated using a time-radioactivity curve from a vena cava ROI. Patlak analysis was performed using the above image derivation input function, and the irreversible acquisition rate K was determined by linear regression between 16 and 40 minutes. i Patlak K made the decision. i Values ​​were calculated at the ROI level for quantification and at the voxel level for visualization. Saturated glucose consumption is as follows:

number

[0082] Echocardiography. For echocardiography, mice are subjected to 0.8 L·min. -1 Subjects were sedated with 1.5-2% isoflurane in O2(g) and placed on a temperature-controlled table. Chest hair was removed using depilatory cream. Respiration and ECG were monitored during the scan, and anesthesia was adjusted to avoid respiratory depression. The total scan time did not exceed 15 minutes. Stroke volume, cardiac output, and wall thickness were measured in a parasternal long-axis view using B-mode and M-mode images. Diastolic left ventricular inflow was measured using mitral valve inflow Doppler in a apical four-chamber view. Offline analysis was performed blinded.

[0083] Mitochondrial respiration analysis. C2C12 myoblasts were cultured in growth medium containing 10% fetal bovine serum and 20 U / ml penicillin-streptomycin. To obtain myotubes, 7500 cells / cm³ of C2C12 myoblasts were placed on poly-L-lysine-coated XF96 plates. 2The myotubes were seeded and cultured in growth medium for 3 days until 80% confluence, then switched to differentiation medium (DM; FBS was replaced with 2% horse serum for 6 days, with the medium changed every 2 days). Respiratory assays were performed using an extracellular flux analyzer according to the manufacturer's "Mito-stress" protocol and mitochondrial parameters. Briefly, differentiated myotubes were pre-treated for 4 hours in growth medium containing 1% horse serum along with the compound of formula (I) (sodium salt preparation). The myotubes were then equilibrated for 1 hour in Seahorse assay medium (1 mM Na-pyruvate, 10 mM glucose, 2 mM L-glutamine) adjusted to pH 7.4 and supplemented with the compound of formula (I) as in the pre-treatment. As baseline, oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) measurements were collected, followed by sequential addition of 1 μM oligomycin, 2 μM FCCP, and 0.5 μM rotenone + 0.5 μM antimycin A. Mitochondrial function parameters were calculated according to the manufacturer's recommendations.

[0084] Western blot analysis. Vastus muscle was isolated from non-fasted mice and lysed in a mortar and pestle using liquid nitrogen. Vastus muscle and INS-1E cell samples were homogenized in ice-cold protein lysis buffer (100 mM Tris pH 6.8, 2% SDS) with a protease inhibitor cocktail and a phosphatase inhibitor cocktail, and then sonicated. After centrifugation at 14,000 rpm for 10 minutes, the vastus muscle supernatant was collected. Protein concentrations were measured using a BCA kit, and then the samples were diluted in Laemmli buffer and denatured. 20 μg of vastus muscle and 7 μg of INS-1E cell protein samples were separated on 4–15% Criterion® TGX Stain-Free® protein gel and blotted to low-fluorescence PVDF or nitrocellulose membranes, respectively. Primary and secondary antibodies are listed in Table 2. Values ​​were normalized to stain-free total protein signal or β-actin. [Table 1]

[0085] qRT PCR. Total RNA was prepared from isolated pancreatic islets using the RNeasy Micro Kit. Total RNA from gastrocnemius muscle and left ventricle was prepared using the RNeasy Fibrous Tissue Mini Kit after first disrupting the tissue in liquid nitrogen using a pestle. First-strand cDNA synthesis was performed using SuperScript III according to the manufacturer's instructions. The primers used for qRT-PCR are listed in Table 3. Tbp expression was used to normalize samples from pancreatic islets, gastrocnemius muscle, and left ventricle. Tbp normalization was validated with Rpl32 in left ventricular samples and gastrocnemius muscle. [Table 2]

[0086] Isolation and culture of pancreatic islets. Mouse pancreatic islets were essentially isolated and cultured for 48 hours in medium supplemented with 11 mM glucose or 22 mM glucose + / - 5 μM of the compound of formula (I) (RPMI 1640 medium, 1% fetal bovine serum, 10 mM HEPES, 1 mM sodium pyruvate, 50 μM 2-mercaptoethanol, 50 U / ml Pen:Strep). Human pancreatic islets from non-diabetic donors were cultured for 48 hours in medium supplemented with 5.5 mM glucose or 25 mM glucose + / - 5 μM of the compound of formula (I) (CMRL medium, 10% fetal bovine serum, 20 U / ml Pen:Strep and 1X GlutaMax).

[0087] RNA-seq was performed. An RNA-seq library was prepared from 150 ng of total RNA using the Illumina TruSeq stranded mRNA Library Kit, followed by 100 bp paired-end sequencing using a NovaSeq 6000 Illumina Sequencer. RNA-seq reads were processed. Fastq files containing 100 nt paired-end sequencing reads were quality-checked and aligned to mouse or human genomes (GRCm39 or GRCh38). Fragment-gene hits were counted. Subsequent analysis was performed in R using DESeq2 and the clusterProfiler package. Normalization and differential expression analysis were performed using DESeq2 with batch correction (referring to islet preparation batches and human donors, respectively), independent filtering (α=0.05), and false discovery rate (FDR) < 0.05. Genes whose baseMean expression exceeded the filter threshold were considered expressed, and for the overexpressed gene category, pvalueCutoff=0.05 and qvalueCutoff=0.2 were used to consider them as constituting the gene background for subsequent ORA and GSEA analyses using clusterProfiler.

[0088] Insulin secretion assay. GSIS was performed in UB buffer (125 mM NaCl, 6 mM KCl, 1.3 mM CaCl2, 1.2 mM MgCl2, 25 mM Hepes (pH 7.3), 2 mM glucose, 0.1% BSA) at 37°C under normal atmospheric conditions. Briefly, INS-1E cells were washed, equilibrated in UB buffer for 1 hour, transferred to UB buffer containing either 2 or 20 mM glucose for 30 minutes, and insulin levels in the supernatant were measured by ELISA. Insulin secretion was normalized to protein content.

[0089] Cell lines. INS-1E and C2C12 were commercially available and therefore not certified after purchase. PCR testing confirmed that these cells did not contain mycoplasma.

[0090] Statistical analysis. The experimental design consisted of a non-diabetic control group (BKS or control, i.e., vehicle-injected F1 mice), a diabetic control group (db / db mice or STZ mice, i.e., STZ-injected F1 mice), and a group(s) treated with the compound of formula (I) (db / db mice or STZ mice continuously fed a diet formulated with the compound of formula (I)). First, the validity of the diabetes model was tested by comparing the control group with the diabetic group. Second, the effect of the diet with the compound of formula (I) was evaluated by comparing the group(s) treated with the compound of formula (I) with the diabetic group. The parametricity and heterogeneity of the data were assessed using the Shapiro-Wilk test and the Rubine test, and used to select the appropriate statistical test. For two-group comparisons, the unequal t-test or Wilcoxon test was used, along with Holm correction for multiple testing. For multiple groups, ANOVA analysis was used, followed by appropriate post-hoc analysis (see legend in the figure). A p-value of <0.05 was considered statistically significant. Data analysis, statistical analysis, and visualization were performed using the tidyverse, rstatix, and ggpubr packages in R.

[0091] Example 1: The compound of formula (I) prevents the development of diabetes in STZ mice. To evaluate whether the compound of formula (I) can improve diabetes in insulin-deficient conditions, β-cell function was resected in mice by streptozotocin (STZ) injection. STZ mice were treated with a diet formulated with 0.25 or 0.5 mg / g of the compound of formula (I), indicated as Cmpd-(I)-(0.25) and Cmpd-(I)-(0.5), starting from day 5 or day 15. In untreated STZ mice, insulin levels rapidly decreased while glucose levels increased (Figure 1(a), 1(b)). In contrast, treatment with the compound of formula (I) starting from day 5 normalized glucose levels (Figure 1(a)) without increasing plasma insulin levels (Figure 1(b)). Pancreatic insulin content was 10 to 20 times lower in mice treated with the compound of formula (I) compared to non-diabetic control mice (Figure 1(c), 8(b)). In both untreated STZ mice and STZ mice treated with the compound of formula (I), the islet cell area and the percentage of Ins+ cells decreased, while the percentage of Glu+ islet cells increased (Figures 1(d)-1(f), 8(a), 8(c), 8(d)). The compound of formula (I) rapidly reduced glucose levels in STZ mice without increasing plasma insulin levels, even when treatment was first initiated on day 15, i.e., when the mice had already developed apparent diabetes (Figures 1(g), 1(h), 8(e)). Pancreatic insulin content was also approximately 20 times lower in mice treated with the compound of formula (I) compared to control mice (Figures 1(i), 8(g)). Similarly, in untreated mice and STZ mice treated with the compound of formula (I), the islet cell area and the percentage of Ins+ islet cells decreased, while the percentage of Glu+ islet cells increased (Figures 1(j)-1(l), 8(f), 8(h), 8(i)). In summary, these findings indicate that the compound of formula (I) potently and insulin-independently reverses diabetes in a mouse model of insulin-deficiency diabetes.

[0092] Overall, this embodiment unexpectedly demonstrated that initiating treatment with the compound of formula (I) after STZ administration did not prevent β-cell destruction (insulin levels decreased over time, as shown in Figure 1(b)), but it did prevent the onset of diabetes (glucose levels remained normal, as shown in Figure 1(a)). Furthermore, even when mice were already diabetic in response to STZ-induced β-cell destruction, treatment with the compound of formula (I) improved glucose control without restoring insulin levels (Figure 1(h)) (Figure 1(g)). Thus, the compound of formula (I) unexpectedly improved glucose effects independently of insulin. In addition, it was observed that administration of the compound of formula (I) did not cause hypoglycemia, as shown in Figures 1(a) and 1(g).

[0093] Example 2: The compound of formula (I) improves hyperglycemia in STZ mice by stimulating muscle glucose uptake. The potent reduction in glucose levels in STZ mice treated with the compound of formula (I) provides evidence that the compound of formula (I) promotes insulin-independent glucose uptake in vivo. Continuous dynamic [ 18 [F]-Fluorodeoxyglucose PET analysis reveals the maximum metabolic rate of glucose in skeletal muscle (MR). glu It was shown that MR was significantly enhanced in STZ mice one week after treatment with the compound of formula (I) (Scan 3) compared to both pre-treatment (Scan 2) and untreated STZ mice at 22 days (Scan 3) (Figures 2(a)-2(c), 9). Furthermore, MR gluIn the hearts of diabetic STZ mice, glucose decreased at days 14-15, i.e., scan 2, but normalized to baseline levels one week after treatment with the compound of formula (I) (Figure 2(c)). To address the metabolic fate of glucose in STZ mice treated with the compound of formula (I), glycogen content in skeletal muscle and cardiac muscle was analyzed. Glycogen content in muscle and cardiac muscle was increased in the muscles and hearts of untreated STZ mice compared to control mice, but decreased in STZ mice treated with the compound of formula (I) compared to untreated STZ mice (Figure 2(d)). Therefore, under hyperglycemic conditions, treatment with the compound of formula (I) stimulates insulin-independent glucose uptake and glucose utilization, but does not stimulate glycogen storage.

[0094] Example 3: The compound of formula (I) promotes a gene expression profile that facilitates glucose oxidation in the muscle and heart of STZ mice. Skeletal muscle TXNIP expression levels are negatively correlated with glucose uptake. Simultaneously with enhanced glucose uptake in skeletal muscle, Txnip mRNA and protein levels were decreased in the skeletal muscle of STZ mice treated with the compound of formula (I) compared to untreated STZ mice (Figure 2(e), 10). Furthermore, the expression of Slc2a4, which encodes Glut4, in skeletal muscle was decreased in STZ mice compared to controls, but normalized in STZ mice treated with the compound of formula (I), and the expression of Slc2a1, which encodes Glut1, in skeletal muscle tended to increase in STZ mice treated with the compound of formula (I) (p=0.059) (Figure 2(e)). Similarly, in STZ mice, increased expression of pyruvate dehydrogenase (PDH) and pyruvate dehydrogenase kinase 4 (Pdk4), a negative regulator of oxidative glucose metabolism, and uncoupling protein 3 (Ucp3), which favors lipids as fuel substrates, was decreased in mice treated with compound (I) (Figure 2(e)), suggesting that the compound of formula (I) counteracts metabolic inflexibility in the skeletal muscle of STZ diabetic mice. Txnip expression was also increased in the hearts of STZ mice, but relatively decreased in STZ mice treated with the compound of formula (I) (Figure 2(f)). Furthermore, cardiac expression of Slc2a1 and Slc2a4 was decreased in untreated STZ mice, but tended to increase in mice treated with the compound of formula (I) (both p=0.088) (Figure 2(f)). Pdk4, Ucp2, and Ucp3 expression was increased in the hearts of untreated STZ mice but attenuated in STZ mice treated with the compound of formula (I) (p=0.055 for Ucp2) (Figure 2(f)). Therefore, the compound of formula (I) promoted changes in gene expression that promote glucose uptake, oxidative glucose metabolism, and ATP production in the skeletal muscle and heart of STZ mice (Figures 2(e), 2(f)). Hyperglycemia has been shown to rapidly increase cardiac expression of Pdk4 and Ucp3 and induce metabolic inflexibility and cardiac dysfunction in mice

[34] . In untreated STZ mice, E peak velocity was gradually decreased and isovolumetric relaxation time (IVRT) was increased, indicating impaired diastolic function.On the other hand, a one-week treatment with the compound of formula (I) reduced IVRT and increased peak E velocity and consequently the E / A ratio (Figure 2(g), Table 4). Improved left ventricular filling in mice treated with the compound of formula (I) also resulted in increased stroke volume and cardiac output (Table 4). Overall, these findings indicate that in diabetic STZ mice, the compound of formula (I) improved hyperglycemia, reduced glycogen accumulation, and reversed diabetic cardiomyopathy by stimulating insulin-independent glucose uptake and utilization. [Table 3]

[0095] Table 4 shows echocardiographic measurements of left ventricular dimensions in parasternal long axis (PLAX) B-mode or M-mode. HR: Heart rate; SV: Stroke volume; CO: Cardiac output; EDV: End-diastolic volume; ESV: End-systolic volume; AWd / s: Diastolic / systolic anterior wall thickness; LVID d / s: Diastolic / systolic left ventricular diameter; PWd / s: Diastolic / systolic posterior wall thickness; EF: Ejection fraction; FS: Shortening fraction; E / A: Ratio of peak wave velocities of E and A; Deceleration time: Deceleration time from peak of E wave to estimated baseline; IVRT: Isovolumetric relaxation time. Statistics were performed using one-way repeated ANOVA with Tukey's post-hoc test. *P<0.05 between RD and the compound of formula (I) at the same time point compared to baseline. # P<0.05. Compared to scan 2. ○ P<0.05. Data are mean ± SD.

[0096] Example 4: The compound of formula (I) improves hyperglycemia and promotes a gene expression profile that facilitates glucose oxidation in the muscle of db / db mice. To investigate the potential of the compound of formula (I) to improve hyperglycemia in association with severe insulin resistance, db / db mice were treated for 9 weeks with a diet formulated with the compound of formula (I) at concentrations of 0.5 or 1.0 mg / g, denoted as (Cmpd-(I)-(0.5) and Cmpd-(I)-(1.0). BKS mice were used as controls. Compensatory hyperinsulinemia was evident in db / db mice at 6 weeks of age (Figure 3(a), 3(b)). Untreated db / db mice subsequently developed insulin resistance. The mice were unable to compensate for this, and as a result of the decrease in insulin levels, blood glucose levels rose rapidly (Figure 3(a), 3(b)). The compound of formula (I) dose-dependently increased insulin levels and consequently attenuated the increase in blood glucose levels compared to both baseline and untreated db / db mice (Figure 3(a), 3(b)). These findings provide evidence that the compound of formula (I) maintained the compensatory insulin secretion response of β-cells, and calculations of the homeostasis model assessment of β-cell function (HOMA-β) showed that the decrease in β-cell function was associated with the compound of formula (I). Attenuation was shown in db / db mice treated with formula (I) (Figure 3(c)). The attenuation of hyperglycemia by the compound of formula (I) in db / db mice was in parallel with a reduction in glycogen storage in skeletal muscle and heart (Figure 3(d)), suggesting that the compound of formula (I) stimulated glucose utilization even in diabetic db / db mice. Compared to untreated db / db mice, the expression of Txnip, Pdk4, and Ucp3 was reduced in the skeletal muscle of db / db mice treated with the compound of formula (I), and Slc2a4 and mitochondrial expression were reduced. This idea is supported by the increased expression of peroxisome proliferator-activated receptor gamma coactivator (PGC)-1α (Ppargc1a), a positive regulator of biosynthesis and respiration, and Cox8b, a driver of oxidative phosphorylation (Figure 3(f)). Furthermore, the increased cardiac expression of Slc2a1 and the attenuation of Txnip, Pdk4, and Ucp3 in treated mice compared to untreated db / db mice (Figure 3(g)) are consistent with those observed in STZ mice treated with compound (I) of formula.Notably, the compound of formula (I) did not increase serum lactate levels in STZ mice and db / db mice treated with the compound of formula (I) (Figure 11). In summary, these findings provide evidence that the compound of formula (I) evades the genetic signature associated with metabolic inflexibility related to diabetes and diabetic cardiomyopathy in both STZ and db / db diabetic mice.

[0097] Example 5: The compound of formula (I) stimulates mitochondrial uncoupling in myotubes. The increased glucose utilization and decreased glycogen content observed in the skeletal muscle and heart of STZ and db / db mice treated with the compound of formula (I) suggest that the compound of formula (I) generates metabolic demands and increases energy expenditure via unproductive cycles and / or mitochondrial uncoupling. To clarify the potential uncoupling ability of the compound of formula (I), mitochondrial and glycolytic functions were tested in intact differentiated C2C12 myotubes. The compound of formula (I) dose-dependently increased the oxygen consumption rate (OCR), an indicator of oxidative phosphorylation, and the ratio of basal OCR to basal ECAR (extracellular acidification rate) in C2C12 myotubes (Figure 4(a)-4(c)), indicating that the compound of formula (I) enhanced the cell's preference for oxidative metabolism. The compound of formula (I) also showed a reduced decrease in OCR in response to oligomycin, which blocks ATP synthase, and consequently an increase in proton leak (Figures 4(a), 4(d)). However, consistent with our previously published findings, the compound of formula (I) did not significantly reduce cellular ATP levels (Figure 4(d)). Overall, these results indicate that the compound of formula (I) increases cellular respiration by acting as a mitochondrial uncoupling agent, thereby providing evidence that the compound of formula (I) induces metabolic demands that enhance energy expenditure by stimulating TCA flux and oxidative metabolism.

[0098] Example 6: The compound of formula (I) maintains the function and mass of β-cells in db / db mice.Consistent with the initial compensatory increase in β-cell volume in the described db / db mice, islet cell area increased at 6 weeks of age in db / db mice compared to BKS mice (Figure 5(a), 5(b)). In untreated db / db mice, islet cell area relatively decreased by 15 weeks of age compared to 6 weeks of age (p=0.09), but was maintained in a dose-dependent manner in mice treated with the compound of formula (I) (Figure 5(a), 5(b)). The proportion of Ins+ cells increased in mice treated with the compound of formula (I) compared to untreated db / db mice, but the proportion of cells positive for other pancreatic endocrine hormones tended to decrease in db / db mice treated with the compound of formula (I) (Figure 5(c), Figure 12(a)). Consequently, pancreatic insulin and proinsulin content, as well as the pancreatic insulin:proinsulin ratio, a measure of the efficiency of proinsulin processing, were increased in db / db mice treated with the compound of formula (I) compared to untreated db / db mice (Figures 5(d), 5(e), 12(b)). Overall, these data provide evidence that the compound of formula (I) avoids β-cell loss and maintains β-cell function in db / db mice. Protein expression of the glucose transporter Glut2 encoded by Src2a2 was partially restored in the pancreatic islets of db / db mice treated with the compound of formula (I) (Figures 5(a), 5(f)). Expression of the transcription factors Nkx6.1 and MafA, markers of mature β-cell identity, increased, and Ipf1 / Pdx1 tended to increase in the pancreatic islets of db / db mice treated with the compound of formula (I) (Figures 5(a), 5(f)). The expression of Raldh3, encoded by aldehyde dehydrogenase 1a3 (Aldh1a3), which is upregulated in diabetic islets, remained unchanged (Figure 5(a), 5(f)). mRNA expression analysis of isolated islets showed increased expression of urocortin 3 (Ucn3), a marker of mature β-cells and a regulator of insulin secretion, and Nkx6.1 in islets of db / db mice treated with compound (I) of formula (Figure 5(g)).Treatment with the compound of formula (I) did not alter Aldh1a3 expression, but the expression of Txnip, which adversely affects insulin transcription and induces oxidative stress in β cells, was reduced in the pancreatic islets of db / db mice treated with Cmpd-(I)-(1.0) compared to untreated db / db mice (Figure 5(g)). Similarly, the expression of endoplasmic reticulum protein 29 (Erp29), an ER stress-related gene, and α-mannosidase-like 2 (Edem2), which enhances ER degradation, was reduced in the pancreatic islets of db / db mice treated with the compound of formula (I) (Figure 5(g)). These data suggest that the reduction in the expression of Txnip and ER stress genes, along with the increase in the expression of genes that regulate β cell identity and insulin secretion, contributes to compensatory β cell function, GSIS, and maintenance of β cell volume in insulin-resistant db / db mice via the compound of formula (I). The maintained compensatory insulin secretion in db / db mice treated with the compound of formula (I) may explain the lack of altered Aldh1a3 expression, suggesting that increased Aldh1a3 expression is a marker of metabolically problematic β-cells.

[0099] Example 7: The compound of formula (I) mitigates changes in pancreatic islet gene expression caused by acute hyperglycemia. To investigate the ability of compounds of formula (I) to directly modulate the pancreatic islet response to glucotoxic conditions, the inventors performed ex vivo RNA-seq analysis of mouse and human pancreatic islets cultured at low glucose (11 mM) and high glucose (22 mM) + / - 5 μM with compounds of formula (I). Mouse pancreatic islets responded to high glucose, with 1924 differentially expressed genes (DEGs) upregulated and 1568 downregulated (Figure 6(a), 13(a)-13(c)). At high glucose, compounds of formula (I) reduced the number of DEGs to 143 upregulated and 208 downregulated (Figure 6(a), 13(a)-13(c)). As expected, exposure of mouse pancreatic islets to high glucose significantly increased Txnip expression, which was attenuated by compounds of formula (I) (Figure 6(b)). Overexpression analysis (ORA) of the Molecular signature Hallmark, KEGG, and Wiki-pathways gene sets revealed that gene sets associated with enhancing β-cell activity, such as "Protein Secretion," "Unfolded protein response," and "Pancreas beta-cells," were upregulated by high glucose but overexpressed in DEGs avoided by the compound of formula (I) (Figure 6(c)). The metabolic pathways "Fatty acid metabolism," "Glycolysis," and "Hypoxia" were also regulated by high glucose and overexpressed in DEGs deregulated by the compound of formula (I), along with the "KEGG:TCA cycle" (P=0.073). Hierarchical clustering of the top 20 most significant DEGs of these pathways, regardless of direction, showed opposite regulation by the addition of high glucose and the compound of formula (I) (Figure 6(d)). These results demonstrate the ability of the compound of formula (I) to directly conserve important metabolic gene expression patterns in β-cells under glucotoxic conditions. The gene expression differences in human pancreatic islets were dominated by the donor effect, and DEG was hardly detected even after batch correction (Figures 13(d)-13(f)).Nevertheless, gene set enrichment analysis (GSEA) showed that high glucose induces genes in the "Pancreatic beta-cells" gene set, which are reversed by the compound of formula (I), and that the compound of formula (I) induces upregulation of genes in the "Hypoxia" and "Glycolysis" gene sets, similar to what was observed in mouse pancreatic islets (Figure 13(g)).

[0100] Example 8: The compound of formula (I) prevents the adverse effects of hyperglycemia on GSIS in INS-1E cells. Chronic hyperglycemia activates mTORC1 signaling and suppresses AMPK signaling in β-cells in vitro, resulting in decreased glucose metabolism and impaired glucose-stimulated insulin secretion (GSIS). Consistently, our RNA-seq analysis has confirmed that mTORC1 signaling is upregulated in pancreatic islets exposed to high glucose (22 mM), and this was prevented by exposure to the compound of formula (I) (Figure 6(c)). To investigate the effect of the compound of formula (I) on GSIS under chronic hyperglycemia conditions, INS-1E cells were cultured under 25 mM glucose for 4 days (days), resulting in impaired GSIS. However, these adverse effects were avoided when cells were simultaneously exposed to the compound of formula (I) throughout the entire 4-day culture period (Figure 7(a)). Short-term (2-hour) exposure of INS-1E cells to the compound of formula (I) at the end of a 4-day culture period also partially reversed the adverse effects of hyperglycemia on GSIS (Figure 7(b)). Thus, the compound of formula (I) prevented and reversed the adverse effects of hyperglycemia on GSIS. Analysis of the downstream targets of mTORC1, namely ribosomal protein 6 (pS6), and the downstream targets of AMPK, namely acetyl-CoA carboxylase (ACC) and Raptor, showed that long-term and short-term exposure of INS1-E cells to the compound of formula (I) under high glucose conditions resulted in decreased pS6-Ser240 / 244 levels, but increased pACC-Ser79 and pRaptor-Ser792 levels (Figures 7(c), 7(d), 14). In summary, these findings provide evidence that the compound of formula (I) conserves β-cell function under hyperglycemic conditions by antagonizing mTORC1 and maintaining AMPK signaling.

Claims

1. A method for regulating blood glucose levels in a subject requiring it by an effect that manifests as an increase in the glucose effect, wherein the subject is in a state of insulin deficiency and is taking or needs to take exogenous insulin, and the method is The above subject includes compounds having the following structure: 【Chemistry 1】 Administering a pharmaceutically acceptable salt, solvate, or prodrug thereof; Monitoring the blood glucose levels of the subject after administration of the compound; and The method comprises delaying the need for the use of exogenous insulin, or reducing or discontinuing the dose of exogenous insulin administered to the subject in order to maintain the subject's blood glucose level within the normal range.

2. The aforementioned normal range is, (i) AIC less than 7%; (ii) A1C less than 154 mg / dL (sometimes reported as eAG); (iii) Pre-meal plasma glucose of 80-130 mg / dL (iv) Postprandial plasma glucose less than 180 mg / dL 1 to 2 hours after the start of a meal The method according to claim 1, or any combination of (i) to (iv).

3. The method according to claim 1 or 2, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is repeatedly administered to the subject over a period of time.

4. The method according to claim 3, wherein the dose of exogenous insulin administered is gradually reduced over the period during which the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered.

5. The method according to claim 4, wherein the dose of exogenous insulin is discontinued after repeated administration of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, to the subject for about two weeks.

6. The method according to claim 5, wherein the dose of exogenous insulin remains discontinued as long as the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered to the subject.

7. The method according to any one of claims 1 to 4, wherein, in order to delay the need for the use of exogenous insulin, the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered to the subject before the administration of exogenous insulin to the subject.

8. A method for treating type 1 diabetes (T1D) in a subject requiring treatment, wherein the subject experiences high and low glucose levels despite using exogenous insulin, The above subject includes compounds having the following structure: 【Chemistry 2】 The method comprising administering a pharmaceutically acceptable salt, solvate, or prodrug thereof.

9. The method according to any one of the preceding claims, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered once daily.

10. The method according to any one of the preceding claims, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered orally.

11. The method according to any one of the preceding claims, wherein an alkali metal salt of the compound is administered.

12. The method according to any one of the preceding claims, wherein the sodium salt of the compound is administered.

13. The method according to any one of claims 1 to 12, wherein the subject is suffering from absolute insulin deficiency.

14. The method according to claim 13, wherein the subject is suffering from type 1 diabetes (T1D), chronic pancreatitis, pancreatectomy, cystic fibrosis, monogenic diabetes, or a post-infectious condition following severe acute respiratory syndrome (SARS) infection or another viral infection.

15. The method according to claim 14, wherein the subject is suffering from juvenile mature diabetes mellitus (MODY) or adult latent autoimmune diabetes mellitus (LADA).

16. The method according to any one of claims 1 to 12, wherein the subject is suffering from acute insulin deficiency.

17. The method according to any one of claims 1 to 12, wherein the subject is suffering from diabetic ketoacidosis.

18. The method according to any one of claims 1 to 12, wherein the subject is suffering from unstable diabetes.

19. The method according to any one of claims 1 to 12, wherein the subject is suffering from T1D and the subject's β cells are lost.

20. The method according to any one of claims 1 to 12, wherein the subject suffers from a genetic condition in which insulin secretion is lost.

21. The method according to any one of claims 1 to 12, wherein the subject suffers from a genetic defect in the insulin response capacity of the tissue.

22. The method according to any one of claims 1 to 12, wherein the subject is suffering from congenital hyperinsulinism.

23. The method according to any one of the preceding claims, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered as a pharmaceutical composition.

24. The method according to claim 23, wherein the pharmaceutical composition is a tablet or a capsule.

25. The method according to any one of the preceding claims, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is orally administered to a patient once daily in a dose of about 100 mg to about 1,000 mg.

26. The method according to claim 25, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is orally administered to the patient once daily in a dose of about 200 mg to about 600 mg.

27. The method according to claim 25, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is orally administered to the patient once daily in a dose of about 400 mg to about 800 mg.

28. The method according to claim 25, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is orally administered to the patient once daily at a dose of about 400 mg.

29. The method according to any one of claims 1 to 28, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is orally administered once daily to the subject, and the steady-state plasma concentration of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is about 70 μg / mL to about 120 μg / mL.

30. The method according to any one of claims 1 to 28, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is orally administered once daily to the subject, and the steady-state plasma concentration of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is about 90 μg / mL to about 160 μg / mL.

31. The method according to claim 30, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is orally administered once daily to the subject, and the steady-state plasma concentration of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is about 100 μg / mL to about 150 μg / mL.

32. The method according to claim 30, wherein the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is orally administered to the subject once daily, and the steady-state plasma concentration of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof is approximately 120 μg / mL to approximately 140 μg / mL.

33. The compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is administered orally to the subject once daily, and the steady state AUC of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof ０－２４ The method according to any one of claims 1 to 32, wherein the concentration is approximately 1,500 h*μg / mL to approximately 4,000 h*μg / mL.

34. The steady-state AUC of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. ０－２４ The method according to claim 33, wherein the concentration is approximately 1,800 h*μg / mL to approximately 3,100 h*μg / mL.

35. The steady-state AUC of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. ０－２４ The method according to claim 33, wherein the concentration is approximately 1,500 h*μg / mL to approximately 2,000 h*μg / mL.

36. The steady-state AUC of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. ０－２４ The method according to claim 33, wherein the concentration is approximately 2,500 h*μg / mL to approximately 4,000 h*μg / mL.

37. The steady-state AUC of the compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. ０－２４ The method according to claim 33, wherein the concentration is approximately 3,000 h*μg / mL to approximately 3,500 h*μg / mL.

38. The compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, C ｍａｘ The method according to any one of claims 33 to 37, wherein the concentration is approximately 70 μg / mL to approximately 200 μg / mL.

39. The compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, C ｍａｘ The method according to any one of claims 33 to 37, wherein the concentration is approximately 80 μg / mL to approximately 140 μg / mL.

40. The compound, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, C ｍａｘ The method according to any one of claims 33 to 37, wherein the concentration is approximately 70 μg / mL to approximately 100 μg / mL.

41. wherein the C of the compound or a pharmaceutically acceptable salt, solvate, or prodrug thereof ｍａｘ is from about 120 μg / mL to about 150 μg / mL, the method according to any one of claims 33 to 37.

42. The method according to any one of the prior claims, wherein the subject is a human.

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