Treatment of cancer with PDK inhibitor 4-chloro-n-(2-(4-chlorobenzyl)-3-oxo-2, 3-dihydro-1, 2, 4-thiadiazol-5-yl) benzamide

Abstract

Provided herein are methods of treating cancer by administering a therapeutic compound to a subject in need thereof. The disclosure also generally relates to methods of using the therapeutic compounds to inhibit the activity of pyruvate dehydrogenase kinase (PDK) in a subject in need thereof. The therapeutic compound is a compound of formula (I): or a pharmaceutically acceptable salt, solvate or prodrug thereof.

Description

Cross-references to related applications

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 517,525, filed August 3, 2023, which is incorporated herein by reference in its entirety.

[0002] Reference to the electronic sequence list The contents of the electronic serial number (286502001040SEQLIST.xml; size: 43,276 bytes; and creation date: July 24, 2024) are incorporated herein by reference in their entirety. Technical Field

[0003] This disclosure generally relates to methods of treating cancer, preventing or delaying its onset by administering the compounds provided herein. This disclosure also provides combination therapies that can be used to treat cancer. Background Technology

[0004] Many human cancers shift their metabolism from oxidative phosphorylation to glycolysis by increasing PDK expression / activity. This phenomenon, commonly known as the Warburg effect, has been shown to protect cancer cells from apoptosis and increase cell proliferation. See Atas E., Oberhuber M., and Kenner L., The Implications of PDK1–4 on Tumor Energy Metabolism, Aggressiveness, and Therapy Resistance. Front Oncol, 2020. 10: p.583217.

[0005] Current research aims to develop pharmacological interventions to counteract the Wahlberg effect and restore metabolic flexibility in cancer cells. The expression and increased activity of pyruvate dehydrogenase kinase (PDK) contribute to increased cell proliferation, invasiveness, and resistance to chemotherapy drugs in certain types of cancer. See Wang J, Qian Y, Gao M. Overexpression of PDK4 is associated with cell proliferation, drug resistance, and poor prognosis in ovarian cancer. Cancer Manag Res2019;11:251-262. Inhibition of PDK through various mechanisms can potentially reverse these effects, as shown in several studies. See Tiersma, JF; Evers, B.; Bakker, BM; Jalving, M.; de Jong, S. Pyruvate Dehydrogenase Kinase Inhibition by Dichloroacetate in Melanoma Cells Unveils Metabolic Vulnerabilities. Int. J. Mol. Sci. 2022, 23 , 3745 and Kim CJ, Terado T, Tambe Y, Mukaisho K, Kageyama S, Kawauchi A, and Inoue H: Cryptotanshinone, a novel PDK 4inhibitor, suppresses bladder cancer cell invasiveness via the mTOR / β-catenin / N-cadherin axis. Int J Oncol 2021, 59: 40. However, highly effective PDK inhibitors exhibiting favorable toxicological characteristics have not yet been developed.

[0006] Therefore, there is an urgent need to develop new PDK inhibitors as anticancer agents. Summary of the Invention

[0007] It has been surprisingly discovered that compounds of formula (I) (I), Or a pharmaceutically acceptable salt, solvate, or prodrug thereof can restore the metabolic flexibility of muscle tissue and cells in subjects suffering from various diseases or complications, particularly cancer. Compounds of formula (I) are also known as 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. In some variants, the subject is a human (e.g., a human patient). The restoration of metabolic flexibility is partly attributed to the discovery that compounds of formula (I) are inhibitors of pyruvate dehydrogenase kinase (PDK) expression and are able to increase tumor oxygenation and oxidative phosphorylation upon administration to subjects in need (e.g., cancer patients), which can lead to reduced proliferation and / or increased apoptosis in certain cancer cells.

[0008] In some aspects, this article provides methods and compositions for treating cancers associated with pyruvate dehydrogenase kinase (PDK) overexpression or activation in subjects of need (e.g., human patients), including methods comprising administering a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the PDK is pyruvate dehydrogenase kinase 4 (PDK4).

[0009] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat bladder cancer. In some embodiments, the bladder cancer is metastatic bladder cancer.

[0010] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat colorectal cancer. In some such embodiments, the colorectal cancer is associated with a KRAS mutation. In some embodiments, the colorectal cancer is metastatic colorectal cancer.

[0011] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat lung cancer. In some such embodiments, the lung cancer is associated with a KRAS mutation. In other such embodiments, the lung cancer is epidermal growth factor receptor (EGFR) positive lung cancer. In some embodiments, the lung cancer is metastatic lung cancer. In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC). In some embodiments, NSCLC is associated with an EGFR mutation.

[0012] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat breast cancer. In some such embodiments, the breast cancer is tamoxifen-resistant breast cancer. In some embodiments, the breast cancer is metastatic breast cancer. In some embodiments, the breast cancer is triple-negative breast cancer.

[0013] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat bladder cancer. In some embodiments, the bladder cancer is metastatic bladder cancer.

[0014] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat melanoma. In some embodiments, the melanoma is metastatic melanoma. In some embodiments, the subject (e.g., a patient) has previously been treated with immunotherapy (e.g., an anti-PD1 or anti-CTLA4 antibody). In some such embodiments, the metastatic melanoma has no response or a limited response to immunotherapy.

[0015] In some implementations, the compound of formula (I) or its pharmaceutically acceptable salt, solvate or prodrug is used to treat hepatocellular carcinoma (HCC).

[0016] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat cervical cancer. In some embodiments, the subject's (e.g., a patient's) cervical cancer has become resistant to doxorubicin. In some embodiments, the subject's (e.g., a patient's) cervical cancer has become resistant to etoposide.

[0017] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to target cancer stem cells. In some such embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to target cancer stem cells resistant to radiotherapy (radiotherapy).

[0018] On the other hand, this article also provides a method for inducing apoptosis in apoptosis-inducing cancer cells, the method comprising administering an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof to the apoptosis-inducing cancer cells. In some embodiments, the cancer cells are bladder cancer cells, gastric cancer cells, colon cancer cells, breast cancer cells, ovarian cancer cells, melanoma cells, or lung cancer cells.

[0019] On the other hand, a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof is administered to a subject (e.g., a patient) with cancer that is resistant to one or more chemotherapeutic agents and / or resistant to radiotherapy (radiotherapy).

[0020] On the other hand, this disclosure provides a method for reducing resistance to chemotherapeutic agents, said method comprising administering to a subject (e.g., a cancer patient) a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the chemotherapeutic agent is cisplatin. In some embodiments, the chemotherapeutic agent is 5-fluorouracil. In some embodiments, the chemotherapeutic agent is erlotinib. In some embodiments, the chemotherapeutic agent is paclitaxel. In some embodiments, the chemotherapeutic agent is doxorubicin.

[0021] In some embodiments, a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with cisplatin to treat a subject in need, such as a cancer patient. In some embodiments, the cancer is bladder cancer, gastric cancer, breast cancer, lung cancer, HCC, cervical cancer, ovarian cancer, or colon cancer.

[0022] In some embodiments, a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with an EGFR inhibitor to treat a subject (e.g., a patient) with lung cancer (e.g., NSCLC). In some embodiments, the EGFR inhibitor is erlotinib, gefitinib, lapatinib, lazatinib, or nexituzumab.

[0023] In some implementations, a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof is administered in combination with paclitaxel to treat lung cancer (e.g., NSCLC) in a subject (e.g., a patient) in need.

[0024] In some implementations, the compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof is administered in combination with doxorubicin to treat a subject (e.g., a patient) with hepatocellular carcinoma (HCC) or cervical cancer.

[0025] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with recombinant arginase. In some such embodiments, the cancer is breast cancer (e.g., triple-negative breast cancer). In other such embodiments, the cancer is HCC. In other embodiments, the cancer is cervical cancer.

[0026] In some implementations, the compound of formula (I) or its pharmaceutically acceptable salt, solvate or prodrug is administered in combination with a Cox2 inhibitor, propranolol, metformin, salinomycin or phenformin.

[0027] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor (e.g., cimiprizumab, nivolumab, or pembrolizumab). In some embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor (e.g., atezolizumab, avermab, or durvalumab). In some embodiments, the immune checkpoint inhibitor is a CTLA-4 inhibitor (e.g., ipilimumab or trimemumab).

[0028] On the other hand, this disclosure provides a method for overcoming tumor radioresistance by administering a pharmaceutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof to a subject suffering from cancer (e.g., a cancer patient). In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered to the subject prior to radiotherapy.

[0029] In any of the foregoing embodiments, the compound of formula (I) may be applied as a salt. In some embodiments, the salt is an alkali metal salt. In some embodiments, the salt is a sodium salt. In some embodiments, the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide has the following formula: , Where X + It represents an alkali metal cation (such as lithium, rubidium, cesium, sodium, or potassium).

[0030] In some embodiments, the compound applied according to this disclosure is a prodrug of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. In some embodiments, the prodrug has the following structure: , or its salt, wherein R 1 Choose from the group consisting of -C(O)-C2H4-CO2H and -PO3H2, or their salts or solvates.

[0031] In any of the foregoing embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject in need (e.g., a patient) at a dose of about 100 mg to about 1,000 mg. All references to doses discussed herein refer to the free acid (protonated) form. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily at a dose of about 200 mg to about 1,000 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily at a dose of about 400 mg to about 800 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily at a dose of about 100 mg to about 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.

[0032] In any of the foregoing embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 40 µg / mL to about 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 in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 40 µg / mL to about 80 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 in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 70 µg / mL to about 120 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 in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 90 µg / mL to about 160 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 in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 100 µg / mL to about 150 µg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered daily to a human subject, resulting in a steady-state plasma concentration of the compound of formula (I) of about 120 µg / mL to about 140 µg / mL.

[0033] In any of the foregoing embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject in need (e.g., a patient), wherein such administration results in a steady-state area under the curve (AUC) of the compound of formula (I). 0-24 The concentration is approximately 1,000 h*µg / mL to approximately 4,000 h*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 in need or a subject (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 1,000 h*µg / mL to approximately 2,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24The concentration is approximately 1,500 h*µg / mL to approximately 4,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 1,800 h*µg / mL to approximately 3,100 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 1,500 h*µg / mL to approximately 2,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 2,500 h*µg / mL to approximately 4,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 It ranges from approximately 3,000 h*µg / mL to approximately 3,500 h*µg / mL.

[0034] In any of the foregoing embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject in need (e.g., a patient), wherein such administration results in the Ci 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 in need (e.g., a patient), wherein such administration results in the C1 of the compound of formula (I) being... 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 in need (e.g., a patient), wherein such administration results in the C1 of the compound of formula (I) being... 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 in need (e.g., a patient), wherein such administration results in the C1 of the compound of formula (I) being... maxThe 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 in need (e.g., a patient), wherein such administration results in the C1 of the compound of formula (I) being... max It is approximately 1200 µg / mL to approximately 150 µg / mL.

[0035] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in an oral dosage form. In some embodiments, the oral dosage form is a capsule. In other embodiments, the oral dosage form is a tablet.

[0036] In other respects, the use of compounds of formula (I) or pharmaceutically acceptable salts, solvates, or prodrugs thereof in therapies (including any of the methods described herein) is also provided. For example, in some embodiments, the use of compounds of formula (I) or pharmaceutically acceptable salts, solvates, or prodrugs thereof is provided for: treating cancer associated with pyruvate dehydrogenase kinase (PDK) overexpression or activation in subjects of need (e.g., human patients); reducing resistance to chemotherapeutic agents; and inducing apoptosis in apoptosis-inducing cancer cells.

[0037] In some aspects, the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof in the preparation of a medicament is also provided. In some embodiments, the medicament is used in 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 preparation of a medicament intended for: treating cancer associated with pyruvate dehydrogenase kinase (PDK) overexpression or activation in a subject of need (e.g., a human patient); reducing resistance to chemotherapeutic agents; and inducing apoptosis in apoptosis-inducing cancer cells. Attached Figure Description

[0038] This application can be understood by referring to the following description in conjunction with the accompanying drawings.

[0039] Figure 1 The stimulation of skeletal muscle glucose uptake after administration of compound of formula (I) to STZ mice was described. Figure 1 (a): Timeline of STZ treatment and FDG-PET scan (in days). Figure 1 (b) Representative PET / CT images of FDG uptake in untreated mice (top panel) and mice treated with the compound of formula (I) (bottom panel). Figure 1(c) Estimated maximum muscle and cardiac glucose uptake (MR) at baseline (scan 1), 9–10 days after the last STZ injection (scan 2), and 1 week after treatment with 0.5 mg / g of compound (I) (n=5) (scan 3) or without treatment (n=5). glu ). ( Figure 1 (d) Glycogen content in the muscle and heart of STZ mice in control (n=6-18) and untreated (n=14-36) or treated with compounds of formula (I) from day 15 onwards (n=13-27). Figure 1 (e) In the muscles 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) Txnip , Slc2a1 , Slc2a4 , Hk2 , Pkm , Ppargc1a , Pdk4 , Pdha1 , Shda and Cox8b The relative mRNA level. Figure 1 (f) Hearts of control mice (n=5), untreated STZ mice (n=9), and STZ mice treated with the compound of formula (I) (n=6-7) Txnip , Slc2a1 , Slc2a4 , Pdk4 , Ucp2 and Ucp3 The relative mRNA level. Figure 1 (g) Mitral Doppler ultrasound of STZ-treated mice at baseline (Scan 1), 15 days after STZ initiation (Scan 2), and 1 week after treatment with 0.5 mg / g of compound (I) (n=9) (Scan 3), or without treatment (n=9). Data are presented as mean ± SEM. Statistical significance between untreated STZ mice and STZ-treated mice with compound (I) was determined by Student's t-test ( Figure 1 (c): Muscle, Figure 1 (g) or by Wilcox test ( Figure 1 (c): Heart, Figure 1 (d) to Figure 1 (f))(* p <0.05,** p <0.01, *** p <0.001) was determined, and the statistical significance between control mice and STZ mice was confirmed by the Wilcoxon test ( ). Figure 2 (d) to Figure 2 (f)) ( #p <0.05, ## p <0.01, ### p <0.001) is determined, and ( Figure 1 (c) Figure 1 The statistical significance between scans in (g) was determined by one-way repeated ANOVA followed by paired Student's t-test (p<0.05).

[0040] Figure 2 Western blot analysis was used to quantify protein levels in the femoral muscle extracts of STZ mice, including control (n=5) and untreated (n=9) mice or those treated with a compound of formula (I) at 0.5 mg / g starting from day 15 (n=9).

[0041] Figure 3 The study described how the compound of formula (I) dose-dependently prevented hyperglycemia in db / db mice. Fasting glucose levels in BKS and untreated or treated db / db mice (n=15-20 / group) with the compound of formula (I) at 0.5 and 1.0 mg / g, respectively, were compared. Figure 3 (a) and insulin ( Figure 3 (b) Levels and Area Under the Curve (AUC). HOMA-IR in untreated or db / db mice treated with compounds of formula (I) at 0.5 and 1.0 mg / g. Figure 3 (c)) and HOMA-β ( Figure 3 (d))(by Figure 3 (a) and Figure 3 (b) Calculate the area under the curve (AUC). Figure 3 (e) Glycogen content in the muscle and heart of BKS (n=5-9) and untreated (n=9-10) or treated with compounds of formula (I) (n=6-7) db / db mice. Figure 3 (f) Muscle samples from db / db mice that were BKS (n=6-13) and untreated (n=15-22) or treated with a compound of formula (I) at 1.0 g / kg (n=8-15) Txnip , Slc2a1 , Slc2a4 , Hk2 , Pkm , Ppargc1a , Pdk4 , Pdha1 , Sdha , Cox8b , Ucp2 and Ucp3 The relative mRNA level. Figure 3 (g) Hearts of BKS (n=4-5), untreated db / db mice (n=8-9), and db / db mice (n=6-7) treated with compounds of formula (I). Txnip , Slc2a1 , Slc2a4 , Pdk4 , Ucp2 and Ucp3 The relative mRNA levels. Data are presented as mean ± SEM. 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. Figure 3 (a) to Figure 3 (d) Student's t-test Figure 3 (e) or Wilcoxon test ( Figure 3 (f) Figure 3 (g)) (* p <0.05,** p <0.01, *** p <0.001) was determined, and the statistical significance between BKS and db / db mice was confirmed by the Wilcoxon test ( ). # p <0.05, ## p <0.01, ### p <0.001)( Figure 3 (a) to Figure 3 (d) Figure 3 (f) Figure 3 (g) or by student t-test ( Figure 3 (e) is confirmed.

[0042] Figure 4 Treatment with the compound of formula (I) did not increase serum lactate levels in STZ and db / db mice. Fasting glucose levels in STZ mice were measured on day 15 (before treatment) and day 22 after one week of treatment with the compound of formula (I). Figure 4 (a)) and lactic acid ( Figure 4 (b) Levels (n=9 / group). Fasting glucose levels in BKS and untreated or treated db / db mice (n=7-9 / group) with compounds of formula (I) at 0.5 and 1.0 mg / g, respectively. Figure 4 (c)) and lactic acid ( Figure 4 (d) Horizontal lines and area under the curve (AUC). Data are expressed as mean ± SEM. Figure 4 The statistical significance between time points in (a) was demonstrated by the Student's t-test.P <0.05 was determined, and statistical significance between untreated and treated db / db mice with the compound of formula (I) was confirmed by Welch's ANOVA followed by Games-Howell post-hoc test (*). P <0.05,** P <0.01, *** P <0.001) is determined, and ( Figure 4 (c) Figure 4 The statistical significance between BKS and db / db mice in (d) was verified by the Wilcoxon test. ## P <0.01) confirmed.

[0043] Figure 5 The mitochondrial uncoupling induced by compounds of formula (I) was depicted. Respiratory chromatograms of intact C2C12 myotubes treated with compounds of formula (I) at 0.625, 1.25, and 2.5 µM for + / - 4 hours followed by sequential injections of oligomycin, FCCP, and a mixture of rotenone and antimycin are shown, illustrating the oxygen consumption rate (OCR). Figure 5 (a) and extracellular acidification rate (ECAR) Figure 5 (b)). Figure 5 (c): From the last baseline measurement ( Figure 5 (a) and Figure 5 (b) Measurement 3) OCR vs ECAR plot. Figure 5 (d): According to Figure 5 Mitochondrial functional parameters calculated from OCR data in (a). Data are expressed as mean ± SEM. n=5 / condition. Figure 5 The statistical significance of the difference between untreated cells and cells treated with the compound of formula (I) in (d) was determined by one-way ANOVA followed by Tukey post-hoc test. ¤ p <0.05, ¤¤ p <0.01). Detailed Implementation

[0044] The following description illustrates exemplary compositions, methods, parameters, etc. However, it should be understood that this description is not intended to limit the scope of this disclosure, but is provided as a description of exemplary embodiments.

[0045] Treatment This article provides information on the application of an effective amount of the compound of formula (I): (I), Methods and compositions for treating and / or preventing cancer, or pharmaceutically acceptable salts, solvates, or prodrugs thereof. The term “effective amount” as used herein refers to an amount of compound or composition sufficient to treat a particular cancer or other cell proliferation (such as improving, alleviating, reducing, and / or delaying one or more of its symptoms). Generally, an effective amount includes an amount sufficient to shrink a tumor and / or reduce the rate of tumor growth (such as inhibiting tumor growth) or to prevent or delay the proliferation of another type of cell. In some embodiments, an effective amount is an amount sufficient to delay development. In some embodiments, an effective amount is an amount sufficient to prevent or delay occurrence and / or recurrence. An effective amount may be administered in one or more administrations. An effective amount of a drug or composition may: (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) to a certain extent inhibit, delay, slow, and preferably prevent the infiltration of cancer cells into peripheral organs; (iv) inhibit (i.e., to a certain extent slow and preferably stop) tumor metastasis; (v) inhibit tumor growth; (vi) prevent or delay the occurrence and / or recurrence of tumors; and / or (vii) to a certain extent alleviate one or more symptoms associated with cancer. In the following disclosure, references to the application of compounds of formula (I) will include various salt forms, solvates and prodrugs as described herein.

[0046] In some aspects, this document provides methods and compositions for treating cancers associated with pyruvate dehydrogenase kinase (PDK) overexpression or activation in subjects of need (e.g., human patients), methods comprising administering a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the PDK is pyruvate dehydrogenase kinase 1 (PDK1). In some embodiments, the PDK is pyruvate dehydrogenase kinase 2 (PDK2). In some embodiments, the PDK is pyruvate dehydrogenase kinase 3 (PDK3). In some embodiments, the PDK is pyruvate dehydrogenase kinase 4 (PDK4). In some embodiments, the PDK is two or more of PDK1, PDK2, PDK3, and PDK4.

[0047] Compounds of formula (I) can reduce the metabolic inflexibility of cancer cells in subjects (e.g., patients), a phenomenon attributed in part to the presence of compounds of formula (I) that inhibit PDK (e.g., PDK4) and increase glucose oxidation in cancer cells. Therefore, compounds of formula (I) can reduce the proliferative anabolic pathways that cancer cells depend on. By inhibiting PDK, compounds of formula (I) can induce apoptosis in cancer cells, thus potentially serving as an effective anticancer therapeutic agent. Furthermore, compounds of formula (I) can act synergistically with other anticancer therapies. Therefore, compounds of formula (I) can be used in combination with other anticancer agents, as disclosed herein, or as adjuvants to other anticancer therapies. As described in the examples, the compounds of formula (I) stimulate glucose uptake and glucose utilization in cells. Furthermore, as shown herein, the compounds of formula (I) promote gene expression profiles that favor glucose oxidation rather than glycogen storage. Therefore, the compounds of formula (I) behave differently from pan-AMPK activators known in the art, which promote glycogen storage rather than glucose oxidation.

[0048] Increased glucose uptake in muscle tissue without an increase in glycogen, and increased energy expenditure observed in animal models, prompted in vitro experiments on mitochondrial function. As shown in the examples, in isolated muscle tubes, compounds of formula (I) increased basal oxygen consumption and extracellular acidification rates, indicating increased glucose utilization through increased flux through the tricarboxylic acid (TCA) cycle and through mitochondrial uncoupling oxidative phosphorylation. Therefore, the current data suggest that compounds of formula (I) are dual AMPK activators and mitochondrial uncoupling agents.

[0049] Furthermore, the compounds of formula (I) promote gene expression profiles favorable for glucose oxidation in muscle and heart. Skeletal muscle thioredoxin inhibitor (TXNIP) expression levels are negatively correlated with glucose uptake. As described herein, administration of the compounds of formula (I) reduces cardiac TXN1P levels. Specifically, compared to hyperglycemic mice, skeletal muscle treated with the compounds of formula (I) showed significantly reduced TXN1P levels. Txnip Decreased mRNA and protein levels (see also...) Figure 1 (e) and Figure 2 Furthermore, compared to controls, STZ mice contained Glut4-encoding... Slc2a4 Skeletal muscle expression was decreased, but normalized in hyperglycemic mice treated with the compound of formula (I), and the expression encoding Glut1 was reduced. Slc2a1 Skeletal muscle expression tends to increase. Similarly, in hyperglycemic mice, Pyruvate dehydrogenase kinase 4 (Pdk4) Pyruvate dehydrogenase (PDH) is a negative regulator of oxidative glucose metabolism and therefore a factor in pyruvate dehydrogenase (PDH) metabolism. Uncoupling protein 3 (Ucp3, Increased expression of the compound (which is beneficial for lipids as fuel substrates) was reduced in mice treated with the compound of formula (I). Figure 1 (e) indicates that the compound of formula (I) counteracts metabolic inflexibility in the skeletal muscle of hyperglycemic diabetic mice. Txnip Expression was also increased in the hearts of hyperglycemic mice, but relatively decreased in hyperglycemic mice treated with compounds of formula (I). Furthermore, Slc2a1 and Slc2a4 Cardiac expression of [the compound] was decreased in untreated hyperglycemic mice, but tended to increase in mice treated with the compound of formula (I). Figure 1(f) Therefore, the compound of formula (I) promotes changes in gene expression that favor glucose uptake, oxidative glucose metabolism and ATP production.

[0050] High blood sugar has shown a rapid increase. Pdk4 and Ucp3 The compound was expressed in the heart and induced metabolic inflexibility and cardiac dysfunction in mice. Untreated STZ mice showed a gradually decreasing peak E velocity and an increased isovolumetric relaxation time (IVRT), indicating impaired diastolic function, while treatment with the compound of formula (I) for 1 week reduced IVRT and increased peak E velocity, thereby increasing the E / A ratio. Figure 1 (g), Table 4).

[0051] In one aspect, this article provides methods and compositions for treating cancers associated with pyruvate dehydrogenase kinase (PDK) overexpression or activation in subjects of need (e.g., human patients), methods comprising administering a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the PDK is pyruvate dehydrogenase kinase 4 (PDK4).

[0052] In some implementations, the cancer is a solid tumor. In some such implementations, solid tumors include breast cancer, ovarian cancer, esophageal cancer, melanoma, brain cancer, lung cancer, head and neck cancer, oral cancer, colorectal cancer, breast cancer, prostate cancer, pancreatic cancer, liver cancer, testicular cancer, endometrial cancer, or uterine cancer. Other types of solid tumors are named according to the specific cells that form them, such as sarcomas formed from connective tissue cells (e.g., bone, cartilage, fat), carcinomas formed from epithelial tissue cells (e.g., breast, colon, pancreas), and lymphomas formed from lymphoid tissue cells (e.g., lymph nodes, spleen, thymus). In some of the aforementioned implementations, the cancer is metastatic cancer.

[0053] In some implementations, the cancer is a blood cancer. In some such implementations, the blood cancer is acute lymphoblastic leukemia, chronic lymphoblastic leukemia, or multiple myeloma.

[0054] In some embodiments, this document provides a method of treating cancer, comprising administering a compound of formula (I), wherein the cancer is associated with a hypoxic tumor. In some embodiments, the cancer is associated with a hypoxic tumor, which is leukemia, breast cancer, cervical cancer, brain cancer, kidney cancer, liver cancer, lung cancer, pancreatic cancer, colorectal cancer, head and neck cancer, prostate cancer, vulvar cancer, skin cancer, or sarcoma. In some embodiments, the hypoxic tumor has a median oxygen level of less than 1%. In some embodiments, the hypoxic tumor has a median oxygen level of less than 1% or less than 0.5%.

[0055] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat bladder cancer. In some embodiments, the bladder cancer is metastatic bladder cancer.

[0056] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat colorectal cancer. In some such embodiments, the colorectal cancer is associated with a KRAS mutation. In some embodiments, the colorectal cancer is metastatic colorectal cancer.

[0057] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat lung cancer. In some such embodiments, the lung cancer is associated with a KRAS mutation. In other such embodiments, the lung cancer is epidermal growth factor receptor (EGFR) positive lung cancer. In some embodiments, the lung cancer is metastatic lung cancer. In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC). In some embodiments, NSCLC is associated with an EGFR mutation.

[0058] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat breast cancer. In some such embodiments, the breast cancer is tamoxifen-resistant breast cancer. In some embodiments, the breast cancer is metastatic breast cancer. In some embodiments, the breast cancer is triple-negative breast cancer. In some embodiments, the breast cancer is hormone receptor-positive human epidermal growth factor receptor 2-negative breast cancer (HR+HER2-mBC).

[0059] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat melanoma. In some embodiments, the melanoma is metastatic melanoma. In some embodiments, the melanoma is associated with activating mutations in the BRAF oncogene. In some embodiments, the subject (e.g., a patient) has previously been treated with immunotherapy (e.g., an anti-PD1 or anti-CTLA4 antibody). In some such embodiments, the metastatic melanoma is unresponsive or has a limited response to immunotherapy. In some embodiments, the metastatic tumor is resistant to or has become resistant to at least one BRAF inhibitor. In some such embodiments, the BRAF inhibitor is selected from vemurafenib, dabrafenib, cannefenib, trametinib, bimetinib, and cobimetinib.

[0060] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat bladder cancer. In some embodiments, the bladder cancer is metastatic bladder cancer.

[0061] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to treat hepatocellular carcinoma (HCC). In some embodiments, the subject (e.g., a patient) has cervical cancer that has become resistant to doxorubicin. In some embodiments, the subject (e.g., a patient) has cervical cancer that has become resistant to etoposide.

[0062] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to target cancer stem cells. In some such embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is used to target cancer stem cells resistant to radiotherapy (radiotherapy).

[0063] This article also provides a method for inducing apoptosis in apoptosis-inducing cancer cells, the method comprising administering an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof to the apoptosis-inducing cancer cells. In some embodiments, the cancer cells are bladder cancer cells, gastric cancer cells, colon cancer cells, breast cancer cells, ovarian cancer cells, melanoma cells, or lung cancer cells.

[0064] On the other hand, a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered to a subject (e.g., a patient) with cancer that is resistant to one or more prior treatments (e.g., chemotherapeutic agents) and / or resistant to radiotherapy (radiotherapy). In some embodiments, the prior treatment is with a platinum-based agent, taxane, nucleoside analog, immune checkpoint inhibitor, Cox-2 inhibitor, anthracycline, pyrimidine analog, topoisomerase inhibitor, mTOR inhibitor, proteasome inhibitor, angiogenesis inhibitor, B-Raf inhibitor, or tyrosine kinase inhibitor. In some embodiments, the cancer is resistant to docetaxel, paclitaxel, cisplatin, or gemcitabine.

[0065] On the other hand, this disclosure provides a method for reducing resistance to chemotherapeutic agents, the method comprising administering to a subject with cancer (e.g., a cancer patient) a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In some embodiments, the chemotherapeutic agent is cisplatin. In some embodiments, the chemotherapeutic agent is 5-fluorouracil. In some embodiments, the chemotherapeutic agent is erlotinib. In some embodiments, the chemotherapeutic agent is paclitaxel. In some embodiments, the chemotherapeutic agent is doxorubicin. In some embodiments, the chemotherapeutic agent is gemcitabine. In some embodiments, the chemotherapeutic agent is an immune checkpoint inhibitor.

[0066] Therapeutic compounds In some of the foregoing embodiments, the therapeutic compound includes exemplary compounds described in further detail below. Compounds used in the methods provided herein may include their salts, solvates, or prodrugs.

[0067] In some respects, this disclosure provides a method of treating cancer in a subject (e.g., a patient) of need, including administering an effective amount of a compound of formula (I) (4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide or a salt, solvate or prodrug thereof.

[0068] In some embodiments, the compound is an alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. An "alkali metal" is a metal present in Group I of the periodic table along with hydrogen. Alkali metals are lithium, sodium, potassium, rubidium, cesium, and francium. Therefore, it should be understood that an "alkali metal salt" is a compound consisting of an aggregate of one or more alkali metal cations and associated anions. Thus, the term "alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide" refers to a compound 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-thiadiazol-5-yl]benzamide. For example, the alkali metal salts of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide are shown below: .

[0069] Where X + It represents an alkali metal cation (such as lithium, rubidium, cesium, sodium, or potassium).

[0070] It should be understood that a "sodium salt" is a compound composed of an aggregate of sodium cations and associated anions. Therefore, the term "sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide" refers to a compound containing both sodium cations and anions of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. For example: , Na + It represents sodium cation.

[0071] Technicians 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-thiadiazol-5-yl]benzamide can dissociate into its anionic and cationic components.

[0072] Throughout this specification, structures may or may not be presented by chemical name. In case of any naming issues, the structure shall prevail. Where compounds may exist as tautomers (e.g., in alternative resonance forms), the depicted structure represents one of the possible tautomer forms, wherein the actual tautomer form 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: To avoid any doubt, the alkali metal salts of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide are solids under ambient conditions, and therefore the scope of this invention includes all its amorphous, crystalline and partially crystalline forms.

[0073] Alkali metal salts of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be prepared using techniques well known to those skilled in the art. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be reacted with a suitable alkali metal hydroxide or an alternative alkali metal compound. Salt conversion techniques can also be used to convert one salt into another.

[0074] Sodium salts of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be prepared using techniques well known to those skilled in the art. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide can be reacted with sodium hydroxide or alternative sodium base compounds. Salt conversion techniques can also be used to convert one salt into another.

[0075] 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 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 can be prepared according to the technique described in International Patent Application WO 2011 / 004162.

[0076] Unless otherwise stated, all technical and scientific terms used herein shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0077] In a specific embodiment, the alkali metal salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide is a 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 a sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0078] In some embodiments, the compounds used in the methods provided herein are compounds of formula (II): (II), or its salt, wherein 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 is capable of being metabolized in vivo to form 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide. In some embodiments, the compound is a salt. For example, the salts of the compound include: , Where X + The cation represents an alkali metal, alkaline earth metal, or quaternary ammonium (e.g., lithium, magnesium, calcium, ammonium, tetramethylammonium, especially sodium and potassium) cation, with appropriate stoichiometric adjustments made according to the charge of the ion. In some embodiments, X + It represents an alkali metal cation (e.g., lithium, rubidium, cesium, or especially sodium or potassium).

[0079] Drug dosage form Therapeutic compounds are administered to human subjects in need in the form of pharmaceutical formulations, which are also referred to herein as pharmaceutical dosage forms.

[0080] In one embodiment, the therapeutic compound is the only active pharmaceutical ingredient present in the dosage form. In another embodiment, the therapeutic compound is present in the dosage form along with one or more other active pharmaceutical ingredients, or may be administered as part of a combination therapy with one or more other active pharmaceutical ingredients.

[0081] In specific embodiments, the therapeutic compound is provided in the form of particles having a particle size distribution defined by a D90 of less than about 10 µm (e.g., as measured using laser diffraction). In one embodiment, particles containing the therapeutic compound may have a particle size distribution defined by a D90 of less than about 10 µm (e.g., from about 5 µm to about 10 µm) (e.g., as measured using laser diffraction). The particle size distribution may alternatively be defined by a D90 of less than about 8 µm (e.g., from about 5 µm to about 8 µm). In another embodiment, particles composed of the therapeutic compound may have a particle size distribution defined by a D50 of less than about 6 µm (e.g., from about 0.5 µm to about 6 µm). In yet another embodiment, the particle size distribution of particles composed of the therapeutic compound may be further defined by a D10 of less than about 2 µm (e.g., from about 0.2 µm to about 2 µm). The above particle size distribution parameters may be applied individually or in combination. For example, in a specific embodiment, the dosage form comprises particles containing a therapeutic compound having a particle size distribution defined by a D90 of less than about 10 µm and a D50 of less than about 6 µm. More specifically, the particles may have a particle size distribution defined by a D90 of less than 9 µm, a D50 of less than 6 µm or less than 5 µm, and a D10 of less than 2 µm or less than 1.5 µm. The particle size distribution of the particles containing the therapeutic compound can be measured using, for example, a commercially available particle size analyzer via laser diffraction.

[0082] oral dosage form In some variations, the therapeutic compound may be provided in tablet or capsule form. For example, capsules containing a single therapeutic compound or in combination with a suitable medium (e.g., vegetable oil, fat, etc.), such as soft gelatin capsules, may be prepared. Similarly, hard gelatin capsules may contain a single therapeutic compound or in combination with solid powder ingredients such as disaccharides (e.g., lactose or sucrose), alcoholic sugars (e.g., sorbitol or mannitol), plant starches (e.g., potato starch or corn starch), polysaccharides (e.g., amylopectin or cellulose derivatives), or gelling agents (e.g., gelatin).

[0083] The drug formulations described herein can be prepared according to standard and / or recognized pharmaceutical practices. Drug formulations are typically provided as mixtures comprising a therapeutic compound and one or more pharmaceutically acceptable excipients. One or more pharmaceutically acceptable excipients can be selected based on standard pharmaceutical practices, taking into account the intended route of administration. Such pharmaceutically acceptable excipients are preferably chemically inert to the active compound and preferably do not have harmful side effects or toxicity under the conditions of use. Suitable drug formulations can be found, for example, in Remington, The Science and Practice of Pharmacy, 19th edition, 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).

[0084] Those skilled in the art will understand that the pharmaceutical dosage forms described herein can act systemically and therefore can be administered accordingly using suitable techniques known to those skilled in the art. Pharmaceutical dosage forms as described herein are typically administered orally, for example, as oral pharmaceutical dosage forms. Therefore, in some variations, an oral pharmaceutical dosage form is provided comprising about 100 to about 1000 mg of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide or a pharmaceutically acceptable salt, solvate, or prodrug thereof. In one variation, an oral pharmaceutical dosage form is provided comprising about 200 to about 1000 mg of a sodium salt of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide.

[0085] Dosage forms intended for oral administration may also include an enteric coating to prevent or minimize dissolution or disintegration in the gastric environment. Therefore, oral formulations (e.g., capsules or tablets) coated with an enteric coating can provide 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., on the surface of a tablet or capsule), or each particle containing the therapeutic compound may be coated with an enteric coating. Therefore, in specific embodiments, pharmaceutical dosage forms used in the methods of the present invention further include an enteric coating.

[0086] In some implementations, the enteric coating is present on the drug dosage form, and in some variations, the coating may be provided as an outer layer on the drug dosage form.

[0087] Alternatively, the particles containing the therapeutic compound can be coated individually with an enteric coating, and the coated particles can be formulated into a pharmaceutical dosage form. Therefore, in a specific embodiment, the pharmaceutical dosage form contains particles comprising the therapeutic compound, and each particle is coated with an enteric coating.

[0088] The term "enteric coating" refers to a substance (e.g., a polymer) incorporated into an orally administered drug (e.g., applied to the surface of tablets, capsules, granules, or pills) and inhibiting the dissolution or disintegration of the drug in the gastric environment. Enteric coatings are generally stable at the highly acidic pH of the stomach but rapidly decompose at the relatively alkaline pH of the small intestine. Therefore, enteric coatings prevent the release of the active ingredient in the drug until it reaches the small intestine.

[0089] Any enteric coating known to those skilled in the art can be used in this invention. Specific enteric coating materials that may be mentioned include those comprising beeswax, shellac, alkyl cellulose polymer resins (e.g., ethyl cellulose polymer, carboxymethyl ethyl cellulose, or hydroxypropyl methyl cellulose phthalate), or acrylic polymer resins (e.g., acrylic and methacrylic acid copolymers, methyl methacrylate copolymers, ethoxyethyl methacrylate, cyanoethyl methacrylate, methacrylate copolymers, methacrylic acid copolymers, aminoalkyl methacrylate copolymers, poly(acrylic acid), poly(methacrylic acid), alkylamide methacrylate copolymers, poly(methyl methacrylate), poly(methacrylic acid) (anhydride), polymethacrylate, methyl methacrylate copolymers, poly(methyl methacrylate) copolymers, polyacrylamide, poly(methacrylic anhydride) and glycidyl methacrylate copolymers), cellulose acetate phthalate, and polyvinyl acetate phthalate.

[0090] In some variations, 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. Specifically, the at least one pharmaceutically acceptable excipient may be a lubricant, binder, filler, surfactant, diluent, anti-adhesion agent, coating agent, flavoring agent, coloring agent, flow aid, preservative, sweetener, disintegrant, adsorbent, buffer, antioxidant, chelating agent, solubility enhancer, solubility inhibitor, or wetting agent.

[0091] Specific pharmaceutically acceptable excipients that may be mentioned include mannitol, PVP (polyvinylpyrrolidone) K30, lactose, sucrose, sorbitol, starch, amylopectin, cellulose derivatives, gelatin or other suitable ingredients, as well as disintegrants and lubricants such as sodium lauryl sulfate, sodium docusate, magnesium stearate, calcium stearate, sodium stearoyl fumarate, and polyethylene glycol wax. In preparing pharmaceutical dosage forms of therapeutic compounds for oral administration, particles (preferably ground) containing the therapeutic compound may be mixed together with or separately from mannitol, PVP (polyvinylpyrrolidone) K30, and sodium lauryl sulfate.

[0092] In preparing a pharmaceutical dosage form, a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof may be mixed together or separately with one or more of the pharmaceutical excipients listed above (including basic excipients).

[0093] A mixture of a therapeutic compound and one or more pharmaceutically acceptable excipients can be processed into pills or granules, or compressed into tablets. Therefore, the pharmaceutical dosage form of the method of the present invention can be a tablet, microtablet, block, pill, granule, powder, or granule for oral administration.

[0094] The pharmaceutical formulations that may be mentioned include those in which the compound of formula (I) or its pharmaceutically acceptable salt, solvate or prodrug is present in a total amount of at least 1% (or at least 10%, at least 30% or at least 50%) by weight of the formulation. That is, the weight ratio of the therapeutic compound to all components of the pharmaceutical formulation (i.e., the therapeutic compound and all pharmaceutical excipients, such as adjuvants, diluents and carriers) is at least 1:99 (or at least 10:90, at least 30:70 or at least 50:50).

[0095] Dosage of oral formulations As used herein, “therapeutic effective amount,” “effective amount,” or “dosage” means the amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof sufficient to produce a desired effect, which may be a therapeutic effect and / or a beneficial effect. Effective amounts or dosages will vary with individual or subject (e.g., human) age or general condition, 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 knowledge and expertise of those skilled in the art.

[0096] As used herein, when referring to measurable values ​​such as the amount of a compound, dosage, time, temperature, etc., the term "about" refers to a variation of 20%, 10%, 5%, 1%, 0.5%, or even 0.1% of the specified amount. It is anticipated that, in each case, such a term could be replaced by a symbol such as "±10%" (or by indicating the variance of a specific amount calculated based on the relevant value). It is also anticipated that, in each case, such a term could be removed.

[0097] It should be understood that the dosage and pharmacokinetic parameters described below relate to the treatment of patients with specific diseases, particularly heart diseases as described herein. The dosage and pharmacokinetic parameters may also be applied to human subjects taking the drug to improve physical condition or cardiac function. As mentioned above, in some embodiments, lower amounts of the compound of formula (I) may be required to improve physical condition or cardiac function compared to administration of the compound to a subject with heart disease (e.g., a patient).

[0098] In any of the foregoing embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject in need (e.g., a patient) at a dose of about 100 mg to about 1,000 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily at a dose of about 200 mg to about 1,000 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily at a dose of about 400 mg to about 800 mg. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily at a dose of about 100 mg to about 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 at a dose of about 100 mg, about 200 mg, about 300 mg, about 400 mg, or about 500 mg.

[0099] In any of the foregoing embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 40 µg / mL to about 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 in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 40 µg / mL to about 80 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 in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 70 µg / mL to about 120 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 in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 90 µg / mL to about 160 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 in need (e.g., a patient), wherein the administration results in a steady-state plasma concentration of the compound of formula (I) of about 100 µg / mL to about 150 µg / mL. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered daily to a human subject, resulting in a steady-state plasma concentration of the compound of formula (I) of about 120 µg / mL to about 140 µg / mL.

[0100] In any of the foregoing embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 1,000 h*µg / mL to approximately 4,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 1,000 h*µg / mL to approximately 2,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24The concentration is approximately 1,500 h*µg / mL to approximately 4,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 1,800 h*µg / mL to approximately 3,100 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 1,500 h*µg / mL to approximately 2,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 The concentration is approximately 2,500 h*µg / mL to approximately 4,000 h*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 in need (e.g., a patient), wherein such administration results in a steady-state AUC of the compound of formula (I). 0-24 It ranges from approximately 3,000 h*µg / mL to approximately 3,500 h*µg / mL.

[0101] In any of the foregoing embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof may be administered orally once daily to a subject in need (e.g., a patient), wherein such administration results in the Ci 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 in need (e.g., a patient), wherein such administration results in the C1 of the compound of formula (I) being... 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 in need (e.g., a patient), wherein such administration results in the C1 of the compound of formula (I) being... 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 in need (e.g., a patient), wherein such administration results in the C1 of the compound of formula (I) being... maxThe 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 in need (e.g., a patient), wherein such administration results in the C1 of the compound of formula (I) being... max It is approximately 1200 µg / mL to approximately 150 µg / mL.

[0102] In some embodiments, the compound of formula (I) is provided in tablet form having the components shown in Table 1.

[0103] Table 1: Exemplary composition of sodium salt tablets containing formula (I)

[0104] *The free acid form corresponding to the compound of formula (I) in 400 mg form. Combination therapy Those skilled in the art will understand that the methods of the present invention may include further treatments (e.g., combinations thereof) for the same condition. Such combinations are selected based on the condition to be treated, the cross-reactivity of the components, and the pharmacological properties of the combination. In one embodiment, a compound as described herein or a pharmaceutically acceptable salt thereof is provided for use in combination with another anticancer therapy (such as a chemotherapy agent, immunotherapy agent, gene therapy agent, or a combination thereof). For example, when treating cancer, the compounds and compositions provided herein may be combined with other anticancer therapies, surgical procedures, radiation procedures, or any of the foregoing combinations. Combination therapies are also contemplated in the treatment methods described herein.

[0105] As used herein, “combination therapy” means a therapy comprising two or more different anticancer compounds or therapeutic agents. Therefore, in one aspect, a combination therapy comprising a compound detailed herein and another compound or therapeutic agent is provided. In some variations, the combination therapy optionally comprises one or more pharmaceutically acceptable carriers or excipients, non-pharmaceutical active compounds, and / or inert substances. In various embodiments, treatment with the combination therapy may result in additive or even synergistic (e.g., greater than additive) results compared to the administration of a single compound of this disclosure alone. In some embodiments, a lower amount of each compound is used as part of the combination therapy compared to the amount typically used for a single therapy. Preferably, the combination therapy achieves the same or greater therapeutic benefit compared to the use of any single compound alone. In some embodiments, the use of a smaller amount (e.g., a lower dose or a lower frequency of dosing) of the compound in the combination therapy achieves the same or greater therapeutic benefit compared to the amount typically used for a single compound or therapy. Preferably, the use of a smaller amount of the compound results in a reduction in the number, severity, frequency, and / or duration of one or more compound-related side effects.

[0106] The compounds of this disclosure can also be combined with one or more other active ingredients in a single dosage form for simultaneous or sequential administration to a subject (e.g., a patient). Combination therapy can be administered as a simultaneous or sequential regimen. When administered sequentially, the combination can be administered two or more times.

[0107] When treating a disease or condition that is improved by activating AMPK, the salt of the present invention may be administered in combination with one or more other (i.e. different) therapeutic agents that can be used to treat the disease or condition.

[0108] Such combination therapy may involve administering the salt of the invention in combination with different therapeutic agents in the same formulation or preferably in separate formulations (e.g., sequentially or simultaneously) to a subject. The term "administration in conjunction with" (and similarly "administered in conjunction with") includes the sequential or simultaneous administration of the respective active ingredients as part of a medical intervention for the treatment of the relevant condition. "Simultaneously" means the side-by-side administration of the salt of the invention and different therapeutic agents in a single pharmaceutical dosage form comprising both active ingredients or in separate dosage forms for simultaneous administration.

[0109] Therefore, for the purposes of this invention, the term "administration in conjunction with" (and similarly "administered in conjunction with") includes administering the salt of this invention together with different therapeutic agents, or at sufficiently close times, to produce a greater beneficial effect on a subject (e.g., a patient) during the treatment of the relevant condition than either agent alone in the absence of the other component during the same treatment. Determining whether the combination provides a greater beneficial effect in the treatment of a particular condition and during the treatment of that condition will depend on the condition to be treated, but can be conventionally performed by those skilled in the art.

[0110] Furthermore, in the context of this invention, the term "in combination with" includes the possibility that one or the other of two active ingredients may be applied before, after, and / or simultaneously with the other (optionally repeatedly). When used in this context, the terms "simultaneous application" and "simultaneous application with" include the application of individual doses of the salt of the invention and different therapeutic agents within 6 hours, 3 hours, 2 hours, 1 hour, 45 minutes, 30 minutes, 20 minutes, or 10 minutes of each other.

[0111] The methods of the present invention disclosed herein (and the oral dosage forms used in such methods) also have the following advantages: dose-effective methods using the salts of the present invention can be more effective, less toxic, have a longer duration of action, be more potent, produce fewer side effects, be more easily absorbed, and / or have better pharmacokinetic characteristics (e.g., higher oral bioavailability and / or lower clearance) than other therapies known in the prior art, whether for the aforementioned indications or other indications. Specifically, due to the dose-effective properties of the salts of the present invention, the methods of the present invention can have the following advantages: they are more effective in vivo and / or exhibit advantageous properties, such as fewer side effects.

[0112] Specific examples of other anticancer agents that can be used in combination with compounds of formula (I) include, but are not limited to, platinum-based chemotherapeutic agents (such as cisplatin, carboplatin, or oxaliplatin), kinase inhibitors (such as bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, vemodilpirkone, ibrutinib), topoisomerase inhibitors (such as topoisomerase I inhibitors (e.g., irinotecan or topotecan)) or topoisomerase II inhibitors (e.g., etoposide or teniposide), anthracyclines (such as daunorubicin, ... Doxorubicin, epirubicin, idarubicin, mitoxantrone or pentorubicin), histone deacetylase inhibitors (such as vorinostatin or romidesin), bromodomain inhibitors, other epigenetic inhibitors, taxanes (such as paclitaxel or docetaxel), anti-angiogenic inhibitors, nucleotide analogs or precursor analogs (such as azacitidine, azathioprine, capecitabine, cytarabine, deoxyfluorouridine, 5-fluorouracil, gemcitabine, hydroxyurea, mercaptopurine, methotrexate or thioguanine) or pemetrexed.

[0113] In some embodiments, a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with cisplatin to treat a subject (e.g., a patient) with cancer. In some embodiments, the cancer is bladder cancer, gastric cancer, breast cancer, lung cancer, HCC, ovarian cancer, or colon cancer.

[0114] In some embodiments, a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with an EGFR inhibitor to treat a subject (e.g., a patient) with lung cancer (e.g., NSCLC). In some embodiments, the EGFR inhibitor is erlotinib, gefitinib, lapatinib, lazetinib, or nexituzumab.

[0115] In some implementations, a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof is administered in combination with paclitaxel to treat lung cancer (e.g., NSCLC) in a subject (e.g., a patient) in need.

[0116] In some embodiments, a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with doxorubicin to treat a subject in need (e.g., a patient) with HCC. In some embodiments, a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with doxorubicin to treat a subject in need (e.g., a patient) with cervical cancer.

[0117] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with recombinant arginase. In some such embodiments, the cancer is breast cancer (e.g., triple-negative breast cancer). In other such embodiments, the cancer is HCC.

[0118] In some implementations, the compound of formula (I) or its pharmaceutically acceptable salt, solvate or prodrug is administered in combination with a COX2 inhibitor, propranolol, metformin, salinomycin or phenformin.

[0119] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered in combination with an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor (e.g., cimiprizumab, nivolumab, or pembrolizumab). In some embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor (e.g., atezolizumab, avermab, or durvalumab). In some embodiments, the immune checkpoint inhibitor is a CTLA-4 inhibitor (e.g., ipilimumab or trimemumab).

[0120] On the other hand, this disclosure provides a method for overcoming tumor radioresistance by administering a pharmaceutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof to a subject suffering from cancer (e.g., a cancer patient). In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof is administered to the subject (e.g., a patient) prior to receiving radiotherapy.

[0121] Uses of compounds In some aspects, the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof in a therapy (including 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 treating a subject of need (e.g., a human patient) with cancer associated with pyruvate dehydrogenase kinase (PDK) overexpression or activation 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 reducing resistance to chemotherapeutic agents. 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 inducing apoptosis in apoptosis-inducing cancer cells is provided.

[0122] In some aspects, the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof in the preparation of a medicament is also provided. In some embodiments, the medicament is used in 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 preparation of a medicament for treating a subject of need (e.g., a human patient) with cancer associated with pyruvate dehydrogenase kinase (PDK) overexpression or activation is provided. In other embodiments, the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof in the preparation of a medicament for reducing resistance to chemotherapeutic agents is provided. In other embodiments, the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, or prodrug thereof in the preparation of a medicament for inducing apoptosis in apoptosis-inducing cancer cells is provided.

[0123] Example The subject matter disclosed herein will be better understood by referring to the following embodiments, which are provided as examples of the invention and not as limitations.

[0124] In this document, unless otherwise stated, “Cmpd-(I)” or “compound of formula (I)” is mentioned in the experiments detailed in the following examples and is 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide.

[0125] Mice. Mice were housed in certified animal facilities with a 12-hour light / dark cycle and free access to appropriate food. Male BKS.Cg-Dock7m + / + Leprdb / J (db / db) and C57BLKS / J (BKS) mice with leptin receptor deficiency were obtained. F1 mice were obtained by crossing male C57BL / 6J mice with female CBA / CaCrl. Nine-week-old male F1 mice were treated with multiple low-dose streptozotocin (50 mg / kg*day for 5 consecutive days; freshly prepared in 0.1 mM sodium citrate, pH 4.5) to induce diabetes. Mice were randomly fed a D10001 diet or D10001 formulated with compounds of formula (I) at 0.25 mg / g, 0.5 mg / g, and 1 mg / g (CAS # 1261289-04-6). BKS, db / db, and F1 mice were housed in cohorts of 4–5 mice per cage. Mice with significant health problems, such as weight loss >10% or fighting, were excluded; no differences were observed between groups. Within each cohort, mice were randomly assigned to cages based on weight and fasting blood glucose, either by cage or, if possible, individually to different treatments to minimize the impact of starting weight and glucose homeostasis. For in vitro analyses, such as Western blotting, qPCR, histological, and immunohistochemical work, samples from 5–9 mice per diet were randomly selected. All in vivo analyses were performed between 9:00 AM and 3:00 PM.

[0126] Echocardiography. For echocardiography, use 1.5%-2% isoflurane (0.8L min). -1 Mice were sedated with O2(g) and placed on a temperature-controlled platform. Chest hair was removed using depilatory cream. Respiration and ECG were monitored during scanning, and anesthesia was adjusted to avoid respiratory depression. The total scanning time did not exceed 15 minutes. Stroke volume, cardiac output, and wall thickness were measured in the parasternal long-axis section using B-mode and M-mode images. Diastolic left ventricular inflow was observed in the apical four-chamber view using mitral Doppler. Offline analysis was performed blinded.

[0127] Mitochondrial respiration analysis. C2C12 myoblasts were cultured in a medium containing 10% fetal bovine serum and 20 U / ml penicillin-streptomycin. To obtain myotubes, C2C12 myoblasts were cultured at 7500 cells / cm². 2Cells were seeded in poly-L-lysine-coated XF96 culture dishes and cultured in growth medium for 3 days until 80% confluence, after which they were switched to differentiation medium (DM; growth medium with 2% horse serum instead of FBS, cultured for 6 days, with medium changed every 2 days). Respiration was measured using a Seahorse XFe96 extracellular flux analyzer according to the manufacturer's "Mitochondrial Stress" protocol and mitochondrial parameters. Briefly, differentiated myotube cells were pretreated for 4 hours by switching to growth medium containing 1% horse serum and the compound of formula (I) (sodium salt preparation). Myotube cells were then equilibrated for 1 hour in hippocampal assay medium (1 mM sodium pyruvate, 10 mM glucose, 2 mM L-glutamine) adjusted to pH 7.4, supplemented with the compound of formula (I) during pretreatment. Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) measurements were collected as baseline values, followed by the sequential addition of 1 µM oligomycin, 2 µM FCCP, and 0.5 µM rotenone + 0.5 µM antimycin A. Mitochondrial functional parameters were calculated according to the manufacturer's recommendations.

[0128] Western blot analysis. Femoral muscle was isolated from non-fasted mice and crushed in a mortar using a pestle and liquid nitrogen. Femoral muscle and INS-1E cell samples were homogenized in ice-cold protein lysis buffer (100 mM Tris pH 6.8, 2% SDS), with a mixture of protease inhibitors and phosphatase inhibitors added, and then sonicated. The femoral muscle supernatant was collected after 10 minutes at 14,000 rpm. Protein concentration was measured using a BCA kit, followed by dilution and denaturation in Laemmli buffer. Protein samples of 20 µg Vastus and 7 µg INS-1E cells were separated on 4%–15% Criterion™ TGX Stain-Free™ protein gels and blotted onto low-fluorescence PVDF or nitrocellulose membranes, respectively. Primary and secondary antibodies are listed in Table 2. Values ​​were normalized to unstained total protein signal or β-actin.

[0129] Table 2 qRT-PCR. Total RNA from isolated pancreatic islets was prepared using the RNeasy Micro Kit. Total RNA from the gastrocnemius muscle and left ventricle was prepared using the RNeasy Fibrous Tissue Micro Kit after the tissue was first crushed in liquid nitrogen using a pestle. First-strand cDNA synthesis was performed using SuperScript III according to the manufacturer's instructions. Primers used for qRT-PCR are listed in Table 3. Tbp The expression was used to normalize samples from the pancreas, gastrocnemius muscle, and left ventricle. Through Rpl32 Validated in left ventricular samples and gastrocnemius muscle. Tbp Normalization.

[0130] Table 3 Islet isolation and culture. Mouse islets were substantially isolated and cultured for 48 h in a medium supplemented with 11 mM glucose or 22 mM glucose plus / min 5 µM of compound (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 islets from non-diabetic donors were cultured for 48 h in a medium supplemented with 5.5 mM glucose or 25 mM glucose plus / min 5 µM of compound (I) (CMRL medium, 10% fetal bovine serum, 20 U / ml Pen:Strep and 1X GlutaMax).

[0131] RNA-seq. Perform RNA-seq. Prepare an RNA-seq library from 150 ng of total RNA using the Illumina TruSeq Chain mRNA Library Kit, and then perform 100 bp paired-end sequencing on a NovaSeq 6000 Illumina sequencer. Process RNA-seq reads. Perform quality checks on Fastq files with 100-nt paired-end sequencing reads, aligning them to mouse or human genomes (GRCm39 or GRCh38). Count fragment-gene hits. Use software packages. DESeq2 Subsequent analyses were performed in R using clusterProfiler. A batch-corrected profile (referencing islet preparation batches and human donors respectively), independent filtering (α=0.05), and false detection rate (FDR) <0.05 were used. DESeq2 Normalization and differential expression analysis were performed. Genes with baseMean expression > filterThreshold were considered expressed and constituted subsequently used. clusterProfiler For the genetic background used in ORA and GSEA analyses, pvalueCutoff=0.05 and qvalueCutoff=0.2 for overrepresented gene classes.

[0132] Cell lines. INS-1E and C2C12 are commercially available and therefore not certified after purchase. The cells are mycoplasma-free, as determined by PCR.

[0133] Statistical Analysis. The experimental design consisted of a non-diabetic control group (BKS or control (i.e., F1 mice injected with the medium), a diabetic control group (db / db mice or STZ mice, i.e., F1 mice injected with STZ), and a compound treatment group of formula (I) (db / db mice or STZ mice maintained on a diet formulated with the compound of formula (I)). First, the effectiveness of the diabetes model was tested by comparing the control and diabetic groups. Second, the effect of the compound diet of formula (I) was evaluated by comparing the compound treatment group of formula (I) with the diabetic group. The parametric nature and heteroscedasticity of the data were assessed using the Shapiro-Wilk test and the Leven test, and appropriate statistical tests were selected. For comparisons between two groups, the heteroscedasticity t-test or Wilcoxon test was used for multiple tests with Holm correction. For multiple groups, ANOVA analysis was used, followed by appropriate post-hoc analyses (see figure). P A value <0.05 was considered statistically significant. tidyverse , rstatix and ggpubr The software package performs data analysis, statistical analysis, and visualization in R.

[0134] Example 1: The compound of formula (I) promotes gene expression profiles that favor glucose oxidation.

[0135] Skeletal muscle TXNIP expression levels were negatively correlated with glucose uptake. While skeletal muscle glucose uptake was enhanced, TXNIP expression in the skeletal muscle of STZ mice treated with the compound of formula (I) was significantly higher than that in untreated STZ mice. Txnip Decreased mRNA and protein levels Figure 1 (e) Figure 2 Furthermore, compared to the control, STZ mice contained Glut4-encoding... Slc2a4 Skeletal muscle expression was decreased, but normalized in STZ mice treated with the compound of formula (I), and the expression of Glut1-encoding Glut1 was reduced in STZ mice treated with the compound of formula (I). Slc2a1 Skeletal muscle expression tends to increase ( p =0.059) ( Figure 1 (e)). Similarly, in STZ mice, Pyruvate dehydrogenase kinase 4 ( Pdk4 pyruvate dehydrogenase (PDH) and therefore a negative regulator of glucose oxidative metabolism) and untie Coupling protein 3 ( Ucp3 Increased expression of (which is beneficial for lipids as fuel substrates) was reduced in mice treated with compounds of formula (I). Figure 1 (e) indicates that the compound of formula (I) counteracts metabolic inflexibility in the skeletal muscle of STZ diabetic mice. TxnipExpression was also increased in the hearts of STZ mice, but relatively decreased in STZ mice treated with compounds of formula (I). Figure 1 (f)). Furthermore... Slc2a1 and Slc2a4 Cardiac expression of [the compound] was decreased in untreated STZ mice, but tended to increase in mice treated with compounds of formula (I) (both...). p =0.088) ( Figure 1 (f)). Pdk4 , Ucp2 and Ucp3 The expression of [the compound] was increased in the hearts of untreated STZ mice, but decreased in STZ mice treated with compounds of formula (I) (for Ucp2, [the expression was] attenuated). p =0.055) ( Figure 1 (f)). Therefore, the compound of formula (I) promoted changes in gene expression that favored glucose uptake, oxidative glucose metabolism, and ATP production in the skeletal muscle and heart of STZ mice. Figure 1 (e) Figure 1 (f)). High blood sugar has shown a rapid increase. Pdk4 and Ucp3 The compound expresses in the heart and induces metabolic inflexibility and cardiac dysfunction in mice

[34] . Untreated STZ mice showed a gradually decreasing peak E velocity and an increased isovolumetric relaxation time (IVRT), indicating impaired diastolic function, while treatment with the compound of formula (I) for 1 week reduced IVRT and increased peak E velocity, thereby increasing the E / A ratio ( Figure 1 (g), Table 4). Improved left ventricular filling in mice treated with compounds of formula (I) also led to increased stroke volume and cardiac output (Table 4). In summary, these findings suggest that compounds of formula (I) improve hyperglycemia, reduce glycogen buildup, and reverse diabetic cardiomyopathy in diabetic STZ mice by stimulating insulin-independent glucose uptake and utilization.

[0136] Table 4 Table 4 shows the echocardiographic measurements of left ventricular dimensions in parasternal long axis (PLAX) mode B 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; LVIDd / s, diastolic / systolic left ventricular diameter; PWd / s, diastolic / systolic posterior wall thickness; EF, ejection fraction; FS, fractional shortening; E / A, peak wave velocity ratio of E to A; deceleration time, deceleration time of the E wave from peak to predicted baseline; IVRT, isovolumetric relaxation time. Statistical analysis was performed using one-way repeated ANOVA and Tukey. afterwardsTest. The relationship between RD and the compound of formula (I) at the same time point* P <0.05. Compared to baseline, # P <0.05. Compared to scan 2, ¤ P <0.05. Data are mean ± SD.

[0137] Example 2: The compound of formula (I) improves hyperglycemia and promotes gene expression profiles that favor glucose oxidation in the muscles of db / db mice.

[0138] To explore the potential of the compound of formula (I) to improve hyperglycemia in cases of severe insulin resistance, db / db mice were treated 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)) for 9 weeks. BKS mice were used as controls. Compensatory hyperinsulinemia (CMP) was observed in db / db mice at 6 weeks of age. Figure 3 (a) Figure 3 (b)). Untreated db / db mice were unable to compensate for the resulting insulin resistance, and due to the decrease in insulin levels, blood glucose levels rose rapidly. Figure 3 (a) Figure 3 (b)). Compared with starting values ​​and untreated db / db mice, the compound of formula (I) dose-dependently increased insulin levels, thus attenuating the increase in blood glucose levels. Figure 3 (a) Figure 3 (b)). These findings provide evidence that the compound of formula (I) maintains the compensatory β-cell insulin secretion response, and HOMA-β calculations of β-cell function homeostasis modeling show that the decline in β-cell function is attenuated in db / db mice treated with the compound of formula (I). Figure 3 (c)). The reduction in hyperglycemia mediated by the compound of formula (I) in db / db mice paralleled the decrease in glycogen accumulation in skeletal muscle and heart. Figure 3 (d) indicates that the compound of formula (I) also stimulates glucose utilization in diabetic db / db mice. Compared with untreated db / db mice, skeletal muscle of db / db mice treated with the compound of formula (I) showed increased glucose utilization. Txnip , Pdk4 and Ucp3 Decreased expression of Slc2a4, Peroxisome proliferator-activated receptor γ-coactivator (PGC)-1α ( Ppargc1a Increased expression of Cox8b (a positive regulator of mitochondrial biogenesis and respiration) and Cox8b (a driver of oxidative phosphorylation) supports this view. Figure 3(f)). Compared with untreated db / db mice, treated db / db mice showed increased cardiac expression of Slc2a1 and Txnip , Pdk4 and Ucp3 The weakening expression ( Figure 3 (g) is also consistent with what was observed in STZ mice treated with compounds of formula (I). Notably, in STZ mice and db / db mice treated with compounds of formula (I), compounds of formula (I) did not increase serum lactate levels. Figure 4 In summary, these findings provide evidence that compounds of formula (I) avoid genetic traits associated with metabolic inflexibility, which is linked to diabetes and diabetic cardiomyopathy, in STZ and db / db diabetic mice.

[0139] Example 3: The compound of formula (I) stimulates the uncoupling of mitochondria in myotubules.

[0140] Increased glucose utilization and decreased glycogen content observed in the skeletal muscle and heart of STZ and db / db mice treated with compounds of formula (I) suggest that compounds of formula (I) increase energy expenditure by generating metabolic demand via inefficient cycling and / or mitochondrial uncoupling. Studies of mitochondrial and glycolytic function in intact, differentiated C2C12 myotubes were conducted to elucidate the potential uncoupling potential of compounds of formula (I). Compounds of formula (I) dose-dependently increased the oxygen consumption rate (OCR) (a measure of oxidative phosphorylation) and the ratio of basal OCR to basal ECAR (extracellular acidification rate) in C2C12 myotubes. Figure 5 (a) to Figure 5 (c) indicates that the compounds of formula (I) increase the cell's preference for oxidative metabolism. The compounds of formula (I) also show a reduced OCR in response to oligomycin (which blocks ATP synthase), thus increasing proton leakage. Figure 5 (a) Figure 5 (d)). However, consistent with our previously published findings, the compound of formula (I) did not significantly reduce cellular ATP levels ( Figure 5 (d)). In summary, these results indicate that compounds of formula (I) increase cellular respiration by acting as mitochondrial uncoupling agents, providing evidence that compounds of formula (I) induce enhanced metabolic demand for energy expenditure by stimulating TCA flux and oxidative metabolism.

Claims

1. A method for treating cancers associated with pyruvate dehydrogenase kinase (PDK) overexpression or activation in persons of need, comprising administering an effective amount of a compound of formula (I): (I)， Or its pharmaceutically acceptable salt.

2. The method of claim 1, wherein the administration restores the metabolic flexibility of the human cancer cells.

3. The method of claim 1 or 2, wherein the PDK is PDK4.

4. The method of any one of claims 1-3, wherein the cancer in the person is resistant to one or more anticancer agents.

5. The method of claim 4, wherein the one or more anticancer agents are platinum-based agents, taxanes, nucleoside analogs, immune checkpoint inhibitors, Cox-2 inhibitors, anthracyclines, pyrimidine analogs, topoisomerase inhibitors, mTOR inhibitors, proteasome inhibitors, angiogenesis inhibitors, β-Raf inhibitors, or tyrosine kinase inhibitors, or any combination thereof.

6. The method of any one of claims 1 to 5, wherein the cancer in the person is resistant to radiotherapy.

7. The method of any one of claims 1 to 6, wherein the administration increases tumor oxygenation.

8. The method of claim 7, wherein the tumor is a hypoxic tumor.

9. The method of any one of claims 1 to 8, wherein the cancer has metastasized.

10. The method of any one of claims 1 to 9, wherein the cancer is bladder cancer, colon cancer with a KRAS mutation, lung cancer with a KRAS mutation, stomach cancer, breast cancer, or ovarian cancer, or any combination thereof.

11. The method of claim 10, wherein the cancer is colon cancer with a KRAS mutation.

12. The method of claim 11, further comprising administering 5-fluorouracil to the person.

13. The method of claim 10, wherein the cancer is tamoxifen-resistant breast cancer.

14. The method of claim 13, further comprising administering tamoxifen to the person.

15. The method of claim 10, wherein the cancer is bladder cancer.

16. A method for reducing resistance to chemotherapeutic agents, the method comprising administering a compound of formula (I) to a person with cancer: (I)， Or its pharmaceutically acceptable salt.

17. The method of claim 16, wherein the chemotherapeutic agent is cisplatin.

18. The method of claim 16 or 17, wherein the cancer is bladder cancer, stomach cancer, breast cancer, lung cancer, HCC, ovarian cancer, or colon cancer, or any combination thereof.

19. The method of claim 16, wherein the chemotherapeutic agent is an EGFR inhibitor.

20. The method of claim 19, wherein the EGFR inhibitor is erlotinib, gefitinib, lapatinib, lazetinib, or nexituzumab or any combination thereof.

21. The method of claim 19 or 20, wherein the cancer is non-small cell lung cancer (NSCLC).

22. The method of claim 16, wherein the chemotherapeutic agent is paclitaxel.

23. The method of claim 22, wherein the cancer is non-small cell lung cancer (NSCLC).

24. The method of claim 16, wherein the chemotherapeutic agent is doxorubicin.

25. The method of claim 24, wherein the cancer is hepatocellular carcinoma (HCC) or cervical cancer.

26. The method of claim 16, wherein the chemotherapeutic agent is a Cox2 inhibitor, propranolol, metformin, salinomycin, or phenformin, or any combination thereof.

27. The method of claim 16, wherein the chemotherapeutic agent is an immune checkpoint inhibitor.

28. The method of claim 27, wherein the immune checkpoint inhibitor is an anti-PD1 antibody.

29. The method of claim 27, wherein the immune checkpoint inhibitor is an anti-PDL1 antibody.

30. The method of claim 27, wherein the immune checkpoint inhibitor is an anti-CTLA4 antibody.

31. A method for inducing apoptosis in apoptosis-inducing cancer cells, comprising administering an effective amount of a compound of formula (I) to the apoptosis-inducing cancer cells: (I)， Or its pharmaceutically acceptable salt.

32. The method of any one of claims 1 to 31, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally to the person once daily at a dose of about 100 mg to about 1,000 mg.

33. The method of any one of claims 1 to 31, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally to the person once daily at a dose of about 100 mg to about 500 mg.

34. The method of any one of claims 1 to 31, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally to the person once daily at a dose of about 200 mg to about 400 mg.

35. The method of any one of claims 1 to 31, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered to the person daily, resulting in a steady-state plasma concentration of the compound of formula (I) of about 40 μg / mL to about 120 μg / mL.

36. The method of any one of claims 1 to 35, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered daily to a human subject, resulting in a steady-state AUC of the compound of formula (I). 0-24 It ranges from approximately 1,000 h*μg / mL to approximately 4,000 h*μg / mL.

37. The method according to any one of claims 1 to 36, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is an alkali metal salt.

38. The method of claim 37, wherein the alkali metal salt is a sodium salt.

39. The method of any one of claims 1 to 38, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in the form of a pharmaceutical composition.

40. The method of claim 39, wherein the pharmaceutical composition is a tablet or capsule.

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