Use of danshensu or a pharmaceutically acceptable salt thereof or a danshensu derivative in the preparation of a PDK4 agonist

Abstract

This invention discloses the use of tanshinone or its pharmaceutically acceptable salts or derivatives in the preparation of PDK4 agonists. Tanshinone reverses EndMT and inhibits the progression of coronary artery disease atherosclerosis and endothelial dysfunction by restoring the metabolic regulator PDK4. It effectively prevents the progression of coronary artery disease by targeting anti-EndMT and metabolic regulation to improve abnormal endothelial function. The use of tanshinone in the preparation of drugs for treating angina pectoris in coronary artery disease opens up new application areas for tanshinone.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and more specifically to the use of tanshinone or its pharmaceutically acceptable salts or tanshinone derivatives in the preparation of PDK4 agonists. Background Technology

[0002] Coronary heart disease (CHD) is a common chronic cardiovascular disease. Endothelial cell dysfunction plays a crucial role in the occurrence and development of CHD and is also an important factor in the occurrence of adverse cardiovascular events. Vascular endothelial cells play an important role in inhibiting inflammatory responses, regulating vasoconstriction, and inhibiting platelet aggregation. Therefore, protecting coronary artery endothelial cells and preventing their functional damage is an important pathway and initiating factor for the prevention and control of CHD. Vascular endothelial cells have a metabolic characteristic that is highly dependent on glycolysis for energy, with ATP utilization from glycolysis reaching over 85%. Glycolysis refers to the process by which cells break down glucose in the cytoplasm to produce pyruvate. Pyruvate is further catalyzed by pyruvate dehydrogenase complex (PDC) to produce acetyl-CoA, which enters the tricarboxylic acid cycle (TCA). Pyruvate dehydrogenase kinase (PDK) phosphorylates PDC, inhibiting its activity, thereby regulating the conversion of pyruvate produced from glucose and amino acid oxidation into acetyl-CoA, maintaining a dynamic balance between glycolysis and TCA, and helping to regulate glucose metabolism and ATP production. PDK4 is a key enzyme regulating PDC activity and a crucial regulator of pyruvate oxidation and glucose homeostasis. Therefore, developing agonists to activate the PDK4 signaling pathway to regulate glycolysis holds great promise for protecting coronary endothelial cell function and preventing and controlling CHD. Summary of the Invention

[0003] The present invention aims to develop a PDK4 agonist and its application in the preparation of a drug for treating coronary heart disease, thereby regulating mitochondrial metabolism and protecting endothelial vascular cell function by activating PDK4 enzyme activity.

[0004] Through extensive and in-depth research and screening of numerous compounds, the inventors have studied the ApoE activity of tanshinone in the treatment of coronary artery disease and atherosclerosis. - / - In immunofluorescence staining experiments on mouse models, tanshinone was found to activate PDK4 enzyme activity. Experimental results indicate that the tanshinone of this invention can reverse ApoE in a dose-dependent manner. - / - The expression of PDK4, a regulatory factor for protein metabolism in the mouse aorta, was investigated. Based on this, the present invention was completed.

[0005] This invention relates to the use of tanshinone or its pharmaceutically acceptable salts or tanshinone derivatives in the preparation of PDK4 agonists.

[0006] Danshensu, chemically named [D-(+)-β-(3,4-dihydroxyphenyl)lactic acid], is usually preserved as a pharmaceutically acceptable salt due to its unstable structure.

[0007] As used herein, the term "pharmaceutically acceptable salt of tanshinone" refers to acidic and / or basic salts formed by tanshinone with inorganic acids and / or bases, organic acids and / or bases, including zwitterionic salts (internal salts), and quaternary ammonium salts, such as alkyl ammonium salts. The pharmaceutically acceptable salts of tanshinone described in this invention may be, for example, tanshinone hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoric acid, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate, or trifluoroacetate.

[0008] As an example, the pharmaceutically acceptable salts of tanshinone described in this invention are selected from: sodium tanshinone, potassium tanshinone, calcium tanshinone, lithium tanshinone, magnesium tanshinone, ammonium tanshinone, meglumine tanshinone, amine tanshinone, arginine tanshinone, and lysine tanshinone. For example, sodium tanshinone is tanshinone sodium, with the molecular formula C9H9NaO5 and a molecular weight of 220.15, having the structure shown in structural formula (1):

[0009]

[0010] The tanshinone derivatives mentioned above can be referenced in CN112830884B and CN113336704B.

[0011] The present invention also relates to tanshinone or a pharmaceutically acceptable salt or derivative thereof, and to the use of pharmaceutical compositions comprising tanshinone or a pharmaceutically acceptable salt or derivative thereof in the preparation of a medicament as a PDK4 agonist.

[0012] In the above applications, the pharmaceutical composition further includes pharmaceutically acceptable excipients and / or carriers, and the pharmaceutical composition uses tanshinone or its pharmaceutically acceptable salts or tanshinone derivatives as active ingredients.

[0013] The excipients include any one or a combination of at least two of the following: diluents, excipients, fillers, stabilizers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, pH adjusters, antioxidants, antibacterial agents, colorants, or buffers.

[0014] Furthermore, in the above applications, the drug is used to treat endothelial dysfunction.

[0015] Furthermore, the drug is used to treat coronary heart disease, including but not limited to coronary atherosclerosis and angina pectoris.

[0016] This invention also aims to provide the use of tanshinone or its pharmaceutically acceptable salts or tanshinone derivatives in the preparation of medicaments for treating coronary heart disease.

[0017] This invention allows for the administration of tanshinone, a pharmaceutically acceptable salt of tanshinone, a tanshinone derivative, or a pharmaceutical composition to patients. Tanshinone, its pharmaceutically acceptable salt, or a tanshinone derivative must be mixed with a suitable carrier or excipient to form a pharmaceutical composition to ensure an effective therapeutic dose. "Effective therapeutic dose" refers to the dose of tanshinone, its pharmaceutically acceptable salt, or a tanshinone derivative necessary to achieve a therapeutic effect.

[0018] Drugs acting as PDK4 agonists can be formulated in various dosage forms, including solid, semi-solid, liquid, and aerosol formulations. Specific dosage forms within these categories include tablets, pills, lozenges, granules, gels, ointments, solutions, suppositories, injections, inhalers, and sprays. These dosage forms can be used for both local and systemic administration, as well as for immediate-release or sustained-release administration.

[0019] For example, the drug formulation may be an ointment, oral formulation, injectable formulation, suppository, inhaler, or spray. Further, the oral formulation may be an oral tablet.

[0020] When PDK4 agonists are administered as injectable formulations, i.e., when tanshinone or its pharmaceutically acceptable salts or derivatives are injected, these compounds can be formulated into solutions, suspensions, and emulsions using water-soluble or lipid-soluble solvents. Lipid-soluble solvents specifically include vegetable oils and similar oils, synthetic fatty acid glycerides, higher fatty acid esters, and proylene glycol esters. These compounds are more readily soluble in ethanol solutions and trace amounts of DMSO solutions.

[0021] When drugs acting as PDK4 agonists are administered orally, specifically when tanshinone or its pharmaceutically acceptable salts or derivatives are given orally, they can be compounded with pharmaceutically acceptable excipients using common techniques. These excipients can formulate these compounds into various dosage forms that can be taken orally by patients, such as tablets, pills, suspensions, and gels. There are several methods for preparing oral formulations, such as first mixing the compound and solid excipients, thoroughly grinding the mixture, adding appropriate excipients, and then processing it into granules. Excipients that can be used to formulate oral dosage forms include: sugars such as lactose, sucrose, mannitol, or sorbitol; and celluloses such as corn starch, wheat starch, potato starch, gelatin, sweet potato gum, methylcellulose, hydroxymethylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidone.

[0022] The tanshinone or its pharmaceutically acceptable salts or derivatives involved in this invention can also be formulated into a spray, which is achieved through a pressurizer and a sprayer or a dry powder inhalation device. Suitable propellants that can be used in the sprayer include dichlorodifluoromethane, chloroform, dichlorotetrafluoroethane, carbon dioxide, and dimethyl ether. The dosage of the spray can be adjusted by a valve on the sprayer.

[0023] The various dosage forms involved in this invention relate to the effective therapeutic dose of tanshinone or its pharmaceutically acceptable salts. The effective therapeutic dose of these compounds depends on the patient receiving treatment. In determining the appropriate dose, the patient's weight, condition, method of administration, and the prescribing physician's subjective judgment must be taken into account. The therapeutically effective dose of tanshinone derivatives and compositions containing these compounds should be determined by a competent and experienced prescribing physician.

[0024] Although the effective therapeutic dose of tanshinone or its pharmaceutically acceptable salts or tanshinone derivatives can vary depending on the patient’s condition, the effective therapeutic dose is usually 1-1000 mg / kg.

[0025] Pharmaceutically acceptable salts. The advantages of this invention compared to existing technologies:

[0026] Pharmacological experiments have shown that tanshinone reverses EndMT and inhibits the progression of coronary artery atherosclerosis and endothelial dysfunction by restoring the metabolic regulator PDK4. By targeting anti-EndMT and regulating metabolism to improve abnormal endothelial function, it effectively prevents the progression of coronary artery disease. The use of tanshinone in the preparation of drugs for treating angina pectoris in coronary artery disease opens up new application areas for tanshinone. Attached Figure Description

[0027] Figure 1 To reverse ApoE in a dose-dependent manner using tanshinone - / - Immunofluorescence staining of EndMT markers in aortic root plaques in mice; compared with control group A, *P<0.05,**P<0.01,***P<0.0001,****P<0.0001.

[0028] Figure 2 Tanshinone dose-dependently reversed endothelial dysfunction in HUVEC cells; compared with control group B and model group B, **P<0.01, ****P<0.0001.

[0029] Figure 3 Tanshinone dose-dependently reversed the immunoblotting of EndMT markers in HUVEC cells; compared with control group B and model group B, *P<0.05,**P<0.01,***P<0.0001,****P<0.0001.

[0030] Figure 4 Tanshinone could not reverse PDK4-siRNA-induced endothelial dysfunction in HUVEC cells; compared with the control group C and the model group C, ****P<0.0001.

[0031] Figure 5 The immunoblot of PDK expression in HUVEC cells induced by PDK4-siRNA that could not be reversed by tanshinone; compared with the control group C and the model group C, **P<0.01.

[0032] Figure 6 To reverse ApoE in a dose-dependent manner using tanshinone - / - Immunofluorescence staining of the expression of the metabolic regulator PDK4 in the aortic root plaque of mice; compared with control group A, *P<0.05,***P<0.001,****P<0.0001. Detailed Implementation

[0033] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0034] This invention is based on transcriptomics sequencing analysis to predict the potential action pathways and targets of tanshinone. It uses HUVECs induced by combined TGF-β1 and IL-1β as a cell model, and ApoE knockout induced by a high-cholesterol, high-fat diet. - / - A mouse model was used to verify whether tanshinone has anti-atherosclerotic and anti-endothelial dysfunction effects in coronary heart disease and to explore its molecular mechanism of action. This study not only discovers potential new drugs for the treatment of coronary heart disease but also broadens the medical applications and fields of tanshinone, which has significant clinical and scientific research value.

[0035] The following are the examples and the drugs and reagents used in the experiments:

[0036] Biological reagents and consumables were purchased commercially (R&D system, PeproTech, Abcam, Sigma-Aldrich, Shanghai Titan Technology Co., Ltd., Sinopharm Chemical Reagent Co., Ltd., Beyotime Biotechnology Co., Ltd., Suzhou Xinsaimei Biotechnology Co., Ltd., Wuhan Sanying Biotechnology Co., Ltd., Wuhan Aiboteke Biotechnology Co., Ltd., Shanghai Heyuan Liji Biotechnology Co., Ltd., etc.). Immunoblotting and RIPA cell lysis buffer, protein quantification kits, and other reagents were purchased from Beyotime Biotechnology Co., Ltd., while antibodies were purchased from Abcam, Wuhan Aiboteke Biotechnology Co., Ltd., Wuhan Sanying Biotechnology Co., Ltd., etc.

[0037] Example 1

[0038] 1 Method

[0039] 1.1 Cell Culture

[0040] Immortalized HUVEC cell lines were purchased from the American Type Culture Collection (ATCC). HUVEC cells were cultured in endothelial cell-specific ECM medium, supplemented with 5% fetal bovine serum, 1% penicillin-streptomycin, and 1% endothelial cell ECGS growth supplement, at 37°C and 5% CO2.

[0041] 1.2 Cell Model Induction

[0042] 1.2.1 In the cytokine-induced EndMT assay, the cell grouping and treatment methods are as follows:

[0043] Control group B: When HUVEC cells reached approximately 60% fusion, fetal bovine albumin was added and cultured for 48 hours.

[0044] Model group B: When HUVEC cells reached approximately 60% confluence, they were co-cultured with 10 ng / mL TGFβ-1 and 1 ng / mL IL-1β for 48 hours.

[0045] Tanshinone-low dose group B: When HUVEC cells reached approximately 60% confluence, they were pretreated with 50 μM tanshinone for 24 hours, and then cultured for another 48 hours with 10 ng / mL TGFβ-1 and 1 ng / mL IL-1β added.

[0046] Tanshinone-high-dose group B: The treatment method was the same as that of tanshinone-low-dose group B, but the concentration of tanshinone was 200 μM.

[0047] 1.2.2 In the PDK4-siRNA-induced EndMT assay, the cell grouping and treatment methods are as follows:

[0048] Control group C: When HUVEC cells reached approximately 40% confluence, the cells were starved in serum-free medium for 8 hours, and then NC-siRNA and Lipofectamine 2000 were co-incubated to form a complex. This complex was then added to the cell culture medium and seeded into the cells for transfection.

[0049] Model group C: When HUVEC cells reached approximately 40% confluence, the cells were starved in serum-free medium for 8 hours, and then PDK4-siRNA and Lipofectamine 2000 were co-incubated to form a complex. This complex was then added to the cell culture medium and seeded into the cells for transfection.

[0050] When the C:HUVEC cells in the low-dose tanshinone group reached approximately 40% confluence, the cells were starved in serum-free medium for 8 hours, pretreated with 50 μM tanshinone for 24 hours, and then PDK4-siRNA and Lipofectamine 2000 were added to the cell culture medium and seeded into the cells for transfection.

[0051] Tanshinone-high dose group C: The treatment method was the same as that of tanshinone-low dose group C, but the concentration of tanshinone was 200 μM.

[0052] The PDK4 target sequences are as follows: forward 5'-CCGCCUUUAGUUAUACAUATT-3' and reverse 5'-UAUUAACUAAAGAGGCGGTT-3'.

[0053] 1.3 Cell scratches

[0054] Cells were induced to complete confluence according to experimental method 1.2.1 or 1.2.2. Uniform scratches were made on the cell monolayer using a sterile pipette tip, followed by washing with PBS. Migration distances were recorded by photographing at 0, 6, 12, and 24 hours. Migration rate was determined by calculating changes in wound area at 0 and 24 hours.

[0055] 1.4 Immunoblotting

[0056] Proteins were extracted from cell and tissue samples using RIPA strong lysis buffer containing protease and phosphatase inhibitors, and protein content was determined using a BCA protein assay kit. Equal volumes of protein were separated on SDS-PAGE gels and transferred to PVDF membranes. The PVDF membranes were blocked with 10% skim milk and incubated with primary antibodies (CD31, α-SMA, PDK4) at 4°C for 12–16 hours, followed by incubation with HRP-conjugated secondary antibodies for 1 hour. Protein blots were visualized using ECL chemiluminescence buffer.

[0057] 1.5 Statistical Analysis

[0058] All statistical analyses were performed using GraphPad Prism 7.0 software. The Kolmogorov-Smirnov normality test was used to test the normality of continuous variables. The central tendency and dispersion of continuous variables were described using mean ± standard deviation (mean ± SD), and categorical variables were described using frequency and percentage. Statistical significance was defined as P < 0.05.

[0059] 2 Experimental Results

[0060] 2.1 Tanshinone inhibits TGF-β1 and IL-1β-induced EndMT in HUVEC cells in vitro

[0061] Because mesenchymal cells have a higher migration capacity than endothelial cells, a cell scratch assay was used to test the cell migration ability of HUVEC cells pretreated with tanshinone. The results showed that tanshinone pretreatment dose-dependently inhibited the migration rate of HUVEC cells, such as... Figure 2 As shown. Western blot results revealed that tanshinone induced an increase in CD31 protein expression and a decrease in α-SMA protein expression in EndMT-induced HUVEC cells. Figure 3 These results indicate that tanshinone pretreatment can effectively inhibit the TGF-β1 and IL-1β co-induced EndMT process in HUVEC cells. Pretreatment of HUVEC cells with pharmaceutically acceptable tanshinone salts or tanshinone derivatives can also effectively inhibit the TGF-β1 and IL-1β co-induced EndMT process in HUVEC cells.

[0062] Therefore, the tanshinone or its pharmaceutically acceptable salt or tanshinone derivatives, through dose-dependent reversal of TGF-β1 and IL-1β co-induced EndMT in HUVEC cells, exhibited decreased cell migration ability, increased expression of endothelial cell marker CD31, and decreased expression of mesenchymal cell marker α-SMA.

[0063] 2.2 Tanshinone inhibits EndMT by restoring the expression level of the metabolic regulator PDK4.

[0064] Recent studies have shown that the metabolic basis of EndMT (End-Metabolic Transformation) includes fatty acid β-oxidation metabolism, glycolysis, aspartate metabolism, and pyruvate-derived metabolites. Notably, downregulation or inhibition of PDK4 enhances EndMT by boosting glycolysis and impairing fatty acid oxidation, thereby inducing metabolic stress in endothelial cells. Therefore, regulation of PDK4 activity provides a promising therapeutic strategy for reversing endothelial metabolic dysfunction and the EndMT process in the progression of coronary atherosclerosis.

[0065] We constructed PDK4-siRNA to knock down mitochondrial matrix enzymes to observe the regulatory effect of tanshinone on PDK4. The results showed that PDK4 knockdown significantly enhanced the migration ability of EndMT-induced HUVEC cells, indicating that HUVEC cells had acquired mesenchymal characteristics. After treatment with low and high doses of tanshinone, HUVEC cells did not show significant inhibition of the EndMT process. Figure 4 Simultaneously, in HUVEC cells treated with PDK4-siRNA, significant downregulation of the endothelial cell marker CD31 protein and upregulation of the mesenchymal cell marker α-SMA protein were also detected. Figure 5 However, both low and high doses of tanshinone failed to inhibit the occurrence of EndMT. The results indicate that inhibition of PDK4 leads to the induction of EndMT, and the lack of PDK4 counteracts the protective effect of tanshinone on EndMT. This results in increased HUVEC cell migration and significant mesenchymal cell characteristic changes, suggesting that the inhibitory effect of tanshinone on EndMT depends on the metabolic regulator PDK4.

[0066] Therefore, the tanshinone or its pharmaceutically acceptable salt or tanshinone derivatives cannot reverse the PDK4-siRNA-induced increase in HUVEC cell migration and decrease in PDK4 expression.

[0067] Example 2

[0068] Preparation of tanshinone sodium injection: Dissolve tanshinone sodium in water for injection to prepare a 15 mg / mL tanshinone sodium injection solution. This injection solution is used for the following ApoE model in patients with high cholesterol and high fat. - / - Treatment of mice.

[0069] 1. Materials

[0070] 1.1 Animals

[0071] Male ApoE - / - Mice and C57BL / 6J mice (6-8 weeks old, weighing 18-22g) were provided by Jiangsu Huachuang Biotechnology Co., Ltd. Feed was purchased from Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd. These mice were housed in the SPF-grade animal experimental center of Shanghai University School of Medicine, living in an environment with a 12-hour day-night cycle and free access to water.

[0072] 2 methods

[0073] 2.1 Experimental grouping, modeling, and drug administration

[0074] Mice were randomly assigned to control group A, model group A, low-dose tanshinone group A (tanshinone sodium active ingredient 37.5 mg / kg / d), high-dose tanshinone group A (tanshinone sodium active ingredient 150 mg / kg / d), and simvastatin group A (simvastatin active ingredient 3 mg / kg / d).

[0075] Control group A consisted of normally fed mice. Model group A consisted of mice subjected to a high-cholesterol, high-fat diet to induce ApoE2. - / - Mice, and did not receive further treatment. Tanshinone-low-dose group A and tanshinone-high-dose group A were mice subjected to the following high-cholesterol, high-fat ApoE model. - / - Mice were subjected to low-dose and high-dose tanshinone sodium treatment, respectively. Simvastatin group A consisted of mice receiving the following high-cholesterol, high-fat ApoE model. - / - Mice were used, and later treated with simvastatin. The drug was administered daily via intraperitoneal injection. ApoE model was established in mice with high cholesterol and high fat. - / - Mouse: ApoE - / - Mice were fed a Western diet containing 0.12% cholesterol and 20.06% fat for 12 weeks.

[0076] 2.2 Sample Collection

[0077] Mice weight was recorded weekly to adjust injection dosage. At the end of the 12-week treatment period, mice were anesthetized with a mild dose of sodium pentobarbital, serum samples were collected from the posterior orbital sinus, and the mice were euthanized with an overdose of sodium pentobarbital. Tissue samples, including those from the entire aorta, heart, liver, spleen, lungs, and kidneys, were then rapidly collected for subsequent analysis. The aorta was stored in 4% paraformaldehyde for 24 hours for histological examination or at -80°C for Western blot analysis.

[0078] 2.3ApoE - / - Analysis of mouse aortic root tissue sections

[0079] Pre-fixed tissue from the heart to the aorta was rapidly embedded in an OCT embedding compound for cryosectioning. Sections were prepared at a thickness of 8 μm and stained with Oil Red O, hematoxylin-eosin, masson, and immunofluorescence for histological examination.

[0080] 2.4 Immunofluorescence staining

[0081] Aortic sections were fixed in 4% paraformaldehyde, infiltrated with 0.1% PBS Triton X-100, and blocked with 10% goat serum. Primary antibody (CD31, α-SMA, PDK4) was incubated at 4°C for 12-16 hours, followed by incubation at 25°C with Alexa Fluor-conjugated secondary antibody for 1 hour. Cell nuclei were counterstained with DAPI.

[0082] 2.5 Statistical Analysis

[0083] All statistical analyses were performed using GraphPad Prism 7.0 software. The Kolmogorov-Smirnov normality test was used to test the normality of continuous variables. The central tendency and dispersion of continuous variables were described using mean ± standard deviation (mean ± SD), and categorical variables were described using frequency and percentage. Statistical significance was defined as P < 0.05.

[0084] 3 Experimental Results

[0085] 3.1 Tanshinone alleviates ApoE - / - Lipid accumulation in the aortic root of mice and inhibition of EndMT

[0086] like Figure 1 As shown, compared with model group A, tanshinone dose-dependently increased the fluorescence expression of CD31 in the aortic root lining and decreased the fluorescence expression of α-SMA in the aortic root lining. This change was reversed after treatment with low-dose and high-dose tanshinone sodium. Treatment with other injections containing tanshinone or its acceptable salts or tanshinone derivatives also increased CD31 protein expression and decreased α-SMA protein expression in ApoE- / - mice.

[0087] Therefore, the tanshinone or its pharmaceutically acceptable salt or tanshinone derivatives ameliorate the abnormal expression of endothelial-mesenchymal transition (EndMT) markers CD31 and α-SMA in atherosclerotic plaques of the aortic root in ApoE- / - mice.

[0088] 3.2 Tanshinone improves ApoE by mediating PDK4 upregulation in vivo. - / - EndMT process in the aorta of mice

[0089] Given that in vitro data indicate that tanshinone's regulation of EndMT is PDK4-dependent, we evaluated ApoE. - / - Changes in PDK4 expression in the EndMT of the mouse aorta. We further found that, compared with control group A, the fluorescence intensity of PDK4 in model group A was significantly reduced (…). Figure 6 Tanshinone strongly upregulates ApoE in a dose-dependent manner. - / - Fluorescent expression of PDK4 in mouse aortic root sections. This indicates that tanshinone treatment does indeed upregulate PDK4, reverse the progression of EndMT, and thus promote the relief of coronary atherosclerosis. Tanshinone may have good therapeutic potential as a PDK4 agonist in angina pectoris of coronary heart disease.

[0090] The tanshinone or its pharmaceutically acceptable salt or tanshinone derivative restores ApoE in a dose-dependent manner. - / - Increased expression of the metabolic regulator PDK4 in atherosclerotic plaques of the aortic root in mice.

[0091] This document describes the compounds, their preparation methods, and applications of the present invention in conjunction with specific embodiments and examples, and also sets forth and explains many details. However, it should be understood that the specific embodiments and examples provided herein are merely exemplary and do not limit the scope of protection of the present invention. In fact, those skilled in the art will recognize that the present invention can be implemented in other specific ways, and any modifications, alterations, or adjustments made accordingly do not depart from the spirit and intent of the present invention, and therefore should all be considered to be included within the scope of the present invention.

Claims

1. The use of tanshinone or its pharmaceutically acceptable salts or tanshinone derivatives in the preparation of PDK4 agonists.

2. The use of tanshinone or a pharmaceutically acceptable salt or derivative thereof, or a pharmaceutical composition comprising tanshinone or a pharmaceutically acceptable salt or derivative thereof, in the preparation of a medicament as a PDK4 agonist, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable excipient and / or carrier, and wherein the pharmaceutical composition has tanshinone or a pharmaceutically acceptable salt or derivative thereof as the active ingredient.

3. The application according to claim 2, characterized in that, The excipients include any one or a combination of at least two of the following: diluents, excipients, fillers, stabilizers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, pH adjusters, antioxidants, antibacterial agents, colorants, or buffers.

4. The application according to claim 2, characterized in that, The drug is used to treat endothelial dysfunction.

5. The application according to claim 2, characterized in that, The drug is used to treat coronary heart disease.

6. The application according to claim 5, characterized in that, The drug is used to treat coronary artery disease, atherosclerosis, and angina pectoris.

7. The application according to claim 2, characterized in that, The drug is prepared as an ointment, oral preparation, injectable preparation, suppository, inhaler, or spray.

8. The application according to claim 2, characterized in that, The effective therapeutic dose of the tanshinone or its pharmaceutically acceptable salt or tanshinone derivative is 1-1000 mg / kg.

9. The use of tanshinone or its pharmaceutically acceptable salts or tanshinone derivatives in the preparation of drugs for the treatment of coronary heart disease.

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