Use of guaiacol or its derivatives for the preparation of a medicament for the prevention or treatment of metabolic syndrome

Abstract

The application relates to application of guaiacol or a derivative thereof in preparation of a medicine for preventing or treating metabolic syndrome, and belongs to the technical field of medicines. The guaiacol and the derivative thereof provided by the application can significantly improve insulin resistance conditions by regulating a fat metabolism channel, and can simultaneously reduce inflammatory reaction and oxidative stress damage, and can realize prevention and treatment of metabolic associated steatohepatitis (MASLD), hyperlipidemia, hypercholesterolemia, obesity and diabetes through synergistic action in multiple dimensions.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, and particularly relates to application of guaiacol or a derivative thereof in preparation of a medicine for preventing or treating metabolic syndrome. BACKGROUND

[0002] With the improvement of living standards and the change of lifestyle, a series of health problems have come one after another. Global analysis shows that by 2022, the total number of obese children, adolescents and adults worldwide has exceeded 1 billion (Ng, M., et al., Global, regional, and national prevalence of overweight and obesity in children and adults during 1980-2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet, 2014. 384(9945): p. 766-81.). Lancet . 2024; 403(10431): 1027-1050.). Obesity not only directly affects the body shape and quality of life of individuals, but also is an important source of causing a variety of metabolic diseases.

[0003] Hyperlipidemia, as a common symptom of obesity, its incidence is also increasing year by year. A large number of clinical studies have shown that the incidence of hyperlipidemia in obese people is as high as 40%, and the high lipid content in the blood can cause atherosclerosis, significantly increase the risk of cardiovascular diseases such as coronary heart disease and cerebral infarction, and seriously threaten people's life and health.

[0004] The incidence of metabolic associated steatohepatitis (MASLD) is closely related to obesity and hyperlipidemia. Due to the increase of high-calorie and high-fat diet intake and the decrease of exercise, the incidence of MASLD has increased explosively. At present, its global prevalence rate has reached about 25%~30%, and gradually shows a trend of youth. Clinically, there is no specific drug for MASLD, and it mainly relies on lifestyle intervention, such as controlling diet and increasing exercise, but it is difficult to adhere to for a long time, and most patients have poor treatment effect. Its pathogenesis is extremely complex, and insulin resistance, oxidative stress and inflammatory response interact with each other, continuously promoting the progression of the disease. If it is not effectively controlled, it may develop into severe liver diseases such as cirrhosis and liver cancer. On March 14, 2024, the U.S. Food and Drug Administration (FDA) approved Resmetirom for marketing, which is used for the treatment of adult patients with metabolic associated steatohepatitis accompanied by liver fibrosis. Resmetirom becomes the first drug approved by FDA for the treatment of metabolic associated steatohepatitis (MASH), but the data shows that only 30% of patients achieve clinical remission. Therefore, from the clinical point of view, there is still a lack of safe and effective drugs for treating MASLD.

[0005] Diabetes is a common metabolic disease. In recent years, the number of diabetic patients has increased dramatically worldwide due to changes in lifestyle and accelerated aging process, and has now exceeded 800 million. Long-term hyperglycemia can cause a series of serious complications, such as diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, etc. These complications seriously reduce the quality of life of patients and even endanger their lives. The existing treatment drugs are mainly aimed at controlling blood glucose levels and delaying the occurrence of complications, but they cannot fundamentally prevent the deterioration of the disease. Therefore, it is urgent to find more effective treatment methods.

[0006] Therefore, there is an urgent need in the clinic for a safe and effective drug that can simultaneously improve metabolic syndrome-related diseases.

[0007] Glycerol guaiacol ether was approved for marketing in Japan in 1949. As a classic expectorant, although the mechanism of action of the drug has not been fully elucidated, existing research suggests that it mainly dilutes airway mucus and reduces its viscosity, while relaxing the adhesion of mucus to the airway wall, so that sputum is more easily coughed out, effectively relieving respiratory obstruction and hyperemia symptoms. It has been widely used in clinical practice, including in children. However, there is still a lack of relevant research on the use of glycerol guaiacol ether in the prevention and treatment of metabolic-related fatty liver disease, hyperlipidemia, hypercholesterolemia, obesity, and diabetes. SUMMARY

[0008] In view of the lack of research on the use of guaiacol or its derivatives in improving metabolic syndrome in the existing disclosed technologies, the present application provides the use of guaiacol or its derivatives in the preparation of a drug for preventing or treating metabolic syndrome to solve the above problems.

[0009] The technical scheme of the present application is as follows:

[0010] The present application provides the use of guaiacol or its derivatives in the preparation of a drug for preventing or treating metabolic syndrome.

[0011] The guaiacol derivative is glycerol guaiacol ether and guaiacol glycerol-β-guaiacyl propyl ether. The structure of guaiacol is shown in formula I, the structure of glycerol guaiacol ether is shown in formula II, and the structure of guaiacol glycerol-β-guaiacyl propyl ether is shown in formula III:

[0012] ; ; .

[0013] Further, metabolic syndrome includes metabolic-related fatty liver disease, hyperlipidemia, hypercholesterolemia, obesity, and diabetes.

[0014] Further, the metabolic-related fatty liver disease includes metabolic-related fatty liver (MASL) and metabolic-related fatty hepatitis (MASH).

[0015] Further, the guaiacol or its derivative is the only active ingredient in the medicine.

[0016] Further, the effective concentration of the guaiacol or its derivative is greater than or equal to 100 mg / kg. Here, the effective concentration refers to the minimum concentration that can achieve a therapeutic effect in a mouse experiment.

[0017] Further, the guaiacol or its derivative also includes a pharmaceutically acceptable salt.

[0018] Further, the medicine for preventing or treating metabolic syndrome includes a pharmaceutically acceptable excipient.

[0019] Further, the acceptable excipient is selected from one or more of a diluent, a disintegrant, a precipitation inhibitor, a glidant, a binder, a dispersant, a suspending agent, an isotonic agent, a thickening agent, an emulsifying agent, a preservative, a stabilizer, a hydrating agent, an ion exchange agent, a flavoring agent, or an antioxidant.

[0020] The beneficial effects of the present application are:

[0021] The guaiacol and its derivative provided by the present application significantly improve the insulin resistance condition by regulating the fat metabolism pathway, while reducing the inflammatory response and oxidative stress damage, and achieve the prevention and treatment of metabolic-related fatty liver disease (MASLD), hyperlipidemia, hypercholesterolemia, obesity, and diabetes from multiple dimensions. BRIEF DESCRIPTION OF DRAWINGS

[0022] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments or prior art description will be briefly introduced below. Obviously, for those skilled in the art, other drawings can also be obtained without creative labor based on these drawings.

[0023] Figure 1 is a graph of the guaiacol or its derivative toxicity test results on HepG2 cells. Among them, A is the effect of different concentrations of guaiacol on the activity of HepG2 cells; B is the effect of different concentrations of guaiacol glycerol ether on the activity of HepG2 cells; C is the effect of different concentrations of guaiacol glycerol-β-guaiacol propyl ether on the activity of HepG2 cells.

[0024] Figure 2Figure 6 is a graph showing the change in the level of triglyceride in HepG2 cells after intervention of guaiacol or its derivatives. Among them, A is the normal control group; B is the fatty acid model group (oleic acid-palmitic acid); C is the guaiacol intervention group; D is the guaiacol glycerol ether intervention group; E is the guaiacol glycerol ether intervention group.

[0025] Figure 3 Figure 7 is a graph showing the lipid deposition in HepG2 cells after intervention of guaiacol or its derivatives. Among them, A is the normal control group; B is the fatty acid model group (oleic acid-palmitic acid); C is the guaiacol intervention group; D is the guaiacol glycerol ether intervention group; E is the guaiacol glycerol ether intervention group.

[0026] Figure 4 Figure 8 is the effect of guaiacol glycerol ether on the expression of lipid metabolism related protein molecules. Among them, A is the immunoblotting experiment showing the effect of guaiacol glycerol ether on the expression of ACC, FASN, SCD1, P-AMPK and AMPK proteins; B is the gray scale analysis result of ACC, FASN, SCD1, P-AMPK and AMPK protein expression, HSP90 protein as control.

[0027] Figure 5 Figure 9 is a graph showing the experimental results of guaiacol glycerol ether improving insulin resistance and reducing blood sugar in obese mice. Among them, A is the fasting blood glucose level of mice in each experimental group; B is the glucose tolerance test (GTT) curve of mice in each experimental group; C is the area under the curve of mice in each experimental group; D is the insulin tolerance test (ITT) curve of mice in each experimental group; E is the area under the curve of mice in each experimental group.

[0028] Figure 6 Figure 10 is a graph showing the experimental results of guaiacol glycerol ether improving obesity in mice. Among them, A is the body weight change of mice in each experimental group; B is the morphological photograph of mice in each experimental group; C is the ratio of fat weight / body weight of mice in each experimental group; D is the ratio of lean body weight / body weight of mice in each experimental group; E is the morphology of brown adipose tissue (BAT), epididymal white adipose tissue (eWAT) and inguinal white adipose tissue (iWAT) of mice in each experimental group.

[0029] Figure 7 Figure 11 is a graph showing the experimental results of guaiacol glycerol ether improving metabolic associated steatohepatitis (MASLD). Among them, A is the liver morphology of mice in each experimental group; B is the liver weight of mice in each experimental group; C is the liver index of mice in each experimental group; D is the liver TG content of mice in each experimental group; E is the liver TC content of mice in each experimental group; F is the serum ALT of mice in each experimental group; G is the serum AST of mice in each experimental group; H is the AST / ALT ratio of mice in each experimental group.

[0030] Figure 8 Figure 7 is a graph showing the effect of guaiacol glycerol ether on the liver pathology of MASLD mice. A is the HE staining of the liver of mice in each experimental group; B is the oil red O staining of the liver of mice in each experimental group.

[0031] Figure 9 Figure 8 is an experimental result of guaiacol glycerol ether improving MASH in mice Figure 1 . A is the liver morphology of mice in each experimental group; B is the body weight of mice in each experimental group; C is the liver weight of mice in each experimental group; D is the liver index of mice in each experimental group.

[0032] Figure 10 Figure 9 is an experimental result of guaiacol glycerol ether improving MASH in mice Figure 2 . A is the serum ALP of mice in each experimental group; B is the serum ALT of mice in each experimental group; C is the serum AST of mice in each experimental group.

[0033] Figure 11 Figure 10 is a graph showing the liver injury and fibrosis pathology of MASLD mice improved by guaiacol glycerol ether. A is the HE staining of the liver of mice in each experimental group; B is the Masson staining of the liver of mice in each experimental group. DETAILED DESCRIPTION

[0034] In order to enable those skilled in the art to better understand the technical solutions in the present application, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor should fall within the scope of protection of the present application.

[0035] Embodiment 1

[0036] Guaiacol and its derivatives alleviate fatty acid-induced lipid deposition and triglyceride content in HepG2 cells

[0037] 1. Materials and methods:

[0038] (1) Human HepG2 cells were used for in vitro experiments, and the culture medium was DMEM low-glucose medium containing NEEA (non-essential amino acids), 10% fetal bovine serum, penicillin 100 units / ml and streptomycin 0.1 mg / ml, and was cultured in a 37°C incubator.

[0039] (2) Compound stock solution preparation: Guaifenesin (CAS: 93-14-1), Guaiacol (CAS: 90-05-1) and Guaiacylglycerol-β-guaiacylpropanediol (CAS: 7382-59-4) were purchased from MCE China (website: https: / / www.medchemexpress.cn). The above three compounds were dissolved in DMSO to prepare a stock solution with a concentration of 40 mM. Then the DMSO solutions of the above three compounds were filtered through a 0.22 μm sterile filter membrane to obtain the stock solution for use.

[0040] (3) Compound toxicity determination: HepG2 cells in the logarithmic growth phase were inoculated in a 96-well plate (5000 cells / well). After overnight culture, different final concentrations of Guaifenesin, Guaiacol and Guaiacylglycerol-β-guaiacylpropanediol were added and cultured for 24 hours. The final concentrations of the above three compounds were 0 μM, 10 μM, 20 μM, 40 μM, 80 μM, 160 μM and 320 μM, respectively. The preparation method was to dilute the stock solution to the predetermined concentration. Then CCK8 working solution was added according to the kit instructions to determine the toxicity of the compounds.

[0041] (4) Cell lipid deposition determination: HepG2 cells in the logarithmic growth phase were inoculated in a 12-well plate containing cell slides. After overnight culture, OA-PA (oleic acid-palmitic acid: 250 μM - 125 μM, referred to as FFA) was added to a final concentration and cultured for 4 hours. Then Guaifenesin, Guaiacol and Guaiacylglycerol-β-guaiacylpropanediol were added to a final concentration of 80 μM and cultured for 24 hours. The following operations were performed in turn: after cell treatment, the cells were first washed slowly with pre-cooled PBS buffer for 2-3 times. Discard the PBS buffer, add 4 % paraformaldehyde, and incubate on ice for 15 minutes. Then wash slowly with PBS buffer for 3 times, each for 3 minutes. Take out the cell slides and place them in a clean 6-well plate. Add 60 % isopropanol aqueous solution, and soak for 5 minutes. After aspirating the 60 % isopropanol aqueous solution, add oil red staining solution, and avoid light for 20 minutes. Aspirate the oil red staining solution, add 60 % isopropanol aqueous solution, and differentiate for 3-5 seconds. Discard the 60 % isopropanol aqueous solution, and wash slowly with PBS buffer for 3 times. Stain with hematoxylin for about 2 seconds (the staining time should not be too long to avoid making the color of hematoxylin deeper than that of oil red). Wash with PBS buffer, then differentiate with 1 % hydrochloric acid alcohol solution for 1-3 seconds. Then wash with water to return blue. Finally, use glycerol gelatin for mounting, and take pictures under a microscope for observation.

[0042] (5) Triglyceride (TG) assay in cells: HepG2 cells in the logarithmic growth phase were inoculated in a 6-well plate, cultured overnight, and then OA-PA (250 μM-125 μM, referred to as FFA) was added at a final concentration to continue culturing for 4 hours. Guaifenesin, guaiacol, and guaiacol glycerol-β-guaiacol propanediol were added at a final concentration of 80 μM to continue culturing for 24 hours. The following operations were performed: the triglyceride content in the cell sample was determined using a tissue cell triglyceride lipase assay kit produced by Beijing Puli Lei Company. The cells were washed twice with pre-cooled PBS buffer, and then a lysis solution (200 μL of cell lysis solution provided by the kit was added to each well of the 6-well plate) was added, and the plate was placed at room temperature for 10 minutes. The cells in the culture plate were scraped off with a cell-specific scraper, and the entire lysis solution was transferred to a new 1.5 ml EP tube. An appropriate amount of lysis solution was taken from the EP tube and placed on ice, and the protein concentration of the sample was determined using a BCA protein quantification kit. The remaining lysis solution was heated in a 70 °C metal bath for 10 minutes, and then placed in a centrifuge at room temperature at 2000 rpm for 5 minutes. The upper clear liquid can be used for TG determination, and the operation is performed according to the instructions.

[0043] (6) Guaifenesin determination of lipid metabolism-related protein molecular expression: HepG2 cells in the logarithmic growth phase were inoculated in a 6-well plate, cultured overnight, and then guaiacol glycerol ether (100 μM) was added to continue culturing for 24 hours. The cells were washed twice with pre-cooled PBS buffer, and then 200 μL of RIPA cell lysis solution (Biyun Tian, P0013B) was added and placed on ice for 10 minutes. The cells in the culture plate were scraped off with a cell-specific scraper, and the entire lysis solution was transferred to a new 1.5 ml EP tube, which was centrifuged at 12000 rpm / minute for 10 minutes. The supernatant was taken, and SDS-PAGE protein loading buffer (Bi Yun Tian, P0015) was added, and the mixture was heated at 100 °C for 10 minutes. Then Western Blot operation was performed, and gel imaging instrument exposure development and gray scale analysis were performed.

[0044] 2. Test results

[0045] The cytotoxicity of guaiacol and its derivatives was evaluated by in vitro experiments, and the results showed that the compound concentration within 320 μM had no significant effect on the activity of HepG2 cells, such as Figure 1TG levels and lipid droplet accumulation in HepG2 cells, as shown in FIGS. 1A-1C. Figure 2 and Figure 3 as shown in FIGS. 2A-2C.

[0046] Western Blot results showed that guaifenesin significantly down-regulated the expression of ACC, FASN and SCD1 proteins, and had no significant effect on P-AMPK expression, as shown in FIG. 3. Figure 4

[0047] Example 2

[0048] Guaifenesin alleviates high-fat diet-induced insulin resistance, obesity and MASLD in mice

[0049] 1. Materials and methods

[0050] Forty 7-week-old male C57BL / 6J mice were first adapted to the diet for 1 week, and then randomly divided into 4 groups: normal diet control group (CD), high-fat diet group (HFD), high-fat diet-guaifenesin low-dose group (Guaifenesin-L, referred to as Gua-L) and high-fat diet-guaifenesin high-dose group (Guaifenesin-H, referred to as Gua-H). The mice in the normal diet control group were fed with normal feed, and the mice in the other groups were fed with 60% high-fat diet (Research Diet, D12492). After 9 weeks of feeding, the mice in the Gua-L group were given 100 mg / kg guaifenesin by gavage daily, and the mice in the Gua-H group were given 300 mg / kg guaifenesin by gavage daily. Guaifenesin was dissolved in normal saline, and the mice in the other groups were given normal saline by gavage at the same time, and the drug intervention was continued for 8 weeks.

[0051] After 5 weeks of drug intervention, glucose tolerance test (GTT) was performed: the mice were fasted for 16 hours without water, the mice were weighed and recorded, the required injection dose of 20% glucose for the mice was calculated according to 1 g glucose / kg body weight, the blood was taken from the tail tip of the mice, the first drop of blood was wiped off, the blood glucose at 0 minutes was measured with a blood glucose meter, the mice were adapted for 10 minutes, and then the calculated dose of glucose was injected intraperitoneally, respectively, and the blood glucose was detected at 30 minutes, 60 minutes, 90 minutes and 120 minutes after intraperitoneal injection of glucose, respectively, and the area under the curve was analyzed by GraphPad Prism.

[0052] ​After 6 weeks of drug intervention, the mouse insulin tolerance test (ITT) was performed: all mice were fasted for 4 hours in advance, the fasting blood glucose level of the mice was measured and recorded (0 minutes), then insulin was injected at a ratio of 0.75 units per kilogram of body weight, and the blood glucose levels of the mice at 15 minutes, 30 minutes, 60 minutes, 90 minutes and 120 minutes after injection of insulin were recorded respectively, and the area under the curve was analyzed by GraphPad Prism.

[0053] After 8 weeks of drug intervention, the mice were fasted for 12 hours without water, and the next morning after weighing, the mice were placed in a fixed tube to restrict their activity, and the Fat mass (fat weight) and Lean mass (lean body weight) of the mice were analyzed and recorded using the Bruker Minispec LF90II system. Then anesthetized with avertin intraperitoneal injection, blood was taken from the retro-orbital plexus, the mouse was fixed, the abdominal cavity was opened, the liver was quickly isolated and removed, washed with physiological saline at 4 ℃, the film was removed, the wet weight of the liver was weighed, and the liver index was calculated. At a distance of 1 cm from the edge of the right lobe of the liver, two pieces of liver tissue of appropriate size were separated: one piece of liver tissue was transversely cut, fixed with 4% paraformaldehyde, routinely dehydrated, paraffin-embedded, sectioned, and HE stained; one piece of liver tissue was frozen sectioned for oil red O staining; the rest of the liver tissue was stored at -80 ℃. At the same time, the inguinal white adipose tissue (iWAT), epididymal white adipose tissue (eWAT), and brown adipose tissue (BAT, between the shoulder blades) were quickly isolated and removed, a portion was fixed with 4% paraformaldehyde and HE stained, and the rest was stored at -80 ℃. After standing at 4 ℃ for 2 hours, the blood sample was centrifuged at 3000 rpm for 10 minutes, the serum was separated, aliquoted into EP tubes, and a portion of the sample was immediately subjected to serum index detection, and the rest was stored at -80 ℃ for detection of other blood biochemical indicators.

[0054] 2. Test results:

[0055] The results of the mouse fasting blood glucose test showed that the blood glucose level of the HFD group mice was significantly higher than that of the CD group mice, and the fasting blood glucose level of the Gua-H and Gua-L group mice was significantly lower than that of the HFD group mice, as shown in Figure 5 A. The results of the GTT experiment showed that compared with the CD group mice, the glucose tolerance of the HFD group mice was significantly impaired, and compared with the HFD group mice, the glucose tolerance of the Gua-H and Gua-L group mice was significantly improved, as shown in Figure 5 B and Figure 5 C. The results of the ITT experiment showed that compared with the CD group mice, the insulin sensitivity of the HFD group mice was significantly reduced, and after intervention with guaiacol glycerol ether, the insulin sensitivity of the Gua-H and Gua-L group mice was significantly improved, as shown in Figure 5 D and Figure 5Gua-H and Gua-L groups, as shown in Fig. E. Therefore, guaiacol glyceryl ether has the pharmacological effect of improving the glucose tolerance and insulin sensitivity of mice and reducing the blood glucose of mice.

[0056] The body weight of mice was recorded every week. The body weight of mice in the HFD group showed a continuous increasing trend, and the body weight of mice in the Gua-H and Gua-L groups was significantly lower than that in the HFD group, as shown in Fig. Figure 6 A. The mice in the HFD group were larger than the mice in the CD group, and the mice in the Gua-H and Gua-L groups were smaller than the mice in the HFD group, as shown in Fig. Figure 6 B. Animal composition analysis showed that, compared with the CD mice, the fat mass / body weight ratio of the HFD mice was significantly increased, and the lean mass / body weight ratio was significantly decreased. After the intervention of guaiacol glyceryl ether, the fat mass / body weight and lean mass / body weight of mice were obviously improved, as shown in Fig. Figure 6 C and Figure 6 D. The pictures of epididymal white adipose tissue (eWAT) and inguinal white adipose tissue (iWAT) of mice showed that the volume of eWAT and iWAT of mice in the HFD group was significantly larger than that of mice in the CD group, and the volume of eWAT and iWAT of mice in the Gua-H and Gua-L groups was significantly reduced, as shown in Fig. Figure 6 E. Therefore, guaiacol glyceryl ether has the pharmacological effect of reducing the obesity of mice.

[0057] The comparison of liver pictures of mice in each group found that the liver of mice in the HFD group was larger in volume and the color was yellow than that of mice in the CD group; the liver of mice in the Gua-H and Gua-L groups was smaller in volume and the color was light yellow than that of mice in the HFD group, as shown in Fig. Figure 7 A. The statistical results of liver weight of mice in each group showed that, compared with the CD group, the liver weight of mice in the HFD group was significantly increased, and the liver weight and the ratio of liver weight to body weight of mice in the Gua-H and Gua-L groups were significantly reduced, as shown in Fig. Figure 7 B and Figure 7 C. At the same time, the TG and TC contents in the liver of mice in each group were determined. Compared with the CD mice, the TG and TC contents in the liver of mice in the HFD group were significantly increased, and the TG and TC contents in the liver of mice in the Gua-H and Gua-L groups were significantly reduced after the intervention of guaiacol glyceryl ether, as shown in Fig. Figure 7 D and Figure 7 E. Through the detection of representative indicators of liver function, it was found that the serum ALT level of mice in the HFD group was significantly increased, and the AST / ALT ratio was significantly reduced. After the intervention of guaiacol glyceryl ether, the ALT level and AST / ALT ratio of mice on high-fat diet were significantly improved, as shown in Fig. Figure 7 F,Figure 7 G and Figure 7 H as shown.

[0058] To clarify the effect of guaiacol glyceryl ether on the pathological changes of mouse liver, HE staining results showed that a large number of fat vacuoles appeared in the liver of HFD group mice, and guaiacol glyceryl ether administration could significantly reduce the liver fat vacuoles of mice, as shown in Figure 8 A. Oil red O staining results showed that the liver cell lipid deposition of HFD group mice increased significantly, and guaiacol glyceryl ether intervention reduced the liver lipid deposition of mice, as shown in Figure 8 B. The above results showed that guaiacol glyceryl ether had the pharmacological effect of reducing liver damage and reducing liver lipid deposition, and improving MASLD.

[0059] Example 3

[0060] Guaiacol glyceryl ether relieves choline-methionine deficiency (MCD) diet-induced MASH in mice

[0061] 1. Materials and methods:

[0062] 32, 7-week-old male C57BL / 6J mice were first adapted for 1 week, and the mice were randomly divided into 4 groups: normal diet control group (CD), choline-methionine deficiency diet group (MCD), MCD diet-guaiacol glyceryl ether high dose group (Gua-H) and MCD diet-guaiacol glyceryl ether low dose group (Gua-L). The CD group of mice was fed with ordinary feed, and the other groups of mice were fed with MCD feed (Synergy Biologics, XTMCD). At the same time, the Gua-H group of mice was given 300 mg / kg guaiacol glyceryl ether by gavage every day, and the Gua-L group of mice was given 100 mg / kg guaiacol glyceryl ether by gavage every day, and guaiacol glyceryl ether was dissolved with normal saline, and the other groups of mice were given blank solvent by gavage at the same time, and the drug intervention was continuously given for 3 weeks.

[0063] After 3 weeks of drug intervention, the mice were fasted for 12 hours without water, and the next morning after weighing, the mice were anesthetized by intraperitoneal injection of avertin, weighed, and blood was collected from the retro-orbital plexus, the mouse was fixed, the abdominal cavity was opened, the liver was quickly separated and removed, washed with 4 ℃ normal saline, the film was removed, and the liver wet weight was weighed, and the liver index was calculated. At the edge of the right lobe of the liver 1 cm, a piece of liver tissue was transversely cut, 4% paraformaldehyde was fixed, conventional dehydration, paraffin embedding, sectioning, HE and Masson staining, and the rest of the liver tissue was stored at-80 ℃. After the blood sample was placed at 4 ℃ for 2 hours, it was centrifuged at 3000 rpm for 10 minutes, the serum was separated, and the EP tube was divided, part of the sample was immediately subjected to blood index detection, and the rest was stored at-80 ℃ for detection of other blood biochemical indexes.

[0064] 2. Test results:

[0065] The present study found that the liver of the MCD group mice was smaller in volume and darker in color than that of the CD group mice. After guaiacol glycerol ether intervention, the liver of the Gua-H group mice was larger in volume than that of the MCD group, as shown in Figure 9 A. Compared with the CD group, the body weight of the MCD group mice decreased. After guaiacol glycerol ether intervention, no significant change in body weight was observed in the mice, as shown in Figure 9 B. Compared with the CD group, the liver weight and the ratio of liver weight to body weight of the MCD group mice significantly decreased, while the liver weight and the ratio of liver weight to body weight of the Gua-H group mice significantly increased, as shown in Figure 9 C and Figure 9 D.

[0066] Through detection of representative indicators of liver function, it was found that the serum ALP, ALT and AST levels of the MCD group mice significantly increased, while after high-dose guaiacol glycerol ether intervention, the ALP, ALT and AST levels of the mice significantly decreased, as shown in Figure 10 A, Figure 10 B and Figure 10 C.

[0067] In order to clarify the effect of guaiacol glycerol ether on the pathological changes of the liver of mice, the HE staining results showed that a large number of fat vacuoles appeared in the liver of the MCD group mice and the morphology of hepatocytes changed, and after guaiacol glycerol ether administration, the fat vacuoles in the liver of the mice and the morphological changes of hepatocytes were significantly reduced, as shown in Figure 11 A. The Masson staining results showed that the liver fibrosis of the MCD group mice significantly increased, and guaiacol glycerol ether intervention reduced the degree of liver fibrosis in the mice, as shown in Figure 11 B. The above results show that guaiacol glycerol ether has the pharmacological effects of reducing liver damage and reducing the degree of liver fibrosis, and plays a role in improving and treating MASH.

[0068] Although the present application has been described in detail with reference to the preferred embodiments thereof, it is apparent that the present application is not limited to the foregoing embodiments. Various modifications or replacements can be made to the embodiments of the present application by those skilled in the art without departing from the spirit and essence of the present application, and these modifications or replacements should be within the scope of the present application. Any person skilled in the art within the technical scope disclosed by the present application can easily think of changes or replacements, and these changes or replacements should be within the protection scope of the present application.

Claims

1. Use of guaiacol glyceryl ether or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating metabolic syndrome, characterized in that, The metabolic syndrome is metabolic-related fatty liver disease, hyperlipidemia, hypercholesteremia, obesity and diabetes; the guaiacol glyceryl ether is the only active ingredient in the medicine.

2. Use according to claim 1, wherein The metabolic-related fatty liver disease includes metabolic-related fatty liver and metabolic-related fatty hepatitis.

3. The use according to claim 1, wherein The medicine for treating metabolic syndrome includes pharmaceutically acceptable adjuvants.

4. Use according to claim 3, wherein the compound is ###0002### The acceptable adjuvant is selected from one or more of diluents, disintegrants, precipitation inhibitors, glidants, binders, dispersants, suspending agents, isotonic agents, thickening agents, emulsifiers, preservatives, hydrating agents, ion exchange agents, flavoring agents or antioxidants. The acceptable adjuvant is selected from one or more of diluents, disintegrants, precipitation inhibitors, glidants, binders, dispersants, suspending agents, isotonic agents, thickening agents, emulsifiers, preservatives, hydrating agents, ion exchange agents, flavoring agents or antioxidants.

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