Gentianone or its phosphate derivatives for the treatment and / or prevention of lipid-related metabolic diseases.

Abstract

This invention discloses a second use of genistein or its phosphate derivative for the treatment and / or improvement of metabolic diseases. Specifically, by administering an effective amount of genistein or its phosphate derivative to an individual, it is possible to inhibit fat accumulation in the individual's body or visceral areas, thereby achieving the therapeutic and / or preventative effects on diseases related to abnormal fat metabolism.

Description

Technical Field

[0001] This invention relates to a second use of a compound, particularly the use of genistein or its phosphate derivative for the treatment and / or prevention of lipid-related metabolic diseases. Prior Technology

[0002] Note that the World Health Organization defines obesity as a chronic disease, increasing the risk of diabetes, metabolic syndrome, high cholesterol, high blood pressure, cardiovascular disease, and joint problems. The primary cause of obesity is consuming more calories than you burn, leading to the accumulation of calories as fat in the body. Factors influencing individual obesity, besides diet and exercise habits, are also related to work stress and genetics.

[0003] As modern people increasingly value their health and physique, they try to avoid obesity by controlling their diet and increasing exercise. However, many still rely heavily on eating out and find it difficult to maintain a consistent exercise routine, making it difficult to effectively curb fat accumulation. Another method is fasting for rapid weight loss, but this method has drawbacks such as decreased metabolic rate, muscle loss, and nutritional deficiencies. Alternatively, weight loss products can be used to improve obesity symptoms, but many on the market contain illegal ingredients, leading to adverse reactions and even irreversible health damage. Summary of the Invention

[0004] The main objective of this invention is to provide the use of genistein or its phosphate derivative for the treatment and / or prevention of fat-related metabolic diseases. It is possible to reduce or improve the accumulation of fat in an individual by administering a low dose of genistein or its phosphate derivative or a composition containing the above ingredients to the individual.

[0005] Therefore, in order to achieve the above-mentioned objectives, this invention discloses the use of genistein or its phosphate derivative for the preparation of a water-soluble epoxide hydrolase (sEH) inhibitor. That is, by administering an effective amount of genistein or its phosphate derivative, or a composition containing it, to an individual, the genistein (GEN) can bind to the water-soluble epoxide hydrolase and inhibit the activity of the water-soluble epoxide hydrolase in the individual, thereby achieving the effect of improving metabolic-related diseases.

[0006] Furthermore, since genistein-7-O-phosphate (G7P) has a better gastrointestinal absorption rate than genistein, and since genistein-7-O-phosphate is metabolized and reduced to genistein in the small intestine epithelial cells after entering the small intestine, the effective dose of genistein-7-O-phosphate for an individual is significantly lower than that of genistein.

[0007] This metabolic-related disease is related to lipid metabolism.

[0008] The half-maximal inhibitory concentration (IC50) of this genistein against water-soluble epoxide hydrolase is 10⁻¹² nM.

[0009] In another embodiment of the present invention, the use of genistein or its phosphate derivatives in the preparation of compositions for the treatment and / or prevention of diseases related to lipid metabolism disorders is disclosed; that is, by administering an effective amount of a genistein phosphate derivative or a composition containing it to an individual, the genistein phosphate derivative can reduce the fat content in the individual's body and tissues, or prevent the continuous accumulation of fat in the body or tissues, thereby achieving the effect of preventing and / or treating diseases related to lipid metabolism disorders.

[0010] Among them, diseases related to abnormal lipid metabolism include fatty liver, obesity, hyperlipidemia or related diseases, high body fat or related diseases, and inflammation caused by fatty liver.

[0011] The composition may be a pharmaceutical, a food, or a nutritional supplement. Simple Explanation of the Diagram

[0012] Figure 1 shows the results of detecting the changes in body weight of mice in each group during the experiment. Figure 2 shows the morphology of the liver, spleen, and kidneys of mice in each group. Figure 3 shows the results of analyzing the weight of the liver, spleen, and kidneys of mice in each group. Figure 4 shows the morphology of abdominal fat, intestinal fold fat, and perirenal fat in mice of each group. Figure 5 shows the results of analyzing abdominal fat, intestinal membrane fat, perirenal fat, and total fat weight in mice of each group. Figure 6 shows the results of H&E staining of liver tissue sections from mice in each group. Figure 7 shows the results of H&E staining of perirenal adipose tissue sections from each group of mice. Figure 8 shows the results of detecting and analyzing serum triglycerides, cholesterol, high-density lipoprotein, and low-density lipoprotein in the serum of mice in each group. Figure 9 shows the results of detecting and analyzing the triglyceride and total cholesterol content in the livers of mice in each group. Figure 10 shows the results of detecting and analyzing the levels of IL-6, TSLP, and TNF-α in the livers of mice in each group. Figure 11 shows the results of the analysis of the docking between the control drug Compound33 and the sEH protein molecule. Figure 12 shows the results of the analysis of the docking between genistein and sEH protein molecules. Implementation

[0013] This invention discloses a second use of genistein or its phosphate derivative for treating and / or improving metabolic diseases. Specifically, the genistein disclosed in this invention can bind to water-soluble epoxide hydrolase in vivo, thereby inhibiting its activity and reducing body fat content, visceral fat content, liver fat size and quantity, blood triglycerides, cholesterol, low-density lipoprotein cholesterol, and inflammatory hormone expression. Furthermore, the genistein phosphate derivative can be highly absorbed by the digestive tract and metabolized into genistein after entering the body. Therefore, the genistein or its phosphate derivative disclosed in this invention can serve as a water-soluble epoxide hydrolase inhibitor or a composition for treating and / or preventing diseases related to abnormal lipid metabolism. In other words, by administering an effective amount of genistein or its phosphate derivative to an individual, it is possible to inhibit fat accumulation in the individual's body or visceral areas, thereby achieving the effect of treating and / or preventing diseases related to abnormal lipid metabolism.

[0014] Furthermore, the absorption rate of the genistein phosphate derivative disclosed in this invention in the digestive tract is approximately 80% higher than that of genistein. Therefore, the genistein phosphate derivative can exert therapeutic and / or preventive effects on diseases related to lipid metabolism disorders in vivo at lower doses. Compared to genistein, the genistein phosphate derivative has better absorption and bioavailability, thus significantly improving its industrial applicability and economic value.

[0015] This system includes humans, dogs, cats, pigs, or other animals.

[0016] Among them, diseases related to abnormal lipid metabolism include obesity, fatty liver, hepatitis caused by fatty liver, hyperlipidemia, hypercholesterolemia, or complications of at least one of the above conditions.

[0017] The term "genistein-7-O-phosphate (G7P)" refers to a compound derived from genistein, whose structure is shown in formula (I).

[0018] (I)

[0019] The term "genistein (GEN)" refers to one of the most abundant isoflavones in soybeans, and its structure is shown in formula (II). Because the structure of this compound is similar to that of estrogen, it is often used as a phytoestrogen. Studies have shown that genistein has functions in preventing osteoporosis, cardiovascular disease, breast cancer, and prostate cancer.

[0020] (II)

[0021] The term "water-soluble epoxide hydrolase (sEH)" refers to an important dual-terminal functional enzyme widely distributed in organs such as the liver, kidneys, and brain. Studies have shown that inhibiting sEH can maintain the expression of endogenous epoxydienetrienoic acids (EETs) and reduce the expression of dihydroxydienetrienoic acids (DHETs), which may help treat or improve cardiovascular, central nervous system, or metabolic diseases.

[0022] The term "Compound 33", also known as sEH inhibitor-14, has the following structural formula (III).

[0023] (III)

[0024] The term "disorders of lipid metabolism" refers to diseases caused by excessive accumulation of fat in the body or excessive fat intake. Generally, patients typically have body fat and / or blood lipid levels higher than the clinically normal range. Disorders of lipid metabolism include hyperlipidemia, hypertension, cardiovascular disease, obesity, metabolic syndrome, diabetes, fatty liver, and hepatitis.

[0025] The term "dosage" or any other numerical value used in this invention shall be interpreted to include the numerical value itself and the acceptable error range in the technical field. The acceptable error range in the technical field to which this invention pertains is the value ± 10%. For example, if the dosage is 0.9 nM, the interpretation should cover the range of 0.9 ± 0.09 nM.

[0026] To illustrate the technical features and effects of this invention, several examples will be provided below, along with accompanying drawings, for detailed explanation.

[0027] The animal experiments in the following examples all comply with relevant ethical guidelines.

[0028] The animal experiments in the following examples use mice as an example, so the dosage used is a dosage that can be used by mice. This dosage will change depending on the subject or method of administration, which can be calculated by those skilled in the art based on their common knowledge.

[0029] Example 1: Animal Experiments

[0030] Multiple male C57BL / 6 mice were divided into 4 groups and housed under the following different conditions:

[0031] Group 1: Feed and water as usual;

[0032] Group 2: Feed 60% high-fat diet and 30% fructose in drinking water, and provide sterile distilled water daily via tube feeding (10 mL / kg / day);

[0033] Group 3: Feeded with 60% high-fat diet and 30% fructose in drinking water, and given genistein isoflavone phosphate derivative (0.1 mg / kg / day) via tube feeding daily;

[0034] Group 4: Feed 60% high-fat diet and 30% fructose in drinking water, and administer genistein (1.0 mg / kg / day) daily via tube.

[0035] The mice were housed for 19 weeks, during which their weight was measured weekly, as shown in Figure 1. At the end of the housing period, the mice were sacrificed, and their liver, kidneys, spleen, abdominal fat, intestinal fat, and perirenal fat were photographed, as shown in Figures 2 and 4. The weight of the organs, the weight of adipose tissue in each location, and the total fat mass were also measured, as shown in Figures 3 and 5. After completion, the liver and adipose tissue were paraffin-embedded for subsequent histological staining analysis.

[0036] As shown in Figures 1 to 5, compared to Group 1 mice, Group 2 mice exhibited a significant increase in body weight, and the weight and / or size of the liver, spleen, kidneys, and visceral fat were also significantly increased, all of which increased with the duration of rearing. This indicates that a high-fat diet and high-sugar water intake do indeed lead to fat accumulation in organs and tissues, resulting in obesity. Furthermore, compared to Group 2 mice, Groups 3 and 4 mice showed a decrease in body weight, and the size of fat in various organs and body parts was significantly reduced.

[0037] The above results show that a high-fat or / and high-sugar diet can indeed lead to obesity. However, if genistein phosphate derivatives or genistein are administered simultaneously, the increase in individual weight and the accumulation of body fat can be inhibited or slowed down. This means that the genistein and its phosphate derivatives disclosed in this invention can both achieve the effect of improving fat-related metabolic diseases. Furthermore, the genistein phosphate derivative only requires 0.1 times the dose of genistein to achieve the effect of improving fat accumulation, indicating that the genistein phosphate derivatives disclosed in this invention have significantly better weight loss or fat accumulation inhibition effects than genistein.

[0038] Example 2: Staining of tissue sections

[0039] The liver and adipose tissue sections from Example 1 were stained with hematoxylin and eosin (H&E stain), and the results are shown in Figures 6 and 7.

[0040] As can be seen from the slice results of groups 1 and 2 in Figures 6 and 7, a high-fat, high-fructose diet will cause fat to accumulate in the liver, resulting in fatty liver, and will also cause fat cells in adipose tissue to enlarge, resulting in a significant increase in body fat.

[0041] Furthermore, as shown in Figures 6 and 7, compared to the second group of mice, the livers of the third and fourth groups of mice showed no significant accumulation of fat, and the size of the fat cells was significantly reduced.

[0042] The above results show that administration of genistein or genistein phosphate derivatives can effectively improve the accumulation of fat in the body, thereby preventing or improving fatty liver, obesity, or lipid metabolism-related diseases. Furthermore, genistein phosphate derivatives can achieve the same efficacy as genistein at lower doses. In other words, the genistein or its phosphate derivatives disclosed in this invention can improve or prevent fat accumulation or lipid metabolism-related diseases.

[0043] Example 3: Blood Biochemistry Test

[0044] Whole blood was collected from each group of mice in Example 1 and centrifuged (10,000 xg, 10 minutes). Serum was collected and serum levels of triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were measured using a fully automated serum biochemistry analyzer (CoBAS MIRA PLUS, Basel, Switzerland). The results are shown in Figure 8.

[0045] As shown in Figure 8, compared with the first group of mice, the levels of total cholesterol and low-density lipoprotein cholesterol in the blood of the second group of mice were significantly increased, indicating that a high-fat, high-fructose diet can lead to the risk of hyperlipidemia, hypercholesterolemia or related diseases. Compared with the second group of mice, the levels of total cholesterol and low-density lipoprotein cholesterol in the blood of the third and fourth groups of mice were significantly decreased.

[0046] The above results show that genistein or genistein phosphate derivatives can inhibit or improve hyperlipidemia and hypercholesterolemia caused by high-fat and / or high-sugar diets, thereby achieving the effects of treating or preventing hyperlipidemia or related diseases, as well as hypercholesterolemia or related diseases. Furthermore, due to the better bioavailability of genistein phosphate derivatives, genistein phosphate derivatives can achieve the same effects as genistein at lower doses.

[0047] Example 4: Detection of triglycerides and cholesterol in the liver

[0048] The largest lobe of the liver of each group of mice in Example 1 was homogenized and centrifuged (10,000 xg, 10 minutes). The contents of triglycerides and total cholesterol in the liver were then detected using a fully automated serum biochemistry analyzer (CoBAS MIRA PLUS, Basel, Switzerland). The results are shown in Figure 9.

[0049] As shown in Figure 9, compared to Group 1 mice, Group 2 mice had significantly increased levels of triglycerides and total cholesterol in their livers, indicating that a high-fat, high-fructose diet can cause fatty liver or related diseases. Compared to Group 2 mice, Groups 3 and 4 mice had significantly decreased levels of total cholesterol and triglycerides in their livers.

[0050] The above results show that genistein or genistein phosphate derivatives can inhibit or improve fatty liver or related diseases caused by high-fat and / or high-sugar diets, and the genistein phosphate derivatives disclosed in this invention can achieve the effect of improving or preventing fatty liver or related diseases at extremely low doses.

[0051] Example 5: Analysis of liver inflammation-related cytokines

[0052] Liver tissues from each group of mice in Example 1 were taken, and an appropriate amount of phosphate buffer solution was added. The tissues were homogenized and centrifuged (10,000 xg, 10 minutes). The supernatant was collected, and the contents of IL-6, TSLP (Thymic stromal lymphopoietin) and TNF-α in the liver supernatant were analyzed using a commercially available ELISA kit. The results are shown in Figure 10.

[0053] As shown in Figure 10, compared with the first group of mice, the levels of IL-6, TSLP and TNF-α in the liver of the second group of mice were significantly increased, indicating that the liver of the second group of mice was induced by a high-fat and high-sugar diet. Compared with the second group of mice, the levels of inflammatory hormones in the liver of the third and fourth groups of mice were significantly reduced, indicating that the liver of the third and fourth groups of mice did not develop an inflammatory response due to the continuous administration of a high-fat and high-sugar diet.

[0054] The results in Figure 10 confirm that the genistein or its phosphate derivative disclosed in this invention can inhibit or improve liver inflammation or related diseases caused by high-fat and / or high-sugar diets, and the dosage of the genistein phosphate derivative used is 0.1 times that of genistein.

[0055] Example 6: Structural Analysis

[0056] In this example, protein analysis techniques were used to analyze the docking of genistein and compound 33 with the sEH (Soluble epoxide hydrolase) protein molecule. The results are shown in Figures 11 and 12. 3D spatial coordinates and bond energies were also analyzed, and the results are shown in Table 1 below.

[0057] The results in Table 1, Figure 11 and Figure 12 show that genistein can bind to sEH enzyme like commercially available sEH inhibitors (compound 33), indicating that the genistein disclosed in this invention can indeed inhibit sEH activity. Furthermore, since the genistein phosphate derivative is metabolized into genistein in the small intestine, the genistein phosphate derivative disclosed in this invention has the ability to inhibit sEH enzyme activity in vivo.

[0058] Table 1: Docking analysis of compound 33 and genistein with sEH protein molecules Compound 33 Gentian isoflavones 3D spatial coordinates X=15.357481 Y=10.613963 Z=16.643778 X=17.745773 Y=7.967000 Z=10.490318 Bonding energy (kcal / mol) -10.3 -8.7

[0059] Furthermore, analysis of the half-maximal inhibitory concentration (IC50) of genistein for sEH enzyme bonding as disclosed in this invention reveals that the IC50 of genistein is 11.45 nM.

[0060] none

Claims

1. The use of a genistein phosphate derivative in the preparation of a water-soluble epoxide hydrolase (sEH) inhibitor, wherein, The genistein-7-O-phosphate derivative is named genistein-7-O-phosphate (G7P), and this water-soluble epoxide hydrolase inhibitor is used to treat a lipid metabolism disorder.

2. The use of a genistein phosphate derivative in the preparation of a TSLP (Thymic stromal lymphopoietin) inhibitory composition, wherein, The genistein-7-O-phosphate derivative is genistein-7-O-phosphate (G7P), and the TSLP (Thymic stromal lymphopoietin) inhibitory composition is used to treat hepatic lipid metabolism disorders.

3. The use as described in claim 2, wherein, This liver lipid metabolism disorder is either fatty liver or hepatitis.

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