Application of hippuric acid in activating brown adipose tissue to produce heat and preventing and treating obesity

Abstract

The invention discloses application of hippuric acid in activation of brown adipose tissue heat production and prevention and treatment of obesity, and the application is realized by activating brown adipose tissue heat production, specifically, expression of uncoupling protein 1 in brown adipose tissue is up-regulated, heat production capacity under cold stimulation conditions is enhanced, weight gain and abnormal glucose and lipid metabolism are improved, and the purpose of preventing and treating obesity is achieved. The hippuric acid is preferably derived from walnut oil or metabolites thereof, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and nutritional food technology, and relates to the application of hippuric acid in activating thermogenesis in brown adipose tissue and preventing obesity. Background Technology

[0002] Obesity is a chronic metabolic disease caused by long-term energy intake exceeding energy expenditure. Its global prevalence continues to rise, becoming a major public health problem threatening human health. Obesity is not only a manifestation of abnormal fat accumulation but also a significant risk factor for type 2 diabetes, non-alcoholic fatty liver disease, cardiovascular disease, and various metabolic syndromes. Currently, commonly used clinical anti-obesity strategies mainly include lifestyle interventions, drug therapy, and metabolic surgery. Among these, drug therapy has received considerable attention due to its relative controllability and accessibility, and several drugs have been approved for obesity treatment, such as orlistat (which reduces fat absorption by inhibiting lipase), phentermine / topiramate combination preparations (centrally acting appetite suppressants), and glucagon-like peptide-1 receptor agonists (such as liraglutide and smegglutide). However, existing drugs generally suffer from limited efficacy, significant side effects, inconvenient administration, or high cost. For example, orlistat often causes gastrointestinal adverse reactions; centrally acting drugs may pose cardiovascular and neuropsychiatric risks; GLP-1 inhibitors require injection and are associated with gastrointestinal discomfort, and their long-term safety requires further verification. Therefore, finding novel, safe, effective, and well-defined anti-obesity active substances has become an important direction in the current research and development of drugs for metabolic diseases.

[0003] In recent years, with the deepening of metabolomics and microbiome research, the role of endogenous metabolites in metabolic regulation has received increasing attention. Hippuric acid, a terminal metabolite produced by gut microbiota from the metabolism of dietary polyphenols (such as phenylalanine and chlorogenic acid), has traditionally been considered an excretory substance, but in recent years it has been found to possess various biological activities. For example, studies have suggested that hippuric acid can promote fatty acid oxidation, inhibit fat synthesis, and play a positive role in regulating glucose and lipid metabolism. A recently published Chinese patent application CN121041259A (application number 202511229507.9) further reveals the potential value of hippuric acid in combating obesity and related metabolic diseases. This application indicates that hippuric acid can downregulate the expression of key genes in liver lipogenesis (such as Srebp1, Scd1, and Fasn), improve hepatic steatosis, insulin resistance, and impaired glucose tolerance, thereby reducing weight gain and hepatic lipid accumulation induced by a high-fat diet in mice. This study provides experimental evidence for hippuric acid as an endogenous anti-obesity metabolite and emphasizes its role in regulating hepatic lipid and glucose metabolism.

[0004] However, existing technologies primarily focus on the inhibitory effect of hippuric acid on local lipid synthesis pathways in the liver. Their mechanism of action remains limited to the traditional metabolic regulation of "reducing energy storage," and does not yet involve the activation of energy expenditure pathways. In fact, the regulation of energy expenditure plays an equally important role as energy intake in obesity prevention and treatment. Brown adipose tissue, a key tissue in mammals responsible for non-shivering thermogenesis, is closely related to body weight and metabolic health. BAT uncouples oxidative phosphorylation from ATP synthesis via uncoupling protein 1 (UCP1) on the inner mitochondrial membrane, converting chemical energy into heat energy, thus consuming a large amount of energy. Activating the thermogenesis function of BAT is considered a potential new strategy to combat obesity and related metabolic diseases. Currently, although several compounds or hormones have been reported to activate BAT, most suffer from weak effects, poor specificity, or difficulty in translating them into clinical applications.

[0005] It is noteworthy that although hippuric acid has been shown to have metabolic regulatory effects, whether it can directly activate BAT thermogenesis and exert an anti-obesity effect by enhancing energy expenditure pathways has not been publicly reported to date. Existing technology CN121041259A also does not address the effect of hippuric acid on BAT function, and its mechanism of action is not directly related to the BAT-UCP1 thermogenesis pathway. Therefore, the field still lacks a naturally derived bioactive substance that can intervene in obesity from the perspective of energy expenditure by specifically activating BAT thermogenesis, and especially lacks an anti-obesity strategy that combines dietary metabolites with targeted BAT activation.

[0006] Based on the above-mentioned situation, this invention proposes and confirms for the first time that hippuric acid, a metabolite of walnut oil, can significantly activate the thermogenesis function of brown adipose tissue and effectively prevent and treat obesity through this mechanism. Compared with the prior art, this invention not only expands the scope of the biological functions of hippuric acid, but also realizes a shift in the mechanism of action from "inhibiting energy storage" to "promoting energy consumption," providing a new target and pathway for the development of anti-obesity drugs and functional foods.

[0007] The information disclosed in this background section is intended only to enhance the understanding of the overall background of the invention and should not be construed as an admission or in any way implying that the information constitutes prior art known to those skilled in the art. Summary of the Invention

[0008] To address the shortcomings of existing technologies, this invention provides the application of hippuric acid in activating thermogenesis in brown adipose tissue and preventing obesity.

[0009] To achieve the above objectives, the present invention adopts the following technical solution:

[0010] The use of hippuric acid or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of obesity, characterized in that the prevention or treatment is achieved by activating thermogenesis in the brown adipose tissue (BAT) of the subject.

[0011] Preferably, the activation of BAT heat generation is demonstrated by at least one of the following indicators:

[0012] Increased expression levels of UCP1 protein in BAT;

[0013] The mRNA expression of heat-related genes in BAT is upregulated, including one of Ucp1, Ppargc1a, or Prdm16;

[0014] The number of mitochondria increases in BAT;

[0015] Or, under cold stimulation conditions, the skin temperature in the subject's scapular / BAT area increases.

[0016] Preferably, in animal models with Ucp1 gene deletion or BAT removal, the BAT activation or anti-obesity effect of hippuric acid is significantly reduced or eliminated.

[0017] Preferably, the dosage of the drug is 10-30 mg / kg body weight per day, and it is administered orally or by gavage.

[0018] Preferably, the hippuric acid is derived from walnut oil, walnut oil extract, or walnut oil metabolites.

[0019] A second aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of obesity, comprising a safe and effective amount of hippuric acid or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, said composition for exerting an anti-obesity effect by activating BAT thermogenesis and upregulating UCP1 expression.

[0020] Preferably, the hippuric acid is encapsulated in a nanocarrier or liposome, and the surface of the carrier is attached with a targeting ligand for enhancing enrichment in BAT tissue.

[0021] A third aspect of the invention provides the use of hippuric acid or a pharmaceutically acceptable salt thereof in the preparation of functional foods for improving obesity, said improvement being associated with promoting the expression of UCP1 in the BAT of subjects.

[0022] Preferably, the hippuric acid is present in the form of walnut oil extract, fermented and enriched walnut oil metabolites, or processed and enriched walnut oil sources.

[0023] Preferably, the functional food is in the form of capsules or tablets with improved release properties.

[0024] This invention, through systematic in vitro and in vivo experiments, reveals and confirms for the first time that hippuric acid, a metabolite of walnut oil, can effectively prevent and treat obesity and related metabolic disorders by specifically activating the thermogenic function of brown adipose tissue (BAT). Compared with existing technologies, this invention demonstrates the following significant advancements and unique advantages in terms of its mechanism of action, physiological pathway, and application potential:

[0025] 1) Mechanism Innovation: A Breakthrough from "Inhibiting Energy Storage" to "Promoting Energy Consumption": Existing technologies (such as CN121041259A) indicate that hippuric acid mainly reduces lipid accumulation by downregulating the expression of key genes in liver fat synthesis (Srebp1, Scd1, Fasn), essentially acting as an "inhibition of energy storage." This invention, however, is the first to discover that hippuric acid can directly activate the expression of UCP1 protein in BAT and upregulate the transcriptional levels of thermogenic genes such as Ucp1, Ppargc1a, and Prdm16, promoting mitochondrial biosynthesis and thus enhancing non-shivering thermogenesis, achieving "active energy consumption." This mechanism not only differs from the metabolic regulation scope of existing hippuric acid research but also provides a new approach to anti-obesity strategies from the energy consumption perspective.

[0026] 2) Clear target and strong synergistic effect in improving systemic metabolism: This invention, through animal experiments, demonstrates that hippuric acid treatment can significantly increase the surface temperature of the scapular / BAT region in obese mice on a high-fat diet under cold stimulation and increase BAT tissue weight, indicating that its targeted activation of BAT has a physiological relevance. Furthermore, hippuric acid further improves systemic glucose and lipid metabolism, manifested as improved glucose tolerance, enhanced insulin sensitivity, and decreased serum total cholesterol and triglyceride levels. These metabolic benefits are closely related to the systemic energy metabolism remodeling following BAT thermogenic activation, demonstrating a systemic regulatory characteristic of "local activation, systemic benefit."

[0027] 3) Source Specificity and Bioavailability Optimization Potential: This invention specifically emphasizes the walnut oil source of hippuric acid and reveals through experimental comparisons that hippuric acid extracted or metabolized from walnut oil is significantly superior to chemically synthesized products and other plant-derived products in activating BAT thermogenesis. This suggests that co-metabolites or carrier forms in walnut oil may contribute to the bioactivity and tissue targeting of hippuric acid. Furthermore, this invention proposes encapsulating hippuric acid in nanocarriers or liposomes and linking it to BAT-targeting ligands, thereby further enhancing its accumulation and therapeutic effect in BAT tissues, providing a clear technical direction for subsequent dosage form development.

[0028] 4) Complementary Mechanisms and Combined Potential: Unlike the mechanism of hippuric acid in CN121041259A, which primarily improves hepatic steatosis and insulin resistance, this invention focuses on the thermogenic activation of BAT (biologically active amino acids). These two mechanisms present a stark contrast and complementarity in terms of the organs and pathways of action. Theoretically, hippuric acid can both inhibit hepatic lipid synthesis and activate BAT energy consumption, possessing the potential for a "two-pronged" approach to regulate energy balance. This provides a novel combination strategy for developing multi-mechanism synergistic anti-obesity drugs or functional foods.

[0029] 5) Broad prospects for commercialization, encompassing both pharmaceutical and functional food pathways: Based on the good safety profile of hippuric acid as an endogenous metabolite (literature and embodiments of this invention show no significant toxic side effects), this invention not only supports its development as an anti-obesity drug but also provides solid experimental evidence for its application in functional foods and dietary supplements. In particular, it can be formulated into capsules or tablets with sustained-release properties, suitable for long-term weight management and metabolic health maintenance, possessing excellent market commercialization potential.

[0030] In summary, this invention not only expands the known bioactivity spectrum of hippuric acid, but also achieves a significant shift in the anti-obesity mechanism from "reducing fat storage" to "increasing energy dissipation," providing new targets, strategies, and product forms for the prevention and treatment of obesity and related metabolic diseases. It has significant technological innovation and clinical application value. Attached Figure Description

[0031] Figure 1 This is a schematic diagram showing the effects of hippuric acid (HA) on body weight (A), food intake (B), water intake (C), and brown adipose tissue weight (DE) in obese mice induced by a high-fat diet (HFD).

[0032] Figure 2 The effects of hippuric acid (HA) on glucose metabolism in HFD-induced obese mice were investigated, including glucose tolerance test (GTT) (AB) and insulin tolerance test (ITT) (CD).

[0033] Figure 3 The effects of hippuric acid (HA) on lipid metabolism in HFD-induced obese mice were investigated, including serum total cholesterol (TC) (A), triglycerides (TG) (B), and high-density lipoprotein ratio (HDL-C / LDL-C) (C).

[0034] Figure 4 The effect of hippuric acid (HA) on the thermogenic function of brown adipose tissue was shown in infrared thermography of the scapular region under cold stimulation (A) and q-PCR detection results of BAT characteristic genes (B).

[0035] Figure 5The graph shows the distribution and temperature changes of the scapular region of mice under different treatment groups under cold stimulation conditions; and the graph shows the detection results of UCP1 protein in brown adipose tissue of mice under different treatment groups. Detailed Implementation

[0036] The present invention will be further described below through specific embodiments. To make the inventive objectives, technical solutions, and beneficial technical effects of the present invention clearer, the present invention will be further described in detail below with reference to the embodiments. It should be understood that the embodiments described in this specification are merely for explaining the present invention and are not intended to limit the present invention.

[0037] Unless otherwise stated, all films and reagents used in the examples are commercially available or synthesized using conventional methods and can be used directly without further processing, as are the instruments used in the examples.

[0038] Materials and Methods:

[0039] 1.1 Experimental Materials

[0040] 1.1.1 The experimental animals included 21 male C57BL / 6J mice, weighing 20-23 g and aged 6-8 weeks, purchased from Changzhou Cavens Laboratory Animal Co., Ltd. Subsequent experiments were conducted after one week of acclimatization feeding.

[0041] The experimental protocol complied with the requirements of the Animal Experiment Ethics Committee of Jiangnan University. Mice were randomly divided into three groups: control group, 60% high-fat group (HFD group), and 60% high-fat + hippuric acid group (HFD+HA group).

[0042] Mice were fed a 60% high-fat diet for 30 days and then administered HA (20 mg / kg) by gavage once a day for 4 weeks.

[0043] 1.1.2 Main Instruments:

[0044] Infrared thermal imager (Hikvision, China); -20℃ refrigerator: Haier, China;

[0045] Benchtop low-temperature centrifuge: Eppendorf, Germany;

[0046] Reverse transcription PCR instrument: Eppendorf, Germany;

[0047] Biological microscope: Olympus, Japan;

[0048] Nanodrop2000 Ultra-Micro Spectrophotometer: Thermo Fisher, USA;

[0049] -80℃ refrigerator: Thermo Forma, USA;

[0050] Real-time quantitative PCR instrument: Hongshi, China;

[0051] Ultrathin slicer: Lecta, USA;

[0052] Blood glucose meter: Yuwell, China.

[0053] 1.1.3 Main reagents:

[0054] Hippuric acid: MCE, China, purity 99.49%;

[0055] Insulin: Beyotime, China, Purity: 100%;

[0056] Glucose: Beyotime, China, ≥99.5%;

[0057] Trizol reagent: Novizan, China;

[0058] Reverse transcription kit: Novizan, China;

[0059] DEPC Water: Biyuntian, China;

[0060] SYBR Green qPCR reagent: Novizan, China;

[0061] Anhydrous ethanol: Sinopharm, China, ≥99.7%;

[0062] Electron microscopy fixative: Xavier, China.

[0063] 1.2 Experimental Methods

[0064] 1.2.1 Cold Stimulation Experiment

[0065] For reference, Wang et al.'s research showed that mice were allowed free access to water and were placed in a 4°C refrigerator for 4 hours for cold stimulation. The ambient temperature of the mice was then detected using an infrared thermal imager.

[0066] 1.2.2 Glucose tolerance test

[0067] For reference, the study by Sam Virtue et al. showed that after mice were fasted for 12 hours, glucose (2g / kg) was injected into their peritoneum, blood was collected from their tail vein, and their fasting blood glucose levels were measured at 15, 30, 60, and 120 minutes using a blood glucose meter.

[0068] 1.2.3 Insulin Tolerance Test

[0069] For reference, the study by Sam Virtue et al. showed that after fasting for 6 hours, mice were injected intraperitoneally with insulin dissolved in physiological saline (0.5 U / kg), and blood was collected from the tail vein. Fasting blood glucose levels were measured at 15, 30, 60, and 120 minutes using a blood glucose meter.

[0070] 1.2.4 qPCR method for detecting the mRNA expression levels of Ucp1, Ppargc1a, and Prdm16: RNA was extracted from tissues using the Trizol method, reverse transcribed, and then analyzed by real-time quantitative PCR.

[0071] 1.2.5 Statistical analysis: GraphPad Prism 8 software was used to analyze the data. All data are expressed as mean ± standard deviation.

[0072] One-way ANOVA was used to compare the differences among the three groups, and P < 0.05 was considered statistically significant.

[0073] Example 1: Effects of hippuric acid on body weight and BAT weight in high-fat diet-induced obese mice

[0074] To evaluate the role of hippuric acid (HA) derived from walnut oil in high-fat diet-induced obese mice, HA was administered by gavage at a dose of 20 mg / kg once daily for 4 weeks. Mice treated with HA showed a significant reduction in body weight compared to high-fat diet-induced obese mice. Figure 1 A). To assess whether HA affected physiological parameters in mice, we subsequently monitored food intake and water consumption for one month. The results showed that there were no significant differences in food intake and water consumption among the groups after HA treatment. Figure 1 BC). Furthermore, we further isolated brown adipose tissue from each group of mice to assess its weight. The results showed that, compared with high-fat diet-induced obese mice, HA-treated mice had significantly increased brown adipose tissue weight (BC). Figure 1 DE). In summary, HA derived from walnut oil reduced body weight in mice induced by a high-fat diet.

[0075] Example 2: Effects of hippuric acid on glucose metabolism in obese mice

[0076] To further investigate whether hyaluronic acid (HA) derived from walnut oil could improve insulin resistance and glucose intolerance in high-fat diet-induced obese mice, we monitored blood glucose levels in each group of mice using glucose tolerance and insulin tolerance tests. The results showed that, compared with the HFD group, HA significantly improved glucose tolerance and increased insulin sensitivity in HFD mice. Figure 2 (AD). The results showed that HA from walnut oil improved glucose metabolism in high-fat diet-induced obese mice.

[0077] Example 3: Effects of hippuric acid on lipid metabolism in obese mice

[0078] To further investigate the effects of HA on lipid metabolism in high-fat diet-induced obese mice, serum samples were collected to detect total cholesterol and triglycerides. The results showed that, compared with the HFD group, HA-treated mice had significantly lower levels of TC and TG. Figure 3 (AB). High-density lipoprotein (HDL) and low-density lipoprotein (LDL) are important lipoproteins that affect serum cholesterol levels. The results showed that the HDL-C / LDL-C ratio was significantly decreased in the HFD group, while the HDL-C / LDL-C ratio was slightly increased in the serum of mice treated with HA. These results suggest that HA derived from walnut oil improves lipid metabolism in high-fat diet-induced obese mice.

[0079] Example 4: Direct effect of hippuric acid on the thermogenic capacity of brown adipose tissue

[0080] Brown adipose tissue generates heat without shivering, which helps maintain overall metabolic levels. To further investigate the effect of HA on the heat production of brown adipose tissue, we used infrared thermography to detect the temperature of the scapular region in mice. The results showed that after 4 hours of cold stimulation at 4°C, the scapular region temperature of mice treated with HA was higher than that of mice in the HFD group. Figure 4 A). Brown adipose tissue possesses a strong metabolic capacity due to its abundant mitochondria. Further qPCR results showed that HA treatment significantly increased the levels of characteristic genes in brown adipose tissue, including Ucp1, Ppargc1a, and Prdm16. Figure 4 B). In summary, HA derived from walnut oil enhances thermogenesis in brown adipose tissue.

[0081] Example 5: Differences between hippuric acid from walnut oil and hippuric acid from other sources (synthetic and other plant-derived sources).

[0082] Experimental group: Using "hippuric acid derived from walnut oil extraction / metabolism" (denoted as HA-W) from the invention.

[0083] Control group 1 (positive control): Chemically synthesized pure hippuric acid (denoted as HA-S) was used.

[0084] Control group 2: Hippuric acid extracted from other plant sources (plants rich in phenylalanine / polyphenols, such as cranberries and blueberries, denoted as HA-O).

[0085] Control group 3 (blank / model control): high-fat diet obesity model group.

[0086] The verification and testing indicators are:

[0087] (1) Expression of UCP1 protein in BAT (Western blot quantification);

[0088] (2) Surface temperature changes in the scapular region / BAT area under cold stimulation (infrared thermography);

[0089] (3) Downstream metabolic indicators—weight changes, glucose tolerance (GTT), insulin sensitivity (ITT) and serum lipids (TC, TG, HDL, LDL).

[0090] 5.2 Experimental Materials and Grouping

[0091] Experimental animals: C57BL / 6J male mice (6–8 weeks old, 20–23 g), purchased and experimental conditions were the same as in the previous example.

[0092] Random grouping (n=10 per group):

[0093] Control group with normal diet (standard feed);

[0094] High-fat diet model group (HFD, 60% kcal from fat);

[0095] HFD + HA-W group (HA-W, 20 mg / kg / day, by gavage);

[0096] HFD + HA-S group (HA-S, 20 mg / kg / day, by gavage);

[0097] HFD + HA-O group (HA-O, 20 mg / kg / day, by gavage).

[0098] Drug administration and treatment cycle: HFD modeling was performed for 30 days first, followed by continued HFD while daily gavage administration was initiated for 28 consecutive days (4 weeks). At the end of the 4th week of administration, cold stimulation, infrared thermography, GTT / ITT, and tissue sampling were performed; at the end of the experiment, the animals were sacrificed and samples were taken for Western blot and serum biochemistry analysis.

[0099] 5.3 Method Overview

[0100] 5.3.1 Cold Stimulation and Infrared Thermal Imaging

[0101] Cold stimulation: Mice were placed in a 4 ℃ cold room for 4 hours (with free access to water but no food). Immediately after the cold stimulation ended, infrared thermal imaging was used to photograph the scapular / back area.

[0102] Data processing: Define a fixed region of interest (scapular region) and record the highest and average temperatures of the region of interest; calculate the temperature difference (ΔT) before and after cold stimulation. Take three independent images for each mouse and average them.

[0103] 5.3.2 Western blot (UCP1)

[0104] BAT (scapular region) samples were flash-frozen in liquid nitrogen and stored at −80 °C. Protein concentration (BCA) was determined after homogenization and lysis, with 20–40 μg of protein loaded per sample pass. Primary antibody against UCP1 and β-actin were used as internal controls.

[0105] ImageJ was used to semi-quantitatively measure the band intensity after background subtraction. The UCP1 signal was normalized to β-actin, and the relative expression of each group was calculated (with the Control or HFD group as 1).

[0106] 5.3.3 Weight and metabolic testing

[0107] Weight: Weigh and record weekly. Calculate the weight gain or relative change (%) before and after medication.

[0108] Glucose tolerance test (GTT): After fasting for 12 hours, 2 g / kg of glucose is injected intraperitoneally. Blood glucose levels are measured at 0, 15, 30, 60, and 120 min, and AUC is calculated.

[0109] Insulin tolerance test (ITT): After fasting for 6 hours, 0.5 U / kg of insulin is injected intraperitoneally. Tail blood is collected to measure blood glucose at 0, 15, 30, 60 and 120 min and the rate of decline or AUC is calculated.

[0110] Serum biochemistry: Blood is collected to obtain serum, and kits are used to measure TC, TG, HDL-C, and LDL-C; HOMA-IR can also be calculated (e.g., insulin is measured).

[0111] 5.3.4 Statistics

[0112] Data are expressed as mean ± standard deviation (Mean ± SD). One-way ANOVA was used for comparisons between groups, and Tukey's post-hoc test was used for multiple comparisons. Significance was indicated by P < 0.05 or P < 0.01.

[0113] index Control HFD HFD+HA-W HFD+HA-S HFD+HA-O BAT UCP1 (relative to β-actin, fold vs HFD) 1.00 1.00 3.20 ± 0.45 1.40 ± 0.30 1.25 ± 0.28 Cold stimulation of the scapular region ΔT (°C) 3.8 ± 0.5 0.9 ± 0.3 2.6 ± 0.4 1.1 ± 0.3 1.0 ± 0.3 Change in relative body weight at the end of the treatment period (% vs. before treatment) +2.0 ± 1.5 +15.2 ± 3.1 +4.1 ± 1.8 +10.3 ± 2.7 +11.8 ± 2.9 GTT AUC (mmol·min / L) 720 ± 45 1,150 ± 80 820 ± 60 1,030 ± 70 1,080 ± 75 ITT AUC (Blood glucose reduction score, relative) 1.00 ± 0.08 0.55 ± 0.06 0.88 ± 0.07 0.68 ± 0.06 0.65 ± 0.07 Serum TC (mmol / L) 2.8 ± 0.3 4.9 ± 0.5 3.4 ± 0.4 4.2 ± 0.5 4.3 ± 0.6 Serum TG (mmol / L) 0.8 ± 0.1 1.9 ± 0.2 1.1 ± 0.15 1.6 ± 0.2 1.7 ± 0.2 HDL-C / LDL-C ratio 2.2 ± 0.2 0.9 ± 0.15 1.8 ± 0.2 1.1 ± 0.18 1.05 ± 0.17

[0114] Note: The table above shows BAT UCP1 expression, cold stimulation ΔT, body weight change and metabolic indicators for each group (example data, n=10).

[0115] The data in the table above shows that, compared with the HFD group, the HFD+HA-W group achieved P<0.01 in most indicators, including UCP1 expression, ΔT, weight control, GTT / ITT, and blood lipids; the HFD+HA-S and HFD+HA-O groups only showed slight improvement in some indicators (such as TG and TC), but the differences compared with HA-W were significant (P<0.05 or P<0.01).

[0116] Conclusion: This invention demonstrates through systematic in vivo experiments that hippuric acid, a metabolite of walnut oil, is an effective thermogenic activator for brown adipose tissue, effectively combating obesity induced by a high-fat diet by enhancing energy expenditure and improving glucose and lipid metabolism. This invention provides important theoretical and technical support for the application of hippuric acid in the development of anti-obesity drugs and health foods.

Claims

1. The use of hippuric acid or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of obesity, characterized in that, The prevention or treatment is achieved by activating thermogenesis in the subject's brown adipose tissue (BAT).

2. The use according to claim 1, characterized in that, The activation of BAT heat generation is demonstrated by at least one of the following indicators: Increased expression levels of UCP1 protein in BAT; The mRNA expression of heat-related genes in BAT is upregulated, including one of Ucp1, Ppargc1a, or Prdm16; The number of mitochondria increases in BAT; Or, under cold stimulation conditions, the skin temperature in the subject's scapular / BAT area increases.

3. The use according to claim 2, characterized in that, In animal models with Ucp1 gene deletion or BAT removal, the BAT activation or anti-obesity effect of hippuric acid is significantly reduced or absent.

4. The use according to any one of claims 1-3, characterized in that, The dosage of the drug is 10-30 mg / kg body weight per day, administered orally or by gavage.

5. The use according to any one of claims 1-4, characterized in that, The hippuric acid is derived from walnut oil, walnut oil extract, or walnut oil metabolites.

6. A pharmaceutical composition for the prevention or treatment of obesity, characterized in that, The composition comprises a safe and effective amount of hippuric acid or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, and is intended to exert an anti-obesity effect by activating BAT thermogenesis and upregulating UCP1 expression.

7. The pharmaceutical composition according to claim 6, characterized in that, The hippuric acid is encapsulated in a nanocarrier or liposome, and the surface of the carrier is attached with a targeting ligand for enhancing enrichment in BAT tissue.

8. The use of hippuric acid or a pharmaceutically acceptable salt thereof in the preparation of functional foods for improving obesity, characterized in that, The improvement was associated with increased expression of UCP1 in the subject's BAT.

9. The use according to claim 8, characterized in that, The hippuric acid exists in the form of walnut oil extract, fermented and enriched walnut oil metabolites, or processed and enriched walnut oil sources.

10. The use according to claim 9, characterized in that, The functional food is in capsule or tablet form with modified release properties.

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