Application of compound and composition thereof in preparation of medicine for treating non-alcoholic fatty liver disease

Abstract

The invention relates to the technical field of medicines for treating non-alcoholic fatty liver diseases, in particular to an application of a compound and a composition thereof in preparation of medicines for treating non-alcoholic fatty liver diseases. The compound used for preparing the medicine for treating the non-alcoholic fatty liver disease or having the body weight reducing effect or the weight losing effect comprises podocarpus macrophyllus flavone A and pharmaceutically acceptable salt thereof, hevea brasiliensis biflavone and pharmaceutically acceptable salt thereof, 3 beta-O-trans-p-coumaroyl maslinic acid and pharmaceutically acceptable salt thereof, 1, 3, 4-trimethyl-1, 3, 4-trimethyl-1, 3, 4-trimethyl-1, 3, 4-trimethyl-1, 3-trimethyl-1, 3-trimethyl-1, 3-trimethyl-1, 3-trimethyl-1, 3-trimethyl-1 The compound is selected from at least one of 3, 6-di-O-caffeoyl-beta-D-glucopyranoside and a pharmaceutically acceptable salt thereof, and 3, beta-O-trans-p-coumaroyl-2alpha-hydroxy-ursoene-12-ene-28-acid and a pharmaceutically acceptable salt thereof. The compound is selected from at least one of 3, 6-di-O-caffeoyl-beta-D-glucopyranoside and a pharmaceutically acceptable salt thereof. According to the technical scheme, the technical problem that drugs for comprehensively adjusting lipid metabolism, improving liver functions and effectively preventing NAFLD development are lacked in the prior art can be solved, and the pharmaceutical composition has ideal application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of non-alcoholic fatty liver drugs, and specifically relates to the application of a compound and its composition in the preparation of a drug for treating non-alcoholic fatty liver. Background Art

[0002] Non-alcoholic fatty liver disease (NAFLD) is a chronic progressive liver disease caused by overnutrition and insulin resistance, which mainly occurs in genetically susceptible individuals. The clinical spectrum of NAFLD includes non-alcoholic fatty liver (NAFL), non-alcoholic steatohepatitis (NASH), and its associated liver fibrosis and cirrhosis. With the changes in global lifestyle and eating habits, the incidence of NAFLD shows an upward trend. Currently, the global prevalence has reached about 30%, and it has become one of the main causes of liver diseases and liver-related deaths. In China, the total prevalence of NAFLD has increased significantly in the past 20 years, reaching 32.9% in 2018, which has become a major challenge in the national health field.

[0003] The pathogenesis of NAFLD is not yet clear, but abnormal lipid metabolism is considered to be one of the key pathological processes. The liver plays an important role in lipid metabolism, responsible for the storage, transport, synthesis, and decomposition of lipids. Imbalance of lipid metabolism will lead to excessive deposition of fat in the liver, which is not only the core mechanism of the occurrence of NAFLD, but also a driving factor for the progression of its more severe stages (such as NASH, cirrhosis, and hepatocellular carcinoma). In addition, abnormal lipid metabolism is also closely related to the occurrence of diseases such as atherosclerosis and diabetes, further aggravating the health burden of patients and significantly affecting the quality of life.

[0004] Currently, the treatment options for NAFLD are limited. Although there are already certain drugs clinically available for treating dyslipidemia at present, such as statins, fibrates, cholesterol absorption inhibitors, and PCSK-9 inhibitors, etc., these drugs can improve blood lipid levels to a certain extent, but they fail to effectively address the complex pathological process of NAFLD. Statins inhibit HMG-CoA reductase to reduce cholesterol synthesis, thereby reducing the level of Low-Density Lipoprotein Cholesterol (LDL-C). Fibrates activate Peroxisome Proliferator-Activated Receptor α (PPARα) to promote fatty acid oxidation and improve dyslipidemia by inhibiting the activation of macrophages and Kupffer cells. Cholesterol absorption inhibitors, such as ezetimibe, can play a role in reducing blood lipids by reducing cholesterol absorption. As a new type of lipid-lowering drug, PCSK-9 inhibitors bind to PCSK9, inhibit its binding to the Low-Density Lipoprotein Receptor (LDL-R), reduce the degradation of LDL-R, thereby increasing the number of LDL-R on the surface of hepatocytes and improving the efficiency of the liver to clear LDL-C.

[0005] One of the most common side effects of statins is muscle pain and injury, manifested as muscle soreness, fatigue or weakness. In rare cases, the use of statins may lead to rhabdomyolysis, a serious muscle injury that can cause renal failure and death. The side effects of fibrates include abnormal liver function such as elevated transaminases and jaundice, and in severe cases, it can lead to liver failure. When fibrates are used in combination with statins, it may increase the risk of myopathy and induce rhabdomyolysis in severe cases. The common adverse reactions of ezetimibe are mainly problems in the digestive system, such as abdominal pain, diarrhea, and flatulence, and it may also cause abnormal neurological symptoms such as headache and fatigue. The most common side effect of PCSK9 inhibitors is local reactions at the injection site, including erythema, pain, swelling, or itching. At the same time, as a new type of lipid-lowering drug, the long-term safety of PCSK9 inhibitors has not been fully clarified, and their high price limits their widespread use.

[0006] Thus, these existing treatment methods have limited effects in improving liver function and preventing disease progression, especially there are still significant deficiencies in the deep regulation of liver lipid metabolism imbalance. Therefore, there is an urgent need to develop new drugs, especially treatment strategies that can more comprehensively regulate lipid metabolism, improve liver function, and effectively prevent the progression of NAFLD. Summary of the Invention

[0007] The object of the present invention is to provide the use of a compound in the preparation of a medicament for treating non-alcoholic fatty liver, so as to solve the technical problem in the prior art of lacking a medicament that comprehensively regulates lipid metabolism, improves liver function and effectively prevents the progression of NAFLD.

[0008] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0009] The use of a compound in the preparation of a medicament for treating non-alcoholic fatty liver or a medicament with weight loss efficacy or a medicament with slimming efficacy, wherein the compound comprises at least one of podocarpusflavone A and its pharmaceutically acceptable salts, heveaflavone and its pharmaceutically acceptable salts, 3β-O-trans-p-coumaroylmaslinic acid and its pharmaceutically acceptable salts, 1,6-di-O-caffeoyl-β-D-glucopyranoside and its pharmaceutically acceptable salts, and 3β-O-trans-p-coumaroyl-2α-hydroxy-urs-12-ene-28-oic acid and its pharmaceutically acceptable salts.

[0010] Further, the compound comprises at least one of heveaflavone and its pharmaceutically acceptable salts, and 1,6-di-O-caffeoyl-β-D-glucopyranoside and its pharmaceutically acceptable salts.

[0011] Further, the compound is composed of heveaflavone or its pharmaceutically acceptable salt and 1,6-di-O-caffeoyl-β-D-glucopyranoside or its pharmaceutically acceptable salt.

[0012] Further, the molar ratio of heveaflavone or its pharmaceutically acceptable salt to 1,6-di-O-caffeoyl-β-D-glucopyranoside or its pharmaceutically acceptable salt is 1:1 - 1:2.

[0013] Further, the compound is used for compounding with at least one additional component selected from the following: a pharmaceutically acceptable carrier, a pharmaceutically acceptable vehicle, and a pharmaceutically acceptable excipient.

[0014] Further, the compound is used to be made into tablets, capsules, oral liquids, injections, powders or ointments.

[0015] The present technical solution also provides a composition formed by a flavonoid substance and a glucopyranoside substance, which is composed of heveaflavone or its pharmaceutically acceptable salt and 1,6-di-O-caffeoyl-β-D-glucopyranoside or its pharmaceutically acceptable salt.

[0016] Further, the molar ratio of heveaflavone or its pharmaceutically acceptable salt to 1,6-di-O-caffeoyl-β-D-glucopyranoside or its pharmaceutically acceptable salt is 1:1 - 1:2.

[0017] Furthermore, a composition formed by a flavonoid substance and a glucopyranoside substance is used to reduce liver cholesterol and triglyceride levels.

[0018] Furthermore, a composition formed by a flavonoid substance and a glucopyranoside substance is used to reduce body weight, reduce BMI, and suppress appetite.

[0019] In summary, the beneficial effects of this technical solution are as follows:

[0020] The pathogenesis of non-alcoholic fatty liver disease (NAFLD) is closely related to abnormal lipid metabolism. The imbalance of lipid metabolism causes excessive fat deposition in the liver, promoting the progression of the disease. At least one compound among podocarpusflavone A and its pharmaceutically acceptable salts, heveaflavone and its pharmaceutically acceptable salts, 3β-O-trans-p-coumaroylmaslinic acid and its pharmaceutically acceptable salts, 1,6-di-O-caffeoyl-β-D-glucopyranoside and its pharmaceutically acceptable salts, and 3β-O-trans-p-coumaroyl-2α-hydroxy-urs-12-ene-28-oic acid and its pharmaceutically acceptable salts involved in the present invention can act on the lipid metabolism process and serve as candidate drugs for the treatment of NAFLD. Among them, heveaflavone and 1,6-di-O-caffeoyl-β-D-glucopyranoside respectively show significant inhibitory effects on triglyceride (TG) and total cholesterol (TC). If the two compounds are used in combination, it has been experimentally verified that the ability to regulate lipid metabolism can be significantly enhanced, effectively reducing liver fat deposition, and thus improving the condition of non-alcoholic fatty liver. At the same time, these compounds may also play a role in reducing body weight, BMI, and suppressing appetite, and have a positive impact on health problems related to NAFLD in multiple dimensions.

[0021] The beneficial effects of this technical solution are as follows:

[0022] (1) Enhanced efficacy: Existing drugs for the treatment of NAFLD have limited effects in regulating lipid metabolism, improving liver function, and preventing the progression of the disease. The compounds screened in the present invention, especially when heveaflavone and 1,6-di-O-caffeoyl-β-D-glucopyranoside are used in combination, exhibit a synergistic effect, can significantly enhance the ability to regulate lipid metabolism, more effectively reduce liver fat deposition, and the overall treatment effect is significantly better than that of using a single drug alone, greatly improving the treatment effect on non-alcoholic fatty liver.

[0023] (2) Reduced dosage: Due to the synergistic effect between the compounds, when achieving the same treatment effect, compared with using a single drug alone, the dosage of each drug required for combined medication may be lower, which helps to reduce the overall drug load in the body.

[0024] (3) Reducing side effects: Currently available drugs for treating dyslipidemia, such as statins, fibrates, cholesterol absorption inhibitors, and PCSK-9 inhibitors, all have varying degrees of side effects. The compounds and their compositions of the present invention provide a new option for the treatment of NAFLD, and are expected to reduce the risk of side effects caused by the use of traditional drugs. For example, it can avoid muscle pain and injury caused by statins, abnormal liver function caused by fibrates, digestive system problems caused by ezetimibe, and injection site reactions caused by PCSK-9 inhibitors. Moreover, the long-term safety may be more advantageous.

[0025] (4) Providing a new strategy: It provides a completely new combination drug strategy for the clinical intervention of NAFLD, enriches the treatment means, lays an important foundation for further in-depth research and development of treatment programs for NAFLD, and is of great significance for solving the current problems in the field of NAFLD treatment. Description of the Drawings

[0026] Figure 1 Experimental results of the lipid-lowering effect evaluation of 5 compounds in Example 1 (A: TC level in HpeG2 cells; B: TG level in HpeG2 cells; n = 5; "*" indicates p < 0.05, "**" indicates p < 0.01; FFA indicates the experimental group treated with fatty acids).

[0027] Figure 2 Experimental results of the lipid-lowering effect evaluation of Compound B, Compound D, the combination of Compound B and Compound B in Example 2 (A: TG level in HpeG2 cells; B: TC level in HpeG2 cells; n = 5; "*" indicates p < 0.05, "**" indicates p < 0.01; FFA indicates the experimental group treated with fatty acids).

[0028] Figure 3 Effect of drug treatment in Example 3 on the body weight and liver weight of HFD-fed mice (A: Gross morphology of the liver; B: Liver weight; n = 5; "*" indicates p < 0.05, "**" indicates p < 0.01; HFD indicates the experimental group with a high-fat diet).

[0029] Figure 4 Research results of the effect of drug treatment in Example 3 on the liver TG and TC levels (A: Liver TC level; B: Liver TG level; n = 5; "**" indicates p < 0.01; HFD indicates the experimental group with a high-fat diet).

[0030] Figure 5Research results on the inhibitory effects of the drug treatment in Example 3 on the body weight and food intake of HFD mice (A: Changes in mouse body weight; B: Body weight at the 12th week; C: Mouse BMI; D: Mouse food intake; n = 5; "*" indicates p < 0.05, "**" indicates p < 0.01; HFD represents the experimental group with a high-fat diet). Detailed implementation manners

[0031] The present invention will be further described in detail below in conjunction with embodiments, but the implementation manners of the present invention are not limited thereto. Unless otherwise specified, the technical means used in the following embodiments and experimental examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used can all be obtained from commercial channels.

[0032] Example 1

[0033] (1) Establishment of a non-alcoholic fatty liver disease (NAFLD) cell model and treatment with the drug to be tested

[0034] A mixture of oleic acid (OA) and palmitic acid (PA) (OA:PA = 2:1, total concentration 1 mM) was used to induce HepG2 cells to establish an in vitro NAFLD model. While adding OA and PA, the corresponding compound to be tested was also added respectively (the final concentration of each compound to be tested was 50 μM). After co-treating the cells for 24 h, the samples were collected, and the changes in TC and TG were measured using a total cholesterol test kit and a triglyceride test kit to screen for compounds for the treatment of NAFLD. Among them, HepG2 is a commonly used human hepatocellular carcinoma cell line, which is easy to culture and operate. It is generally cultured in DMEM medium and can induce lipid accumulation by adding free fatty acids (such as oleic acid and palmitic acid) to mimic the pathological state of NAFLD. In the model group (Model), the compound to be tested was not added, and only free fatty acids were used to establish the model; in the blank group (Control), neither free fatty acids nor drugs were added to the cells, and only routine cell culture was carried out. Sample collection and processing were performed at the same time points as other experimental groups.

[0035] Details of the compounds to be tested for the experiment are shown in Table 1.

[0036] Table 1: Compounds to be tested

[0037]

[0038]

[0039] After taking out the treated cells, discard the supernatant, wash them with PBS 1-2 times. After adding 1 mL of PBS, use a cell scraper to scrape the cells in the same direction, transfer them to a 1.5 mL EP tube, centrifuge at 3000 r / min for 5 min, discard the supernatant, and keep the cell pellet. Add 0.2 - 0.3 mL of PBS to the cell pellet for homogenization, and perform ultrasonic fragmentation under ice-water bath conditions (power 300 W, 3 - 5 s / time, interval 30 s, repeat 3 - 5 times). The prepared homogenate is directly measured for total cholesterol and triglyceride without centrifugation. Use the Nanjing Jiancheng TC / TG kit (product number A111-1-1 / A110-1-1) to detect total cholesterol (TC) and triglyceride (TG). The detection results are expressed in "mmol / gprot", that is: the number of millimoles of TC or TG contained in each g of total protein. In biological samples, TC and TG usually exist in the form of complexes such as lipoproteins bound to proteins. In different individuals or different physiological and pathological states, the protein content in the samples may vary. Using the unit of mmol / gprot is to correlate the content of TC and TG with the protein content, standardize based on the protein, so as to more accurately reflect the relative relationship between TC and TG and proteins, eliminate the interference caused by differences in protein content, and make the comparison between different samples more scientific and comparable. Cell experiments usually involve different numbers of cells or different volumes of samples. The directly measured TG content (such as mmol / L or mmol / mL) may deviate due to different cell numbers or sample volumes. By dividing the TG content by the protein concentration, these differences can be eliminated and the results can be made more comparable. See Table 2 for the sample addition situation.

[0040] Table 2: Sample addition situation for TC and TG detection (the sample is the homogenate mentioned above)

[0041]

[0042] (2) Experimental results

[0043] Specific results of TC and TG concentration measurements are shown in Figure 1 . Preliminary screening of the lipid-lowering effects of 5 compounds found that compared with the Model group, all 5 compounds had a certain ability to reduce TC and TG (p < 0.05). However, the effects of Compound A, Compound C, and Compound D in reducing TG and TC were weaker. At the same time, Compound B had the most significant effect on reducing TG, while Compound D had the most significant effect on reducing TC. Therefore, Compound B and Compound D were selected for subsequent research.

[0044] Example 2

[0045] The inventors attempted to conduct experimental studies on Compound B and Compound D and also tried the effect of using the two substances in combination. The experimental method was carried out with reference to Example 1, with the difference that the total final concentration of the drugs in the Compound B and Compound D combination group (Compound B+D) was 50 μM, and the molar ratio of the two drugs was 1:1.

[0046] The experimental results are shown in Figure 2 . Among them, for the test results of TG content: the average TG content in the Control group was 0.204±0.005 mmol / gprot, the average TG content in the Model group was 1.196±0.014 mmol / gprot, the average TG content in the Compound B group was 0.758±0.009 mmol / gprot, the average TG content in the Compound D group was 0.830±0.005 mmol / gprot, and the average TG content in the Compound B+D group was 0.396±0.037 mmol / gprot.

[0047] The Bliss independence model was used to evaluate the mode of action between the two drugs. When screening drug combinations, the Bliss independence model can quickly determine whether there is a synergistic effect, an antagonistic effect, or an additive effect when two or more drugs are used in combination. For example, in the research and development of anti-cancer drugs, it can evaluate the combined effects of different chemotherapy drugs or targeted drugs to help determine the most promising drug combination plan and provide a basis for clinical trials. In this model, the expected effect of the combined action is calculated based on the effect data of each drug acting alone. For example: the effect produced by drug A acting alone is E A , the effect produced by drug B acting alone is E B , and in the Bliss independence model, the calculation method of the expected effect E AB of the combined action is generally E AB = E A + E B - E A × E B . If the expected effect of the combined use of the two drugs is greater than the actual effect, or the expected effect of the combined use of the two drugs is basically equal to the actual effect, it indicates that there is no synergistic effect between the two drugs, or there is an antagonistic effect; if the expected effect of the combined use of the two drugs is less than the actual effect, it indicates that there is a synergistic effect between the two drugs.

[0048] According to the Bliss independence model, using the TG content as an index, the theoretical combined effect E 理论 = E Compound B + E Compound D - ECompound B ×E Compound D Among them, the treatment effect (E) is calculated as follows: Treatment effect (E) = (TG in the Model group - TG in the Compound group) / (TG in the Model group - TG in the Control group) × 100%. The treatment effect (E) is the proportion by which the treatment reduces TG.

[0049] The treatment effect (E Compound B ) of the Compound B group = (1.196 - 0.758) / (1.196 - 0.204) × 100% = 44.2%;

[0050] The treatment effect (E Compound D ) of the Compound D group = (1.196 - 0.830) / (1.196 - 0.204) × 100% = 37.0%;

[0051] The treatment effect (E Compound B+D ) of the Compound B+D group = (1.196 - 0.396) / (1.196 - 0.204) × 100% = 80.7%;

[0052] The theoretical effect value of the Compound B+D group: E theory = 0.442 + 0.370 - (0.442 × 0.370) = 0.649 (64.9%) < 80.7% (Compound B and D have a synergistic effect in reducing the TG level in the cell model).

[0053] Compared with the Model group, the Compound B group, and the Compound D group, after treatment with the Compound B+D group, the intracellular TG content decreased significantly, and the actual combined effect was significantly higher than the theoretical combined effect, indicating that there is a significant synergistic effect between Compound B and Compound D in combination.

[0054] Among them, for the test results of TC content: the average TC content of the Control group was 0.153 ± 0.006 mmol / g prot, the average TC content of the Model group was 0.811 ± 0.009 mmol / g prot, the average TC content of the Compound B group was 0.674 ± 0.004 mmol / g prot, the average TC content of the Compound D group was 0.602 ± 0.007 mmol / g prot, and the average TC content of the Compound B+D group was 0.399 ± 0.006 mmol / g prot. Calculated according to the aforementioned formula, the effects of each experimental group are as follows:

[0055] The therapeutic effect (E Compound B ) of the Compound B group = (0.811 - 0.674) / (0.811 - 0.153)×100% = 20.8%;

[0056] The therapeutic effect (E Compound D ) of the Compound D group = (0.811 - 0.602) / (0.811 - 0.153)×100% = 31.8%;

[0057] The therapeutic effect (E Compound B+D ) of the Compound B+D group = (0.811 - 0.399) / (0.811 - 0.153)×100% = 62.6%;

[0058] The theoretical effect value of the Compound B+D group: E 理论 = 0.208 + 0.318 - (0.208×0.318) = 0.460 (46.0%) < 62.6% (Compound B and D have a synergistic effect in reducing the TC level of the cell model).

[0059] It can be seen that compared with the Model group, the Compound B group, and the Compound D group, after treatment with the Compound B+D group, the intracellular TC content decreased significantly, and the actual combined effect was significantly higher than the theoretical combined effect, indicating that there is a significant synergistic effect between Compound B and Compound D in combination.

[0060] Example 3

[0061] This example is used to test the effect of the drug in a non-alcoholic fatty liver disease (NAFLD) animal model induced by a high-fat diet (HFD). The NAFLD animal model was constructed by the following method: Male C57BL / 6 mice at 6 weeks of age were continuously fed a HFD for 3 months to induce an in vivo NAFLD model. During this period, Compound B and Compound D or Compound B + Compound D were administered by gavage every two days. The blank control group and the model group were respectively gavaged with an equal volume of corn oil, and the blank control group was not fed a high-fat diet. More specifically, Compound B and / or Compound D were dissolved in corn oil, and the drug was delivered into the mice by gavage to evaluate the effect of the compound on non-alcoholic fatty liver. The dosing regimen is shown in Table 3 for details.

[0062] Table 3: Dosing regimen for C57 mice with non-alcoholic fatty liver

[0063]

[0064] Experimental result 1: The effect of the drug on the liver

[0065] After the experiment, the mice were sacrificed, and the livers of the sacrificed mice were weighed and the morphology of the livers was recorded. The experimental results of the effect of drug treatment on the liver are shown in Figure 3 . Compared with the Control group induced by HFD, the liver weight / body weight in the Model group was significantly increased. The livers of the mice in the Control group were dark red-brown in appearance, with uniform and firm texture, smooth surface, and no swelling; the livers of the mice in the Model group induced by HFD were soft in texture, pink in color, significantly swollen, and the liver weight was significantly increased. The livers of the mice in the Compound B and Compound D groups were soft in texture, and most of the livers were still pink. The liver weight was lower than that in the Model group; the livers of the mice in the Compound B + D group returned to dark red-brown, the texture became firmer, the surface was smooth, no significant swelling was observed, and the liver weight decreased significantly ( Figure 3 A and B, p < 0.05).

[0066] Experimental result 2: The effect of the drug on liver TC and TG

[0067] The levels of TG and TC in the liver were analyzed, and the results are shown in Figure 4 (A: Liver TC level; B: Liver TG level).

[0068] Regarding the TC level in the liver, the average TG content in the Control group was 0.232 ± 0.030 mmol / gprot, the average TC content in the Model group was 0.868 ± 0.111 mmol / gprot, the average TC content in the Compound B group was 0.594 ± 0.051 mmol / gprot, the average TC content in the Compound D group was 0.602 ± 0.096 mmol / gprot, and the average TC content in the Compound B+D group was 0.266 ± 0.024 mmol / gprot.

[0069] The therapeutic effect (E Compound B ) of the Compound B group = (0.868 - 0.594) / (0.868 - 0.232) × 100% = 43.1%;

[0070] The therapeutic effect (E Compound D ) of the Compound D group = (0.868 - 0.602) / (0.868 - 0.232) × 100% = 41.8%;

[0071] The therapeutic effect (E Compound B+D ) of the Compound B+D group = (0.868 - 0.266) / (0.868 - 0.232) × 100% = 94.7%;

[0072] The theoretical effect value of the Compound B+D group: E theory = 0.431 + 0.418 - (0.431 × 0.418) = 0.669 (66.9%) < 94.7% (Compound B and D have a synergistic effect in reducing the TC level in the liver).

[0073] Regarding the TG level in the liver, the average TG content in the Control group was 0.292 ± 0.093 mmol / gprot, the average TG content in the Model group was 1.246 ± 0.164 mmol / gprot, the average TG content in the Compound B group was 0.876 ± 0.092 mmol / gprot, the average TG content in the Compound D group was 0.868 ± 0.168 mmol / gprot, and the average TG content in the Compound B+D group was 0.423 ± 0.084 mmol / gprot.

[0074] The therapeutic effect (E Compound B) = (1.246 - 0.876) / (1.246 - 0.292) × 100% = 38.8%;

[0075] Therapeutic effect of Compound D group (E Compound D ) = (1.246 - 0.868) / (1.246 - 0.292) × 100% = 39.6%;

[0076] Therapeutic effect of Compound B + D group (E Compound B+D ) = (1.246 - 0.423) / (1.246 - 0.292) × 100% = 86.3%;

[0077] Theoretical effect value of Compound B + D group: E theory = 0.388 + 0.396 - (0.388 × 0.396) = 0.630 (63.0%) < 86.3% (Compound B and D have a synergistic effect in reducing liver TG level).

[0078] From the above experimental results, it can be seen that after the combined use of Compound B and Compound D, the effect of reducing the contents of TG and TC is significantly better than that of single drug use, and there is a significant synergistic effect, indicating that the combined drug use is a more effective treatment strategy.

[0079] Experimental result three: Effects of drugs on mouse body weight, BMI and food intake

[0080] See details of the experimental results in Figure 5 (A: Changes in mouse body weight; B: Body weight at the 12th week; C: Mouse BMI; D: Mouse food intake).

[0081] From Figure 5 A, it can be seen that after the mice were fed with HFD, their body weight increased linearly; after giving Compound B and Compound D, the body weight of the mice increased more slowly than that of the Model group; after giving Compound B + D, the body weight gain of the mice was significantly slower than that of the Model group.

[0082] In Figure 5 B, the average body weight of the Control group was 27.76 ± 0.82 g, the average body weight of the Model group was 35.28 ± 0.79 g, the average body weight of the Compound B group was 33.02 ± 0.79 g, the average body weight of the Compound D group was 31.64 ± 0.67 g, and the average body weight of the Compound B + D group was 29.56 ± 0.36 g.

[0083] Therapeutic effect of Compound B group (ECompound B ) = (35.28 - 33.02) / (35.28 - 27.76) × 100% = 30.5%;

[0084] Therapeutic effect of Compound D group (E Compound D ) = (35.28 - 31.64) / (35.28 - 27.76) × 100% = 48.4%;

[0085] Therapeutic effect of Compound B + D group (E Compound B+D ) = (35.28 - 29.56) / (35.28 - 27.76) × 100% = 76.06%;

[0086] Theoretical effect value of Compound B + D group: E 理论 = 0.305 + 0.484 - (0.305 × 0.484) = 0.6391 (63.91%) < 76.06% (Compound B and D have a synergistic effect in reducing body weight).

[0087] In Figure 5 C, the average BMI of the Control group is 0.256 ± 0.024 g / cm 2 , the average BMI of the Model group is 0.378 ± 0.033 g / cm 2 , the average BMI of the Compound B group is 0.334 ± 0.027 g / cm 2 , the average BMI of the Compound D group is 0.342 ± 0.016 g / cm 2 , the average BMI of the Compound B + D group is 0.304 ± 0.011 g / cm 2 .

[0088] Therapeutic effect of Compound B group (E Compound B ) = (0.378 - 0.334) / (0.378 - 0.256) × 100% = 36.07%;

[0089] Therapeutic effect of Compound D group (E Compound D ) = (0.378 - 0.342) / (0.378 - 0.256) × 100% = 29.51%;

[0090] Therapeutic effect of Compound B + D group (E Compound B+D) = (0.378 - 0.304) / (0.378 - 0.256) × 100% = 60.66%;

[0091] The theoretical effect value of Compound B + D group: E 理论 = 0.3607 + 0.2951 - (0.3607 × 0.2951) = 0.5493 (54.93%) < 60.66% (Compound B and D have a synergistic effect in reducing BMI).

[0092] In Figure 5 D, the average food intake of the Control group was 3.10 ± 0.31 g / day, the average food intake of the Model group was 5.60 ± 0.59 g / day, the average food intake of the Compound B group was 5.34 ± 0.40 g / day, the average food intake of the Compound D group was 5.24 ± 0.63 g / day, and the average food intake of the Compound B + D group was 4.36 ± 0.84 g / day.

[0093] The treatment effect of the Compound B group (E Compound B ) = (5.60 - 5.34) / (5.60 - 3.10) × 100% = 10.40%;

[0094] The treatment effect of the Compound D group (E Compound D ) = (5.60 - 5.24) / (5.60 - 3.10) × 100% = 14.40%;

[0095] The treatment effect of the Compound B + D group (E Compound B+D ) = (5.60 - 4.36) / (5.60 - 3.10) × 100% = 49.60%;

[0096] The theoretical effect value of the Compound B + D group: E 理论 = 0.104 + 0.144 - (0.104 × 0.144) = 0.233 (23.30%) < 49.60% (Compound B and D have a synergistic effect in suppressing appetite).

[0097] Experimental data show that both Compound B and Compound D have the effects of reducing body weight, reducing BMI, and inhibiting food intake (i.e., suppressing the appetite of model animals, having an appetite-suppressing effect), and can be used as a weight loss drug or a drug for reducing body weight. Moreover, the combined use of Compound B and Compound D can produce a synergistic effect in terms of reducing body weight, reducing BMI, and inhibiting food intake. The combined use of the two drugs can achieve a more ideal weight loss function.

[0098] The above are only embodiments of the present invention, and common knowledge such as specific structures and characteristics known in the art are not described in detail herein. It should be noted that for those skilled in the art, without departing from the structure of the present invention, several modifications and improvements can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the implementation effect of the present invention and the practicality of the patent. The protection scope required by this application shall be subject to the content of its claims, and the specific implementation manners described in the specification can be used to interpret the content of the claims.

Claims

1. Use of a compound in the preparation of a drug for treating non-alcoholic fatty liver disease or a drug having a weight loss effect or a drug having a weight loss effect, characterized in that: The compound includes at least one of Podocarpus flavonoids A and pharmaceutically acceptable salts thereof, Hevea brasiliensis biflavonoids and pharmaceutically acceptable salts thereof, 3β-O-trans-p-coumaroylmaslinic acid and pharmaceutically acceptable salts thereof, 1,6-di-O-caffeoyl-β-D-glucopyranoside and pharmaceutically acceptable salts thereof, and 3β-O-trans-p-coumaroyl-2α-hydroxy-ursene-12-ene-28-acid and pharmaceutically acceptable salts thereof.

2. Use of the compound according to claim 1 in the preparation of a drug for treating non-alcoholic fatty liver disease or a drug having a weight loss effect or a drug having a weight loss effect, characterized in that: The compound comprises at least one of rubber tree biflavonoids and pharmaceutically acceptable salts thereof, 1,6-di-O-caffeoyl-β-D-glucopyranoside and pharmaceutically acceptable salts thereof.

3. Use of the compound according to claim 2 in the preparation of a drug for treating non-alcoholic fatty liver disease or a drug having a weight loss effect or a drug having a weight loss effect, characterized in that: The compound consists of rubber tree biflavonoids or a pharmaceutically acceptable salt thereof and 1,6-di-O-caffeoyl-β-D-glucopyranoside or a pharmaceutically acceptable salt thereof.

4. Use of the compound according to claim 3 in the preparation of a drug for treating non-alcoholic fatty liver disease or a drug having a weight loss effect or a drug having a weight loss effect, characterized in that: The molar ratio of the rubber tree biflavonoids or a pharmaceutically acceptable salt thereof to the 1,6-di-O-caffeoyl-β-D-glucopyranoside or a pharmaceutically acceptable salt thereof is 1:1-1:

2.

5. Use of the compound according to claim 4 in the preparation of a drug for treating non-alcoholic fatty liver disease or a drug having a weight loss effect or a drug having a weight loss effect, characterized in that: The compound is used to be formulated with at least one additional component selected from the group consisting of a pharmaceutically acceptable carrier, a pharmaceutically acceptable vehicle and a pharmaceutically acceptable excipient.

6. Use of the compound according to claim 5 in the preparation of a drug for treating non-alcoholic fatty liver disease or a drug having a weight loss effect or a drug having a weight loss effect, characterized in that: The compound is used to prepare tablets, capsules, oral liquids, injections, powders or pastes.

7. A composition of flavonoids and glucopyranoside substances, characterized in that: The invention consists of rubber tree biflavonoids or a pharmaceutically acceptable salt thereof and 1,6-di-O-caffeoyl-β-D-glucopyranoside or a pharmaceutically acceptable salt thereof.

8. The composition of flavonoids and glucopyranoside according to claim 7, characterized in that: The molar ratio of the rubber tree biflavonoids or a pharmaceutically acceptable salt thereof to the 1,6-di-O-caffeoyl-β-D-glucopyranoside or a pharmaceutically acceptable salt thereof is 1:1-1:

2.

9. The composition of flavonoids and glucopyranoside according to claim 8, characterized in that: It is used to lower liver cholesterol and triglyceride levels.

10. The composition of flavonoids and glucopyranoside according to claim 9, characterized in that: It is used to reduce body weight, lower BMI, and suppress appetite.

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