A class of lipid-lowering and anti-obesity cadinane dimer sesquiterpenoid compounds and their application

Abstract

The present invention discloses the use of a cadinane dimer sesquiterpene compound or a derivative thereof in the preparation of a lipid-lowering or weight-loss composition. The present invention provides a class of cadinane dimer sesquiterpene compounds, which have lipid-lowering and weight-loss functions, and in particular, some cadinane dimer sesquiterpene compounds have strong lipid-lowering and anti-obesity activities and good safety, and are expected to be developed into lipid-lowering, weight-loss, and blood sugar-lowering products.

Description

Technical Field

[0001] The invention belongs to the technical field of anti-obesity drugs, and in particular relates to a class of cadinane dimer sesquiterpene compounds and applications thereof. Background Art

[0002] Obesity often induces and increases the risk of many chronic diseases, such as hypertension, type II diabetes, and cardiovascular and cerebrovascular diseases, which seriously reduce life expectancy and quality of life. According to the World Obesity Federation, by 2035, 24% of the world's population (about 1.914 billion) will suffer from obesity. The prevalence of obesity in adults, adolescents and children in my country has increased year by year, and the number of obese patients currently exceeds 100 million. There are currently five types of anti-obesity drugs in clinical use, which are divided into lipase inhibitors (including orlistat) and appetite suppressants (including phentermine-topiramate, naltrexone-bupropion, liraglutide and semaglutide). However, the long-term use of existing anti-obesity drugs has prominent toxic and side effects, so it is very necessary to develop safe and effective new anti-obesity drugs, which has important clinical significance. Plants of the genus Tutianqi in the ginger family are rich in novel structural cadinane-type dimerized sesquiterpenes, which have diverse polymerization modes and are prone to rearrangement and degradation, forming complex structures with multiple rings, cages and multiple chiral carbons. At present, there are no studies and reports on the lipid-lowering and anti-obesity activities of cadinane-type dimerized sesquiterpenes. Summary of the invention

[0003] The technical problem to be solved by the present invention is to provide a class of cadinane dimer sesquiterpene compounds and their applications, which have strong lipid-lowering and anti-obesity activities and good safety, and are expected to be developed into lipid-lowering, weight-loss and blood sugar-lowering products.

[0004] Application of a cadinane dimer sesquiterpene compound or a derivative thereof in preparing a lipid-lowering or weight-loss composition.

[0005] Furthermore, the cadinane dimer sesquiterpene compound is of C-2 and C-2' connection type.

[0006] Furthermore, the general structural formula of the cadinane dimer sesquiterpene compound is as shown in Formula I, Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII or Formula VIII:

[0007]

[0008] In Formula I: C-4 and C-11 are connected by an oxygen bridge or not; R 1 and R 2 is selected from hydrogen or hydroxyl;

[0009] In formula III: R 1 Selected from methyl or -CH 2 OH;

[0010] In Formula IV: C-4 and C-11 are connected by an oxygen bridge or not; C-1' and C-2' are connected by a double bond or not; R 1 and R 2 is selected from hydrogen or hydroxyl; R 3 is selected from hydrogen or hydroxyl; R 4 is selected from keto, acetyl or acetoxy; R 5 Selected from hydroxyl or methoxy;

[0011] In Formula V: C-1 and C-2' are connected or not connected by a five-membered oxygen ring; C-4 and C-11 are connected or not connected by an oxygen bridge; R 1 is selected from hydrogen or acetyl; R 2 and R 3 is selected from hydrogen or hydroxyl; R 4 Selected from hydrogen, keto or =COHCH 3 ;

[0012] In Formula VI: R 1 is selected from keto or acetyl; R2 is selected from hydrogen or hydroxyl;

[0013] In Formula VII: C-4 and C-11 are connected by an oxygen bridge or not; R 1 and R 2 is selected from hydrogen or hydroxy.

[0014] Furthermore, the cadinane dimer sesquiterpene compound is selected from:

[0015]

[0016] Furthermore, the cadinane dimer sesquiterpene compound is selected from at least one of compounds 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.

[0017] The present invention also provides the use of a cadinane dimer sesquiterpene compound or a derivative thereof in preparing a product for preventing or treating diabetes or lowering blood sugar.

[0018] The present invention also provides the above-mentioned cadinane dimer sesquiterpene compound, whose general structural formula is shown in Formula I, Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII or Formula VIII:

[0019]

[0020] In Formula I: C-4 and C-11 are connected by an oxygen bridge or not; R 1 and R 2 is selected from hydrogen or hydroxyl;

[0021] In formula III: R1 Selected from methyl or -CH 2 OH;

[0022] In Formula IV: C-4 and C-11 are connected by an oxygen bridge or not; C-1' and C-2' are connected by a double bond or not; R 1 and R 2 is selected from hydrogen or hydroxyl; R 3 is selected from hydrogen or hydroxyl; R 4 is selected from keto, acetyl or acetoxy; R 5 Selected from hydroxyl or methoxy;

[0023] In Formula V: C-1 and C-2' are connected or not connected by a five-membered oxygen ring; C-4 and C-11 are connected or not connected by an oxygen bridge; R 1 is selected from hydrogen or acetyl; R 2 and R 3 is selected from hydrogen or hydroxyl; R 4 Selected from hydrogen, keto or =COHCH 3 ;

[0024] In Formula VI: R 1 is selected from keto or acetyl; R2 is selected from hydrogen or hydroxyl;

[0025] In Formula VII: C-4 and C-11 are connected by an oxygen bridge or not; R 1 and R 2 is selected from hydrogen or hydroxy.

[0026] Furthermore, the cadinane dimer sesquiterpene compound is selected from:

[0027]

[0028] Furthermore, the cadinane dimer sesquiterpene compound is selected from at least one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 14, 15, 16, 18, 19, 20, 21, and 23.

[0029] The present invention also provides a lipid-lowering or weight-loss composition, wherein the active ingredients of the composition include a cadinane dimer sesquiterpene compound or a derivative thereof.

[0030] Beneficial effects of the present invention:

[0031] The present invention provides a class of cadinane dimer sesquiterpene compounds that can significantly reduce the content of triglycerides in 3T3-L1 cells, and the cytotoxicity is relatively low. Among them, the abundant compound 24 shows good lipid-lowering and anti-obesity activities at the cell and animal levels: experiments have found that it can inhibit the expression of important transcription factors for adipocyte differentiation and the synthesis of related fat synthases, thereby inhibiting lipid synthesis and accumulation in 3T3-L1 cells; it can also significantly inhibit the increase in weight of obese mice induced by a high-fat diet without affecting the appetite of mice; it can significantly reduce fat accumulation in the liver, reduce the volume of fat cells in adipose tissue, and has no obvious toxicity to the liver and kidneys; it can significantly reduce blood lipids (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) in obese mice; it can also improve glucose tolerance damage caused by a high-fat diet, improve insulin sensitivity, reduce the increase in fasting blood sugar caused by obesity, and make it tend to normal. Caducane dimer sesquiterpenoid compounds have strong lipid-lowering and anti-obesity activities and good safety, and are expected to be developed into lipid-lowering, weight-loss and blood sugar-lowering products. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 Compound 24 affects the differentiation of 3T3-L1 cells into lipid-related factors and the transcription level of liposynthetic enzymes;

[0033] Figure 2 Compound 24 affects the differentiation of 3T3-L1 cells into lipid-related factors and lipase protein levels;

[0034] Figure 3 is a curve of the effect of compound 24 on the body weight of mice induced by a high-fat diet (HFD) over time;

[0035] Figure 4 This is a statistical graph of the daily food intake of mice;

[0036] Figure 5 This is a statistical chart of the weight of mouse organs (heart, lung, liver, spleen, and kidney);

[0037] Figure 6 It is a statistical chart of the adipose tissue weight of various parts of mice (inguinal white adipose IWAT, epididymal adipose EWAT, perirenal adipose PWAT);

[0038] Figure 7 This is a graph showing changes in glucose tolerance (OGTT) in mice;

[0039] Figure 8 This is the curve diagram of the change of insulin sensitivity (ITT) of mice;

[0040] Fig. 9 This is a statistical chart of fasting blood glucose values ​​in mice;

[0041] Fig.10 These are H&E staining images of mouse liver, gonadal fat and kidney tissue pathological sections, as well as Oil Red O staining images of liver tissue pathological sections. DETAILED DESCRIPTION

[0042] The specific embodiments of the present invention are further described below in conjunction with the accompanying drawings. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0043] Example 1

[0044] Application of a cadinane dimer sesquiterpene compound or a derivative thereof in preparing a lipid-lowering or weight-loss composition.

[0045] Furthermore, the cadinane dimer sesquiterpene compound is of C-2 and C-2' connection type.

[0046] Furthermore, the general structural formula of the cadinane dimer sesquiterpene compound is as shown in Formula I, Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII or Formula VIII:

[0047]

[0048] In Formula I: C-4 and C-11 are connected by an oxygen bridge or not; R 1 and R 2 is selected from hydrogen or hydroxyl;

[0049] In formula III: R 1 Selected from methyl or -CH 2 OH;

[0050] In Formula IV: C-4 and C-11 are connected by an oxygen bridge or not; C-1' and C-2' are connected by a double bond or not; R 1 and R 2 is selected from hydrogen or hydroxyl; R 3 is selected from hydrogen or hydroxyl; R 4 is selected from keto, acetyl or acetoxy; R 5 Selected from hydroxyl or methoxy;

[0051] In Formula V: C-1 and C-2' are connected or not connected by a five-membered oxygen ring; C-4 and C-11 are connected or not connected by an oxygen bridge; R 1 is selected from hydrogen or acetyl; R 2 and R 3 is selected from hydrogen or hydroxyl; R 4 Selected from hydrogen, keto or =COHCH3 ;

[0052] In Formula VI: R 1 is selected from keto or acetyl; R2 is selected from hydrogen or hydroxyl;

[0053] In Formula VII: C-4 and C-11 are connected by an oxygen bridge or not; R 1 and R 2 is selected from hydrogen or hydroxy.

[0054] Furthermore, the cadinane dimer sesquiterpene compound is selected from:

[0055]

[0056] Furthermore, the cadinane dimer sesquiterpene compound is selected from at least one of compounds 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.

[0057] Example 2

[0058] The present invention also provides the use of a cadinane dimer sesquiterpene compound or a derivative thereof in preparing a product for preventing or treating diabetes or lowering blood sugar.

[0059] Example 3

[0060] The present invention also provides the above-mentioned cadinane dimer sesquiterpene compound, whose general structural formula is shown in Formula I, Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII or Formula VIII:

[0061]

[0062] In Formula I: C-4 and C-11 are connected by an oxygen bridge or not; R 1 and R 2 is selected from hydrogen or hydroxyl;

[0063] In formula III: R 1 Selected from methyl or -CH 2 OH;

[0064] In Formula IV: C-4 and C-11 are connected by an oxygen bridge or not; C-1' and C-2' are connected by a double bond or not; R 1 and R 2 is selected from hydrogen or hydroxyl; R 3 is selected from hydrogen or hydroxyl; R 4 is selected from keto, acetyl or acetoxy; R 5 Selected from hydroxyl or methoxy;

[0065] In Formula V: C-1 and C-2' are connected or not connected by a five-membered oxygen ring; C-4 and C-11 are connected or not connected by an oxygen bridge; R 1 is selected from hydrogen or acetyl; R 2 and R 3 is selected from hydrogen or hydroxyl; R 4 Selected from hydrogen, keto or =COHCH 3 ;

[0066] In Formula VI: R 1 is selected from keto or acetyl; R2 is selected from hydrogen or hydroxyl;

[0067] In Formula VII: C-4 and C-11 are connected by an oxygen bridge or not; R 1 and R 2 is selected from hydrogen or hydroxy.

[0068] Furthermore, the cadinane dimer sesquiterpene compound is selected from:

[0069]

[0070] Furthermore, the cadinane dimer sesquiterpene compound is selected from at least one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 14, 15, 16, 18, 19, 20, 21, and 23.

[0071] Example 4

[0072] The present invention also provides a lipid-lowering or weight-loss composition, wherein the active ingredients of the composition include a cadinane dimer sesquiterpene compound or a derivative thereof.

[0073] Example 5

[0074] Preparation and structural identification of cadinane dimer sesquiterpenoids

[0075] Chemical research equipment and reagents: NMR data were collected using Bruker AVANCE-500and / or-800MHz NMRspectrometers; mass spectrometry data were collected using Q-TOF Micro LC-MS / MS spectrometer; infrared data were collected using Bruker Tensor-27infrared spectrometer (KBr wafer coating); optical rotation data were collected using JASCOP-1020polarimeter (room temperature); UV data were collected using Shimadzu UV-2401A instrument; high performance liquid chromatography columns YMC-Pack ODS-A (Vimex, Japan) and Lux@Amylose-1 (Phenomenon, USA) (both specifications are 300×10mm, S-5μm); column chromatography silica gel (300-400 mesh, Qingdao Ocean Chemical Plant); GF254TLC thin layer chromatography silica gel plate (Qingdao Ocean Chemical Plant); dextran gel Sephadex LH-20gel (40-70 μmol / L, GE, USA); C-18 reverse phase material RP-18 (150-200 mesh, Merck, Germany); all solvents used (analytical grade and chromatographic grade) were purchased from Shanghai Chemical Reagent Co., Ltd.

[0076] Pharmacological research equipment and reagents: CO 2Incubator (Thermo), inverted microscope (Leica), multifunctional microplate reader (Thermo), high-speed refrigerated centrifuge (Eppendorf), electrophoresis and membrane transfer device (Bio-Rad), qPCR instrument (Bio-Rad), cell culture dish / cell culture plate (Corning), 0.22 μmol / L syringe filter (Millipore), mouse surgical instruments (Shanghai Jinzhong), sodium heparin anticoagulant tube (Shanghai Yuanye Biotechnology), electronic balance (Sartorius), pipette (Eppendorf), blood glucose meter (Roche). DMEM culture medium (Gibco), 0.25% EDTA-Typsin (Gibco), fetal bovine serum (BI), penicillin-streptomycin (Gibco), 3-isobutyl-1-methylxanthine (Sigma), dexamethasone (Sigma), insulin (Sigma), rosiglitazone (Sigma), DMSO (Sigma), Oil Red O (Sigma), 4% paraformaldehyde (Biosharp), triglyceride quantitative detection kit (Nanjing Jiancheng Biological), RIPA lysis buffer (Biyuntian Biological), PMSF (Biyuntian Biological), RNA extraction kit (Tiangen), RNA reverse transcription kit (Takara), BCA protein quantification kit (Biyuntian Biological), antibody (Cell signaling technology), ECL developer (Biyuntian Biological), isopropanol (Sinopharm), glucose (Sinopharm), insulin injection (Novo Nordisk).

[0077] The whole plant of dried ginger notoginseng is taken, dried and crushed, and 95% ethanol is added to extract at room temperature for 3 times, the amount of ethanol added each time is 3 to 5 times the volume of the raw material, the extraction time is 7 days each time, the 3 extracts are combined and concentrated under reduced pressure to obtain ginger notoginseng ethanol extract; the extract is suspended and dissolved with 5 times the volume of distilled water, and an equal volume of ethyl acetate solvent is added to extract 3 times, the ethyl acetate extract is combined and concentrated under reduced pressure to recover the solvent, and ginger notoginseng ethyl acetate extract is obtained. The ethyl acetate extract is segmented using a D101 macroporous resin column, and gradient elution is performed with ethanol-water mixed solvents with volume ratios of 1:1, 4:1, and 19:1 in sequence, and the corresponding proportions of eluents are collected respectively, and the solvent is removed by reduced pressure concentration to obtain 3 elution components in sequence: 50% ethanol macroporous phase, 80% ethanol macroporous phase, and 95% ethanol macroporous phase.

[0078] The above-mentioned medium polar 80% ethanol macroporous phase extract was taken and preliminarily segmented using petroleum ether: ethyl acetate by silica gel column chromatography, and then further separated using RP-18, gel Sephadex LH-20 and silica gel column, and finally purified by semi-preparative high performance liquid chromatography under acetonitrile / water or methanol / water solvent conditions to obtain 25 compounds. The racemic compound was separated using a chiral column under n-hexane / isopropanol solvent conditions. Through structural analysis and identification, it was found that except for compounds 12, 13, 17, 22, 24 and 25 which were known compounds, the rest were new compounds.

[0079] Some physical and physicochemical data of the other new compounds are as follows:

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086] Example 6

[0087] In vitro lipid-lowering activity test of cadinane dimer sesquiterpenoids

[0088] 1. Inhibition of triglyceride (TG) activity detection:

[0089] (1) Culture medium and reagents used in the experiment:

[0090] Basic culture medium: DMEM high-glucose culture containing 10% fetal bovine serum and 1% penicillin-streptomycin.

[0091] Cell differentiation induction solution I: 500 μmol / L 3-isobutyl-1-methylxanthine (IBMX), 100 ng / mL dexamethasone, 2 μg / mL insulin, 1 μmol / L rosiglitazone, 10% fetal bovine serum, and 1% p / s DMEM culture medium.

[0092] Cell differentiation induction medium II: DMEM culture medium containing 2 μg / mL insulin, 10% fetal bovine serum and 1% p / s.

[0093] (2) Experimental group settings:

[0094] 3T3-L1 cells in the logarithmic growth phase were plated, 20,000 cells were inoculated per well of a 48-well plate, and cultured in basal medium at 5% CO2 and 37°C conditions; when the cells grew to 100% complete confluence, an undifferentiated control group (UND), a differentiated control group (Vehicle) and a drug treatment group (25 compounds, each with 6 concentration gradients: 20, 10, 5, 2.5, 1.25, 0.625 μmol / L) were set up and cultured for 9 days respectively.

[0095] Undifferentiated control group (UND): The cells were cultured with basal medium from day 1 to day 9, and the medium was changed every three days.

[0096] Differentiation control group (Vehicle): on days 1-3, use cell differentiation induction solution I; on days 4-6, use cell differentiation induction solution II; on days 7-9, use basal culture medium.

[0097] Drug treatment group: On days 1-3, cells were cultured with different concentrations (20, 10, 5, 2.5, 1.25, 0.625 μmol / L) of the compound (drug solution) prepared in cell differentiation induction solution I; on days 4-6, cells were cultured with different concentrations (20, 10, 5, 2.5, 1.25, 0.625 μmol / L) of the compound (drug solution) prepared in cell differentiation induction solution II; on days 7-9, cells were cultured with different concentrations (20, 10, 5, 2.5, 1.25, 0.625 μmol / L) of the compound (drug solution) prepared in basal culture medium.

[0098] After the cell culture is completed, discard the culture medium, gently rinse once with PBS, add 50μL of 2% Triton X-100 (diluted with ddH2O) to each well, and lyse for 40-60 minutes until the cells are completely lysed. Take a 96-well plate, add 200μL of TG working solution to each well, and then add 10μL of cell sample and 2μL of standard to each well. Set up a blank group, add 200μL of working solution and 10μL of 2% Triton X-100, and mix well. Incubate at 37℃ for 10-15 minutes, and measure OD 510 Absorbance value. The protein concentration of the cell sample was measured with the BCA protein quantification kit, and the TG content was calculated according to the following formula (the standard concentration was 2.26mmol / L). Where: TG content (mmol / gprot) = [(sample OD value - blank OD value) / (calibrator OD value - blank OD value)] × calibration concentration (mmol / L) ÷ protein concentration of the sample to be tested (gprot / L). Finally, Graphpad software was used to calculate EC 50 Value (half effective concentration that exerts lipid-lowering activity in 3T3-L1 cells).

[0099] 2. Cytotoxicity Assay:

[0100] When the density of 3T3-L1 preadipocytes reaches 80%-90%, the plates are inoculated. 5000 3T3-L1 cells are inoculated in each well of a 96-well plate, and the culture system is 100 μL. After 24-36 hours of culture, different concentrations (100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78 μmol / L) of compounds prepared in the culture medium are given after the cells adhere to the wall, and the control group is treated with an equal volume of DMSO. After 48 hours of administration, the supernatant is discarded, and a 0.5 mg / mL MTT solution prepared in serum-free DMEM culture medium is added (5 mg / mL MTT stock solution is obtained by dissolving 50 mg MTT powder in 10 mL 1×PBS (pH 7.4) buffer and mixing thoroughly, and stored at 4°C in the dark), incubated at 37°C for 4 hours, then the supernatant is discarded and 100 μL of DMSO is added to dissolve the formazan precipitate, and the OD is detected. 510 Absorbance value. Inhibition rate of the compound in the drug-added group = 1-OD value of the drug-added group / OD value of the control group. Finally, set up gradient concentration rescreening and use Graphpad software to calculate the half-inhibitory concentration IC 50 Value (half-maximal inhibitory concentration of the compound on 3T3-L1 cell viability).

[0101] The calculated lipid-lowering activities (EC 50 ) and cytotoxic activity (IC 50 ) The result is as follows:

[0102]

[0103] Where SI is the selection index, and its value is IC 50 / EC 50 .

[0104] From the above research results, it can be concluded that among the 25 cadinane-type dimerized sesquiterpenes, except for compounds 3 and 13 (EC 50 >20μmol / L), and the EC of the remaining compounds 50 The value is 1.04~18.64μmol / L, and the selection index SI (IC 50 / EC 50 The calculated values) were all greater than 2.03, indicating that they could significantly reduce the triglyceride content in 3T3-L1 cells and had relatively low cytotoxicity. Among them, compound (+)-4 showed the best lipid-lowering activity (EC 50 =1.04±0.34μmol / L) and had no significant toxicity to cells (IC 50 >100μmol / L). Considering the sample quantity and the factors of the same type of skeleton, the abundant compound 24 also has very good lipid-lowering activity in 3T3-L1 cells (optically pure monomer EC 50The values ​​were 3.19±0.47 and 3.68±1.80 μmol / L, respectively), and the toxicity was also low (SI selection index was greater than 3.66). Therefore, compound 24 was used in subsequent efficacy evaluation experiments.

[0105] Determination of the effect of compound 24 on the differentiation of 3T3-L1 cells into lipid-related factors and the transcription level of liposynthetic enzymes:

[0106] The effect of compound 24 on the mRNA transcription level of 3T3-L1 cell differentiation-related factors was detected: 3T3-L1 cells were treated with different concentrations of compound 24 (1 μmol / L and 10 μmol / L) for 3 days and 6 days, and total RNA was extracted from the cells. The transcription expression level of related genes was detected by real-time fluorescence quantitative PCR (RT-qPCR). The test results are shown in Figure 2. Figure 1 As shown, * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001.

[0107] The data results showed that compound 24 could down-regulate the gene expression levels of adipose differentiation factors (PPARγ, C / EBPα) and related fat synthesis enzymes (FASN, ACACA) in a concentration gradient-dependent manner.

[0108] Determination of the effect of compound 24 on the differentiation of 3T3-L1 cells into lipid-related factors and lipase protein levels:

[0109] The effect of compound 24 on the mRNA transcription level of 3T3-L1 cell differentiation-related factors was detected: 3T3-L1 cells were treated with different concentrations of compound 24 (5 μmol / L and 10 μmol / L) for 3 days and 6 days, and then the total cell protein was extracted. The protein expression level of related genes was detected by Western-blot. The test results are as follows: Figure 2 As shown, * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001.

[0110] The data results show that compound 24 can downregulate the expression of adipose differentiation factors (PPARγ, C / EBPα) and related fat synthesis enzymes (FASN, ACC) at the protein level in a concentration gradient-dependent manner. This is consistent with the result that compound 24 can reduce triglyceride levels, indicating that compound 24 inhibits lipid synthesis and accumulation in 3T3-L1 cells by inhibiting the expression of important transcription factors for cell differentiation and the synthesis of related fat synthesis enzymes.

[0111] Example 7

[0112] In vivo anti-obesity activity study of cadinane dimer sesquiterpenoid compound 24

[0113] 40 C57BL / 6 mice were randomly divided into 3 groups (10 mice in each group) after one week of adaptive feeding: normal diet group (Normal diet), obesity model group (HFD diet), compound 24 treatment group (administered at a dose of 40 mg / kg according to the weight of mice) (HFD+24). Each group of mice had free access to water, the normal diet group was given maintenance feed, and the other groups were given 60% high-fat feed for continuous feeding for 8-9 weeks (at this time, the average weight of mice in the high-fat diet group exceeded that of the normal diet group by more than 20%); the compound 24 treatment group was given compound 24 (administered at a dose of 40 mg / kg according to the weight of mice), and the administration was carried out by gavage once every two days for 40 days. During the entire experiment, the normal diet group and the obesity model group were gavaged with an equal volume of 0.5% CMC-Na solution as the experimental control. The weight and food intake of the mice were recorded every 2 days starting from the day of administration, and the effect of the compound on the weight of obese mice was observed. The curve of weight change over time is shown in Figure 3 As shown, *** represents P < 0.001, ### represents P < 0.001; the statistical results of the daily food intake of mice are shown in Figure 4 As shown, NS means not significant.

[0114] from Figure 3 and Figure 4 The data show that compound 24 can significantly inhibit the weight gain of obese mice induced by a high-fat diet; compared with the mice in the HFD control group, the mice lost 29.32% of their weight after 40 days of administration at a dose of 40 mg / kg; and it did not affect the appetite of the mice.

[0115] After the experiment, the anti-obesity activity of compound 24 was evaluated and tested:

[0116] ① To confirm whether compound 24 can effectively reduce the fat content in mice, the adipose tissues of the main organs of mice: heart, lung, liver, spleen, kidney and various parts (inguinal white adipose tissue IWAT, epididymal adipose tissue EWAT, perirenal adipose tissue PWAT) were separated and weighed. The data results of the main organs are as follows Figure 5 As shown, ### represents P<0.001. Data results of adipose tissue in various parts Figure 6 shown.

[0117] The experimental results showed that compound 24 could inhibit the increase of liver mass induced by high-fat diet in mice, but had no significant effect on the heart, kidney and brain of mice. Figure 5); At the same time, compound 24 can effectively reduce the content of adipose tissue in various parts of the mouse body, such as inguinal white fat IWAT, epididymal fat EWAT, perirenal fat PWAT ( Figure 6 ).

[0118] ② Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were performed 3 and 4 weeks after administration respectively:

[0119] OGTT test: The mice were fasted overnight but not water-deprived. According to the weight of the mice, they were given glucose by gavage at a dose of 2g / kg. The blood glucose levels of the mice were tested at 0min, 15min, 30min, 60min, 90min, and 120min to detect the changes in glucose tolerance of the mice. The data results are as follows Figure 7 As shown, *** represents P < 0.001, # represents P < 0.05, ### represents P < 0.001, and NS means not significant.

[0120] ITT test: After fasting for 6 hours, each mouse was given 0.6U insulin intraperitoneally, and the blood glucose levels of the mice were tested at 0min, 15min, 30min, 60min, 90min, and 120min to detect changes in the insulin sensitivity of the mice. The data results are as follows: Figure 8 As shown, *** represents P<0.001, # represents P<0.05, ## represents P<0.01, and ### represents P<0.001. The fasting blood glucose level of mice was measured, and the data results are shown in Fig. 9 As shown, ## represents P < 0.01.

[0121] The experimental results showed that compound 24 could improve the glucose tolerance impairment caused by high-fat diet ( Figure 7 ), and improve insulin sensitivity ( Figure 8 ), reduce the increase of fasting blood sugar caused by obesity and make it tend to normal ( Fig. 9 ).

[0122] ③ On the day of the experiment, the mouse liver (Liver), gonadal fat (Fat) and kidney tissue (Kidney) obtained by dissection were fixed with 4% paraformaldehyde, and the liver and kidney were stained with H&E to evaluate the toxicity of compound 24 on the liver and kidney; the gonadal fat tissue was stained with H&E to evaluate the effect of compound 24 on fat accumulation; the liver tissue was stained with Oil Red O to evaluate the effect of compound 24 on liver fat accumulation. The staining data results are shown in Fig.10 shown.

[0123] The experimental results showed that compound 24 could significantly reduce fat accumulation in the liver and the volume of fat cells in adipose tissue; compound 24 had no obvious toxicity to the liver and kidneys, and the liver and kidney tissue morphology of the drug-treated group was similar to that of the normal diet group (Normal diet), with no obvious lesions.

[0124] ④ Blood biochemical index detection:

[0125] Mice were fasted 6 hours before the end of the animal experiment. Blood was collected by removing the eyeballs, and the blood was collected in 1.5 mL sodium heparin anticoagulant tubes and stored on ice. Centrifuged at 4°C, 3000 rpm / min for 15 min, and the supernatant was retained and centrifuged again. The levels of triglycerides (TG), cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine (CREA) and urea nitrogen (UREA) in serum were detected by biochemical automatic analyzer to evaluate the therapeutic effect and toxicity of compound 24 on obesity-related metabolic syndrome.

[0126] The test results of mouse serum biochemical indexes are shown in the following table:

[0127] index Normal diet HFD diet HFD+24 (40 mg / kg) TG(mM / L) 0.77±0.06 1.49±0.18 0.82±0.16 TC(mM / L) 2.38±0.10 6.85±0.56 4.51±0.31 HDL-C (mM / L) 1.74±0.07 4.67±0.78 3.41±0.35 LDL-C (mM / L) 0.14±0.02 0.43±0.10 0.26±0.05 AST (IU / L) 144.10±15.65 176.00±19.87 154.10±18.12 ALT (IU / L) 34.33±3.90 33.63±3.55 31.42±3.30 CREA(mM / L) 17.67±2.50 15.45±1.47 16.22±3.81 UREA(mM / L) 10.27±1.58 9.75±0.51 9.36±2.16

[0128] The results showed that compound 24 could significantly reduce blood lipids (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) in obese mice; the levels of ALT and AST, the liver function indicators, CREA and UREA in the treatment group were not significantly different from those in the normal group, indicating that compound 24 had no significant effect on liver function indicators alanine aminotransferase (ALT) and aspartate aminotransferase (AST); and had no significant effect on renal function indicators creatinine (CREA) and urea nitrogen (UREA). This research data is consistent with the H&E staining results of the liver and kidney sections above.

[0129] In summary, cadinane dimer sesquiterpenoid compounds including compound 24 have good anti-obesity activity and good safety, and are expected to be developed into new lipid-lowering, weight-loss or blood sugar-lowering products.

[0130] The embodiments of the present invention are described in detail above with reference to the accompanying drawings, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the scope of protection of the present invention.

Claims

1. The use of cadinane dimer sesquiterpene compounds in the preparation of lipid-lowering or weight-loss products, characterized in that: The cadinane dimer sesquiterpene compound is selected from:

2. The use according to claim 1, characterized in that: The cadinane dimer sesquiterpene compound is selected from at least one of compounds 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.

3. The use of cadinane dimer sesquiterpene compounds in the preparation of products for preventing or treating diabetes and lowering blood sugar, characterized in that: The cadinane dimer sesquiterpene compound is selected from:

4. A cadinane dimer sesquiterpene compound, characterized in that: Selected from:

5. The cadinane dimer sesquiterpene compound according to claim 4, characterized in that: The cadinane dimer sesquiterpene compound is selected from at least one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 14, 15, 16, 18, 19, 20, 21, and 23.

6. A lipid-lowering or weight-loss composition, characterized in that: The active ingredient of the composition comprises the cadinane dimer sesquiterpene compound according to claim 4 or 5.

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