Triterpenoid compounds and preparation methods and applications thereof

Abstract

The present invention relates to the field of medicine, and specifically to new triterpenoid compounds (structures shown in the following formulas 1 to 3) isolated from the branches and leaves of the genus Pinaceae, a plant of the genus Pinaceae, and their preparation methods and uses. Compound 1 is a lanolin triterpene, compound 2 is a methyl-rearranged lanolin triterpene, and compound 3 is a methyl-rearranged product of a cycloartane triterpene, belonging to a cycloartane triterpene. The above-mentioned new triterpene compounds all form an identical furanic acid structural fragment in the side chain. Multiple in vitro activity tests have shown that this triterpene compound has significant ATP-citrate lyase inhibitory activity and can be used in the preparation of drugs for preventing, delaying or treating ACL-mediated glucose and lipid metabolism disorders and other related diseases. The present invention can also provide a lead compound for the development of new drugs for diseases related to glucose and lipid metabolism disorders.

Description

Technical Field

[0001] The present application relates to the field of medical technology, specifically to a Keteleeria ) Plant tung tree[ Keteleeria fortunei A type of triterpenoid compound with furanic acid structural fragment formed in the side chain is isolated from Murr. Carr.] and its preparation method and use. The present invention also relates to the use of the compound in the preparation of drugs for preventing or curing ATP-citrate lyase (ACL)-mediated glucose and lipid metabolism disorder-related diseases (such as hyperlipidemia, non-alcoholic fatty liver disease, type II diabetes, etc.) or the lead compound of the drug. Background Art

[0002] ATP-citrate lyase (ACL) is a key enzyme in glucose metabolism and the biosynthesis of fatty acids and cholesterol. The human ACL enzyme protein is a homotetramer composed of four identical 120 KDa subunits, each polypeptide chain containing 1101 amino residues (Singh et al, J. Biol. Chem. ACL can hydrolyze citrate produced and transported from mitochondria into acetyl-CoA and oxaloacetate (Beigneux et al., J. Biol. Chem. 2004, 279:9557−9564.). Acetyl-CoA is the substrate for the de novo synthesis of cholesterol, and cholesterol is generated through 33 steps of reaction. It can be seen that ACL is closely related to the synthesis of fatty acids and cholesterol, and changes in its expression are related to hyperlipidemia and the cardiovascular diseases (CVDs) caused by it (Granchi, Expert Opin Ther Pat. 2022, 32(7), 731–742; Verbark et al, Trends Mol. Med . 2021, 27(12), 1095–1105; Burke et al., Curr. Opin. Lipidol. 2017,28:193−200.) At the same time, abnormal triglyceride metabolism also increases the risk factors for lipid metabolism disorders such as nonalcoholic fatty liver disease (NAFLD) and type II diabetes mellitus (T2DM). Therefore, ACL can also be used as a potential therapeutic target for nonalcoholic fatty liver disease and type II diabetes (Morrow et al,Cell Metab . 2022, 7, 34(6), 919–936; Armstrong et al., Hepatology 2014, 59:1174−1197; Cohen et al, Science 2011, 332:1519−1523.). Studies have shown that ACL is also closely related to the occurrence of cancer. The increase in lipid synthesis provides essential lipids for cell growth and division, is one of the important signs of cancer occurrence, and is also an early event in tumor occurrence (Migita et al, Cancer Res. 2008, 68:8547−8554.). ACL is the main source of acetyl-CoA (an important component of de novo fatty acid synthesis). Inhibiting its activity can significantly inhibit the proliferation of tumor cells and induce apoptosis. Therefore, ACL has also been widely studied as a potential anti-cancer target in recent years (Singh et al, Oncogene 2021, 40, 592–602; Singh et al, Oncogene 2021, 40, 592–602; Huang et al, Eur. J. Med. Chem. 2021, 212,113036; Granchi et al., Eur. J. Med. Chem. 2018, 157:1276−1291.).

[0003] ACL, as a new drug target, has become a hot topic in the research of innovative drugs for lipid metabolism disorders in recent years. With the widespread use of high-throughput screening technology, a large number of ACL small molecule inhibitors have been discovered. However, only bempedoic acid, a small molecule ACL inhibitor developed by Esperion Therapeutics (ESPR) in the United States, has been approved by the FDA for the treatment of hypercholesterolemia and atherosclerotic cardiovascular disease. Except for bempedoic acid, the development of other ACL inhibitors is limited due to their low cell penetration, low affinity with ACL, and weak specificity. Therefore, it is of great significance to find small molecule ACL inhibitors with high efficiency, high selectivity, and good pharmacokinetic properties, which have broad application prospects in the treatment of diseases such as cardiovascular diseases and cancer.

[0004] Natural products are characterized by structural complexity and diversity and are an important source for new drug discovery. The unique chemical structure of natural products and their derivatives gives them the advantages of low toxicity, high efficacy, high selectivity for specific targets, and potential unique mechanisms of action (Atanasov et al., Nat. Rev. Drug Discov.2021,20, 200–216; Newman et al., J. Nat. Prod. 2020, 83: 770–803; Tiago et al., Nat. Chem. 2016, 8: 531–541). Natural products are produced by long-term selection and evolution of natural laws. They can often effectively bind to biological macromolecules and can be considered as the elite compound library left after years of screening and elimination of biological systems highly related to human proteins. At the same time, single-target drugs often have poor efficacy, while Chinese medicine / plant medicine and its components have novel structures (chemical components), multi-target and multi-pathway characteristics, and have unique advantages in the treatment of complex diseases. Therefore, it is of great research value to find and develop new, efficient and low-toxic ACL inhibitors from chemical components of natural sources (especially plant sources). Natural products with unique sources and structural characteristics are expected to gain an advantage in the current fierce competition in the field of new drug research and development for lipid metabolism disorders. Statistical analysis of the natural sources of drug molecules showed that drug molecules derived from terrestrial plants are concentrated in certain families (active natural products come from 792 of the 6763 families in nature, accounting for only 11.7%), and marketed drugs and candidate drugs in clinical trials are concentrated in 144 of these families, while the vast majority of rare and endangered plants are clearly in these plant families that can produce drugs (Zhu et al., Proc. Natl. Acad. Sci. U. S. A . 2011, 108:12943–12948; Mohamed et al., Proc. Natl. Acad. Sci. 2013, 110:16832—16837.). It can be seen that the novelty and drugability of natural products from rare and endangered plants are much higher than the average level of general plants.

[0005] It is worth mentioning that Pinaceae is among the top 20 biological families that are most capable of producing drug molecules (Zhu et al., Proc. Natl. Acad. Sci. U.S.A. 2011, 108:12943–12948.). Keteleeria fortunei (Murr.) Carr.] belongs to the genus Pinaceae ( Keteleeria) plant, was included in the "Red Data of Chinese Plants - Rare and Endangered Plants" in 1992 and listed as "vulnerable species". If a small amount of samples of Chinese cypress plants (such as renewable branches and leaves) can be collected for protection and systematic research on their chemical composition, it will actively promote scientific understanding, active protection and comprehensive development and utilization of this rare plant, so that its resources can continue to serve mankind. The present invention is the first to isolate rare triterpenoid compounds (structures shown in Formulas 1 to 3) from the methanol extract of Chinese cypress branches and leaves. Compound 1 is a lanostane-type triterpene; Compound 2 is a methyl-rearranged lanostane-type triterpene (Shen et al., Bioorganic Med. Chem. Lett , 2001,3119–3122; Vieira et al., Phytochemistry , 2004, 393–398); Compound 3 is a friedo-cyloartane-type triterpene obtained by rearrangement of the methyl group of a cycloartane-type triterpene. This skeleton is very rare in nature and has only been reported in two papers (Corey et al., J. Am. Chem. Soc. , 1994, 116, 3149–3150; Nguyen et al., Phytochemistry , 2011, 72, 290–295.). Compounds 1, 2 and 3 all form a common furanic acid structural fragment on the side chain. Multiple pharmacological studies have shown that the compounds all have significant ACL inhibitory activity. Summary of the invention

[0006] The compound involved in the present invention is a triterpenoid compound with a furanic acid structural fragment formed in the side chain, which is isolated from the branches and leaves of the Chinese yew. It is found in the in vitro pharmacological activity screening that it has a good ACL inhibitory effect, and thus may have a therapeutic effect on hyperlipidemia, atherosclerosis, fatty liver, type II diabetes, cancer and other ACL-mediated diseases, and thus has great potential uses in the pharmaceutical field. Based on this, the present application provides a new use of triterpenoid compounds in the Chinese yew.

[0007] Another object of the present invention is to provide a novel triterpene compound with medicinal value and a method for extracting the compound of the present invention from the plant Lucerne fir.

[0008] A triterpenoid compound has the structural characteristics that: the triterpenoid compound is any one of compounds 1-3, wherein compound 1 is a lanolin triterpene, compound 2 is a methyl-rearranged lanolin triterpene, and compound 3 is a cycloartane triterpene obtained by methyl rearrangement of a cycloartane triterpene, and the side chain of the triterpene compound has a furanic acid structural fragment. The molecular structural formula 1-3 corresponding to the compound 1-3 is as follows:

[0009]

[0010] The present invention provides a method for preparing the triterpene compound, comprising the following steps:

[0011] S1: The leaves of the dried Chinese fir branches and leaves are dried at room temperature, crushed, extracted once or multiple times with a methanol-water solution with a concentration of more than 70% at room temperature, the extract is concentrated, suspended in water, extracted with petroleum ether, ethyl acetate and n-butanol in sequence, and four components are obtained: petroleum ether extract, ethyl acetate extract, n-butanol extract and water;

[0012] S2: The ethyl acetate extract is concentrated under reduced pressure and then subjected to silica gel column chromatography and MCI column chromatography in a gradient elution with an elution solvent, and then separated by Sephadex LH-20 gel and reversed-phase semi-preparative high performance liquid chromatography to obtain compounds 1-3 shown in formulas 1 to 3. The compounds of the present invention can be obtained by separation and purification from plants; they can also be synthesized by chemical methods well known to those skilled in the art.

[0013] Preferably, the methanol-water solution is a methanol-water solution with a concentration of 90%.

[0014] Preferably, the number of extractions with methanol-water solution at room temperature in S1 is 3 or more times, and may be 5 times. The time of extraction at room temperature is not particularly limited, and may be 12 hours or more per time.

[0015] Preferably, the ethyl acetate extract in S2 is concentrated under reduced pressure and then eluted by silica gel column chromatography with a gradient of petroleum ether-ethyl acetate (15:1 → 10:1 → 5:1 → 3:1 → 1:1 → 0:100, v / v), and then eluted with a gradient of ethyl acetate-methanol (5:1 → 1:1 → 0:100, v / v) to obtain 8 components (Fr.1-Fr.8) and then enter the MCI column chromatography stage.

[0016] Preferably, the elution component (Fr. 5) with a petroleum ether:ethyl acetate ratio of 5:1 is subjected to MCI column chromatography and gradient elution with a water:methanol volume ratio of 50:50-0:100, and the eluents are collected separately; then the elution component with a water:methanol volume ratio of 20:80 is sequentially subjected to Sephadex LH-20 gel chromatography (methanol elution) separation and reversed-phase semi-preparative high performance liquid chromatography separation (MeOH-H2O, 90:10, v / v; flow rate, 3 mL / min, t R = 16.0 min) and compound 1 was obtained.

[0017] Preferably, the elution component (Fr. 7) with a petroleum ether: ethyl acetate ratio of 3:1 is subjected to MCI column chromatography and gradient elution with a water: methanol volume ratio of 50:50-0:100, and the eluents are collected separately; then the elution component with a water: methanol volume ratio of 20:80 is sequentially subjected to Sephadex LH-20 gel chromatography (methanol elution) separation and reversed-phase semi-preparative high performance liquid chromatography separation (MeOH-H2O, 90:10, v / v; flow rate, 3 mL / min, t R = 16.0 min) to obtain compound 3.

[0018] Preferably, the elution component (Fr. 7) with a petroleum ether:ethyl acetate ratio of 3:1 is subjected to MCI column chromatography and gradient elution with a water:methanol volume ratio of 50:50-0:100, and the eluents are collected separately; then the elution component with a water:methanol volume ratio of 10:90 is sequentially subjected to Sephadex LH-20 gel chromatography (methanol elution) separation and reversed-phase semi-preparative high performance liquid chromatography separation (MeOH-H2O, 75:25, v / v; flow rate, 3 mL / min, t R = 22.6 min) and purified to give compound 2.

[0019] A further object of the present invention is to provide the use of the above-mentioned compound or composition in the preparation of drugs for diseases mediated by ACL. The use of a mixture of any one or more of the compounds 1-3 shown in Formula 1 to Formula 3 as a drug and / or health product for preventing, delaying or treating glucose and lipid metabolism disorders, complications of glucose and lipid metabolism disorders or other diseases mediated by ACL. The diseases related to glucose and lipid metabolism disorders include: hyperlipidemia, non-alcoholic steatohepatitis, type 2 diabetes and obesity, etc.

[0020] Preferably, any one or more of the compounds of formula 1 to formula 3 are used in the preparation of ATP-citrate lyase inhibitor drugs. The triterpene compounds of the present invention show a strong inhibitory effect in the ACL inhibitory activity experiment, and the activity is comparable to that of the positive control BMS303141.

[0021] Another aspect of the present invention provides a drug combination, which uses one or more of the above triterpene compounds as raw materials, contains a therapeutically effective amount of one or more selected from the above compounds as active ingredients, and the composition may further include pharmaceutically acceptable pharmaceutical excipients, such as carriers, excipients, adjuvants or diluents. The drug combination can be used to prepare drugs for preventing, delaying or treating ACL-mediated glycolipid disorder-related diseases (especially hyperlipidemia and related cardiovascular diseases) or as a lead compound for such drugs.

[0022] Based on the advantages of this type of compound in terms of novel chemical structure and significant biological activity, it has a good development prospect and is expected to develop into a novel therapeutic drug or lead compound for ACL-mediated diseases.

[0023] The present invention has the following significant advantages: Compound 1 is a lanostane-type triterpene, Compound 2 is a methyl-rearranged lanostane-type triterpene, and Compound 3 is a friedo-cyloartane-type triterpene obtained by methyl rearrangement of a cycloartane-type triterpene, and this skeleton is very rare in nature. Compounds 1, 2 and 3 all form a same furanic acid structural fragment in the side chain; and it is the first time that this type of triterpene compound has significant ACL inhibitory activity. This will have important application prospects for diseases related to glucose and lipid metabolism disorders that are common in modern people, such as hyperlipidemia, non-alcoholic fatty hepatitis, type II diabetes and the like. DETAILED DESCRIPTION

[0024] The following specific examples are used to further illustrate the preparation steps and pharmacological experimental processes of the compounds of the present invention. It should be understood that the following examples are only used to illustrate the present invention and are not used to limit the scope of the present invention, and those skilled in the art may make various modifications and changes thereto.

[0025] Chinese fir Keteleeria fortunei)The branches and leaves were collected from Kunming, Yunnan, dried in the shade and then crushed into powder; the specific rotation test was completed at 25℃ using a Rudolf Autopol IV polarimeter; Hitachi U-2900E UV spectrometer; Thermo Scientific Nicolet Is5 FT-IR infrared spectrometer; ECD spectrum was measured by a JASCO-810 CD spectrometer; ESI-MS was measured by an Agilent 1100 LC-MS instrument, and HR-ESIMS was measured by an AB Sciex TripleTOF 5600 instrument; the silica gel used was produced by Qingdao Ocean Chemical Company; the silica gel thin layer plate was produced by Yantai Jiangyou Silica Gel Development Co., Ltd., with a specification of GF254 / 0.25 mm; MCI gel CHP20P was produced by Mitsubishi Corporation of Japan, with a specification of 75-150 μ m; Sephadex LH-20 gel was produced by GE Healthcare Bio-Sciences, Switzerland; the semi-preparative HPLC was Shimadzu LC-20AT equipped with SPD-M20A PDA detector and Waters XBridge ODS and Cosmosil semi-preparative columns (250 × 10 mm, 5 μ m); All analytical grade reagents were produced by Shanghai Sinopharm Chemical Reagent Co., Ltd.; chromatographic grade solvents were produced by Shanghai Xingke High Purity Solvent Co., Ltd.; deuterated reagents were produced by Sigma-Aldrich.

[0026] Example 1: Preparation of the compounds of the present invention

[0027] After the dried leaves and branches of Chinese fir (10.0 kg) were fully crushed, they were extracted with 90% methanol (10 L) at room temperature for 5 times, each time for 24 hours. The extracts were combined and concentrated under reduced pressure to obtain a total extract of 2.66 kg. The extract was dispersed with 1 L of water and extracted three times with equal volumes of petroleum ether, ethyl acetate and n-butanol, respectively, to obtain four components: petroleum ether, ethyl acetate, n-butanol and water. The ethyl acetate component totaled 134.0 g and was subjected to silica gel column chromatography with a gradient elution of petroleum ether-ethyl acetate (15:1 → 10:1 → 5:1 → 3:1 → 1:1 → 0:100, v / v), followed by ethyl acetate-methanol (5:1 → 1:1 → 0:100) to obtain 8 components (Fr.1-Fr.8).

[0028] Component Fr.5 (15.6 g) was first chromatographed on an MCI column, and seven subcomponents (Fr.5A-Fr.5G) were obtained after gradient elution with methanol-water. Fr.5C was purified by Sephadex LH-20 (MeOH) and semi-preparative HPLC (MeOH-H2O, 88:12, v / v; flow rate, 3 mL / min, t R = 22.0 min) and purified to give compound 1.

[0029] Component Fr.7 (12.6 g) was first crudely fractionated by MCI column chromatography, and 11 subcomponents (Fr.7A-Fr.7K) were obtained by methanol-water gradient elution. Fr.7B was purified by Sephadex LH-20 (MeOH) and semi-preparative HPLC (MeOH-H2O, 90:10, v / v; flow rate, 3 mL / min, t R = 16.0 min) to obtain compound 3. Fr.7F was purified by Sephadex LH-20 (MeOH) and semi-preparative HPLC (MeOH-H2O, 75:25, v / v; flow rate, 3 mL / min, t R = 22.6 min) and purified to give compound 2.

[0030] Compound 1, its NMR and physicochemical data are as follows:

[0031] White powder; α ]25 D –14.8 ( c 0.4, MeOH); UV (MeOH) λ max (log ε ) 246(3.91) nm; 1 H NMR (600 MHz, pyridine— d 6) δ H : 3.66 (1H, t, J = 3.0 Hz, H—3),1.73 (1H, dd, J = 11.0, 6.8 Hz, H—5), 5.72 (1H, m, H—7), 2.37 (1H, dd, J =12.3, 6.8 Hz, H—9), 0.98 (3H, s, H—18), 1.13 (3H, s, H—19), 0.89 (3H, d, J= 6.5 Hz, H—21), 2.78 (1H, dd, J = 14.8, 3.6 Hz, H—22a), 2.30 (1H, dd, J =14.8, 9.6 Hz, H—22b), 6.82 (1H, s, H—24), 8.47 (1H, s, H—27), 1.25 (3H, s, H—28), 0.99 (3H, s, H—29), 1.03 (3H, s, H—30); 13 C NMR (600 MHz, pyridine— d 6) δ c : 30.5 (C—1), 26.9 (C—2), 75.2 (C—3), 37.9 (C—4), 43.0 (C—5), 28.9 (C—6), 122.2 (C—7), 149.0 (C—8), 49.1 (C—9), 36.0 (C—10), 23.2 (C—11), 23.5 (C—12), 43.8 (C—13), 53.0 (C—14), 33.7 (C—15), 35.2 (C—16), 53.6 (C—17), 23.8 (C—18), 24.8 (C—19), 36.5 (C—20), 18.7 (C—21), 34.7 (C—22), 157.3 HRESIMS m / z 467.3165 [M + H] + (calcd for C 30 H 45 O4, 467.3167, Δ = –0.3 ppm).

[0032] Compound 2, its NMR and physicochemical data are as follows:

[0033] Colorless, amorphous powder; [ α ]25 D +31.2 ( c 0.85, CHCl3); UV (MeOH) λ max(log ε ) 246 (3.93) nm; 1 H NMR (600 MHz, CDCl3) δ H : 4.68 (1H, br s, H—3),1.61 (1H, dd, J = 12.0, 5.0 Hz, H—5), 5.59 (1H, br s, H—7), 5.12 (1H, br s,H—15), 0.94 (3H, s, H—18), 1.00 (3H, s, H—19), 0.83 (3H, d, J = 6.8 Hz,H—21), 2.77 (1H, dd, J = 14.3, 3.2 Hz, H—22a), 2.28 (1H, dd, J = 14.3, 10.4Hz, H—22b), 6.37 (1H, s, H—24), 7.98 (1H, s, H—27), 1.01 (3H, s, H—28),0.95 (3H, s, H—29), 1.00 (3H, s, H—30), 2.08 (3H, s, H—2´); 13 C NMR (600MHz, CDCl3) δ c : 27.3 (C—1), 22.3 (C—2), 78.2 (C—3), 35.7 (C—4), 38.2 (C—5), 21.9 (C—6), 120.0 (C—7), 136.1 (C—8), 52.3 (C—9), 34.0 (C—10), 32.9(C—11), 25.6 (C—12), 51.2 (C—13), 152.2 (C—14), 114.5 (C—15), 44.6 (C—16), 50.2 (C—17), 15.9 (C—18), 21.7 (C—19), 37.0 (C—20), 15.1 (C—21),30.9 (C—22), 157.5 (C—23), 104.9 (C—24), 118.3 (C—25), 167.2 (C—26),146.9 (C—27), 27.8 (C—28), 22.6 (C—29), 23.2 (C—30), 170.1 (C—1´), 21.6(C—2´); HRESIMSm / z 507.3189 [M — H] — (calcd for C 32 H 43 O5, 507.3189, Δ = +0.1 ppm).

[0034] Compound 3, its NMR and physicochemical data are as follows:

[0035] Colorless, amorphous powder; [ α ]25 D +23.7 ( c 0.3, MeOH); UV (MeOH) λ max (log ε ) 246 (3.82) nm; 1 H NMR (600 MHz, CDCl3) δ H : 3.50 (1H, t, J = 3.2Hz, H—3), 1.80 (1H, dd, J = 12.1, 4.4 Hz, H—5), 5.46 (1H, dd, J = 8.0, 2.5Hz, H—12), 1.08 (3H, s, H—18), 0.20 (1H, d, J = 4.2 Hz, H—19a), 0.49 (1H,d, J = 4.2 Hz, H—19b), 0.83 (3H, d, J = 6.8 Hz, H—21), 2.97 (1H, dd, J =14.2, 1.6 Hz, H—22a), 2.70 (1H, dd, J = 16.5, 14.2 Hz, H—22b), 6.36 (1H, s,H—24), 7.97 (1H, s, H—27), 0.99 (3H, s, H—28), 0.89 (3H, s, H—29), 0.91 (3H, s, H—30); 13 C NMR (600 MHz, CDCl3) δ c: 28.3 (C—1), 26.3 (C—2), 77.1 (C—3), 39.2 (C—4), 41.1 (C—5), 19.7 (C—6), 25.5 (C—7), 50.2 (C—8), 28.3 (C—9), 26.8 (C—10), 31.4 (C—11), 115.8 (C—12), 159.3 (C—13), 46.5 (C—14), 37.6 (C—15), 32.9 (C—16), 49.1 (C—17), 25.6 (C—18), 27.8 (C—19), 38.1 (C—20), 14.9 (C-21), 30.8 (C-22), 158.4 HRESIMS m / z 465.3004 [M — H] — (calcd for C 30 H 41 O4, 465.3010, Δ = –1.3ppm).

[0036] Example 2: ACL inhibitory activity test of the compounds of the present invention

[0037] Experimental method: In this experiment, ACL can catalyze the conversion of citric acid into acetyl-CoA, thereby producing the precursor molecule for fatty acid synthesis - malonyl-CoA. This reaction is accompanied by the consumption of ATP, so ADP-Glo ​​and kinase detection kits are used to detect the changes in ATP to indirectly reflect the inhibitory effect of the compound on ACL enzyme activity. Specifically, the compound concentration of 20 µ The percentage inhibition rate of ACL enzyme activity was investigated at 100 g / ml. The test results showed that the above-mentioned compounds all had enzyme inhibition effects. Further determination of IC 50 Value: Dissolve the sample in DMSO to a suitable concentration before use, dilute 3 times, 7 dilutions, three replicate wells, take 2 μ 1 L sample solution was added to the standard activity assay system (40 mM Tris, pH 8.0, 10 mM MgCl2, 5 mM DTT, ATP, CoA, sodium citrate and ACL) and incubated at 37°C for 30 min. μL ADP-Glo ​​reagent was added and incubated at room temperature for 30 min to terminate the reaction and consume the remaining ATP. After adding kinase detection reagent and incubating for 30 min, the fluorescence signal was read by EnVision, and the slope of the first-order reaction of the kinetic curve was used as the activity index of the enzyme. The relative activity was plotted against the compound concentration, and the formula v / v 0 = 100 / (1+ b *[I] / IC 50 ) Fitting to get IC 50 The experiment was repeated three times and the results were averaged. The IC value of the positive control BMS303141 50 The value is 0.46 ± 0.2 µ M, the results are shown in Table 1.

[0038] Table 1. ACL inhibitory activity data of triterpenoids from Cotton Tree

[0039] Compound <![CDATA[IC 50 ( µ M)]]> 1 8.8 ± 0.9 2 5.7 ± 1.4 3 6.6 ± 0.2

[0040] The above test results show that the new triterpenoid compounds described in this patent, each of which has three side chains forming a furanic acid structural fragment, all exhibit significant inhibitory activity against ACL, indicating that the compounds described in the present invention can be used to prepare drugs for treating diseases related to lipid metabolism disorders or as lead compounds for such drugs.

Claims

1. A triterpenoid compound, characterized in that: Its structural features are: The triterpenoid compound is any one of compounds 1-3, wherein compound 1 is a lanolin triterpene, compound 2 is a methyl-rearranged lanolin triterpene, and compound 3 is a cycloartane triterpene obtained by methyl rearrangement of a cycloartane triterpene. The side chain of the triterpene compound has a furanic acid structural fragment. The molecular structural formula 1-3 corresponding to the compound 1-3 is as follows:

2. The method for preparing triterpenoids according to claim 1, comprising the following steps: S1: drying and crushing the dried branches and leaves of Chinese fir at room temperature, extracting them once or multiple times with a methanol-water solution with a concentration of more than 70% at room temperature, concentrating the extract, suspending it in water, and extracting it with petroleum ether and ethyl acetate in sequence to obtain a petroleum ether extract and an ethyl acetate extract; S2: The ethyl acetate extract was concentrated under reduced pressure and then subjected to silica gel column chromatography and MCI column chromatography in a gradient elution with an elution solvent, and then separated by Sephadex LH-20 gel and reversed-phase semi-preparative high performance liquid chromatography to obtain compounds 1-3 represented by formulas 1 to 3; The ethyl acetate extract was concentrated under reduced pressure and then eluted by silica gel column chromatography with a gradient of petroleum ether-ethyl acetate 15:1 → 10:1 → 5:1 → 3:1 → 1:1 → 0:100, v / v, and then with a gradient of ethyl acetate-methanol 5:1 → 1:1 → 0:100, v / v, to obtain 8 components Fr.1-Fr.8, and then entered the MCI column chromatography stage; The fraction Fr. 5 with a petroleum ether:ethyl acetate ratio of 5:1 was subjected to MCI column chromatography and gradient eluted with a water:methanol volume ratio of 50:50-0:100, and the eluents were collected separately; then the fraction with a water:methanol volume ratio of 20:80 was subjected to Sephadex LH-20 gel chromatography with methanol elution and reversed-phase semi-preparative high-performance liquid chromatography with MeOH-H2O, 88:12, v / v; flow rate, 3 mL / min, t R = Compound 1 was obtained after 22.0 min; The fraction Fr. 7 eluted with petroleum ether: ethyl acetate in a ratio of 3:1 was subjected to MCI column chromatography and gradient eluted with a water: methanol volume ratio of 50:50-0:100, and the eluents were collected separately; then the fraction eluted with a water: methanol volume ratio of 10:90 was subjected to Sephadex LH-20 gel chromatography with methanol elution and reverse phase semi-preparative high performance liquid chromatography with MeOH-H2O, 75:25, v / v; flow rate, 3 mL / min, t R = 22.6 min After purification, compound 2 was obtained; The fraction Fr. 7 eluted with petroleum ether: ethyl acetate in a ratio of 3:1 was subjected to MCI column chromatography and gradient eluted with a water: methanol volume ratio of 50:50-0:100, and the eluents were collected separately; then the fraction eluted with a water: methanol volume ratio of 20:80 was subjected to Sephadex LH-20 gel chromatography with methanol elution and reversed-phase semi-preparative high-performance liquid chromatography with MeOH-H2O, 90:10, v / v; flow rate, 3 mL / min, t R = 16.0 min later, compound 3 was obtained.

3. The method for preparing triterpenoids according to claim 2, characterized in that: The methanol-water solution described in S1 is a methanol-water solution with a concentration of 90%, and the number of extractions with the methanol-water solution at room temperature is more than 3 times.

4. The use of triterpenoid compounds according to claim 1, characterized in that: A mixture of any one or more of the compounds 1-3 represented by Formula 1 to Formula 3 is used for preparing drugs and / or health products for preventing, delaying or treating glucose and lipid metabolism disorders, complications of glucose and lipid metabolism disorders or other diseases mediated by ACL.

5. The use of triterpenoid compounds according to claim 1, characterized in that: Any one or more of the compounds of formula 1 to formula 3 are used in the preparation of ATP-citrate lyase inhibitor drugs.

6. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises a mixture of any one or more of compounds 1-3 represented by formula 1 to formula 3 as claimed in claim 1 as an active ingredient, and the pharmaceutical composition is used to prepare a drug for preventing, delaying or treating ACL-mediated glycolipid disorder-related diseases.

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