Terpenoid, application and preparation method thereof, and pharmaceutical composition

Abstract

The invention discloses a terpenoid and application and a preparation method thereof and a pharmaceutical composition, and relates to the technical field of medicine, the terpenoid comprises a terpenoid and / or pharmaceutically acceptable salt thereof, and the terpenoid comprises at least one of a structural formula (I), a structural formula (II) and a structural formula (III). The three new terpenoids, namely, the fargesisin A (I), the fargesisin B (II) and the fargesisin C (III), are extracted and separated from branches and leaves of torreya grandis for the first time, all have remarkable inhibition effects on ACL activity, can be used as ACL inhibitors, and can be used for preparing the novel terpenoids, namely, the fargesisin A (I), the fargesisin B (II) and the fargesisin C (III). The compound has a wide application prospect in preparation of drugs for preventing, delaying or treating ACL-mediated related diseases such as hyperlipidemia, atherosclerosis, non-alcoholic fatty liver disease, type 2 diabetes, obesity and cancer.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a terpene compound and an application and preparation method thereof, and a pharmaceutical composition. Background Art

[0002] ATP citrate lyase (ACL) is a key enzyme in cell metabolism, the main source of acetyl-CoA, an important precursor for the biosynthesis of fatty acids, cholesterol and isoprenoids, and is also involved in protein acetylation. Changes in ACL expression are closely related to the occurrence of hyperlipidemia, cardiovascular disease, non-alcoholic fatty liver disease (NALFD), type 2 diabetes, obesity and cancer.

[0003] In recent years, ACL has become a hot topic in biological and innovative drug research as a new drug target. With the widespread application of high-throughput screening technology, a variety of ACL small molecule inhibitors have been discovered. However, to date, no ACL inhibitor has been successfully marketed. Bempedoic acid is an effective ACL small molecule inhibitor used to treat hyperlipidemia and is currently in the clinical trial stage. 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. Summary of the invention

[0004] The main purpose of the present invention is to provide a terpenoid compound and its application, preparation method and pharmaceutical composition, aiming to provide a new ACL inhibitor.

[0005] To achieve the above object, the present invention provides a terpenoid compound, including a terpenoid compound and / or a pharmaceutically acceptable salt thereof, wherein the terpenoid compound includes at least one of the structural formulas of formula (I), formula (II) and formula (III);

[0006]

[0007] The present invention provides an application of the above-mentioned terpenoid compound in preparing an ATP citrate lyase inhibitor.

[0008] The present invention provides an application of the above-mentioned terpenoid compound in the preparation of a medicine for preventing, delaying or treating diseases mediated by ATP citrate lyase.

[0009] In one embodiment, the disease mediated by ATP citrate lyase includes hyperlipidemia, atherosclerosis, non-alcoholic fatty liver disease, type 2 diabetes, obesity or cancer.

[0010] The present invention provides a method for extracting terpenoid compounds, comprising the following steps:

[0011] The dried branches and leaves of Torreya grandis are crushed, extracted and concentrated to obtain an extract;

[0012] After suspending the extract with water, extracting with petroleum ether and ethyl acetate in sequence to obtain an extract;

[0013] The extract is separated and purified to obtain terpenoid compounds.

[0014] In one embodiment, in the step of separating and purifying the extract to obtain terpenoid compounds, the separation and purification process includes:

[0015] S10, sequentially subjecting the extract to silica gel column chromatography and MCI column chromatography to obtain an elution component;

[0016] S20, sequentially subjecting the eluted components to Sephadex LH-20 gel chromatography separation and reversed-phase semi-preparative high performance liquid chromatography separation to obtain terpenoid compounds.

[0017] In one embodiment, step S10 includes:

[0018] S101, subjecting the extract to silica gel column chromatography, eluting with petroleum ether and ethyl acetate in a gradient manner, collecting a first elution component and a second elution component; the volume ratios of petroleum ether and ethyl acetate in the gradient elution are 10:1, 8:1, 6:1, 4:1, 2:1, 1:1, 1:2, and 0:1, respectively; the first elution component is the elution component collected when the volume ratio of petroleum ether to ethyl acetate is 2:1; the second elution component is the elution component collected when the volume ratio of petroleum ether to ethyl acetate is 1:1;

[0019] S102, subjecting the first elution component to MCI column chromatography, eluting with water and methanol in a gradient manner, and collecting a third elution component and a fourth elution component; the volume ratios of water and methanol in the gradient elution are 20:80, 15:85, 10:90, 5:95, and 0:100, respectively; the third elution component is the elution component collected when the volume ratio of water to methanol is 5:95; the fourth elution component is the elution component collected when the volume ratio of water to methanol is 0:100;

[0020] S103. Subject the second elution component to MCI column chromatography, and perform gradient elution with a volume ratio of water to methanol to collect the fifth elution component; the volume ratio gradient of water to methanol in the gradient elution is 30:70, 20:80, 10:90, 5:95, and 0:100; the fifth elution component is the elution component collected when the volume ratio of water to methanol is 5:95.

[0021] In one embodiment, step S20 includes:

[0022] S201, subjecting the third eluted component to Sephadex LH-20 gel chromatography and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpenoid compound having a structural formula as shown in formula (III);

[0023] S202, subjecting the fourth eluted component to Sephadex LH-20 gel chromatography and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpenoid compound having a structural formula as shown in formula (I);

[0024] S203, subjecting the fifth eluted component to Sephadex LH-20 gel chromatography separation and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpene compound having a structural formula as shown in formula (II).

[0025] The present invention also provides a pharmaceutical composition, comprising the above-mentioned terpenoid compound, pharmaceutically acceptable pharmaceutical adjuvants and excipients.

[0026] In one embodiment, the pharmaceutical composition is in the form of tablets, pills, capsules or granules.

[0027] The three new terpenoid compounds fargesisin A (I), fargesisin B (II) and fargesisin C (III) extracted and separated from branches and leaves of Torreya grandis for the first time in the present invention all have a significant inhibitory effect on ACL activity, can be used as ACL inhibitors, and have broad application prospects in the preparation of drugs for preventing, delaying or treating ACL-mediated diseases such as hyperlipidemia, atherosclerosis, non-alcoholic fatty liver disease, type 2 diabetes, obesity and cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the structures shown in these drawings without paying creative work.

[0029] Figure 1 The fargesisin C (III) in Example 1 provided by the present invention 1 H NMR spectrum;

[0030] Figure 2 The fargesisin A (I) in Example 1 provided by the present invention 1 H NMR spectrum;

[0031] Figure 3 The fargesisin B (II) in Example 1 provided by the present invention 1 H NMR spectrum.

[0032] The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0033] To make the purpose, technical scheme and advantages of the embodiment of the present invention clearer, the technical scheme in the embodiment of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiment, it is carried out according to the normal conditions or the conditions recommended by the manufacturer. If the reagents or instruments used do not specify the manufacturer, they are all conventional products that can be purchased commercially. In addition, the meaning of "and / or" appearing in the full text includes three parallel schemes, taking "A and / or B" as an example, including scheme A, or scheme B, or schemes that A and B meet at the same time. In addition, the technical schemes between the various embodiments can be combined with each other, but must be based on the ability of ordinary technicians in the field to achieve. When the combination of technical solutions is contradictory or cannot be achieved, it should be considered that the combination of such technical solutions does not exist, and is not within the scope of protection required by the present invention. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without making creative work belong to the scope of protection of the present invention.

[0034] ATP citrate lyase (ACL) is a key enzyme in cell metabolism, the main source of acetyl-CoA, an important precursor for the biosynthesis of fatty acids, cholesterol and isoprenoids, and is also involved in protein acetylation. Changes in ACL expression are closely related to the occurrence of hyperlipidemia, cardiovascular disease, non-alcoholic fatty liver disease (NALFD), type 2 diabetes, obesity and cancer.

[0035] In recent years, ACL has become a hot topic in biological and innovative drug research as a new drug target. With the widespread application of high-throughput screening technology, a variety of ACL small molecule inhibitors have been discovered. However, to date, no ACL inhibitor has been successfully marketed. Bempedoic acid is an effective ACL small molecule inhibitor used to treat hyperlipidemia and is currently in the clinical trial stage. 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.

[0036] In view of this, the present invention provides a terpenoid compound, including a terpenoid compound and / or a pharmaceutically acceptable salt thereof, wherein the terpenoid compound includes at least one of the structural formulas such as formula (I), formula (II) and formula (III);

[0037]

[0038]

[0039] Torreya fargesii Franch is an evergreen tree of the genus Torreya in the family Taxaceae. It is a species endemic to my country and is mainly distributed in Shennongjia. At present, the chemical composition of Torreya fargesii has only been reported on alkaloid components. The three new terpenoid compounds fargesisin A (I), fargesisin B (II) and fargesisin C (III) extracted and separated from the branches and leaves of Torreya fargesii for the first time in the present invention all have a significant inhibitory effect on ACL activity and can be used as ACL inhibitors. They have broad application prospects in the preparation of drugs for preventing, delaying or treating ACL-mediated diseases such as hyperlipidemia, atherosclerosis, non-alcoholic fatty liver disease, type 2 diabetes, obesity and cancer.

[0040] The present invention provides an application of the above-mentioned terpenoid compound in the preparation of a medicine for preventing, delaying or treating diseases mediated by ATP citrate lyase.

[0041] Since the present application adopts all the technical solutions of all the above embodiments, it has at least all the beneficial effects brought by the technical solutions of the above embodiments, which will not be described one by one here.

[0042] In some embodiments of the present invention, the diseases mediated by ATP citrate lyase include hyperlipidemia, atherosclerosis, non-alcoholic fatty liver disease, type 2 diabetes, obesity or cancer.

[0043] Since ATP citrate lyase is related to the above diseases, the occurrence of the above diseases can be inhibited, prevented or delayed by inhibiting the activity of ATP citrate lyase.

[0044] The terpenoid 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.

[0045] The present invention provides a method for extracting terpenoid compounds, comprising the following steps:

[0046] The dried branches and leaves of Torreya grandis are crushed, extracted and concentrated to obtain an extract;

[0047] After suspending the extract with water, extracting with petroleum ether and ethyl acetate in sequence to obtain an extract;

[0048] The extract is separated and purified to obtain terpenoid compounds.

[0049] In the technical solution of the present invention, petroleum ether is mainly used to extract fat-soluble components, and ethyl acetate is used to extract components with strong polarity such as terpenes. The extract containing terpenes is separated and purified to obtain terpenoid compounds with high purity.

[0050] In some embodiments of the present invention, in the step of separating and purifying the extract to obtain terpenoid compounds, the separation and purification process includes:

[0051] S10, sequentially subjecting the extract to silica gel column chromatography and MCI column chromatography to obtain an elution component;

[0052] S20, sequentially subjecting the eluted components to Sephadex LH-20 gel chromatography separation and reversed-phase semi-preparative high performance liquid chromatography separation to obtain terpenoid compounds.

[0053] In the technical solution of the present invention, silica gel is suitable for separating compounds with strong polarity; MCI column chromatography is an adsorbent based on cellulose modification, which is suitable for separating complex mixtures in natural products, and the purity of the separated components is higher than that of silica gel column chromatography.

[0054] In some embodiments of the present invention, step S10 includes:

[0055] S101, subjecting the extract to silica gel column chromatography, eluting with petroleum ether and ethyl acetate in a gradient manner, collecting a first elution component and a second elution component; the volume ratios of petroleum ether and ethyl acetate in the gradient elution are 10:1, 8:1, 6:1, 4:1, 2:1, 1:1, 1:2, and 0:1, respectively; the first elution component is the elution component collected when the volume ratio of petroleum ether to ethyl acetate is 2:1; the second elution component is the elution component collected when the volume ratio of petroleum ether to ethyl acetate is 1:1;

[0056] S102, subjecting the first elution component to MCI column chromatography, eluting with water and methanol in a gradient manner, and collecting a third elution component and a fourth elution component; the volume ratios of water and methanol in the gradient elution are 20:80, 15:85, 10:90, 5:95, and 0:100, respectively; the third elution component is the elution component collected when the volume ratio of water to methanol is 5:95; the fourth elution component is the elution component collected when the volume ratio of water to methanol is 0:100;

[0057] S103. Subject the second elution component to MCI column chromatography, and perform gradient elution with a volume ratio of water to methanol to collect the fifth elution component; the volume ratio gradient of water to methanol in the gradient elution is 30:70, 20:80, 10:90, 5:95, and 0:100; the fifth elution component is the elution component collected when the volume ratio of water to methanol is 5:95.

[0058] In the technical solution of the present invention, the target compound can be separated more finely by gradually changing the ratio of the elution solvent.

[0059] In some embodiments of the present invention, step S20 includes:

[0060] S201, subjecting the third eluted component to Sephadex LH-20 gel chromatography and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpenoid compound having a structural formula as shown in formula (III);

[0061] S202, subjecting the fourth eluted component to Sephadex LH-20 gel chromatography and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpenoid compound having a structural formula as shown in formula (I);

[0062] S203, subjecting the fifth eluted component to Sephadex LH-20 gel chromatography separation and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpene compound having a structural formula as shown in formula (II).

[0063] In the technical solution of the present invention, the target terpenoid compounds can be separated and purified efficiently and accurately by combining Sephadex LH-20 gel chromatography and reversed-phase semi-preparative high performance liquid chromatography.

[0064] The present invention also provides a pharmaceutical composition, comprising the aforementioned terpenoid compound, a pharmaceutically acceptable pharmaceutical excipient and an excipient. The pharmaceutically acceptable pharmaceutical excipient includes a binder such as sodium carboxymethylcellulose, hydroxypropyl methylcellulose or polyvinyl pyrrolidone, a disintegrant such as cross-linked sodium carboxymethylcellulose, cross-linked polyvinyl pyrrolidone, sodium carboxymethyl starch or microcrystalline cellulose, and a lubricant such as magnesium stearate or talc; the excipient includes lactose, microcrystalline cellulose, starch, pregelatinized starch or dextrin.

[0065] In some embodiments of the present invention, the type of the pharmaceutical composition is tablets, pills, capsules or granules. The type of the pharmaceutical composition can be changed according to the specific usage scenario to achieve the best therapeutic effect.

[0066] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments and drawings. It should be understood that the following embodiments are only used to explain the present invention and are not used to limit the present invention.

[0067] In the following examples, the branches and leaves of Torreya grandis were collected from Shennongjia Forest District, Hubei Province;

[0068] Specific rotation test was completed by JASCO P-1020 polarimeter;

[0069] Ultraviolet and infrared spectral data were obtained by Shimadzu UV-2550 ultraviolet spectrometer and Nicolet AVATAR 360 infrared spectrometer, respectively;

[0070] NMR data were measured by Bruker Avance II 400 / 600 instrument;

[0071] HRESIMS data were measured by AB Sciex TripleTOF 5600 instrument;

[0072] Silica gel and thin layer plates were produced by Yantai Kangbino Company;

[0073] Sephadex LH-20 gel is produced by GE Healthcare Bio-Sciences, Switzerland;

[0074] The semi-preparative HPLC was Waters e2695, and the semi-preparative column model was COSMOSIL Cholester (5 μm, 250 × 10 mm);

[0075] All reagents not otherwise specified were produced by Shanghai Sinopharm Chemical Reagent Co., Ltd.

[0076] Example 1 Extraction of terpenoids

[0077] Take 1kg of Torreya grandis branches and leaves, dry and crush them, and then use 90% methanol aqueous solution to cold-immerse and extract them 4 times at room temperature. Combine the 4 extracts and concentrate them under reduced pressure to obtain 105g of extract. After the extract is dispersed with 500g of water, it is extracted with equal volumes of petroleum ether and ethyl acetate in sequence to obtain 20g of ethyl acetate extract. The ethyl acetate extract is chromatographed on a 100-200 mesh silica gel column, and gradient eluted with a volume ratio of petroleum ether to ethyl acetate of 10:1-0:1 (the specific gradient ratios are 10:1, 8:1, 6:1, 4:1, 2:1, 1:1, 1:2, 0:1, and each gradient eluent is used in an amount of 2-3 times the column volume), collect the obtained lower column liquid, perform color development and identification by TLC thin layer chromatography, and combine the lower column liquids with the same spots to obtain 10 components (Fr.1-Fr.10).

[0078] Component Fr.6 (i.e., the elution component with the eluent being petroleum ether:ethyl acetate=2:1) ​​was subjected to MCI column chromatography and eluted with water:methanol (20:80-0:100, v / v, 20:80, 15:85, 10:90, 5:95, 0:100 gradient); then the 5:95 (water:methanol) elution component was first eluted with Sephadex LH-20 (pure methanol as the eluent, the column liquid was collected and identified by TLC thin layer chromatography, and the components with the same spots were combined to obtain 4 subcomponents (Fr.6.A.1-Fr.6.A.4), and then the subcomponent Fr.6.A.2 was further separated and purified by semi-preparative HPLC (MeCN:H2O:TFA=91:9:0.5, v / v, 3mL / min) to obtain compound fargesisin C(III) (4.9 mg, tR=22.9 min) (see hydrogen spectrum for the spectrum). Figure 1 ); the 0:100 (water: methanol) elution fraction was further purified by Sephadex LH-20 (pure methanol as eluent), the column liquid was collected and identified by TLC thin layer chromatography, and the fractions with the same spots were combined to obtain three subfractions (Fr.6.B.1-Fr.6.B.3), and then the subfraction Fr.6.B.1 was further separated and purified by semi-preparative HPLC (MeOH:H2O=96:4, v / v, 3mL / min) to obtain the compound fargesisin A(I) (2.8mg, tR=27.9min) (the hydrogen spectrum is shown in Figure 2 ).

[0079] Component Fr.7 (i.e., the elution component with petroleum ether: ethyl acetate = 1:1 as the eluent) was subjected to MCI column chromatography using a water: methanol (30:70-100:0, v / v, 30:70, 20:80, 10:90, 0:100) gradient, and then the 5:95 (water: methanol) elution component was first subjected to Sephadex LH-20 (pure methanol as the eluent), the column liquid was collected and identified by TLC thin layer chromatography, and the components with the same spots were combined to obtain 5 subcomponents (Fr.7.A.1-Fr.7.A.5), and then the subcomponent Fr.7.A.2 was further separated and purified by semi-preparative HPLC (MeOH:H2O = 96:4, v / v, 3 mL / min) to obtain compound fargesisin C (Ⅱ) (3.5 mg, tR = 22.4 min) (see hydrogen spectrum for the spectrum) Figure 3 ).

[0080] It was identified that fargesisin A (I) and fargesisin B (II) are new triterpene dimer compounds, and fargesisin C (III) is a myrrhanane-type sesquiterpene tetramer. Their spectral and physicochemical data are as follows:

[0081] Fargesisin A(I):white amorphous powder; [α] 20 D -9(c 0.10,CHCl3); UV(MeOH)λmax(logε)202(3.77)nm; IR(KBr)ν max 3446,2957,2920,2848,1581,1384,771cm -1 ; 1H NMR(600MHz,CDCl3):δ1.59(m,H-1a),1.47(m,H-1b),1.72(m,H-2a),1.66(m,H-2b),3.48(brs,H-3),5.60(d,J=5.8Hz,H-6),2.40(dd,J=19.0,7.6Hz,H-7a),1.82(m,H-7b),1.76(d,J=7.9Hz,H-8),2.27(d,J=12.5Hz,H-10),1.65(m,H-11a),1.41(m,H-11b),1.81(m,H-12a),1.50(m,H-12b),1.21(m,H-15a),1.12(m,H-15b),1.88(m,H-16a),1.31(m,H-16b),1.48(m,H-17),0.86(3H,s,H3-18),0.92(3H,s,H3-19),1.48(m,H-20),0.91(3H,d,J=6.4Hz,H3-21),1.14(m,H-22a),0.97(m,H-22b),1.41(2H,m,H-23),3.35(dd,J=10.1,2.0Hz,H-24),1.15(3H,s,H3-26),1.16(3H,s,H3-27),1.03(3H,s,H3-28),1.14(3H,s,H3-29),0.81(3H,s,H3-30),1.59(m,H-1ˊa),1.47(m,H-1ˊb),1.72(m,H-2ˊa),1.66(m,H-2ˊb),3.48(br s,H-3ˊ),5.60(d,J=5.8Hz,H-6ˊ),2.37(dd,J=19.0,7.6Hz,H-7ˊa),1.82(m,H-7ˊb),1.76(d,J=7.9Hz,H-8ˊ),2.27(d,J=12.5Hz,H-10ˊ),1.65(m,H-11ˊa),1.41(m,H-11ˊb),1.81(m,H-12ˊa),1.50(m,H-12ˊb),1.21(m,H-15ˊa),1.12(m,H-15ˊb),1.88(m,H-16ˊa),1.31(m,H-16ˊb),1.48(m,H-17ˊ),0.84(3H,s,H3-18ˊ),0.92(3H,s,H3-19ˊ),1.48(m,H-20ˊ),0.89(3H,d,J=6.4Hz,H3-21ˊ),1.14(m,H-22ˊa),0.97(m,H-22ˊb),1.41(2H,m,H-23ˊ),3.26(dd,J=6.8,2.0Hz,H-24ˊ),1.16(3H,s,H3-26ˊ),1.17(3H,s,H3-27ˊ),1.03(3H,s,H3-28ˊ),1.14(3H,s,H3-29ˊ),0.81(3H,s,H3-30ˊ). 13 C NMR (150MHz, CDCl3): δ21.1(C-1),28.9(C-2),76.6(C-3),41.4(C-4),141.2(C-5),121.4(C-6),24.4(C- 7),43.6(C-8),34.5(C-9),37.8(C-10),32.3(C-11),30.5(C-12),46.2(C-13),49.2(C-14),34.7(C-15), 28.0(C-16),50.5(C-17),15.4(C-18),27.9(C-19),36.3(C-20),18.9(C-21),34.0(C-22),28.2(C-23), 78.7(C-24),79.0(C-25),21.1(C-26),23.0(C-27),27.3(C-28),25.4(C-29),17.8(C-30),21.1(C-1ˊ),2 8.9(C-2ˊ),76.6(C-3ˊ),41.4(C-4ˊ),141.2(C-5ˊ),121.5(C-6ˊ),24.4(C-7ˊ),43.6(C-8ˊ),34.5(C-9ˊ) ,37.8(C-10ˊ),32.3(C-11ˊ),30.4(C-12ˊ),46.2(C-13ˊ),49.2(C-14ˊ),34.7(C-15ˊ),28.0(C-16ˊ),50.1 (C-17ˊ),15.3(C-18ˊ),28.0(C-19ˊ),36.8(C-20ˊ),18.7(C-21ˊ),33.8(C-22ˊ),30.8(C-23ˊ),78.5(C-2 4ˊ),73.4(C-25ˊ),24.2(C-26ˊ),27.1(C-27ˊ),27.2(C-28ˊ),25.4(C-29ˊ),17.8(C-30ˊ); HRESIMS[M+Na] + m / z925.7623 (calcd for C 60 H 102 O5Na,925.7619,Δ=+0.4ppm).

[0082] Fargesisin B(II): white amorphous powder; [α] 20 D+33(c 0.08,CHCl3);UV(MeOH)λ max (logε)202(3.88)nm;IR(KBr)ν max 3451,2942,2865,1736,1464,1290,1177,1075,983,756cm -1 ; 1H NMR(600MHz,CDCl3):δ1.59(m,H-1a),1.41(m,H-1b),2.01(ddd,J=14.1,2.8,2.8Hz,H-2a),1.72(m,H-2b),3.44(t,J=2.5Hz,H-3),1.33(m,H-5),1.61(m,H-6a),1.47(m,H-6b),1.66(m,H-7a),1.40(m,H-7b),2.19(d,J=11.0Hz,H-8),5.27(d,J=5.8Hz,H-11),2.09(d,J=17.1Hz,H-12a),1.92(m,H-12b),1.32(2H,m,H-15),1.92(m,H-16a),1.31(m,H-16b),1.61(m,H-17),0.67(3H,s,H3-18),1.07(3H,s,H3-19),1.58(m,H-20),0.91(3H,d,J=6.6Hz,H3-21),1.47(2H,m,H-22),1.77(2H,m,H-23),3.14(dd,J=9.5,5.0Hz,H-24),4.29,4.07(d,J=11.4Hz,H-26),1.18(3H,s,H3-27),0.97(3H,s,H3-28),0.89(3H,s,H3-29),0.76(3H,s,H3-30),1.81(m,H-1ˊa),1.50(m,H-1ˊb),2.01(ddd,J=14.1,2.8,2.8Hz,H-2ˊa),1.72(m,H-2ˊb),3.44(t,J=2.5Hz,H-3ˊ),1.33(m,H-5ˊ),1.59(m,H-6ˊa),1.47(m,H-6ˊb),1.66(m,H-7ˊa),1.40(m,H-7ˊb),2.91(d,J=11.0Hz,H-8ˊ),5.27(d,J=5.8Hz,H-11ˊ),2.09(d,J=16.4Hz,H-12ˊa),1.92(m,H-12ˊb),1.32(2H,m,H-15ˊa),1.92(m,H-16ˊa),1.31(m,H-16ˊb),1.61(m,H-17ˊ),0.67(3H,s,H3-18ˊ),1.07(3H,s,H3-19ˊ),1.58(m,H-20ˊ),0.90(3H,d,J=6.6Hz,H3-21ˊ),1.47(2H,m,H-22ˊ),1.77(2H,m,H-23ˊ),3.18(t,J=5.7Hz,H-24ˊ),1.55(3H,s,H3-27ˊ),0.97(3H,s,H3-28ˊ),0.89(3H,s,H3-29ˊ),0.76(3H,s,H3-30ˊ). 13 C NMR (150MHz, CDCl3): δ30.5(C-1),25.7(C-2),76.3(C-3),37.8(C-4),46.7(C-5),21.3(C-6),28.0(C-7) ,41.8(C-8),148.5(C-9),39.3(C-10),114.6(C-11),37.1(C-12),44.3(C-13),47.1(C-14),33.9(C-15), 28.0(C-16),50.9(C-17),14.4(C-18),22.1(C-19),35.7(C-20),18.2(C-21),33.2(C-22),29.7(C-23), 77.2(C-24),73.3(C-25),70.0(C-26),21.0(C-27),28.3(C-28),22.5(C-29),18.5(C-30),30.1(C-1ˊ),2 2.5(C-2ˊ),76.3(C-3ˊ),37.8(C-4ˊ),46.7(C-5ˊ),21.3(C-6ˊ),28.0(C-7ˊ),41.8(C-8ˊ),148.5(C-9ˊ), 39.3(C-10ˊ),114.5(C-11ˊ),37.1(C-12ˊ),44.3(C-13ˊ),47.1(C-14ˊ),33.9(C-15ˊ),28.0(C-16ˊ),50.8 (C-17ˊ),14.4(C-18ˊ),22.1(C-19ˊ),36.0(C-20ˊ),18.2(C-21ˊ),32.3(C-22ˊ),25.7(C-23ˊ),63.0(C-24 HRESIMS[M+Na] + m / z 953.7210 (calcd for C 60 H 98 O7Na,937.7205,Δ=+0.6ppm).

[0083] Fargesisin C(III): white amorphous powder; [α] 20 D +233(c 0.10,CHCl3); UV(MeOH)λ max(logε)206(4.88)nm;IR(KBr)ν max 2925,1690cm -1 ; 1 H NMR(400MHz,CDCl3):δ2.07(m,H-1),1.79(m,H-2a),1.41(m,H-2b),2.49(d,J=16.8Hz,H-3a),2.91(m,H-3b),7.10(br s,H-5),2.26(m,H-6a),1.93(m,H-6b),5.02(d,J=10.2Hz,H-8),3.18(m,H-9),5.15(d,J=5.7Hz,H-10),1.63(s,H-12),2.93(m,H-13a),1.85(m,H-13b),1.65(s,H-14),2.94(m,H-1′),5.63(d,J=10.2Hz,H-2′),5.60(d,J=10.2Hz,H-3′),1.86(m,H-6′a),1.49(m,H-6′b),5.07(d,J=10.2Hz,H-8′),3.16(m,H-9′),5.15(d,J=5.7Hz,H-10′),1.65(s,H-12′),1.93(m,H-13′a),1.85(m,H-13′b),1.65(s,H-14′),2.94(m,H-1″),5.63(d,J=10.2Hz,H-2″),5.60(d,J=10.2Hz,H-3″),1.86(m,H-6″a),1.49(m,H-6″b),5.07(d,J=10.2Hz,H-8″),3.16(m,H-9″),5.15(d,J=5.7Hz,H-10″),1.65(s,H-12″),1.93(m,H-13″a),1.85(m,H-13″b),1.66(s,H-14″),2.97(m,H-1″′),5.53(d,J=10.3Hz,H-2″′),5.60(d,J=10.3Hz,H-3″′),2.56(m,H-5″′),1.93(m,H-6″′a),1.76(m,H-6″′b),5.16(t,J=6.8Hz,H-8″′),2.66(t,J=7.3Hz,H-9″′),5.09(d,J=6.8Hz,H-10″′),1.69(s,H-12″′),1.62(s,H-13″′),1.48(s,H-14″′). 13C NMR (150MHz, CDCl3): δ42.0(C-1),27.1(C-2),24.3(C-3),129.3(C-4),142.4(C-5),31.2(C-6),138.9(C-7), 123.8(C-8),43.4(C-9),120.0(C-10),133.4(C-11),23.4(C-12),31.6(C-13),14.4(C-14),172.9(C-15),42. 0(C-1′),132.5(C-2′),131.0(C-3′),48.3(C-4′),28.2(C-5′),28.8(C-6′),138.1(C-7′),126.1(C-8′),43.5 (C-9′),120.0(C-10′),133.5(C-11′),23.4(C-12′),31.5(C-13′),12.4(C-14′),180.4(C-15′),42.1(C-1″), 132.7(C-2″),131.3(C-3″),48.3(C-4″),28.2(C-5″),28.9(C-6″),138.3(C-7″),126.1(C-8″),43.5(C-9″),1 20.0(C-10″),133.5(C-11″),23.4(C-12″),31.5(C-13″),12.4(C-14″),180.7(C-15″),42.9(C-1″′),132.9(C -2″′),130.4(C-3″′),48.3(C-4″′),28.2(C-5″′),29.8(C-6″′),137.3(C-7″′),124.1(C-8″′),27.1(C-9″′), 123.0(C-10″′), 131.5(C-11″′), 25.4(C-12″′), 17.7(C-13″′), 13.4(C-14″′), 180.4(C-15″′); HRESIMS[M+H] + m / z925.5610(calcd for C 60 H 77 O8,925.5618,Δ=-0.9ppm). Example 2 Determination of ATP-citrate lyase inhibitory activity of terpenoids

[0084] Experimental method: ATP citrate lyase ACL catalyzes citrate to generate acetyl-CoA, and this reaction is accompanied by the consumption of ATP. The ACL activity test relies on the ADP-Glo ​​fluorescent detection reagent, which reflects the enzyme activity of ACL by detecting the amount of ADP generated. The intensity of the fluorescent signal is proportional to the amount of ADP generated. Specifically, 2 μL of enzyme, 2 μL of substrate, and 1 μL of different concentrations of the test compound were added to the 384 reaction plate (ProxiPlateTM-384Plus, PerkinElmer), incubated at 37°C for 0.5 hours, and then 2.5 μL of reagent 1 in the ADP-Glo ​​Kinase Assay Kit (ADP-Glo ​​Kinase Assay, Promega) was added and incubated at room temperature for 1 hour to consume the unreacted substrate ATP, and then 5 μL of the detection reagent 2 in the ADP-Glo ​​Kinase Assay Kit was added and incubated at room temperature for 0.5 hours to convert ADP into ATP. The reading value was then detected with the EnVision instrument to reflect the intensity of the fluorescent signal. At the same time, the reaction set up a positive control group, a blank control group and a solvent control group in which DMSO was substituted for the test compound. Each sample and each concentration were set up with 3 replicates. The final reaction volume was 12.5 μL. The data was processed by plotting the logarithm of the concentration against the activity percentage, and then the fitting curve was calculated using nonlinear regression. The IC was calculated using the software GraphPad Prism 5 formula log (inhibitor) vs. response--Variable slope 50 BMS 303141 was used as a positive control (Koerner SK et al., Eur. J. Med. Chem. 2017, 126: 920-928).

[0085] The initial screening selected the corresponding compounds at a concentration of 20 μg / mL to investigate the percentage inhibition of ACL enzyme activity. The experimental results showed that the inhibition rates of fargesisin A (I), fargesisin B (II) and fargesisin C (III) were 88.7%, 85.3% and 103.6%, respectively. Further determination of IC 50 The relative activity was plotted against the compound concentration using the formula v / v0 = 100 / (1+b*[I] / IC 50 ) is fitted to obtain IC 50 Values, where v / v0 represents relative activity, [I] represents compound concentration, IC 50 represents half-maximal inhibitory concentration, b represents a constant, the experiment was repeated three times, and the results were averaged. The test data are shown in Table 1 below:

[0086] Table 1 Results of terpenoids inhibiting ACL activity

[0087] Compound <![CDATA[IC 50 Value (μM)]]> Fargesisin A(I) 3.5±0.8 Fargesisin B(II) 4.2±0.5 Fargesisin C(III) 1.9±0.4 BMS 303141 0.5±0.1

[0088] As can be seen from Table 1, the above three terpenoid compounds all showed significant inhibitory activity, indicating that the compounds of the present invention can be used to prepare drugs for treating ATP citrate lyase-mediated hyperlipidemia, atherosclerosis, non-alcoholic fatty liver disease, type 2 diabetes, obesity and cancer, or as lead compounds for such drugs.

[0089] The above are only preferred embodiments of the present invention, and do not limit the patent scope of the present invention. For those skilled in the art, the present invention may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the patent protection scope of the present invention.

Claims

1. A terpene compound, characterized in that: Including terpenoid compounds and / or pharmaceutically acceptable salts thereof, wherein the terpenoid compounds include at least one of the structural formulas of formula (I), formula (II) and formula (III); 2. Use of the terpenoid compound as claimed in claim 1 in the preparation of an ATP citrate lyase inhibitor.

3. Use of the terpenoid compound according to claim 1 in the preparation of a drug for preventing, delaying or treating ATP citrate lyase-mediated diseases.

4. The use according to claim 3, characterized in that The diseases mediated by ATP citrate lyase include hyperlipidemia, atherosclerosis, non-alcoholic fatty liver disease, type 2 diabetes, obesity or cancer.

5. A method for extracting terpenoids as claimed in claim 1, characterized in that: The following steps are involved: Crush the dried branches and leaves of Torreya grandis 、 Extraction and concentration to obtain extract; After suspending the extract with water, extracting with petroleum ether and ethyl acetate in sequence to obtain an extract; The extract is separated and purified to obtain terpenoid compounds.

6. The method for extracting terpenoids according to claim 5, characterized in that: In the step of separating and purifying the extract to obtain terpenoid compounds, the separation and purification process includes: S10, sequentially subjecting the extract to silica gel column chromatography and MCI column chromatography to obtain an elution component; S20, sequentially subjecting the eluted components to Sephadex LH-20 gel chromatography separation and reversed-phase semi-preparative high performance liquid chromatography separation to obtain terpenoid compounds.

7. The method for extracting terpenoids according to claim 6, characterized in that: Step S10 includes: S101, subjecting the extract to silica gel column chromatography, eluting with petroleum ether and ethyl acetate in a gradient manner, collecting a first elution component and a second elution component; the volume ratios of petroleum ether and ethyl acetate in the gradient elution are 10:1, 8:1, 6:1, 4:1, 2:1, 1:1, 1:2, and 0:1, respectively; the first elution component is the elution component collected when the volume ratio of petroleum ether to ethyl acetate is 2:1; the second elution component is the elution component collected when the volume ratio of petroleum ether to ethyl acetate is 1:1; S102, subjecting the first elution component to MCI column chromatography, eluting with water and methanol in a gradient manner, and collecting a third elution component and a fourth elution component; the volume ratios of water and methanol in the gradient elution are 20:80, 15:85, 10:90, 5:95, and 0:100, respectively; the third elution component is the elution component collected when the volume ratio of water to methanol is 5:95; the fourth elution component is the elution component collected when the volume ratio of water to methanol is 0:100; S103. Subject the second elution component to MCI column chromatography, and perform gradient elution with a volume ratio of water to methanol to collect the fifth elution component; the volume ratio gradient of water to methanol in the gradient elution is 30:70, 20:80, 10:90, 5:95, and 0:100; the fifth elution component is the elution component collected when the volume ratio of water to methanol is 5:

95.

8. The method for extracting terpenoids according to claim 7, characterized in that: Step S20 includes: S201, subjecting the third eluted component to Sephadex LH-20 gel chromatography and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpenoid compound having a structural formula as shown in formula (III); S202, subjecting the fourth eluted component to Sephadex LH-20 gel chromatography and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpenoid compound having a structural formula as shown in formula (I); S203, subjecting the fifth eluted component to Sephadex LH-20 gel chromatography separation and reversed-phase semi-preparative high performance liquid chromatography separation in sequence to obtain a terpene compound having a structural formula as shown in formula (II).

9. A pharmaceutical composition, characterized in that The invention comprises the terpenoid compound as claimed in claim 1, pharmaceutically acceptable pharmaceutical adjuvants and excipients.

10. The pharmaceutical composition according to claim 9, characterized in that The pharmaceutical composition is in the form of tablets, pills, capsules or granules.

Images