A kind of enantiotesenane type 2-norditerpene lactone compound and its preparation method and application

Abstract

The present invention relates to the technical field of natural medicines, and in particular to an enantiopysenane-type 2-norditerpene lactone compound and a preparation method and application thereof. The structural formula of the enantiopysenane-type 2-norditerpene lactone compound is shown in Formula I; the present invention realizes the extraction of enantiopysenane-type 2-norditerpene lactone compounds with the function of inhibiting heart valve calcification from Euphorbia chamaejasme for the first time, and the compounds have great development value as new drugs for the preparation of prevention or treatment of calcific aortic valve disease, and the design ideas of the compounds also provide new ideas and approaches for the development of new drugs for the preparation of prevention or treatment of calcific aortic valve disease.

Description

Technical Field

[0001] The invention relates to the technical field of natural medicines, and in particular to an enantiotesenane-type 2-norditerpene lactone compound and a preparation method and application thereof. Background Art

[0002] Calcific aortic valve disease (CAVD) is a progressive disease characterized by lipoprotein deposition, chronic inflammation, and thickening of the valve leaflets. CAVD remains one of the most common and expensive heart diseases in developed countries, affecting 4.6% of people over 75 years old and 6% of adults over 65 years old. Currently, there is no effective drug treatment for CAVD except for surgery or interventional valve replacement. Aortic valvular interstitial cells, as the main cell population of the aortic valve, are responsible for tissue homeostasis and valve function, and play a key role in the occurrence and development of CAVD. Therefore, the proliferation and calcification changes of valvular interstitial cells in the early stage of CAVD are important targets for drug intervention, which may effectively prevent the occurrence of heart valve calcification. Therefore, it is necessary to seek treatments and drugs with good efficacy and less toxic side effects. Summary of the invention

[0003] Based on the above content, the present invention provides an enantiotesenane-type 2-norditerpene lactone compound and its preparation method and application. The compound has good heart valve calcification inhibitory activity and can be used to prepare a drug for calcific aortic valve disease.

[0004] To achieve the above object, the present invention provides the following solutions:

[0005] One of the technical solutions of the present invention is an enantiotesenane-type 2-norditerpene lactone compound, the structural formula of which is shown in Formula I;

[0006]

[0007] The second technical solution of the present invention is a method for preparing the above-mentioned enantiotesenane-type 2-norditerpene lactone compound, comprising the following steps:

[0008] The dried root of Euphorbia chamaejasme is used as a raw material, and the extract is obtained after percolation extraction and concentration, and the extract is suspended in water to obtain a suspension, and the extract is obtained after extraction and concentration;

[0009] The extract is subjected to gradient elution by silica gel column chromatography, MCI resin column chromatography, and silica gel column chromatography in sequence, and isocratic elution is performed by Sephadex LH-20 gel column chromatography and ODS medium pressure column chromatography to obtain Fr.1 to Fr.4. Fr.2 is prepared by ODS liquid chromatography to obtain the enantiotherm-type 2-norditerpene lactone compound shown in formula I.

[0010] The third technical solution of the present invention is the use of the above-mentioned enantiotesenane-type 2-norditerpene lactone compounds in the preparation of drugs for preventing or treating calcific aortic valve disease.

[0011] A fourth technical solution of the present invention is a drug for preventing or treating calcific aortic valve disease, the raw materials of which include the above-mentioned enantiotesenane-type 2-norditerpene lactone compounds.

[0012] The present invention discloses the following technical effects:

[0013] The present invention realizes for the first time the extraction of enantiotesenane-type 2-norditerpene lactone compounds capable of inhibiting heart valve calcification from Euphorbia chamaejasma. Such compounds have great development value as new drugs for the prevention or treatment of calcific aortic valve disease. The design ideas of such compounds also provide new ideas and approaches for the development of new drugs for the prevention or treatment of calcific aortic valve disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] 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 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 these drawings without paying creative work.

[0015] Figure 1 1H-NMR spectrum of the enantiopyrane-type 2-norditerpene lactone compound of the present invention;

[0016] Figure 2 is the 13C-NMR spectrum of the enantiopyrane-type 2-norditerpene lactone compound of the present invention;

[0017] Figure 3 The single crystal diffraction structure of the enantiopyrane-type 2-norditerpene lactone compound of the present invention;

[0018] Figure 4 It is a curve diagram of the inhibition rate of the proliferation of valvular interstitial cells (VICs) by the enantiotesenane-type 2-norditerpene lactone of the present invention;

[0019] Figure 5 The immunoblot diagram (a) and the corresponding statistical diagram (b) are the effects of the enantiotesenane-type 2-norditerpene lactone of the present invention on the expression of ALP and Runx2 in VICs cultured in osteogenic medium;

[0020] Figure 6 The staining diagram (a) and the corresponding statistical diagram (b) are the effects of the enantiotesenane-type 2-norditerpene lactone of the present invention on the alkaline phosphatase (ALP) activity in VICs cultured in osteogenic medium;

[0021] Figure 7 This is a staining diagram showing the effect of the enantiotesenane-type 2-norditerpene lactone of the present invention on calcium precipitation in VICs cultured in osteogenic medium. DETAILED DESCRIPTION

[0022] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0023] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0024] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0025] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0026] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0027] The "%" described in the present invention, unless otherwise specified, refers to mass percentage.

[0028] In recent years, traditional Chinese medicine has provided an important source and new opportunity for modern drug development. The active ingredients of natural products have become an important source of new drug research and development due to their multi-target, high efficiency and low toxicity.

[0029] Euphorbia chamaejasme, a perennial herbaceous plant of the Euphorbiaceae family, has a thick fleshy root with white latex and is cylindrical. It is one of the base plants of the Chinese medicine wolfsbane. It tastes spicy, flat, toxic, and enters the liver and spleen meridians. It has the effect of dispersing knots and killing insects, and is widely used in Chinese patent medicines and clinical prescriptions. In traditional medicine, wolfsbane is used for phlegm, food and insect accumulation, edema and abdominal distension, heart and abdominal pain, accumulation of masses, lymph tuberculosis, scabies, etc. Modern research results show that the Chinese medicine wolfsbane has a variety of pharmacological activities, such as anti-tuberculosis, anti-HIV virus, anti-tumor, antibacterial, anti-inflammatory and insecticidal effects. Modern research shows that the chemical components of wolfsbane euphorbia mainly include two major categories: phenolic acid and terpenes. Among them, terpenes are mainly rich and diverse diterpenes, which are also considered to be the main material basis for wolfsbane euphorbia to exert many pharmacological activities. For a new enantiopyran-type 2-norditerpene lactone compound and its heart valve calcification inhibitory activity involved in the present invention, there has been no patent or literature report so far.

[0030] The first aspect of the present invention provides an enantiotesenane-type 2-norditerpene lactone compound, the structural formula of which is shown in Formula I;

[0031]

[0032] The second aspect of the present invention provides a method for preparing the above-mentioned enantiotesenane-type 2-norditerpene lactone compound, comprising the following steps:

[0033] The dried root of Euphorbia chamaejasme is used as a raw material, and the extract is obtained after percolation extraction and concentration, and the extract is suspended in water to obtain a suspension, and the extract is obtained after extraction and concentration;

[0034] The extract is subjected to gradient elution by silica gel column chromatography, MCI resin column chromatography, and silica gel column chromatography in sequence, and isocratic elution is performed by Sephadex LH-20 gel column chromatography and ODS medium pressure column chromatography to obtain Fr.1 to Fr.4. Fr.2 is prepared by ODS liquid chromatography to obtain the enantiotherm-type 2-norditerpene lactone compound shown in formula I.

[0035] In a preferred embodiment of the present invention, before the MCI resin column chromatography, the extract is subjected to gradient elution by silica gel column chromatography to obtain a first eluate, and the eluent is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to ethyl acetate is 10:1 to 1:10. Specifically, the eluent includes eluents with a volume ratio of petroleum ether to ethyl acetate of 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, and 1:10, and 7 eluates are collected segmentally by gradient elution with the eluent.

[0036] In a preferred embodiment of the present invention, the first eluate is subjected to gradient elution through the MCI resin column chromatography to obtain a second eluate, the eluent is methanol-water, and the volume ratio of the methanol to water is 20:80 to 90:10. Specifically, the eluent includes eluents with a volume ratio of methanol to water of 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, and 90:10, and 8 eluates are collected segmentally through gradient elution with the eluent.

[0037] Further preferably, the first eluate is obtained by eluting with an eluent having a volume ratio of petroleum ether to ethyl acetate of 1:1.

[0038] In a preferred embodiment of the present invention, the second eluate is subjected to gradient elution through the silica gel column chromatography to obtain a third eluate, and the eluent is petroleum ether-acetone-methanol, and the volume ratio of the petroleum ether, acetone and methanol is 15:1:1 to 1:1:1. Specifically, the eluent includes eluents with a volume ratio of petroleum ether-acetone-methanol of 15:1:1, 10:1:1, 8:1:1, 5:1:1, 3:1:1, and 1:1:1, and 6 eluates are collected segmentally after gradient elution with the eluent.

[0039] Further preferably, the second eluate is obtained by eluting with an eluent having a volume ratio of methanol to water of 60:40.

[0040] In a preferred embodiment of the present invention, the third eluate is isocratically eluted through the Sephadex LH-20 gel column chromatography to obtain a fourth eluate, the eluent is dichloromethane-methanol, and the volume ratio of dichloromethane to methanol is 1:1; specifically, the elution rate of the eluent is 1 mL / min, the elution time is 7 h, and the eluate is collected every 1 h, and 7 eluates are collected in segments after elution with the eluent.

[0041] Further preferably, the third eluate is obtained by eluting with the eluent having a volume ratio of petroleum ether-acetone-methanol of 3:1:1.

[0042] In a preferred embodiment of the present invention, the fourth eluate is eluted by the ODS medium pressure column chromatography to obtain Fr.1-Fr.4, the mobile phase is methanol-water, and the volume ratio of methanol to water is 60:40 to 80:20. Specifically, the eluent includes a volume ratio of methanol to water of 60:40 to 80:20, the rising speed is 2.5 ratios per hour, and the eluate is collected every 2 hours. The four eluates are collected by the gradient elution of the eluent segment.

[0043] Further preferably, the fourth eluate is obtained by eluting with the eluent in the fifth time period.

[0044] Further preferably, the flow rate of eluting the fourth eluate through the ODS medium pressure column chromatography is 25 mL / min, and the total elution time is 8 hours.

[0045] In a preferred embodiment of the present invention, Fr.2 is eluted by ODS liquid chromatography with a fixed ratio mobile phase to obtain the compound of formula I, the mobile phase is acetonitrile-water, and the volume ratio of acetonitrile to water is 50:50.

[0046] In a preferred embodiment of the present invention, the percolation extraction step is as follows: extracting the dried root of Euphorbia chamaejasme by percolation with ethanol at room temperature, obtaining an extract-like ethanol extract after concentration, and suspending the ethanol extract in water to obtain a suspension; the ethanol concentration is 95%, the percolation extraction time is 120h, and the percolation extraction flow rate of the ethanol is 0.5L / h;

[0047] The extraction step is as follows: adding dichloromethane in an equal volume to the suspension, extracting and concentrating to obtain the extract.

[0048] The third aspect of the present invention provides a use of the above-mentioned enantiotesenane-type 2-norditerpene lactone compound in the preparation of a drug for preventing or treating calcific aortic valve disease.

[0049] In a preferred embodiment of the present invention, the enantiotesenane-type 2-norditerpene lactone compound is used as a raw material to prepare a pharmaceutical composition for preventing or treating calcific aortic valve disease or any pharmaceutically acceptable salt thereof.

[0050] The pharmaceutical composition is added with excipients, and the present invention has no special limitation on the excipients, which may be pharmaceutically acceptable excipients well known to those skilled in the art.

[0051] The pharmaceutical composition or any pharmaceutically acceptable salt is in the form of tablets, capsules, granules, oral liquids, granules, pills or pellets.

[0052] The fourth aspect of the present invention provides a drug for preventing or treating calcific aortic valve disease, the raw materials of which include the above-mentioned enantiotesenane-type 2-norditerpene lactone compound.

[0053] The drug for preventing or treating calcific aortic valve disease also includes pharmaceutically acceptable excipients; the dosage form can be tablets, capsules, granules, oral liquids, granules, pills or micropills.

[0054] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0055] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0056] The present invention provides an enantiotesenane-type 2-norditerpene lactone compound having a structure shown in the following formula I:

[0057]

[0058] The preparation method, structural identification results and pharmacological activity of the enantiotesenane-type 2-norditerpene lactone compound are as follows:

[0059] Example 1

[0060] The dried root (1 kg) of Euphorbia chamaejasma is used as a raw material, and 95% ethanol is used for percolation extraction for 120 hours at a flow rate of 0.5 L / h, and rotary evaporation is performed to obtain an ethanol extract in an extract form, and the ethanol extract is suspended in water to obtain a suspension, and dichloromethane equal to the volume of the suspension is added for extraction, and the extract solution is concentrated to obtain an extract. The dichloromethane extract is subjected to silica gel column chromatography, and gradient elution is performed using a petroleum ether-ethyl acetate solution with a volume ratio of 10:1 to 1:10. Specifically, the eluent includes an eluent with a volume ratio of petroleum ether-ethyl acetate of 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, and 1:10, and 7 eluates are collected in sections after gradient elution with the eluent. The eluate obtained by elution with an eluent having a volume ratio of petroleum ether to ethyl acetate of 1:1 was subjected to MCI resin column chromatography and gradient eluted with a methanol-water solution with a volume ratio of 20:80 to 90:10. Specifically, the eluent included eluents having a volume ratio of methanol to water of 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, and 90:10. Eight eluates were collected in segments after gradient elution with the eluent. The eluate eluted with an eluent of methanol-water in a volume ratio of 60:40 was subjected to silica gel column chromatography, and gradient eluted with a petroleum ether-acetone-methanol solvent in a volume ratio of 15:1:1 to 1:1:1. Specifically, the eluent includes an eluent with a volume ratio of petroleum ether-acetone-methanol of 15:1:1, 10:1:1, 8:1:1, 5:1:1, 3:1:1, and 1:1:1. Six eluates were collected by segmental elution with the eluent. The eluate eluted with an eluent of petroleum ether-acetone-methanol in a volume ratio of 3:1:1 was subjected to Sephadex LH-20 gel column chromatography, and gradient eluted with a dichloromethane-methanol solvent in a volume ratio of 1:1. Specifically, the elution rate of the eluent was 1 mL / min, the elution time was 7 hours, and the eluate was collected every 1 hour. Seven eluates were collected by segmental elution with the eluent. The eluate eluted by the eluent in the fifth time period was subjected to ODS medium pressure column chromatography, and gradient eluted with a methanol-water solvent with a volume ratio of 60:40 to 80:20. Specifically, the eluent includes a volume ratio of methanol and water of 60:40 to 80:20 with a gradient increase, a rising speed of 2.5 ratios per hour, a flow rate of 25 mL / min, and collected once every 2 hours. The total elution time is 8 hours, and four eluates Fr.1 to Fr.4 were collected in segments by eluent gradient elution. The Fr.2 part was prepared by preparative ODS liquid chromatography with acetonitrile-water solution with a volume ratio of 50:50 as the mobile phase to obtain the compound of formula I.

[0061] Example 2

[0062] The structure of the compound of formula I prepared in Example 1 was identified.

[0063] The identification results are as follows:

[0064] The compound shown in formula I is a colorless block crystal (methanol), which is easily soluble in methanol and dichloromethane. A 10% sulfuric acid ethanol solution shows purple red. The compound shown in formula I is tested by nuclear magnetic resonance, mass spectrum, optical rotation, infrared spectrum, ultraviolet spectrum, X-ray single crystal diffraction and other data, thereby confirming that the compound of formula I is a new enantiopyrane-type 2-norditerpene lactone compound of the present invention.

[0065] Compound of formula I: colorless blocky crystals; melting point: 137–138°C; [α] 2 D 5 :+36.5(methanol, c 0.1); UV(methanol)λ max (logε): 202(4.34)nm; IR (potassium bromide)ν max :3426, 2921, 2850, 1715, 1646cm –1 ;(+)-HRESIMS m / z 341.1725[M+Na] + (C 19 H 26 O4Na + The calculated value of m / z is 341.1729); 1 H and 13 C NMR data are shown in Table 1.

[0066] Table 1 Compounds of Formula I 1 H-NMR (600MHz, CDCl3) and 13 C-NMR (150MHz, CDCl3) data

[0067]

[0068] Figure 1 It is the 1H-NMR spectrum of the enantiopyrane-type 2-norditerpene lactone compound of the present invention.

[0069] Figure 2 It is the 13C-NMR spectrum of the enantiopyrane-type 2-norditerpene lactone compound of the present invention.

[0070] Figure 3 It is the single crystal diffraction structure of the enantiopyrane-type 2-norditerpene lactone compound of the present invention.

[0071] Example 3: Pharmacological activity of compounds of formula I

[0072] Test methods and results

[0073] 1. CCK-8 assay to detect the effect of the compound of formula I on the proliferation of valvular interstitial cells (VICs)

[0074] CCK8 kit was purchased from Bio-Tech. Valvular interstitial cells were cultured at 1×10 3 (cells / well) were inoculated in a 96-well plate. After 24 hours, the cells were treated with different concentrations (50-4000 μM) of the compound of formula I for 72 hours, stained according to the instructions of the kit, and the absorbance value at 450 nm was measured. The calculation formula of the valvular interstitial cell growth inhibition rate of the test compound is: Cell growth inhibition rate % = (A (negative control group) - A (drug group)) / A (negative control group) × 100%

[0075] GraphPad Prism 8 statistical software was used for experimental data analysis, and the cell proliferation inhibitory activity of the samples was evaluated using the half-maximal inhibitory concentration.

[0076]

[0077] A G is the average OD value of the drug-treated group, A K is the average OD value of the blank control group, A M is the average OD value of the model group.

[0078] Compared with the control group, the compound of formula I inhibited the growth of valvular interstitial cells in a dose-dependent manner, IC 50 The value was 207.3 μmol / L, and it had no cytotoxicity to valvular interstitial cells within 100 μmol / L.

[0079] 2. Formula I compounds can inhibit the osteogenic differentiation of valvular interstitial cells

[0080] Valvular interstitial cells were collected and homogenized with RIPA lysis buffer containing protease and phosphatase inhibitor cocktails. Equal amounts of protein were taken for SDS-PAGE electrophoresis, electrophoresed in 4-20% Tris-Glycine Mini Gels for 40-60 minutes depending on the target protein, and then transferred to polyvinylidene difluoride membranes. Blocked with TBS-T (50mM Tris-HCl, pH 8.0, 150mM NaCl and 0.1% Tween-20) buffer containing 5% skim milk powder for 1 hour at room temperature, and then incubated with the primary antibody at 4°C overnight. Subsequently, it was washed with TBS-T and incubated with the corresponding secondary antibody labeled with the compound of formula I at room temperature for 1 hour. The immune complex was then visualized using enhanced chemiluminescence (ECL) reagent. GAPDH was used as a standard for total protein determination. Specific bands were quantified by optical density determination using image J1.54. In the alkaline phosphatase (ALP) staining experiment, valvular interstitial cells were incubated with the above interventions in conditioned medium for 7 days. ALP staining was used to detect ALP expression. The cultured valvular interstitial cells were washed twice with PBS and fixed with 4% PEA for 15 min at room temperature. ALP staining was then performed using an alkaline phosphatase staining kit, and the procedure was performed according to the kit instructions. In the Alizarin Red staining experiment, VICs were cultured in different conditioned media for 21 days. To detect calcium deposition, Alizarin Red staining was performed. The specific steps were as follows: VICs were washed twice with PBS and then fixed with 4% paraformaldehyde for 10 min at room temperature. The cells were washed three times with ddH2O, then incubated with 0.2% Alizarin Red solution for 30 min, and then washed three times with ddH2O. Both alkaline phosphatase staining and Alizarin Red staining were visualized using an Olympus BX51 microscope, and image acquisition was performed using an Olympus DP71 camera and cellens software.

[0081] Based on the dose-response curve and IC 50 Value, in the next experiment, the cells were treated with the compound of formula I at a concentration of 4 μmol / L. After 3 days of treatment with the compound of formula I, Western blotting found that the compound of formula I could significantly reduce the expression levels of osteoblast markers ALP and Runx2. In addition, alkaline phosphatase staining showed that after 7 days of culture in osteogenic medium, the compound of formula I reduced the ALP activity of valvular interstitial cells. In addition, Alizarin red staining showed that after 21 days of culture in osteogenic medium, the compound of formula I significantly reduced calcium deposition in valvular interstitial cells.

[0082] Figure 4 The graph is a graph showing the inhibition rate of the proliferation of valvular interstitial cells (VICs) by the enantiotesenane-type 2-norditerpene lactone of the present invention.

[0083] Figure 5The immunoblot diagram (a) and the corresponding statistical diagram (b) show the effects of the enantiotesenane-type 2-norditerpene lactone of the present invention on the expression of ALP and Runx2 in VICs cultured in osteogenic medium.

[0084] Figure 6 The staining diagram (a) and the corresponding statistical diagram (b) show the effect of the enantiotesenane-type 2-norditerpene lactone of the present invention on the alkaline phosphatase (ALP) activity in VICs cultured in osteogenic medium.

[0085] Figure 7 This is a staining diagram showing the effect of the enantiotesenane-type 2-norditerpene lactone of the present invention on calcium precipitation in VICs cultured in osteogenic medium.

[0086] In summary, the enantiotesenane-type 2-norditerpene lactone compound prepared by the present invention has good heart valve calcification inhibitory activity and can be used to prepare drugs for calcific aortic valve disease. Such compounds have great development value as new drugs for the prevention or treatment of calcific aortic valve disease, and the design ideas of such compounds also provide new ideas and approaches for the development of new drugs for the prevention or treatment of calcific aortic valve disease.

[0087] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. An enantiotesenane-type 2-norditerpene lactone compound, characterized in that: The structural formula is shown in Formula I; 2. A method for preparing the enantiotesenane-type 2-norditerpene lactone compound according to claim 1, characterized in that: The following steps are involved: The dried root of Euphorbia chamaejasme is used as a raw material, and the extract is obtained after percolation extraction and concentration, and the extract is suspended in water to obtain a suspension, and the extract is obtained after extraction and concentration; The extract is sequentially subjected to silica gel column chromatography, MCI resin column chromatography, and silica gel column chromatography for gradient elution, and then subjected to isocratic elution through Sephadex LH-20 gel column chromatography and elution through ODS medium pressure column chromatography to obtain Fr.1 to Fr.4, and Fr.2 is subjected to ODS liquid chromatography to obtain an enantiopyrane-type 2-norditerpene lactone compound shown in formula I; Before the MCI resin column chromatography, the extract is subjected to gradient elution through silica gel column chromatography to obtain a first eluate, the eluent is petroleum ether-ethyl acetate, and the volume ratio of the petroleum ether to ethyl acetate is 10:1 to 1:10; Taking the first eluate and subjecting it to gradient elution through the MCI resin column chromatography to obtain a second eluate, wherein the eluent is methanol-water, and the volume ratio of the methanol to water is 20:80 to 90:10; The second eluate is subjected to gradient elution through the silica gel column chromatography to obtain a third eluate, wherein the eluent is petroleum ether-acetone-methanol, and the volume ratio of the petroleum ether, acetone and methanol is 15:1:1 to 1:1:1; The third eluate is subjected to isocratic elution through the Sephadex LH-20 gel column chromatography to obtain a fourth eluate, wherein the eluent is dichloromethane-methanol, and the volume ratio of dichloromethane to methanol is 1:1; The fourth eluate is taken and eluted by the ODS medium pressure column chromatography to obtain Fr.1-Fr.4, the mobile phase is methanol-water, and the volume ratio of the methanol to water is 60:40-80:20; Fr.2 was taken and subjected to ODS liquid chromatography with a fixed ratio mobile phase for elution to obtain a compound of formula I, wherein the mobile phase was acetonitrile-water, and the volume ratio of acetonitrile to water was 50:50; The percolation extraction step is as follows: extracting the dried root of Euphorbia chamaejasme by percolation with ethanol at room temperature, obtaining an ethanol extract in the form of an extract after concentration, and suspending the ethanol extract in water to obtain a suspension; the ethanol concentration is 95%, the percolation extraction time is 120 hours, and the percolation extraction flow rate of the ethanol is 0.5 L / h; The extraction step is as follows: adding dichloromethane in an equal volume to the suspension, extracting and concentrating to obtain the extract.

3. Use of the enantiotesenane-type 2-norditerpene lactone compound according to claim 1 in the preparation of a medicament for preventing or treating calcific aortic valve disease.

4. A drug for preventing or treating calcific aortic valve disease, characterized in that: The raw materials include the enantiotesenane-type 2-norditerpene lactone compound according to claim 1.

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