C13 non-isoprene derivative, extraction method thereof, pharmaceutical composition and application of C13 non-isoprene derivative in preparation of anti-inflammatory drugs or / and anti-tumor drugs

Abstract

The invention relates to the technical field of tobacco extracts, in particular to a C13 non-isoprene derivative, an extraction method thereof, a pharmaceutical composition and application of the C13 non-isoprene derivative in preparation of anti-inflammatory drugs or / and anti-tumor drugs. The C13 nonisoprene derivative is obtained by taking tobacco leaves in southern Anhui Province of Anhui Province as a raw material, extracting by using a mixed solvent of ketone and water, extracting by using an ether solvent, sequentially eluting and separating by using silica gel column chromatography, MCI macroporous adsorption resin column chromatography, Sephadex LH-20 gel column chromatography and preparative thin-layer chromatography, and combining TLC (Thin Layer Chromatography) color development and ultraviolet absorption. A cellular level anti-inflammatory experiment shows that the compound has an obvious anti-inflammatory effect on lipopolysaccharide induced inflammation, and has an obvious proliferation inhibition effect on four cancer cells, namely A549, A549-TAX, HT-29 and SNU-398; the compound can be used for preparing anti-inflammatory drugs or / and anti-tumor drugs.

Description

Technical Field

[0001] This invention relates to the field of tobacco extract technology, and more particularly to C 13 Non-isoprene derivatives and their extraction methods, pharmaceutical compositions and their use in the preparation of anti-inflammatory and / or antitumor drugs. Background Technology

[0002] Tobacco (Nicotiana tabacum L.) is a plant belonging to the Solanaceae family, and its dried leaves have unique medicinal value and potential for industrial application. Anhui Southern Tobacco, belonging to the kingdom Plantae, phylum Angiospermae, class Dicotyledons, order Solanales, family Solanaceae, genus Nicotiana, is a distinctive cultivar selected and cultivated in southern Anhui, China. Under the subtropical monsoon climate and unique red-yellow soil ecological conditions, Anhui Southern Tobacco has, through long-term adaptive domestication, developed abundant secondary metabolites, which are of great significance for its research.

[0003] Modern phytochemical research shows that tobacco leaves are rich in novel secondary metabolites, with over 2,500 compounds identified to date, covering alkaloids, terpenes, phenols, flavonoids, phenylpropanoids, etc. Some components have been shown to possess analgesic, anti-inflammatory, antibacterial, and neuromodulatory activities. For example, patent document CN102675105A discloses a phenolic compound from tobacco, its preparation method, and its applications. The phenolic compound (5-hydroxy-4-oxo-valeric acid)-(3-hydroxy-4,5-dimethoxy)-phenethyl ester isolated from tobacco exhibits good cytotoxic activity against MKN-28 cells and also shows some activity against K562 and Bel-7402 cells. Therefore, identifying new secondary metabolites from tobacco leaves as lead compounds can provide a material basis for drug development. Summary of the Invention

[0004] The present invention aims to solve the above problems by providing a C 13 Non-isoprene derivatives and their extraction methods, pharmaceutical compositions and their use in the preparation of anti-inflammatory and / or antitumor drugs.

[0005] The technical solution to the problem solved by the present invention is, in a first aspect, to provide a C 13 Non-isoprene derivatives, having the following formula The structure shown is as follows:

[0006] Mode ;

[0007] in, Indicates a single or double bond; R1 and R2 are independently selected from hydrogen, hydroxyl, methoxy, ethoxy, amino, and nitro, respectively; R3 is selected from hydrogen, hydroxyl, methoxy, ethoxy, and C1~6 alkoxy.

[0008] As a preferred embodiment of the present invention, the C 13 Non-isoprene derivatives have the following formula The structure shown is as follows:

[0009] Mode ;

[0010] R1 and R2 are independently selected from hydrogen, hydroxyl, methoxy, ethoxy, amino, and nitro, respectively; R3 is selected from hydrogen, hydroxyl, methoxy, ethoxy, and C1~6 alkoxy.

[0011] As a preferred embodiment of the present invention, R1 is a hydroxyl group, R2 is hydrogen, and R3 is a hydroxyl group; or R1 is hydrogen, R2 is hydroxyl, and R3 is hydroxyl; or R1, R2, and R3 are all hydroxyl groups.

[0012] As a preferred embodiment of the present invention, the C 13 Non-isoprene derivatives, having the following formula The structure shown is as follows:

[0013] Mode .

[0014] Secondly, another object of the present invention is to provide the aforementioned C 13 The extraction method for non-isoprene derivatives includes the following steps:

[0015] S1. Using tobacco leaves from southern Anhui as raw material, extract and concentrate the extract using a mixed solvent of ketones and water to obtain an extract;

[0016] S2. The extract is extracted with an ether solvent to obtain an ether phase extract;

[0017] S3. The ether phase extract was separated by silica gel column chromatography, and eluted sequentially with petroleum ether and petroleum ether-ethyl acetate system, and the elution fraction with a volume ratio of petroleum ether to ethyl acetate of 4:1 to 1:10 was collected.

[0018] S4. The collected eluted fraction is subjected to MCI macroporous adsorption resin column chromatography with gradient elution in an alcohol-water system. The secondary components are obtained by combining TLC color development and ultraviolet absorption.

[0019] S5. The obtained sub-fractions were subjected to Sephadex LH-20 gel column chromatography, eluted with a petroleum ether-dichloromethane-methanol system with a volume ratio of 2:1:1, and obtained by TLC color development and ultraviolet absorption.

[0020] S6. The obtained subfractions are separated by preparative thin-layer chromatography, combined with TLC color development and ultraviolet absorption, and eluted with ester solvents to obtain the C. 13 Non-isoprene derivatives.

[0021] In step S1 The chemical composition of tobacco leaves varies depending on the production area. This invention uses tobacco leaves from southern Anhui Province as raw material. Preferably, the lower leaves of tobacco leaves from southern Anhui Province are used as raw material. Due to the shading effect of the upper leaves, the lower leaves grow and mature under conditions of high humidity, poor light, and poor ventilation. Furthermore, their own nutrients are continuously transported upwards to the upper leaves, resulting in thinner leaves, looser tissue, and lower dry matter content. Therefore, their quality as cigarette raw material is poor. Using them for extraction allows for full utilization of their nutrients.

[0022] During extraction, the ketone solvent can, for example, be a moderately polar ketone solvent such as acetone or butanone. Preferably, the mixed solvent used during extraction is a mixture of acetone and water.

[0023] The proportion of acetone in the mixed solvent affects the polarity of the mixed solvent. Preferably, it is a mixed solvent with an acetone to water volume ratio of (2~4):1; more preferably, it is a mixed solvent with an acetone to water volume ratio of 3:1, suitable for including C 13 Dissolution of compounds, including non-isoprene derivatives.

[0024] As a preferred embodiment of the present invention, ultrasonic immersion is employed. Preferably, the immersion time is 20-28 hours, the ultrasonic treatment time is 25-40 minutes, and the extraction temperature is 25-30°C. For example, the immersion time can be 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, or 28 hours, preferably 24 hours; the ultrasonic treatment time can be 25 minutes, 30 minutes, 35 minutes, or 40 minutes, preferably 30 minutes; and the extraction temperature can be 25°C, 26°C, 27°C, 28°C, 29°C, or 30°C, preferably 28°C. Preferably, ultrasonic immersion extraction is performed 4-6 times, and the extracts obtained each time are combined; more preferably, ultrasonic immersion extraction is performed 5 times.

[0025] The method of concentration after extraction is not limited, but as a preferred embodiment of the present invention, the concentration is vacuum concentration.

[0026] In step S2 As a preferred embodiment of the present invention, the extract is suspended in water and then extracted with the ether solvent. The resulting extract is then concentrated to obtain an ether phase extract. Preferably, the volume ratio of water to ether solvent is 1:1. The concentration method is not limited, but preferably, the concentration is performed under reduced pressure.

[0027] Ether solvents may, for example, be diethyl ether, cyclopentylmethyl ether, etc. Preferably, diethyl ether is used as the ether solvent in this invention.

[0028] As a preferred embodiment of the present invention, the extraction is performed 1 to 3 times, and the extracts obtained each time are combined; preferably, the extraction is performed 3 times.

[0029] In step S3 As a preferred embodiment of the present invention, the silicone column is 300-400 mesh.

[0030] As a preferred embodiment of the present invention, in the gradient elution of the petroleum ether-ethyl acetate system, a petroleum ether:ethyl acetate gradient elution of 99:1 to 5:95 (volume ratio) is adopted. Specifically: in the initial stage, elution is carried out with petroleum ether and ethyl acetate in a volume ratio of 99:1; in the final stage, elution is carried out with petroleum ether and ethyl acetate in a volume ratio of 5:95; in the intermediate stage between the initial and final stages, the proportion of ethyl acetate in the eluent gradually increases, for example, elution with petroleum ether and ethyl acetate in volume ratios of 91:9, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, and 9:91, etc. The elution fraction with a petroleum ether:ethyl acetate volume ratio of 4:1 to 1:10 is collected.

[0031] As a preferred embodiment of the present invention, after gradient elution of the petroleum ether-ethyl acetate system, the elution further includes sequential elution of the dichloromethane-methanol system and elution of dichloromethane. Preferably, in the dichloromethane-methanol system, the volume ratio of dichloromethane to methanol is 4:1.

[0032] The elution rate is not limited, but as a preferred embodiment of the present invention, the elution rate is 40-50 mL / min; for example, it can be 40 mL / min, 41 mL / min, 42 mL / min, 43 mL / min, 44 mL / min, 45 mL / min, 46 mL / min, 47 mL / min, 48 mL / min, 49 mL / min, or 50 mL / min, with 45 mL / min being the most preferred.

[0033] In step S4 As a preferred embodiment of the present invention, the gradient elution of the alcohol-water system is performed by sequentially eluting with alcohol solvents of volume concentrations of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100%. Preferably, the alcohol-water system is a methanol-water system.

[0034] The elution rate is not limited, but as a preferred embodiment of the present invention, the elution rate is 40-50 mL / min; for example, it can be 40 mL / min, 41 mL / min, 42 mL / min, 43 mL / min, 44 mL / min, 45 mL / min, 46 mL / min, 47 mL / min, 48 mL / min, 49 mL / min, or 50 mL / min, with 45 mL / min being the most preferred.

[0035] As a preferred embodiment of the present invention, the combination of TLC color development and ultraviolet absorption in step S4 refers to: developing the sample on TLC using a petroleum ether-diethyl ether solvent with a volume ratio of 1:4, and collecting and combining the developed R... f The subcomponent is obtained by selecting a component with a value of 0.4 to 0.6 and exhibiting ultraviolet absorption at a wavelength of 254 nm under ultraviolet light. Preferably, the expanded R... f The component with a value of 0.5 and ultraviolet absorption at a wavelength of 254 nm under ultraviolet light is obtained as the subcomponent.

[0036] In step S5 The elution rate is not limited. As a preferred embodiment of the present invention, the elution rate is 1 to 5 mL / min; for example, it can be 1 mL / min, 2 mL / min, 3 mL / min, 4 mL / min, or 5 mL / min, preferably 3 mL / min.

[0037] As a preferred embodiment of the present invention, the combination of TLC color development and ultraviolet absorption in step S5 refers to: developing the sample on TLC using a petroleum ether-diethyl ether solvent with a volume ratio of 1:4, and collecting and combining the developed R... f The subcomponent is obtained by selecting components with a value of 0.4 to 0.6 and exhibiting ultraviolet absorption at a wavelength of 254 nm under ultraviolet light. Preferably, the expanded R... f The subcomponent is obtained by selecting a component with a value of 0.5 and exhibiting ultraviolet absorption at a wavelength of 254 nm under ultraviolet light.

[0038] In step S6, as a preferred embodiment of the present invention, the silica gel used in the preparative thin-layer chromatography is 6~10 μm.

[0039] As a preferred embodiment of the present invention, step S6, combining TLC color development and ultraviolet absorption, refers to: developing the TLC using a dichloromethane-methanol system with a volume ratio of 20:1, and collecting R... f The component with a value of 0.5 to 0.6 and exhibiting ultraviolet absorption at a wavelength of 254 nm under ultraviolet light is obtained as described above. 13 Non-isoprene derivatives.

[0040] As a preferred embodiment of the present invention, the ester solvent is ethyl acetate. Preferably, the washing is performed 3 to 5 times.

[0041] Thirdly, another object of the present invention is to provide a pharmaceutical composition comprising the aforementioned C 13 Non-isoprene derivatives, or / and the C 13 Stereoisomers of non-isoprene derivatives, the C 13 Tautomers of non-isoprene derivatives, the C 13 Geometric isomers of non-isoprene derivatives, the C 13 At least one of the pharmaceutically acceptable salts of non-isoprene derivatives.

[0042] As a preferred embodiment of the present invention, the pharmaceutical composition further includes a pharmaceutically acceptable carrier and / or excipients.

[0043] As a preferred embodiment of the present invention, the dosage form of the pharmaceutical composition is one of tablets, capsules, granules, oral liquids, and injections.

[0044] Fourthly, another object of the present invention is to provide the C 13 Application of non-isoprene derivatives in the preparation of anti-inflammatory drugs and / or antitumor drugs.

[0045] As a preferred embodiment of the present invention, the C 13 Non-isoprene derivatives can be used in the preparation of anti-inflammatory drugs by inhibiting LPS-induced inflammatory responses. Preferably, the inflammation is at least one of pneumonia, enteritis, hepatitis, or neuroinflammatory diseases.

[0046] As a preferred embodiment of the present invention, the C 13 Non-isoprene derivatives can be used in the preparation of antitumor drugs by inhibiting cancer cell proliferation. Preferably, the C... 13 Non-isoprene derivatives can be used in the preparation of anticancer drugs by inhibiting the proliferation of cancer cells. Preferably, the cancer cells are at least one of A549, A549-TAX, HT-29, and SNU-398. The cancers involved are at least one of lung cancer, paclitaxel-resistant lung cancer, liver cancer, and colorectal cancer.

[0047] The beneficial effects of this invention are:

[0048] This application provides a C 13 This non-isoprene derivative, a novel compound discovered in the lower leaves of tobacco leaves from southern Anhui, China, has shown significant anti-inflammatory effects against lipopolysaccharide-induced inflammation through cellular-level anti-inflammatory experiments. It is a novel drug for the preparation of treatments for inflammatory diseases. Furthermore, it exhibits significant inhibitory effects on the proliferation of four cancer cells: A549, A549-TAX, HT-29, and SNU-398. It can be used to prepare novel drugs for the treatment of inflammatory and anti-tumor diseases. Detailed Implementation

[0049] The following are specific embodiments of the present invention, and the technical solutions of the present invention will be further described, but the present invention is not limited to these embodiments.

[0050] Example 1

[0051] A type of C 13 Non-isoprene derivatives have the structure shown in the following formula (the Arabic numerals in the structural formula are the carbon atom numbers in the chemical structure):

[0052]

[0053] This C 13 Non-isoprene derivatives were obtained through the following steps:

[0054] S1. The lower leaves of *Nicotiana tabacum* L., harvested in 2024 from southern Anhui, China, were air-dried. The viscera and epidermis were carefully peeled off and pulverized. After rinsing with acetone, the leaves were extracted five times with ultrasonication using a 3:1 (acetone to water) mixed solvent. Each extraction lasted 24 hours, with ultrasonication for 30 minutes at 28°C. The extracts were combined and concentrated under reduced pressure to obtain a black extract.

[0055] S2. Suspend the above black extract in water, add an equal volume of anhydrous ether, extract three times, combine the ether extracts, and concentrate under reduced pressure to obtain the ether phase extract.

[0056] S3. The ether phase extract was subjected to silica gel column chromatography with a 300-400 mesh screen and a gradient elution system of pure petroleum ether → petroleum ether: ethyl acetate = 99:1~5:95 → dichloromethane: methanol = 4:1 → pure dichloromethane. The elution fraction C with a volume ratio of petroleum ether / ethyl acetate of 4:1~1:10 was collected.

[0057] S4. The collected elution fraction C was subjected to MCI macroporous adsorption resin column chromatography and eluted with methanol aqueous solutions with alcohol volume fractions of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% in sequence. The fractions were collected and combined and developed on TLC with a petroleum ether-diethyl ether solvent system with a volume ratio of 1:4. The fractions with an Rf value of 0.5 and ultraviolet absorption at a wavelength of 254 nm were obtained as subcomponent CO.

[0058] S5. The subcomponent CO was subjected to Sephadex LH-20 gel column chromatography (Amersham Pharmacia Biotech, Sweden), eluted with a 2:1:1 petroleum ether-dichloromethane-methanol system, and the fractions that were collected and combined and developed on TLC with a 1:4 volume ratio petroleum ether-diethyl ether solvent system, with an Rf value of 0.5 and UV absorption at a wavelength of 254 nm, were obtained as subcomponent COJ.

[0059] S6. Subcomponent COJ was separated by preparative thin-layer chromatography (HSGF254, Yantai Jiangyou Silica Gel Development Co., Ltd.). The mixture was developed using a 20:1 dichloromethane-methanol system. The silica gel adsorption bands that showed UV absorption at a wavelength of 254 nm were marked, scraped off, and the silica gel adsorbate was repeatedly eluted with ethyl acetate and concentrated to obtain tabacumate.

[0060] Structural identification of compounds

[0061] Tabacumate: Green powder; -12 (c 0.40, MeOH); UV (MeOH) λ max (log ε) 323(1.0) nm; ECD (MeOH) λ max (Δε) 200 (–11.9), 230 (+0.4) nm; IR (KBr) 3339,2923, 2853, 1682, 1599, 1447, 1271, 1176 cm -1 ; 1 H and 13 The C NMR data are shown in Table 1 below; HRESIMS m / z 389.1960 [M+H] + (calcd for C 20 H 29 O3, 388.1886).

[0062] Table 1. Tabacumate 1 H (600 MHz) and 13 C (125 MHz) NMR data (CD3OD) No. <![CDATA[δ H mult. (J in Hz)]]> <![CDATA[δ C ]]> 1 - 35.6 s 2 1.46 dd (13.2, 9.3) ,1.69 ddd (13.3, 3.5, 1.5) 43 t 3 4.99 dddd (9.1, 7.6, 5.6, 3.5) 68.9 d 4 1.90dd (14.7, 7.6),2.40ddd, (14.8, 5.5, 1.4) 37.9 t 5 - 67 s 6 - 71.2 s 7 5.95 dd (15.4, 1.4) 125.3 d 8 5.73 dd, (15.4, 5.9) 139.5 d 9 4.32 td (6.2, 1.3) 68.6 d 10 1.24 d (6.4) 23.8 q 11 1.03 s 25.5 q 12 1.20 s 29.2 q 13 1.23 s 20.3 q 14 - 168.6 s 15 6.23 d (15.8 ) 115.4 d 16 7.52 d (15.9 ) 146.8 d 17 - 127.7 s 18 7.04 d (2.1) 115.1 d 19 - 146.8 s 20 - 149.6 s 21 6.79 d (8.1) 116.5 d 22 6.94 dd (8.2, 2.1) 122.9 d

[0063] The chemical structure of the compound tabacumate was determined by nuclear magnetic resonance spectroscopy (Bruker Avance III 500 or 600, Bruker Biospin AG, Germany) and high-resolution mass spectrometry (Agilent G6520 Q-TOF, Agilent Technologies, USA).

[0064] Example 2

[0065] C 13 Application of non-isoprene derivatives in the preparation of anti-inflammatory drugs.

[0066] Using an LPS-induced pro-inflammatory differentiation model of RAW264.7 macrophages, we further screened for anti-inflammatory effects of chemical components in tobacco leaves and tested their anti-inflammatory properties. Details are as follows:

[0067] RAW264.7 cells (Wuhan Pronosei Biotechnology Co., Ltd.) were cultured in DMEM high-glucose (Gibco, USA) medium containing 10% fetal bovine serum FBS (Gibco, USA) and 1% penicillin antibiotics until the cell density reached 80%. Cells were then cultured at 3.0 × 10⁻⁶ cells / year. 4 Raw 264.7 cells were seeded into 96-well plates at a density of 100 μL per well and cultured at 37°C under humidified conditions containing 5% CO2 for 24 h.

[0068] In vitro anti-inflammatory effect: Five concentration gradients (50, 25, 12.5, 6.25, and 3.125 μM) were set up for the treatment group, blank control group, model group, and positive control group (dexamethasone and Adamas). After incubation for 1 h, lipopolysaccharide (LPS) (Gibco, USA) was added to the model group, treatment group, and positive control group to a final concentration of 0.05 μg / mL. After culturing for another 24 h, 70 μL of supernatant from each group was collected and mixed with 70 μL of Greiss buffer (Sigma-Aldrich, St. Louis, MO, USA). After incubation for 15 min, the absorbance (OD) value of the mixture was measured at 546 nm using a microplate reader, and the nitrite level was calculated according to the sodium nitrite standard curve.

[0069] Cytotoxicity: Five concentration gradients (50, 25, 12.5, 6.25, and 3.125 μM) were set up for the treatment groups and a blank control group. C was added to the treatment groups. 13 For non-isoprene derivatives, DMSO was added as a solvent to the blank control group. After 24 hours of drug addition, 20 μL / well of 5 mg / ml MTT [Sangon Biotech (Shanghai) Co., Ltd.] was added to each well of a 96-well plate, and after incubation for 4 hours, the OD value of each group was measured at 570 nm.

[0070] The experimental results are as follows:

[0071] C 13 The non-isoprene derivatives exhibited significant toxicity to RAW264.7 cells at a concentration of 50 μM (cell viability was 56.22%). At concentrations of 25 μM, 12.5 μM, 6.25 μM, and 3.125 μM, the cell viability of RAW264.7 cells was 107.54%, 112.59%, 108.39%, and 108.40%, respectively.

[0072] Within the dose range of 25 μM to 3.125 μM, C 13 Non-isoprene derivatives inhibit LPS-induced NO production in macrophages. 50 The concentration was 11.39 μM, with the IC50 of the positive control drug dexamethasone being [missing value]. 50 The value is 7.62 μM. Therefore, C 13 Non-isoprene derivatives can effectively inhibit LPS-induced inflammatory responses.

[0073] Example 3

[0074] C 13 Application of non-isoprene derivatives in the preparation of antitumor drugs.

[0075] The antitumor activity was tested by inhibiting the proliferation of A549 (human non-small cell lung cancer cells), A549-TAX (human lung adenocarcinoma paclitaxel-resistant cells), HT-29 (human colon cancer cells), and SNU-398 (human liver cancer cells). All cells were obtained from Wuhan Pronosei Biotechnology Co., Ltd. Details are as follows:

[0076] Cells in the logarithmic growth phase were digested with 0.125% trypsin and counted using a cell counter. 3000 cells were added to each well of a 96-well plate with 100 μL of DMEM medium and incubated overnight at 37°C in a 5% CO2 incubator. Five concentration gradients of the compound (3.125, 6.25, 12.5, 25, and 50 μM) were added, with three replicates for each concentration. After 72 h of incubation, 20 μL of MTT reagent was added to each well. After 4 h of incubation, the supernatant was aspirated, and 150 μL of DMSO was added. The absorbance at 490 nm was measured using a microplate reader (Molecular Devices, Sunnyvale, CA, USA). The average absorbance from the three replicates was taken, and cell viability was calculated based on the absorbance values. Cell viability = (OD value of experimental group / OD value of control group) x 100%, ICso value was calculated using GraphPadPrims10 software.

[0077] The experimental results are as follows:

[0078] Table 2. Nicotiana tabacum L. C 13 IC50 of non-isoprene derivatives against the antiproliferative activity of four cancer cells: A549, A549-TAX, HT-29H, and SNU-398. 50 (μM) <![CDATA[A549 IC 50 (μM)]]> <![CDATA[A549-TAX IC 50 (μM)]]> <![CDATA[HT-29 IC 50 (μM)]]> <![CDATA[SNU-398 IC 50 (μM)]]> tabacumate 35.18 19.35 42.12 17.48 Paclitaxel 32.27 8.20 0.0086 >20

[0079] It can be seen that C 13Non-isoprene derivatives showed a significant concentration-dependent inhibitory effect on the proliferation of four cancer cells: A549, A549-TAX, HT-29, and SNU-398. Furthermore, their inhibitory effects on A549 and SNU-398 were comparable to those of the positive control drug paclitaxel. IC 50 The values ​​were 35.18, 19.35, 42.12, and 17.48 μM, respectively.

[0080] The specific embodiments described herein are merely illustrative of the spirit of the invention. Those skilled in the art to which this invention pertains may make various modifications or additions to the described specific embodiments or use similar methods to substitute them, without departing from the spirit of the invention or exceeding the scope defined by the appended claims.

Claims

1. A type of C 13 Non-isoprene derivatives, characterized in that: Having such a formula The structure shown is as follows: Mode ; in, Indicates a single or double bond; R1 and R2 are independently selected from hydrogen, hydroxyl, methoxy, ethoxy, amino, and nitro, respectively; R3 is selected from hydrogen, hydroxyl, methoxy, ethoxy, and C1~6 alkoxy.

2. A C according to claim 1 13 Non-isoprene derivatives, characterized in that: Having such a formula The structure shown is as follows: Mode ; R1 and R2 are independently selected from hydrogen, hydroxyl, methoxy, ethoxy, amino, and nitro, respectively; R3 is selected from hydrogen, hydroxyl, methoxy, ethoxy, and C1~6 alkoxy.

3. A C according to claim 2 13 Non-isoprene derivatives, characterized in that: R1 is a hydroxyl group, R2 is hydrogen, and R3 is a hydroxyl group; or R1 is hydrogen, R2 is a hydroxyl group, and R3 is a hydroxyl group; or R1, R2, and R3 are all hydroxyl groups.

4. A C according to claim 3 13 Non-isoprene derivatives, characterized in that: Having such a formula The structure shown is as follows: Mode .

5. A C as described in any one of claims 1 to 4 13 A method for extracting non-isoprene derivatives, characterized in that: Includes the following steps: S1. Using tobacco leaves from southern Anhui as raw material, extract and concentrate the extract using a mixed solvent of ketones and water to obtain an extract; S2. The extract is extracted with an ether solvent to obtain an ether phase extract; S3. The ether phase extract was separated by silica gel column chromatography, and eluted sequentially with petroleum ether and petroleum ether-ethyl acetate system, and the elution fraction with a volume ratio of petroleum ether to ethyl acetate of 4:1 to 1:10 was collected. S4. The collected eluted fraction is subjected to MCI macroporous adsorption resin column chromatography with gradient elution in an alcohol-water system. The secondary components are obtained by combining TLC color development and ultraviolet absorption. S5. The obtained sub-fractions were subjected to Sephadex LH-20 gel column chromatography, eluted with a petroleum ether-dichloromethane-methanol system with a volume ratio of 2:1:1, and obtained by TLC color development and ultraviolet absorption. S6. The obtained subfractions are separated by preparative thin-layer chromatography, combined with TLC color development and ultraviolet absorption, and eluted with ester solvents to obtain the C. 13 Non-isoprene derivatives.

6. A C according to claim 5 13 A method for extracting non-isoprene derivatives, characterized in that: In step S1, a mixed solvent of acetone and water in a volume ratio of (2~4):1 is used for soaking and ultrasonic extraction; the soaking time is 20~28h, the ultrasonic extraction time is 25~40min, and the extraction temperature is 25~30℃.

7. A pharmaceutical composition, characterized in that: Including C as described in any one of claims 1 to 4 13 Non-isoprene derivatives, or / and the C 13 Stereoisomers of non-isoprene derivatives, the C 13 Tautomers of non-isoprene derivatives, the C 13 Geometric isomers of non-isoprene derivatives, the C 13 At least one of the pharmaceutically acceptable salts of non-isoprene derivatives.

8. Use of the pharmaceutical composition of claim 7 in the preparation of an anti-inflammatory drug and / or an antitumor drug.

9. The application according to claim 8, characterized in that: The inflammation is at least one of pneumonia, enteritis, hepatitis, or neurological inflammation.

10. The application according to claim 8, characterized in that: Inhibiting the proliferation of at least one of A549, A549-TAX, HT-29, and SNU-398 cells can be used in the preparation of antitumor drugs.

Images