CN111995603B - Sesquiterpene compound with antioxidant activity - Google Patents

Abstract

The invention discloses a sesquiterpene compound with antioxidant activity, which has a general formula

Description

Sesquiterpene compound with antioxidant activity

Technical Field

The invention relates to a compound and application thereof, in particular to a sesquiterpene compound with antioxidant activity and application thereof.

Background

At present, the influence of oxidative damage has become an important issue concerning human health. When the body is under stress, free radicals are generated. Failure of endogenous enzymatic and non-enzymatic antioxidant substances to handle free radicals produced under overload conditions can lead to imbalance of oxidative balance, cellular damage and other health problems. Modern pathological studies have shown that a lack of antioxidants can lead to degenerative diseases such as cancer, cardiovascular diseases, alzheimer's disease, neurodegenerative diseases and various inflammatory diseases. According to the World Health Organization (WHO) statistics, 80% of the population worldwide use traditional medical means for primary health care and care, and the treatment is mostly from herbal extracts and their active ingredients. Numerous studies have shown that antioxidants of natural plant origin can reduce the probability of human diseases. Natural products are capable of preventing and scavenging free radical production and are among the most valuable ways to reduce diseases caused by oxidative stress. At present, as some synthetic antioxidants such as BHT, BHA, TBHQ and the like are found to be toxic, finding safe and natural antioxidants is becoming an increasing research focus.

The Chloranthus japonicus is whole plant of Chloranthus japonicus (Chloranthus japonica Seib.) of Chloranthaceae, also called as Chloranthus japonica, ghost, etc., and is mainly produced in China, korea and Japan, and distributed in shady and wet places or furrow grass clusters in mountain slopes, mountain valley miscellaneous trees with elevation of 500-2300 m. The chloranthus japonicus has complex chemical components, mainly comprises sesquiterpenes, volatile oils, coumarins, amides and lignans, and is used for treating traumatic injury, rheumatic arthralgia, cold due to wind-cold, amenorrhea, skin pruritus, carbuncle, swelling, sore and furuncle in folk.

Sesquiterpene compounds are a class of compounds with high content of chloranthus plants and are also abundant in chloranthus japonicus. The sesquiterpenes which are available at present are numerous in types and quantity, mainly including eudesmane type and lindane type, and also include cadinane type, germacrane type and guaiane type. The chemical structure of the sesquiterpenoids determines various biological activities to a certain extent, and the biological activities of different structures are also different. At present, the compounds have the effects of resisting inflammation, tumors, oxidation, bacteria and viruses, and the like, and have the advantages of low toxicity, high activity and the like, and have larger application space. The Chlojaponilactone B is a sesquiterpene compound separated from the chloranthus japonicus, and the research in the earlier stage of the subject group finds that the Chlojaponilactone B has a strong anti-inflammatory effect, and simultaneously, the Chlojaponilactone B plays an anti-oxidation effect by inhibiting the reduction of active oxygen.

Disclosure of Invention

The invention aims to overcome the defect of providing a sesquiterpene compound with antioxidant activity and a preparation method and application thereof.

The technical scheme adopted by the invention is as follows:

in a first aspect of the present invention, there is provided:

sesquiterpene compounds having the general formula shown in formula I:

in the formula: r ₁ ～R ₆ Independently selected from hydrogen, hydroxy, methoxy, ethoxy, methyl ester, ethyl ester, C _1-10 Alkyl radical, C _1-10 Alkenyl radical, C _1-10 Is an ether or alcohol containing one or two oxygens, an amine or cyclic amine containing one or two nitrogens, phenyl, substituted phenyl, naphthyl, pyridyl, thienyl, furyl, methoxy, ethoxy, benzyloxy, formate, acetate, carboxamido, hydroxy, amino, methylamino, dimethylamino, nitro, trifluoromethyl, fluoro, chloro, bromo, and iodo; or R ₁ And R ₂ ，R ₂ And R ₃ Independently constructed as a 3-6 membered saturated carbocyclic ring, a double bond containing carbocyclic nitrogen or oxygen containing heterocyclic ring, or an aromatic ring.

In some examples, R ₂ 、R ₄ 、R ₅ Independently selected from hydrogen.

In some examples, R ₆ Is methyl.

In some examples, R ₂ 、R ₄ 、R ₅ Are all hydrogen, R ₆ Is methyl.

In some examples, the sesquiterpene compound has the structural formula:

in a second aspect of the present invention, there is provided:

use of a sesquiterpene compound according to the first aspect of the invention for the preparation of an agent for combating oxidative damage of experimental cells.

In a third aspect of the present invention, there is provided:

use of a sesquiterpene compound for the preparation of a medicament for the repair of oxidative damage to a cell, the sesquiterpene compound being as described in the first aspect of the invention.

In some examples, the cellular oxidative damage is selected from a neurological disease, cancer, cardiovascular disease, or cellular oxidative damage in inflammation.

In some examples, the neurological disease is alzheimer's disease, parkinson's disease, or the like.

In some examples, the drug is in the form of an oral formulation, an injection, or a mucosal administration.

In a fourth aspect of the present invention, there is provided:

a composition for cellular oxidative damage repair comprising an active ingredient comprising a sesquiterpene compound of the first aspect of the invention together with an acceptable carrier.

In some examples, the composition is a food product.

In a fifth aspect of the present invention, there is provided:

the preparation method comprises the following steps:

s1) dissolving Chlojaponilactone B in a solvent, adding Pd/C for catalytic hydrogenation, and reacting completely;

s2) after full reaction, removing Pd/C, and concentrating;

s3) passing the concentrate through a gel column, and eluting by taking pure methanol as a mobile phase;

s4) dot plate, combining the dots with the same Rf value, and concentrating under reduced pressure to obtain

The invention has the beneficial effects that:

the sesquiterpene compound provided by some embodiments of the invention has good antioxidant activity and good development prospect.

Drawings

FIG. 1 is the H spectrum of Compound 1;

FIG. 2 is a C spectrum of Compound 1;

FIG. 3 DEPT135 ° spectrum of Compound 1

FIG. 4 is a COSY spectrum of Compound 1;

FIG. 5 is the HSQC spectrum of Compound 1;

FIG. 6 is an HMBC spectrum of compound 1;

FIG. 7 is a NOE spectrum of Compound 1;

FIG. 8 is an HR-ESIMS spectrum of Compound 1;

FIG. 9 shows the survival rate of PC12 cells after the administration of Compound 1;

FIG. 10 is a graph of PC12 apoptosis following a dry prognosis with Compound 1 administration;

FIG. 11 is a study of the intervention of Compound 1 administration on H ₂ O ₂ Effect of cellular ROS fluorescence intensity change of stimulated PC 12;

FIG. 12 shows the results of ELISA assay for Compound 1;

FIG. 13 is a graph of oxidized protein mRNA expression.

Detailed Description

The present invention will be further described with reference to the following examples.

Preparation of Compound 1

Chlorojaponicumactone B (100 mg) was dissolved in methanol, and 1% by weight of Pd/C was added to the solution in an amount of 10mM, and the mixture was stirred at room temperature under a hydrogen atmosphere until the reaction was completed, and after 16 hours, the Pd/C catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure. The concentrated product was passed through a gel column with pure methanol as the mobile phase. Spots with the same Rf value were pooled and concentrated under reduced pressure to give Compound 1 (Perhydrochlojaponilactone B,14.5 mg).

Compound 1 is a white powder according to HRESIMS mass spectral data (317.1724, [ M ] +Na)]C + combined with 13C-NMR data (Table 1) to judge that the molecular formula is C ₁₇ H ₂₆ O ₄ The unsaturation degree was 5. ¹ H-NMR data (Table 1) showed that it had 5 methyl groups [ Delta H0.80 (3H, s), 0.86 (3H, d, J = 6.76Hz), 0.92(3H, d, J= 7.00Hz), 1.25 (3H, d, J= 7.36Hz) and 2.02 (3H, s)]And a series of aliphatic hydrogen signals. Analysis of ¹³ C-NMR spectrum and DEPT spectrum, which can be found to have 17 carbons, including: 1 ester group (δ C178.4), 1 acetyl group (δ C169.9), 5 methyl groups (δ C10.4,13.3,16.2,19.2,21.8), 2 sp3 hybridized methylene groups, 7 sp3 hybridized methine groups (1 oxygen linkage) and 1 sp3 hybridized quaternary carbon. Further analysis of the two-dimensional spectrum and ECD of compound 1 confirmed that the structure of compound 1 is as follows:

TABLE 1H NMR (400 MHz) and 13C NMR (100 MHz) data (CDCl 3,. Delta.in ppm) for Compound 1

The structure-related spectrum of the compound 1 is shown in fig. 1 to 8.

Compound 1 has repairing effect on cell oxidative damage

MTT assay

PC12 cells are a cell line cloned from rat adrenal pheochromocytoma, differentiate into sympathetic nerve cells after nerve growth factor stimulation, and are widely used for in vitro studies of neurological diseases. The survival rate of the PC12 cells after the compound 1 is administrated is tested by adopting an MTT method (detection principle: succinate dehydrogenase in mitochondria of living cells can enable exogenous MTT to be reduced into water-insoluble purple blue crystal formazan and be deposited in cells, while dead cells have no function, dimethyl sulfoxide (DMSO) can dissolve the formazan in the cells, an enzyme linked immunosorbent assay detector is used for measuring the light absorption value of the formazan at the wavelength of 450-500nm, the number of the living cells can be indirectly reflected, and the formation amount of MTT crystals is in direct proportion to the number of the cells in a certain cell number range).

The experimental reagent: compound 1 (1) prepared according to the present invention was dissolved in dimethyl sulfoxide (DMSO) to obtain a 10mM stock solution, which was then diluted to the desired concentration with DMEM medium. MTT solution is 5mg/mL, and the positive control drug is VC. The cells used in this experiment were PC12 cells.

The experimental method comprises the following steps: eight groups, compound 1 (40. Mu.M, 20. Mu.M, 10. Mu.M, 5. Mu.M and 2.5. Mu.M) were set up in this experiment as H ₂ O ₂ The treated cells were used as a model group, the DMSO-treated cells were used as a blank control group at a concentration of 0.02 to 0.10% and the DMSO-treated cells were used as a positive control group, and the cell viability was measured by the MTT method. Taking out PC12 cells in logarithmic growth phase from the incubator, digesting the cells growing adherent to the skin with pancreatin, and adding culture medium to stop digestion when the cells can be blown down. Counting with cell counting plate at 5X 10 ⁴ one/mL of the density was seeded in 96-well plates and 100. Mu.L of cell suspension was added per well. By mixing 96-well plates at 37 ℃ and 5% ₂ The culture box is used for culturing for 24 hours. Subsequently, the solvent control group cells were added with 0.02-0.10% DMSO, the administration group was added with different concentrations of Compound 1 (40. Mu.M group, 20. Mu.M group, 10. Mu.M group, 5. Mu.M group and 2.5. Mu.M group) for each group, and the model group was added with H ₂ O ₂ And a positive control group VC (10 mu M) treatment group, continuously culturing the solution for 24h, adding 20 mu L/hole of MTT solution, incubating for 4h in an incubator at 37 ℃ in a dark place, discarding the culture solution, adding 100 mu L DMSO, shaking for 10-20min, fully dissolving the generated crystals, developing, and measuring the absorbance A value of the crystals at the wavelength of 450-500nm by using an enzyme-labeling instrument. Each group was replicated 3 wells, and each group was replicated 3 times.

The inhibition rate calculation formula is as follows:

the experimental results are as follows: compounds 1 to H prepared according to the invention ₂ O ₂ The induced oxidative damage of PC12 cells had a clear repairing effect, as shown in FIG. 9 (in the figure, # indicates the blank ratio p<0.05; * Ratio of representation to model group p<0.05 Shown in (c). This result shows that: the compound 1 of the invention has relatively remarkable antioxidant activity.

Mitochondrial membrane potential detection

The mitochondrial membrane potential detection kit JC-1 is an ideal fluorescent probe widely used for detecting mitochondrial membrane potential. The mitochondrial membrane potential of cells, tissues or purified can be detected. The oxidative damage of the cells can cause apoptosis, the mitochondrial membrane potential reduction is a sign of early apoptosis, and the change of the mitochondrial membrane potential of the cells can be detected by the JC-1 probe.

Experimental reagent: compound 1 prepared according to the present invention was dissolved in dimethyl sulfoxide (DMSO) to obtain a 10mM stock solution, which was then diluted to the desired concentration with DMEM medium. The cells used in this experiment were PC12 cells.

The experimental method comprises the following steps: blank control group and H were set up for this experiment ₂ O ₂ Treatment groups and 3 different concentration administration groups (5. Mu.M, 10. Mu.M and 20. Mu.M). PC12 cells were cultured at 5.0X 10 ⁵ The density of cells/well was cultured in six well plates for 24h. With H ₂ O ₂ Treated cells as model groups, with different concentrations of Compound 1 and H ₂ O ₂ The treated cells were used as the administration group, and the DMSO-treated cells were used as the control group in an amount of 0.02 to 0.10%, and cultured for 24 hours.

Taking out the cells, placing the cells in a super clean bench, sucking out old culture medium, gently washing the cells twice by using ice-cold PBS, adding 1mL of PBS to blow down adherent cells, and centrifuging the cells at 5000rpm for 3-10min to precipitate the cells. The upper PBS layer was aspirated, 1mL serum-free medium was added followed by 1mL JC-1 staining medium, gently mixed by pipetting, and placed in a 37 ℃ cell incubator for further incubation for 20min. After incubation at 37 ℃, cells were pelleted by centrifugation at 5000rpm for 3-10 min. The supernatant was aspirated off and washed 2 times with JC-1 staining buffer (1X). 1mL of cell culture medium was added to resuspend the cells, and the cells were placed in a flow cytometer to observe changes in mitochondrial membrane potential.

The experimental results are as follows: compounds 1 to H prepared according to the invention ₂ O ₂ The induced oxidative damage of PC12 cells has obvious repairing effect, as shown in FIG. 10 (in the figure, A is control group; B is H) ₂ O ₂ Group (d); c is 1 (5. Mu.M) group; d is group 1 (10. Mu.M); e is group 1 (20. Mu.M); f is a histogram of apoptotic cell numbers. # indicates the ratio p to blank<0.005; * And indicate the ratio p to the model group<0.005 and p<0.01 Shown in (c). This result shows that: the compound 1 of the invention has relatively remarkable antioxidant activity.

Reactive Oxygen Species (ROS) detection kit

The Reactive Oxygen Species (ROS) detection kit is a kit for detecting reactive oxygen species by using a fluorescent probe DCFH-DA. DCFH-DA has no fluorescence, can freely pass through cell membranes, and can be hydrolyzed by intracellular esterase to generate DCFH after entering cells. DCFH, however, cannot permeate the cell membrane, thus making the probe easily loaded into the cell. Intracellular reactive oxygen species can oxidize non-fluorescent DCFH to produce fluorescent DCF. The level of reactive oxygen species in the cell can be known by measuring the fluorescence of DCF. The experiment adopts a flow cytometer to detect changes of mitochondrial membrane potential and ROS in PC12 cells, and researches the antioxidant activity of the compound 1.

Experimental reagent: compound 1 prepared according to the present invention was dissolved in dimethyl sulfoxide (DMSO) to obtain a 10mM stock solution, which was then diluted to the desired concentration with DMEM medium. The cells used in this experiment were PC12 cells.

The experimental method comprises the following steps: blank control group and H were set up for this experiment ₂ O ₂ Treatment groups and 3 different concentration administration groups (5. Mu.M, 10. Mu.M and 20. Mu.M). PC12 cells were cultured at 5.0X 10 ⁵ The density of cells/well was cultured in six well plates for 24h. With H ₂ O ₂ Treated cells as model groups, with different concentrations of Compound 1 and H ₂ O ₂ The treated cells were used as the administration group, and the DMSO-treated cells were used as the control group in an amount of 0.02 to 0.10%, and cultured for 24 hours. Taking out the cells, placing the cells in a super clean bench, sucking out old culture medium, gently washing the cells twice by using ice-cold PBS, adding 1mL of PBS to blow down adherent cells, and centrifuging the cells at 5000rpm for 3-10min to precipitate the cells. DCFH-DA was diluted to a final concentration of 10mM in serum-free medium following 1. The supernatant of the cells was removed, the cells were gently washed twice with serum-free medium, and 1mL of diluted DCFH-DA was added. Incubating in a constant-temperature cell incubator at 37 deg.C for 20-30min to allow the probe to fully enter the cell. The cells were gently washed three times with serum-free medium, and the probes that did not enter the cells were sufficiently washed away. 1mL of serum-free medium was added to infiltrate the cells, protected from light, and the active oxygen content was measured by a flow cytometer.

The experimental results are as follows: as shown in FIG. 11 (in the figure, A is a control group; B is H ₂ O ₂ Group (d); c is 1 (5. Mu.M) group; d is 1 (1)0 μ M) group; e is 1 (20. Mu.M) group), and H is higher than that in the control group ₂ O ₂ The DCF fluorescence intensity of the group was significantly increased, indicating a change in the fluorescence intensity over H ₂ O ₂ After treatment, ROS levels in PC12 cells were significantly increased; after the compound 1 is added for administration, the fluorescence intensity of DCF in PC12 cells is obviously reduced, and the ROS content is reduced in a concentration-dependent manner, which shows that the compound 1 prepared by the invention can obviously reduce the cause H ₂ O ₂ ROS production in PC12 cells is stimulated to exert antioxidant activity.

ELISA method for detecting activity of SOD, GSH-Px and GSH

The body produces a large amount of active oxygen and active nitrogen after being harmfully stimulated, which causes imbalance between oxidation state and oxidation resistance state, and causes pathological changes of tissues and cells. Under physiological state, an antioxidant system composed of SOD, GSH-px and GSH maintains the oxidation-reduction steady state of the organism. In order to delve into compound 1 vs H ₂ O ₂ The antioxidant effect in stimulated PC12 cells, we tested the activity of SOD, GSH-Px and GSH by ELISA method.

Experimental reagent: compound 1 prepared according to the present invention was dissolved in dimethyl sulfoxide (DMSO) to obtain a 10mM stock solution, which was then diluted to the desired concentration with DMEM medium. The cells used in this experiment were PC12 cells.

The experimental method comprises the following steps: blank control group and H were set up for this experiment ₂ O ₂ Treatment groups and 3 different concentration administration groups (5. Mu.M, 10. Mu.M and 20. Mu.M). PC12 cells were cultured at 5.0X 10 ⁵ The density of cells/well was cultured in six well plates for 24h. With H ₂ O ₂ Treated cells as model groups, with different concentrations of Compound 1 and H ₂ O ₂ The treated cells were used as the administration group, and the DMSO-treated cells were used as the control group in an amount of 0.02 to 0.10%, and cultured for 24 hours. According to the ELISA detection kit, the activity of detecting SOD, GSH-Px and GSH in the PC12 cells is strong and weak.

The experimental results are as follows: as shown in FIG. 12 (in the figure, # indicates the blank group ratio p<0.05; * Show and model group ratio p<0.05 Shown in (c). As shown in FIG. 12-A, control group comparison, H ₂ O ₂ SOD activity was significantly reduced in the stimulated group (p)<0.05 When the compound 1 is treated with the compound (I),SOD activity was gradually increased, and was significantly increased at a concentration of 20. Mu.M 1, which was statistically significant (p)<0.05). As shown in FIG. 12-B, H ₂ O ₂ Significant reduction of GSH-Px Activity in stimulated groups (p)<0.05 1) concentration of 10. Mu.M, the content thereof is remarkably enhanced (p)<0.05). As shown in FIG. 12-C, control group compares, H ₂ O ₂ Decreased GSH Activity in stimulated groups (p)<0.05 However, after 1 administration, the activity of the GSH is gradually enhanced, and when the concentration of 1 is 20 mu M, the activity of the GSH is most obvious and has statistical significance (p)<0.05). The compound 1 prepared by the invention can obviously increase the enzyme activities of SOD, GSH-Px and GSH so as to play the antioxidation activity.

RT-PCR experiment for detecting expression levels of Nrf2, GCLm, HO-1 and Nqo1 in PC12 cells

The content of antioxidant key protein in cells changes under the oxidative stress state, when the organism is in an oxidative damage state, the expression of antioxidant protein is reduced, and after the regulation treatment by antioxidant drugs, the protein expression is increased. The expression levels of Nrf2, GCLm, HO-1 and Nqo in PC12 cells are detected by RT-PCR experiments, and the antioxidant capacity of the compound 1 is explored.

Experimental reagent: compound 1 prepared according to the present invention was dissolved in dimethyl sulfoxide (DMSO) to obtain a 10mM stock solution, which was then diluted to the desired concentration with DMEM medium. The cells used in this experiment were PC12 cells.

The experimental method comprises the following steps: blank control group and H were set up for this experiment ₂ O ₂ Treatment groups and 3 different concentration administration groups (5. Mu.M, 10. Mu.M and 20. Mu.M). PC12 cells were cultured at 5.0X 10 ⁵ The density of cells/well was cultured in six well plates for 24h. Trizol reagent is added to extract total RNA (1 mL/well) of the cells, and the cells are lysed for 20-40min. Adding 100-300mL chloroform into RNA extract, shaking thoroughly, mixing, standing for 2-5min, and centrifuging at 10000-15000rpm in 4 deg.C centrifuge for 5-20min. Gently take out the sample, carefully suck the upper colorless and transparent layer liquid into a clean EP tube, add isopropanol with equal volume, mix up and down. Standing for 5-20min, and centrifuging at 4 deg.C at 10000-15000rpm for 5-20min. The supernatant was aspirated off, leaving a white precipitate, and 0.5-2mL 75% ethanol was added. Gently bounce off the precipitate and leave at 4 deg.CCentrifuging at 5000-10000rpm in heart machine for 5-20min. Removing supernatant, standing at room temperature for 5-10min to volatilize ethanol completely. Adding 30-100 μ L DEPC water, and storing on ice. The RNA concentration was measured in a Nanodrop 2000 ultramicro spectrophotometer. Based on the RNA concentrations obtained, RNA was loaded onto all the different samples in a uniform amount of 1mg. A sterile 200mL EP tube was added with 1mg of RNA of known concentration, 4. Mu.L of 4 XgDNA wiper Mix and RNase-free ddH was added ₂ And O is increased to 16 mu L, the mixture is lightly blown and uniformly mixed by a pipette gun, and then the mixture is heated in a constant temperature water bath kettle at 42 ℃ for 2min and taken out. Adding 4 μ L of 5 XHiScript II qRT Supermix II, mixing, heating in a 50 deg.C constant temperature water bath for 10-20min, heating in a 85 deg.C constant temperature water bath for 5s, and taking out. Finally, a PCR-dedicated 96-well plate was placed on ice, and the reagents were added to each well according to the ratios in Table 2, with the primer sequences shown in Table 3, in which glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the internal reference. The prepared 96-well plate was placed in a real-time fluorescent quantitative PCR instrument, and an amplification program was set, which is shown in table 4. Each sample was set up with 3 biological replicate wells and 3 technical replicates.

TABLE 2 RT PCR amplification reactant ratio Table

TABLE 3 primer sequence Listing

TABLE 4 RT PCR amplification program Table

Note: after the reaction is finished, an amplification curve and a melting curve are confirmed, and false negative results are eliminated. The standard amplification curve is S-shaped, and when the Ct value falls between 20 and 30, the quantitative analysis is most accurate; a single melting curve shows that the reaction specificity is good and quantitative result analysis can be carried out; if it is meltedIf the curve is bimodal or multimodal, quantitative analysis cannot be performed. The relative mRNA values are related to the Cyclerhreshold (Ct). The Ct relative magnitude value refers to the number of cycles that the fluorescence intensity in each reaction tube has undergone to reach a set threshold value. mRNA relative value =2 ^-ΔΔCt Wherein Δ Ct = sample Ct value-internal reference Ct value, Δ Δ Ct = sample Δ Ct value-blank control Δ Ct.

The experimental results are as follows: the results are shown in FIG. 13 (in which the oxidized protein mRNA expression pattern A (Nrf 2); B (HO-1); C (GCLm); D (Nqo); # # #, # # and #, respectively, indicates and blanks the ratio of p<0.005，p<0.01 and p<0.05; * And indicates and model group ratio is p<0.01 and p<0.05 PC12 cells in H ₂ O ₂ After stimulation and induction, the expression of antioxidant protein is down-regulated, 1 administration is dry, the expression is obviously up-regulated and is improved in a concentration dependence manner, and the compound 1 plays a stronger antioxidant role. Therefore, the invention provides a candidate compound for researching and developing new antioxidant drugs and provides a scientific basis for developing and utilizing natural active substances.

SEQUENCE LISTING

<110> Zhongshan university

<120> sesquiterpene compound with antioxidant activity

<130>

<160> 10

<170> PatentIn version 3.5

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Claims (7)

1. Sesquiterpene compounds having the structural formula:

2. use of a sesquiterpene compound according to claim 1 for the preparation of an agent for combating oxidative damage in experimental cells.

3. Use of a sesquiterpene compound according to claim 1 for the preparation of a medicament for the repair of oxidative damage of cells selected from the group consisting of oxidative damage of cells in neurological diseases, cancer, cardiovascular diseases or inflammation.

4. Use according to claim 3, characterized in that: the dosage form of the medicine is oral preparation, injection or mucosa administration.

5. A composition for cellular oxidative damage repair comprising an active ingredient and an acceptable carrier, wherein: the active ingredient comprises the sesquiterpene compound of claim 1.

6. The composition of claim 5, wherein: the composition is a food product.

7. A method of preparing the sesquiterpene compound of claim 1 comprising:

s1) dissolving Chlojaponilactone B in a solvent, adding Pd/C for catalytic hydrogenation, and reacting completely;

s2) after full reaction, removing Pd/C, and concentrating;

s3) passing the concentrate through a gel column, and eluting by taking pure methanol as a mobile phase;

s4) point plates, combining points with the same Rf value, and concentrating under reduced pressure to obtain

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