阅读说明：本技术 一种生物碱类化合物及其制备方法和应用 (Alkaloid compound and preparation method and application thereof ) 是由 曹亮 肖伟 李海波 杨一帆 顾莎莎 王团结 杨彪 胡晗绯 王振中 于 2020-10-26 设计创作，主要内容包括：本发明公开了一种生物碱类化合物,通过理化性质和现代波谱学手段,对分离得到的化合物进行了结构鉴定。本发明还运用LPS诱导RAW 264.7细胞炎症模型的活性筛选体系进行对其进行活性评价,发现该化合物对小鼠巨噬细胞系RAW 264.7有一定的保护作用,可以显著抑制PGE2,显示出较强的抗炎作用。(The invention discloses an alkaloid compound, which is characterized in that the structure of the separated compound is identified through physicochemical properties and modern spectroscopy means. The invention also applies an activity screening system of an LPS-induced RAW 264.7 cell inflammation model to carry out activity evaluation on the compound, and finds that the compound has a certain protection effect on a mouse macrophage system RAW 264.7, can obviously inhibit PGE2 and shows a stronger anti-inflammatory effect.)

1. An alkaloid compound or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, prodrug molecule, metabolite thereof, wherein the compound has a structure represented by formula I:

2. a process for the preparation of a compound according to claim 1, comprising the steps of:

step 1: reflux-extracting herba Artemisiae Annuae with 3-5 times of water for 2-3 times, mixing extractive solutions, recovering solvent under reduced pressure, adding 90-95% ethanol until alcohol concentration reaches 70-90%, standing, collecting supernatant, and concentrating under reduced pressure to obtain total extract;

step 2: dissolving the total extract in water, separating by macroporous adsorption resin column chromatography, performing gradient elution by sequentially using water, 45-55% ethanol and 90-100% ethanol, respectively collecting eluates, and concentrating under reduced pressure to obtain a water elution part, a 45-55% ethanol elution part and a 90-100% ethanol elution part;

and step 3: and (3) separating the 45-55% ethanol elution part by silica gel column chromatography, performing gradient elution by using dichloromethane-methanol at a volume ratio of 100:0 to 0:100, collecting 11 fractions A-K, performing ODS column chromatography on fraction D, performing gradient elution by using methanol-water at a volume ratio of 90:10 to 80:20 to obtain 3 fractions D1-D3, and performing semi-preparative liquid chromatography on fraction D2.

3. The method of claim 2, wherein the step 1 comprises: reflux-extracting herba Artemisiae Annuae with 3-5 times of water for 2-3 times, each for 1-3 hr, mixing extractive solutions, recovering solvent under reduced pressure to 1/10 of original volume, adding 95% ethanol until alcohol concentration reaches 70-90%, standing overnight, collecting supernatant, and concentrating under reduced pressure to obtain total extract.

4. The preparation method according to claim 2, wherein the gradient elution in the step 2 is gradient elution sequentially with water, 45% ethanol and 90% ethanol aqueous solutions with different concentrations.

5. The method of claim 2, wherein the step 1 comprises extracting with 3-5 times of water under reflux for 2 times each for 2 hours, combining the extractive solutions, recovering solvent under reduced pressure to 1/10, and adding 95% ethanol until the alcohol concentration reaches 80%.

6. The preparation method according to claim 2, wherein the macroporous adsorbent resin is selected from the group consisting of D101 type macroporous adsorbent resin, HP-20 type macroporous adsorbent resin, HPD-100A type macroporous adsorbent resin, and HPD-300 type macroporous adsorbent resin.

7. The method of claim 2, wherein the semi-preparative liquid chromatography conditions include: specification C₁₈A Phenomenex Gemini column with the diameter of 5 mu m and the diameter of 10 multiplied by 250mm, methanol-water with the volume ratio of 10-20: 80-90 as a mobile phase, the detection wavelength of 238-258nm, and the flow rate of 1-5 mL/min.

8. The production method according to claim 3, wherein the conditions of the semi-preparative liquid chromatography include: methanol-water with the volume ratio of 15:85 is used as a mobile phase, the detection wavelength is 248nm, and the flow rate is 4 mL/min.

9. Use of a compound according to claim 1, or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, prodrug molecule, metabolite thereof, for the manufacture of an anti-inflammatory medicament.

10. A medicament comprising a compound of claim 1 or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, prodrug molecule, metabolite thereof.

Technical Field

The invention relates to the technical field of medicines, and particularly relates to a novel compound and a preparation method and application thereof.

Background

Herba Artemisiae Annuae is dried aerial part of Artemisia annua L of Compositae, collected in autumn when the flower is full, removed old stem, and dried in the shade. The bitter, pungent and cold flavor enters liver and gallbladder meridians, and has the effects of clearing deficiency heat, removing bone-steaming, relieving summer-heat, repelling, and eliminating jaundice. Can be used for treating yin impairment due to pathogenic warm, night fever and early coolness, fever due to yin deficiency, hectic fever due to yin deficiency, fever due to summer-heat pathogen, malaria with chills and fever, and jaundice due to damp-heat pathogen.

Anti-inflammatory drugs in clinical treatment are the second largest class of drugs next to anti-infective drugs, including steroidal anti-inflammatory drugs (SAID) and non-steroidal anti-inflammatory drugs (NSAID). However, because of the strong toxic and side effects of many synthetic drugs, people pay more and more attention to the development of anti-inflammatory drugs from natural drugs.

Disclosure of Invention

The invention aims to carry out more intensive research on a novel sweet wormwood anti-inflammatory active ingredient and find an active ingredient which plays an anti-inflammatory role.

In view of the above, the present invention provides an alkaloid compound or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, prodrug molecule, metabolite thereof, wherein the structure of the compound is shown in formula I:

another object of the present invention is to provide a method for preparing the above compound, which comprises the steps of:

step 1: taking dry aerial parts of sweet wormwood, performing reflux extraction for 2-3 times by using 3-5 times of water for 2 hours each time, combining extracting solutions, recovering a solvent to 1/10 of the original volume under reduced pressure, adding 90-95% ethanol until the alcohol concentration reaches 70-90%, standing overnight, collecting supernate, and performing reduced pressure concentration to obtain a total extract;

step 2: dissolving the total extract in water, performing macroporous adsorption resin column chromatographic separation, performing gradient elution by sequentially using water, 45-55% ethanol and 90-100% ethanol, respectively collecting eluates, and performing reduced pressure concentration until no alcohol smell exists to obtain a water elution part, a 45-55% ethanol elution part and a 90-100% ethanol elution part;

and step 3: and (3) separating the 45-55% ethanol elution part by silica gel column chromatography, performing gradient elution with dichloromethane-methanol (100:0 to 0:100, v/v), collecting 11 fractions A-K, performing ODS column chromatography on fraction D, performing gradient elution with methanol-water (90:10 to 80:20, v/v), obtaining 3 fractions D1-D3, and performing semi-preparative liquid chromatography on fraction D2.

Further, the step 1 is as follows: drying aerial parts of herba Artemisiae Annuae, extracting with 3-5 times of water under reflux for 2-3 times, each for 2 hr, mixing extractive solutions, recovering solvent under reduced pressure to 1/10 of original volume, adding 95% ethanol until alcohol concentration reaches 80%, standing overnight, collecting supernatant, and concentrating under reduced pressure to obtain total extract.

Preferably, the gradient elution of step 2 is gradient elution performed by sequentially using water, 45% ethanol and 90% ethanol aqueous solutions with different concentrations.

Specifically, the macroporous adsorption resin is selected from D101 type macroporous adsorption resin, HP-20 type macroporous adsorption resin, HPD-100A type macroporous adsorption resin or HPD-300 type macroporous adsorption resin.

Further, the semi-preparative liquid chromatography conditions are: specification C₁₈A Phenomenex Gemini column with the volume ratio of 5 mu m and 10 multiplied by 250mm, methanol-water with the volume ratio as a mobile phase, the detection wavelength of 238-258nm, and the flow rate of 1-5 mL/min; preferably 246-250nm, at a flow rate of 3-4 mL/min.

Preferably, in the step 1, the extraction is performed by hot reflux extraction for 2 times with 3 times of water, each time lasts for 2 hours, the extracting solutions are combined, the solvent is recovered under reduced pressure to 1/10 of the original volume, 95% ethanol is added until the alcohol concentration reaches 80%, standing overnight is performed, and the supernatant is collected and concentrated under reduced pressure to obtain the total extract.

Alternatively, the conditions of the semi-preparative liquid chromatography are preferably: methanol-water with volume ratio of 15:85 is used as mobile phase, detection wavelength is 248nm, flow rate is 4mL/min, and retention time can be 22.5 min.

The invention also aims to provide application of the compound or pharmaceutically acceptable salts, solvates, tautomers, stereoisomers, prodrug molecules and metabolites thereof in preparing anti-inflammatory drugs. The compound has obvious inhibition effect on LPS induced PGE2 generated by RAW 264.7 cells.

The invention also provides a medicament for treating inflammation, which comprises the compound shown in the formula (I) or pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, prodrug molecule and metabolite thereof. The anti-inflammatory agent is a drug for treating or preventing inflammatory reaction after tissue injury and the like.

Further, the medicament contains a therapeutically effective amount of the compound and one or more pharmaceutically acceptable carriers.

Specifically, the medicament can be any one of the dosage forms in pharmaceutics, including tablets, capsules, soft capsules, gels, oral preparations, suspensions, granules, patches, ointments, pills, powders, injections, infusion solutions, freeze-dried injections, intravenous emulsions, liposome injections, suppositories, sustained-release preparations or controlled-release preparations.

Further, the pharmaceutically acceptable carrier refers to a pharmaceutical carrier conventional in the pharmaceutical field, such as: diluents, excipients, and water, and the like, fillers such as starch, sucrose, lactose, microcrystalline cellulose, and the like; binders such as cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone; humectants such as glycerol; disintegrating agents such as sodium carboxymethyl starch, hydroxypropyl cellulose, crosslinked carboxymethyl cellulose, agar, calcium carbonate and sodium bicarbonate; absorption enhancers such as quaternary ammonium compounds; surfactants such as cetyl alcohol, sodium lauryl sulfate; adsorption carriers such as kaolin and bentonite; lubricants such as talc, calcium and magnesium stearate, micronized silica gel, polyethylene glycol, and the like. Other adjuvants such as flavoring agent, sweetener, etc. can also be added.

The alkaloid compound is a new chemical component found in artemisia apiacea by researchers, and is found to stably exist in different batches of artemisia apiacea. The inventor adopts physical and chemical properties and modern wave spectrum means (HR-MS, MS,¹H-NMR、¹³C-NMR, 2D-NMR and the like), and the compound obtained by the separation method is identified to be a new compound with the structure shown in the formula (I). The invention also utilizes an activity screening system such as an LPS (LPS) -induced RAW 264.7 cell inflammation model and the like to carry out activity evaluation, and finds that the compound has a certain protection effect on a mouse macrophage system RAW 264.7 and can obviously inhibit PGE (platelet-rich antigen)₂Has stronger anti-inflammatory action and good research and development prospect.

Drawings

FIG. 1 is a HR-ESI-Q-TOF-MS spectrum of a compound of the present invention;

FIG. 2 is a UV spectrum of a compound of the present invention;

FIG. 3 is an IR spectrum of a compound of the present invention;

FIG. 4 is a drawing of a compound of the present invention¹H-NMR spectrum;

FIG. 5 is a drawing of a compound of the present invention¹³C-NMR spectrum;

FIG. 6 is a DEPT-135 spectrum of a compound of the present invention;

FIG. 7 is H of a compound of the present invention¹-H¹A COSY spectrum;

FIG. 8 is an HSQC spectrum of a compound of the present invention

FIG. 9 is an HMBC spectrum of a compound of the present invention;

FIG. 10 shows the main HMBC correlation and H for the compounds of the present invention¹-H¹COSY is related.

Detailed Description

The following will specifically describe the contents of the experimental examples.

It is specifically noted that similar alternatives and modifications will be apparent to those skilled in the art, which are also intended to be included within the present invention. It will be apparent to those skilled in the art that the techniques of the present invention may be implemented and applied by modifying or appropriately combining the methods and applications described herein without departing from the spirit, scope, and content of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention.

The present invention is carried out under the conventional conditions or the conditions recommended by the manufacturer if the specific conditions are not specified, and the reagents or apparatuses used are not specified by the manufacturer and are conventional products commercially available.

EXAMPLE 1 preparation of the Compounds of the invention

(1) Drying aerial parts of herba Artemisiae Annuae (Artemisia annua L.), extracting with 3 times of water under reflux for 2 times (each time for 2 hr), mixing extractive solutions, recovering solvent under reduced pressure to 1/10 of original volume, adding 95% ethanol until alcohol concentration reaches 80%, standing overnight, collecting supernatant, and concentrating under reduced pressure to obtain total extract. Dissolving the total extract in water, separating by HP-20 macroporous adsorbent resin column chromatography, sequentially eluting with water, 45% ethanol and 90% ethanol, collecting eluates, respectively, and concentrating under reduced pressure until no ethanol smell exists to obtain water eluate, 45% ethanol eluate and 90% ethanol eluate;

(2) taking the 45% ethanol elution part in the step (1), carrying out silica gel column chromatographic separation, carrying out dichloromethane-methanol gradient elution (100:0 to 0:100, v/v) with 4 column volumes of each gradient elution, collecting 11 fractions A-K, carrying out ODS column chromatographic separation on fraction D, carrying out methanol-water gradient elution (90:10 to 80:20, v/v) with 4 column volumes of each gradient elution, obtaining 3 fractions D1-D3, and carrying out semi-preparative liquid chromatographic separation on fraction D2 to obtain the target product alkaloid.

Wherein, the semi-preparative liquid chromatography conditions in the step (2) are as follows: semi-preparative chromatography column: phenomenex Gemini (C)₁₈5 μm,10 × 250mm), semi-preparative high performance liquid chromatography [ shimadzu, japan pump: LC-6AD (SHIMADZU, LIQUID CHROMATOGRAPH); a detector: SPD-20A (timing UV/VIS DETECTOR); a workstation: LC solution)]Methanol-water with the volume ratio of 15:85 is used as a mobile phase, the detection wavelength is 248nm, the flow rate is 4mL/min, and the retention time on the semi-prepared liquid phase is about 22.5 min.

EXAMPLE 2 preparation of Compounds of the invention

(1) Extracting dry aerial parts of herba Artemisiae Annuae with 4 times of water under reflux for 2 times, each for 2 hr, mixing extractive solutions, recovering solvent under reduced pressure to 1/10 of original volume, adding 95% ethanol until alcohol concentration reaches 80%, standing overnight, collecting supernatant, and concentrating under reduced pressure to obtain total extract. Dissolving the total extract in water, separating by HP-20 macroporous adsorbent resin column chromatography, sequentially eluting with water, 50% ethanol and 95% ethanol, collecting eluates, respectively, and concentrating under reduced pressure until no ethanol smell exists to obtain water eluate, 50% ethanol eluate and 95% ethanol eluate;

(2) separating by silica gel column chromatography, eluting with dichloromethane-methanol gradient (100:0 to 0:100, v/v), each gradient eluting for 4 column volumes, collecting 11 fractions A-K, separating fraction D by ODS column chromatography, eluting with methanol-water (90:10 to 80:20, v/v) gradient, each gradient eluting for 4 column volumes, obtaining 3 fractions D1-D3, and separating fraction D2 by semi-preparative liquid chromatography to obtain the target product alkaloid.

Wherein the semi-preparative liquid chromatography conditions in step (2) are as follows: phenomenex Gemini (C)₁₈5 μm,10 × 250mm), semi-preparative high performance liquid chromatography [ shimadzu, japan pump: LC-6AD (SHIMADZU, LIQUID CHROMATOGRAPH); a detector: SPD-20A (timing UV/VIS DETECTOR); a workstation: LC solution)]Methanol-water with the volume ratio of 15:85 is used as a mobile phase, the detection wavelength is 248nm, the flow rate is 4mL/min, and the retention time on the semi-prepared liquid phase is about 22.5 min.

EXAMPLE 3 preparation of the Compounds of the invention

(1) Extracting dry aerial parts of herba Artemisiae Annuae with 4 times of water under reflux for 3 times, each for 2 hr, mixing extractive solutions, recovering solvent under reduced pressure to 1/10 of original volume, adding 95% ethanol until alcohol concentration reaches 80%, standing overnight, collecting supernatant, and concentrating under reduced pressure to obtain total extract. Dissolving the total extract in water, separating by HP-20 macroporous adsorbent resin column chromatography, sequentially eluting with water, 55% ethanol, and 100% ethanol, collecting eluates, respectively, and concentrating under reduced pressure until no ethanol smell exists to obtain water eluate, 55% ethanol eluate, and 100% ethanol eluate;

(2) separating the 55% ethanol elution part obtained in the step (1) by silica gel column chromatography, performing dichloromethane-methanol gradient elution (100:0 to 0:100, v/v) with 4 column volumes per gradient elution, collecting 11 fractions A-K, performing ODS column chromatography on fraction D, performing methanol-water gradient elution (90:10 to 80:20, v/v) with 4 column volumes per gradient elution to obtain 3 fractions D1-D3, and performing semi-preparative liquid chromatography on fraction D2 to obtain the target product alkaloid.

Wherein the semi-preparative liquid chromatography conditions in step (2) are as follows: phenomenex Gemini (C)₁₈5 μm,10 × 250mm), semi-preparative high performance liquid chromatography [ shimadzu, japan pump: LC-6AD (SHIMADZU, LIQUID CHROMATOGRAPH); a detector: SPD-20A (timing UV/VIS DETECTOR); a workstation: LC solution)]Methanol-water with the volume ratio of 15:85 is used as a mobile phase, the detection wavelength is 248nm, the flow rate is 4mL/min, and the retention time on the semi-prepared liquid phase is about 22.5 min.

EXAMPLE 4 structural characterization of the Compounds of the invention

The compound was a tan gum solid. HR-ESI-MS (FIG. 1) gives M/z 232.1336[ M + H ]]+ (calculated 232.1338), molecular formula C14H17NO2 was determined and unsaturation was calculated as 7. The UV (MeOH) spectrum (FIG. 2) shows λ max (log ε):204(3.47),248(3.34),294(3.28) nm gives the maximum absorption band, and the IR (KBr) spectrum (FIG. 3) shows vmax 3403cm^-1And 1699cm^-1Characteristic absorption peak of carboxyl group.

¹H-NMR(400MHz,in DMSO-d₆) The spectrum (FIG. 4) shows a set of ortho-coupled aromatic hydrogen signals [ delta ]_H 7.54(1H,d,J＝7.9Hz),7.09(1H,d,J＝7.9Hz)]Three methyl hydrogen signals [ delta ]_H 2.43(3H,s),2.25(3H,s),1.20(3H,d,J＝6.9Hz)]And several methylene and methine hydrogen signals.¹³C-NMR(100MHz,in DMSO-d₆) Combined with the DEPT-135 map (FIGS. 5 and 6) showed a total of 14 carbon signals, including 6 quaternary carbon signals (delta)_C172.6,153.7,152.4,136.3,134.6,129.0), 3 methine carbon signals (δ)_C135.5,121.7,31.7), 2 methylene carbon signals (. delta.))_C30.9,27.7), 3 methyl carbon signals (. delta.)_C23.8,21.3,17.7). The directly attached carbons and hydrogens were assigned by HSQC spectroscopy (fig. 8) as shown in table 1.

¹H-¹In the H COSY spectrum (FIG. 7), H-3/H-4 correlation can be seen, in the HMBC spectrum (FIG. 9), H-3 is related to C-2/C-5, H-4 is related to C-2/C-12, the combined molecular formula and the characteristic chemical shifts (delta C153.7, C-2) and (delta C152.4, C-12) of C-2 and C-12 are presumed to contain a pyridine ring segment in the structure.¹H-¹In the H COSY spectrum, H is visible₃-14/H-6/H₂-7/H₂-8 correlation; in HMBC spectrum, H₃14 is related to C-5/C-6/C-7, H-7 is related to C-9, the methylene hydrogen signal H₂8 is related to the two double bond carbon signals C-9/C-11, and in addition, H-4 is clearly related to C-6. Methyl hydrogen signal [ delta ]_H 2.25(3H,s,H₃-15)]In relation to the carbon signal C-9/C-11 of the double bond in the seven-membered ring and the carbon signal C-12 of the pyridine ring, the unsaturation of the binding compound speculates that the pyridine ring is fused with a seven-membered ring. In HMBC spectra, the methyl hydrogen signal [ delta ]_H 2.43(3H,s,H-13)]In relation to C-2/C-3 on the pyridine ring, a methyl group is determined to be attached to the C-2 position. At the same time, the methyl hydrogen signal H₃15 has a clear remote correlation with the carboxyl carbon signal C-10, confirming that a carboxyl group is attached at the C-9 position. The above information was combined to give the compound structure (FIG. 10). The optical rotation of the compound is dextrorotation in chloroform solventDetermining the absolute configuration as S configuration. Other examples were identified as homogeneous compounds.

Table 1 Compounds in DMSO-d₆Nuclear magnetic data of (400MHz for)¹H；100MHz for¹³C)

EXAMPLE 5 in vitro anti-PGE of the Compound of example 1₂Experiment of

1. Material

1.1 pharmaceutical example 1 compound;

1.2 cell model mouse macrophage line RAW 264.7, from Chinese medicine academy of sciences, available from Jiangsu Kangyuan pharmaceutical Co., LtdProviding; the culture conditions are as follows: DMEM + 10% Fetal Bovine Serum (FBS), 37 ℃, 5% CO₂。

2. Principles and methods

2.1 principle of the experiment

Lipopolysaccharide (LPS) of gram-negative outer membrane (Sigma, USA, 114M4009) is one of the most main pathogenic molecules mediating infectious inflammatory lesions, and many diseases are closely related to LPS-induced persistent subclinical inflammation. LPS is widely used to induce inflammation in animals and in cellular experiments.

Macrophages play a crucial role in the inflammatory response, and after stimulation, macrophages produce large amounts of inflammatory factors and mediators, such as: TNF-alpha, IL-1 beta, IL-6, NO and PGE₂And the like. Activation of these inflammatory factors and mediators is a key process of inflammation, and their inhibition is often used as an important index for evaluating the anti-inflammatory activity of drugs.

2.2 drug inhibition assay for secreted PGE2

The method comprises the following steps:

(1) preparing a liquid medicine: the drug was dissolved in 10% FBS DMEM medium to prepare a 2mg/ml stock solution.

(2) The experimental method comprises the following steps: the cells were digested with 0.25% pancreatin (containing 0.02% EDTA), and the cell density was adjusted to 1X 10 in 10% FBS-containing DMEM medium⁵Each/ml, inoculated evenly into a 24-well plate, 400. mu.l per well, and placed into an incubator for 24 hours after plating.

Blank control group (N group): 495 μ l serum-free DMEM medium was added to each well;

vehicle group/solvent control group (RM group): 495 mul serum-free DMEM medium containing one thousandth of DMSO was added to each well;

model group (group M): 495 μ L of 100 μ g/mL LPS per well;

administration sample group: 495 microliter of culture medium containing different concentrations of drugs is added into each well;

simultaneously arranging 6 multiple holes, and putting CO into the 24-hole plate after the medicine is added₂The cell culture box was incubated for 1 hour. After 1 hour, 5. mu.L of 100. mu.g/mL LPS (final concentration of 1. mu.g/mL) was added to each well except for the blank and solvent controlsg/mL), adding 5 mu L of serum-free DMEM medium into each hole of the solvent control group, and putting the 24-hole plate into CO after the medicine is added₂The cell incubator was continued for 18 hours.

After 18 hours, cell culture fluid is collected, and PGE in cell supernatant is detected by ELISA method according to the kit instructions₂The content of (a).

PGE₂Inhibition (%) (model group PGE)₂Average content of-sample group PGE₂Average content of (1)/(PGE of model group)₂Average content of-solvent group PGE₂Average content) x 100%.

3. Results of the experiment

3.1 Effect of drug samples on mouse macrophage line RAW 264.7 cell supernatant PGE2

The result shows that the drug sample can obviously inhibit LPS (LPS) -induced mouse macrophage RAW 264.7PGE₂Shows strong anti-inflammatory action. Data results are shown in table 2.

TABLE 2 Compound (I) concentrations PGE supernatant of mouse macrophage cell line RAW 264.7₂Influence of (A), (B)n＝6)

The compound in the invention is tested by a linear regression analysis method through Graphadprism 7.00 analysis software to inhibit LPS in vitro to induce mouse macrophage RAW 264.7 to secrete inflammatory mediator PGE₂IC of₅₀65.41 μ M.

4. Conclusion

The compound of the invention induces mouse macrophage RAW 264.7 to secrete inflammatory medium PGE by LPS₂Has remarkable inhibitory effect, shows strong anti-inflammatory effect, and can treat PGE with the increase of drug concentration₂The inhibitory effect of secretion is also increased, its IC₅₀65.41 μ M.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.