Polyacetylene compounds, methods for their preparation and use in the preparation of anti-inflammatory and antibacterial drugs

Abstract

This invention discloses polyacetylene compounds, their preparation methods, and their applications in the preparation of anti-inflammatory and antibacterial drugs, belonging to the pharmaceutical field. The polyacetylene compounds of this invention are isolated from Atractylodes lancea plants and possess significant anti-inflammatory and antibacterial activities, making them suitable for the preparation of anti-inflammatory and antibacterial drugs. This invention provides candidate compounds for the development of new anti-inflammatory and antibacterial drugs and is of great significance for the development and utilization of medicinal plant resources. The polyacetylene compounds of this invention can be used alone, in combination, or as pharmaceutical compositions in the preparation of anti-inflammatory or antibacterial drugs, and can be prepared as powders, pills, capsules, tablets, films, ointments, granules, aerosols, or gels, etc.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more particularly to a polyacetylene compound, its preparation method, and its application in the preparation of anti-inflammatory and antibacterial drugs. Background Technology

[0002] Antibiotics help people effectively treat various serious bacterial infections and extend human lifespan. However, the emergence of antibiotic resistance has reduced the use of antibiotics. On the other hand, inflammation plays an important role in the development of many diseases, such as cardiovascular disease, diabetes, Alzheimer's disease, and even cancer. Currently, there are various anti-inflammatory drugs on the market, but long-term use can cause toxic side effects on the liver, kidneys, and gastrointestinal tract. Therefore, finding more structurally novel, highly effective, and low-toxicity antibacterial and anti-inflammatory compounds from traditional Chinese herbal medicines has become a current research hotspot.

[0003] Polyacetylene compounds are found in a variety of medicinal plants, especially abundant in plants of the Asteraceae family such as Atractylodes lancea and Atractylodes macrocephala. These compounds possess a variety of pharmacological activities. Therefore, in-depth research into these chemical components and their biological activities is of great significance. Summary of the Invention

[0004] The present invention aims to at least partially solve one of the technical problems existing in the prior art. Therefore, in a first aspect, the present invention provides a polyacetylene compound, said polyacetylene compound being selected from one of the following compounds:

[0005] .

[0006] Preferably, the polyacetylene compound is selected from one of the following compounds:

[0007] .

[0008] In a second aspect, the present invention provides a method for preparing the above-mentioned polyacetylene compound, wherein the polyacetylene compound is isolated from Atractylodes lancea.

[0009] In some embodiments, the preparation method of the above-mentioned polyacetylene compounds includes the following steps:

[0010] (1) Extraction of extract: Atractylodes lancea root is used as raw material. It is crushed and extracted by reflux or soaking with organic solvent. The extracts are combined, filtered and concentrated into extract. The organic solvent includes one of methanol or ethanol.

[0011] (2) Extraction: The extract obtained in step (1) is suspended in water and extracted with an organic solvent to obtain an extract; the organic solvent includes petroleum ether, dichloromethane, and ethyl acetate;

[0012] (3) Column chromatography separation: The extract obtained in step (2) is separated by column chromatography to obtain the polyacetylene compound; the column chromatography includes at least one of silica gel column chromatography, macroporous resin chromatography, gel column chromatography, reversed-phase silica gel column chromatography, and high performance liquid preparative chromatography, and the column chromatography can be gradient eluted with a mixed solvent consisting of any two of the following solvents: water, methanol, acetonitrile, acetone, ethanol, ethyl acetate, petroleum ether, and dichloromethane.

[0013] Furthermore, the preparation method of the polyacetylene compound includes the following steps:

[0014] (1) Extraction of extract: Atractylodes lancea root is used as raw material. It is crushed and extracted by reflux with methanol or ethanol or by soaking. The extracts are combined, filtered and concentrated into extract.

[0015] (2) Extraction: The extract obtained in step (1) was suspended in water and extracted successively with petroleum ether, dichloromethane and ethyl acetate to obtain petroleum ether extract, dichloromethane extract and ethyl acetate extract, respectively.

[0016] (3) Column chromatography separation: The dichloromethane extract and ethyl acetate extract obtained in step (2) were separated by silica gel column chromatography. They were eluted with a gradient of petroleum ether-ethyl acetate with a volume ratio of 100:1 to 1:1 and detected by TLC. The eluents were combined and obtained in the order of elution: A, B, C, D, E, and F from the dichloromethane extract and I, II, III, IV, and V from the ethyl acetate extract. Compounds 5 and 6 were obtained from the E fraction by gel column chromatography, silica gel column chromatography, and preparative liquid chromatography. Compounds 2 and 3 were obtained from the II fraction and compound 1 and 4 were obtained from the III fraction by gel column chromatography, reversed-phase silica gel column chromatography, and preparative liquid chromatography.

[0017] In a third aspect, the present invention provides the use of the above-mentioned polyacetylene compounds in the preparation of anti-inflammatory and antibacterial drugs.

[0018] The application of the above-mentioned polyacetylene compounds in the preparation of anti-inflammatory and antibacterial drugs includes the combined use of two or more of compounds 1 to 6.

[0019] In a fourth aspect, the present invention provides a medicament comprising one, two, or more of the above-described polyacetylene compounds or pharmaceutically acceptable salts thereof.

[0020] In some implementations, the aforementioned drug also includes pharmaceutically acceptable excipients and carriers.

[0021] In some implementations, the pharmaceutical preparations of the above-mentioned drugs are granules, tablets, pills, solutions, capsules, films, tinctures, creams, ointments, aerosols, suppositories, liniments, gels, or injections.

[0022] The beneficial effects of this invention are as follows:

[0023] 1. This invention provides a polyacetylene compound with significant anti-inflammatory and antibacterial activities and potential application value. It can be used to prepare anti-inflammatory or antibacterial drugs, providing candidate compounds for the development of new anti-inflammatory and antibacterial drugs, and is of great significance to the development and utilization of medicinal plant resources.

[0024] 2. When preparing anti-inflammatory or antibacterial drugs, it can be used alone, in combination, or as a drug composition, and can be prepared as powder, pill, capsule, tablet, film, ointment, granule, aerosol, or gel, etc. Attached Figure Description

[0025] Figure 1 The flowchart shows the separation process for compounds 1–6. Detailed Implementation

[0026] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the art or according to the product instructions. Where specific conditions are not specified in the following embodiments, they are performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the methods used are conventional methods known in the art, and the consumables and reagents used are commercially available unless otherwise specified. Unless otherwise stated, the technical and scientific terms used herein have the same meaning as those familiar to those skilled in the art. Furthermore, any methods or materials similar to or equivalent to those described herein may also be applied to the present invention.

[0027] Example 1

[0028] This embodiment provides a polyacetylene compound, which is as follows:

[0029] .

[0030] The separation process for compounds 1-6 is as follows: Figure 1 As shown, the specific steps include the following:

[0031] Atractylodes lancea ( Atractylodes lancea)After root pulverization, the extract was extracted four times with methanol, and the methanol was recovered. The resulting extract was then extracted successively with petroleum ether, dichloromethane, and ethyl acetate to obtain three solvent extracts. The dichloromethane and ethyl acetate extracts were separated by silica gel column chromatography, using a petroleum ether-ethyl acetate gradient elution with volume ratios of 100:1, 50:1, 10:1, 5:1, 2:1, and 1:1. The eluents were analyzed by TLC and combined. Six fractions (A–F) were obtained from the dichloromethane extract, and five fractions (I–V) were obtained from the ethyl acetate extract, in the order of elution. Fraction E was separated sequentially by gel column chromatography (dichloromethane:methanol = 1:1), silica gel column chromatography (petroleum ether:ethyl acetate = 5:1), and semi-preparative high-performance liquid chromatography (80% methanol-water) to obtain compounds 5 and 6. Compounds 2 and 3 were obtained from fraction II by gel column chromatography (dichloromethane:methanol = 1:1), reversed-phase ODS column chromatography (50%-100% methanol), and semi-preparative solution high-performance liquid chromatography (65% methanol-water), respectively. Compounds 1 and 4 were obtained from fraction III by gel column chromatography (dichloromethane:methanol = 1:1), reversed-phase ODS column chromatography (50%-100% methanol), and semi-preparative solution high-performance liquid chromatography (65% methanol-water), respectively.

[0032] Example 2

[0033] The structures of compounds 1–6 were identified. The structures of the polyacetylene compounds prepared by the above method were determined by high-resolution mass spectrometry (HRESIMS) using molecular ion peaks to determine molecular weight and molecular formula. The structures were also determined by combining the correlations of the two-dimensional spectra with the proton and carbon NMR spectra. Table 1 below shows the proton and carbon NMR data for compound 1, and Table 2 shows the proton NMR data for compounds 2–6.

[0034] Table 1. Compound 1 1 H NMR (400 MHz) δ mult ( J [in Hz)] and 13 C NMR (100MHz) data

[0035]

[0036] Measured in CDCl3.

[0037] Table 2. Compounds 2–6 1 H NMR (400 MHz) δ mult ( J [in Hz)] Data

[0038]

[0039] Measured in CDCl3.

[0040] Example 3

[0041] Anti-inflammatory activity tests of compounds 1–6

[0042] The Griess method was used to assess the anti-inflammatory activity of the samples. RAW 264.7 mouse macrophages were seeded in DMEM medium (containing 10% FBS, 100 IU / mL penicillin, and 100 μg / mL streptomycin) and cultured in a 5% CO2 incubator at 37°C. The cytotoxicity of the samples to RAW 264.7 cells was pre-tested using the MTT assay, and the anti-inflammatory activity was tested within the maximum concentration range where the samples were non-toxic to the cells. The test samples were diluted to appropriate concentrations with DMEM medium, and cells were treated with an equal volume of the diluted sample solution and LPS (1 μg / mL) solution, then cultured in a CO2 incubator for 24 hours. The reacted medium was mixed with Griess reagent and reacted at room temperature for 10 minutes. The absorbance was measured at 540 nm. The experiment included a blank control group, a model group, and a positive control group with indomethacin. Each concentration of sample was tested in triplicate, and each experiment was repeated three times. The NO inhibition rate was calculated using the following formula: NO inhibition rate = (A 造模组 -A 药物组 ) / (A 造模组 -A 空白组 ()×100%. The results are shown in Table 3.

[0043] Table 3. Anti-inflammatory activities of compounds 1–6

[0044]

[0045] The data are expressed as mean ± standard deviation (n = 3).

[0046] Experimental conclusions: Compounds 1–6 significantly inhibited NO release from LPS-stimulated RAW264.7 macrophages, with effects superior to the positive control drug indomethacin. These results indicate that compounds 1–6 have the potential to be developed into anti-inflammatory drugs.

[0047] Example 4

[0048] Antibacterial activity tests of compounds 1-6

[0049] Dilute the bacterial culture to 1.0 × 10⁻⁶. 5 CFU / mL, while simultaneously dissolving the compound in 5% DMSO to obtain a stock solution, which was then serially diluted twice with PBS. 100 μL of sample solution was mixed with 100 μL of microbial suspension in each well. The mixture was then incubated at 37°C. Incubate at C for 24 hours. The solvent used to dissolve the samples was set up as a negative control group, and levofloxacin as a positive control group. The results are shown in Table 4.

[0050] Table 4. Inhibitory effects of compounds 1–6 on Staphylococcus aureus

[0051]

[0052] Table 4 shows that compounds 1-6 have a certain inhibitory effect on Staphylococcus aureus.

[0053] In addition, further testing revealed that compounds 1–6 also had a certain inhibitory effect on Escherichia coli and Pseudomonas aeruginosa.

[0054] Experimental conclusion: Compounds 1-6 have the potential to be developed into antibacterial drugs.

[0055] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A polyacetylene compound, characterized in that, The polyacetylene compound is selected from one of the following compounds: Compound 1 Compound 3 Compound 4.

2. A method for preparing the polyacetylene compound according to claim 1, characterized in that, The polyacetylene compounds were isolated from Atractylodes lancea.

3. The method for preparing polyacetylene compounds according to claim 2, characterized in that, Includes the following steps: (1) Using Atractylodes lancea root as raw material, crush it, extract it by reflux with organic solvent or by soaking, combine the extracts, filter and concentrate it into an extract; (2) The extract obtained in step (1) is suspended in water and extracted with an organic solvent to obtain the extract; (3) The extract obtained in step (2) is separated by column chromatography to obtain the polyacetylene compound.

4. The method for preparing polyacetylene compounds according to claim 3, characterized in that, In step (1), the organic solvent includes one of methanol or ethanol; In step (2), the organic solvent includes petroleum ether, dichloromethane, and ethyl acetate; In step (3), the column chromatography includes at least one of silica gel column chromatography, macroporous resin chromatography, gel column chromatography, reversed-phase silica gel column chromatography, and high-performance liquid preparative chromatography. The column chromatography is performed by gradient elution with a mixed solvent consisting of any two of the following solvents: water, methanol, acetonitrile, acetone, ethanol, ethyl acetate, petroleum ether, and dichloromethane.

5. The use of the polyacetylene compound of claim 1 in the preparation of anti-inflammatory and / or antibacterial drugs.

6. The application according to claim 5, characterized in that, This includes the combined use of two or more of compounds 1, 3, and 4.

7. A drug, characterized in that, This includes one, two, or more of the polyacetylene compounds of claim 1 or their pharmaceutically acceptable salts.

8. The medicament according to claim 7, characterized in that, It also includes pharmaceutically acceptable excipients and carriers.

9. The medicament according to claim 8, characterized in that, The pharmaceutical preparations mentioned are granules, tablets, pills, solutions, capsules, films, tinctures, creams, ointments, aerosols, suppositories, liniments, gels, or injections.

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