阅读说明：本技术 新木脂素类化合物及其制备方法和应用 (New lignan compound and preparation method and application thereof ) 是由 余建清 郑和国 刘星星 于 2021-09-17 设计创作，主要内容包括：本发明提供了一种新木脂素类化合物,所述新木脂素类化合物的结构式选自如下结构中的一种：所述新木脂素类化合物为新化合物。本发明还提供上述新木脂素类化合物的制备方法,其制备工艺简单。本发明还提供上述新木脂素类化合物在制备抗炎药物中的应用。(The invention provides a neolignan compound, wherein the structural formula of the neolignan compound is selected from one of the following structures: the neolignan compound is a novel compound. The invention also provides a preparation method of the neolignan compound, and the preparation process is simple. The invention also provides the novel lignan compoundApplication in preparing anti-inflammatory drugs.)

1. A neolignan compound having a structural formula selected from the group consisting of:

2. a process for the preparation of the neolignan compound of claim 1, wherein the neolignan compound is isolated from an extract of kadsura pepper stem.

3. The method for producing a neolignan compound according to claim 2, comprising the steps of:

step 1): pulverizing caulis Piperis Futokadsurae, extracting with a first solvent to obtain an extractive solution, and concentrating the extractive solution to obtain an extract;

step 2): suspending the extract obtained in the step 1) with water, and extracting with a second solvent to obtain an extract;

step 3): separating the extract obtained in the step 2) by column chromatography to obtain the neolignan compound.

4. The method of producing a neolignan compound according to claim 3, wherein the first solvent is one or two selected from the group consisting of methanol and ethanol in step 1).

5. The method of claim 3, wherein the second solvent is one or more selected from the group consisting of petroleum ether, n-hexane, cyclohexane and ethyl acetate in the step 2).

6. The method according to claim 3, wherein the column chromatography in step 3) is selected from one or more of macroporous resin column chromatography, silica gel column chromatography, reverse phase silica gel column chromatography, and high performance liquid chromatography.

7. The method of preparing a neolignan compound according to claim 3, wherein the step 3) comprises eluting the column chromatography with two or more third solvents selected from the group consisting of water, methanol, acetonitrile, acetone, ethyl acetate, petroleum ether, cyclohexane, and dichloromethane.

8. Use of the neolignan compound of claim 1 in the preparation of an anti-inflammatory agent.

9. A pharmaceutical formulation comprising a compound of formula i and/or a compound of formula ii, wherein the compound of formula i has the following structural formula:

the compound shown in the formula II has the following structural formula:

10. the pharmaceutical formulation of claim 9, wherein the pharmaceutical formulation is an aerosol, gel, film, powder, paste, pill, capsule, tablet, or granule.

Technical Field

The invention relates to the technical field of medicines, in particular to a neolignan compound and a preparation method and application thereof.

Background

Caulis Piperis Futokadsurae is dried stem of caulis Sinomenii (Piper kadsura (Choisy) Ohwi) belonging to Piper genus of Piperaceae family (Piperaceae). It is mainly distributed in Fujian, Zhejiang, Guangdong, Taiwan, Hunan, Hubei, Sichuan and Guizhou provinces, and has the effects of dispelling wind-damp, dredging channels and collaterals and relieving arthralgia. The main types of compounds separated from caulis piperis futokadsurae at present are lignans, alkaloids, flavonoids and the like, wherein the lignans are the main components of the caulis piperis futokadsurae.

Inflammation releases various inflammatory mediators, amplifies various cellular reactions, and causes the loss of functions of tissues or organs, which can be seen in various diseases, such as chronic bronchitis, asthma, nephritis, bacterial and viral infection diseases, myocardial infarction, ischemia-reperfusion injury, perfusion injury and the like. Inflammation is also closely related to the development of cancer. Inhibition of inflammation plays a very important role in the prevention and treatment of a variety of diseases. Some Chinese herbal medicines have good anti-inflammatory effect, and active compounds are mined from the Chinese herbal medicines to develop new anti-inflammatory medicines, so that the Chinese herbal medicines have obvious advantages.

Therefore, there is a great need for the development of new active compounds.

Disclosure of Invention

The present invention is directed to solving at least some of the problems of the prior art, and therefore, in a first aspect of the present invention, the present invention provides a neolignan compound having a structural formula selected from the group consisting of:

in a second aspect of the present invention, there is provided a method for producing the neolignan compound according to the first aspect of the present invention, wherein the neolignan compound is obtained by extracting and separating kadsura pepper stem.

In one or more embodiments of the present invention, the preparation method of the neolignan compound comprises the following steps:

step 1): pulverizing caulis Piperis Futokadsurae, extracting with a first solvent to obtain an extractive solution, and concentrating the extractive solution to obtain an extract;

step 2): suspending the extract obtained in the step 1) with water, and extracting with a second solvent to obtain an extract;

step 3): separating the extract obtained in the step 2) by column chromatography to obtain the neolignan compound.

In one or more embodiments of the present invention, in the step 1), the first solvent is selected from one or two of methanol and ethanol.

In one or more embodiments of the present invention, in the step 2), the second solvent is selected from one or more of petroleum ether, n-hexane, cyclohexane and ethyl acetate.

In one or more embodiments of the present invention, in the step 3), the column chromatography is selected from one or more of macroporous resin column chromatography, silica gel column chromatography, reverse phase silica gel column chromatography, and high performance liquid preparative chromatography.

In one or more embodiments of the present invention, in the step 3), the extract obtained in the step 2) is sequentially separated by macroporous resin column chromatography, silica gel column chromatography, high performance liquid preparative chromatography or silica gel column chromatography.

In one or more embodiments of the present invention, in the step 3), the column chromatography is eluted using two or more third solvents selected from the group consisting of water, methanol, acetonitrile, acetone, ethyl acetate, petroleum ether, cyclohexane and dichloromethane.

In a third aspect of the present invention, there is provided a use of the neolignan compound of the first aspect of the present invention in the manufacture of an anti-inflammatory agent.

In a fourth aspect of the invention, there is provided a pharmaceutical formulation comprising a compound of formula i and/or a compound of formula ii, wherein the compound of formula i has the following structural formula:

the compound shown in the formula II has the following structural formula:

in one or more embodiments of the present invention, the pharmaceutical formulation is an aerosol, a gel, a film, a powder, a paste, a pill, a capsule, a tablet, or a granule.

Compared with the prior art, the invention has the beneficial effects that:

1. the invention provides a neolignan compound, which is a novel compound.

2. The invention provides a preparation method of the neolignan compound, and the preparation process is simple.

3. The invention provides application of the neolignan compound in preparing anti-inflammatory drugs.

Drawings

FIG. 1 is a schematic diagram of a process for the isolation of compounds of formula I and formula II from caulis Piperis Kadsurae.

Detailed Description

The scheme of the invention will be explained with reference to the examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples, where specific techniques or conditions are not indicated, are to be construed according to the techniques or conditions described in the literature in the art or according to the product specifications. The following examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer, by using conventional methods known in the art without specific descriptions, and by using consumables and reagents which were commercially available without specific descriptions. Unless otherwise defined, technical and scientific terms used herein have the same meaning as is familiar to those skilled in the art. In addition, any methods or materials similar or equivalent to those described herein can also be used in the present invention.

Example 1: preparation of Compound 1 (Compound represented by formula I) and Compound 2 (Compound represented by formula II)

FIG. 1 is a schematic diagram of the process for preparing compound 1 (compound shown in formula I) and compound 2 (compound shown in formula II) by separating kadsura pepper stem. Compound 1 (a compound of formula I) has the following structural formula:

the structural formula of the compound 2 (the compound shown in the formula II) is as follows:

the preparation method comprises the following specific steps:

pulverizing dried caulis Piperis Futokadsurae (5.85kg), soaking in 95 (v/v)% ethanol 25.00L at 15-35 deg.C for 3 times, each for 7 days. Mixing extractive solutions, recovering solvent to obtain extract, suspending the extract with 1.0L water, sequentially extracting with petroleum ether and ethyl acetate with equal volume, and removing solvent to obtain 150.67g petroleum ether fraction and 89.52g ethyl acetate fraction.

Separating ethyl acetate part with macroporous resin column, eluting with 30-100% ethanol, sequentially separating into 5 parts (A-E), separating part C with silica gel column chromatography, and gradient eluting with petroleum ether and ethyl acetate (20:1-0:1) to obtain 6 parts (P1-P6); the fraction P4 was gel separated and prepared by HPLC (60% methanol in water) to give compound 1; fractions P3 were chromatographed on silica gel eluting with oil ether and ethyl acetate (5:1) to give compound 2.

Example 2: structural determination of Compound 1 (Compound represented by formula I) and Compound 2 (Compound represented by formula II)

Structural identification of compound 1 (compound shown in formula I) and compound 2 (compound shown in formula II):

the structures of compound 1 (compound represented by formula i) and compound 2 (compound represented by formula ii) obtained in example 1 were determined by data tests such as high-resolution mass spectrometry, ultraviolet spectroscopy, infrared spectroscopy, optical rotation, nuclear magnetic resonance, circular dichroism, and the like.

Compound 1 (compound of formula i): a colorless oily liquid;UV(MeOH)λ_max(logε)201.5(3.93),247.0(3.85)nm；IR(KBr)ν_max 2942,2362,1689,1617,1463,1364,1247,1237,1170,1141,1121,1050,1027,963,816,758,667cm^-1；ECD[λ_max(Δε)]205(+135.28),257(+9.92)nm；¹H and ¹³c NMR data, see table 1; HRESIMSm/z389.1953[ M + H ]]⁺(calcd for C₂₂H₂₉O₆,389.1959)。

Compound 2 (compound of formula ii): a yellow oily liquid;UV(MeOH)λ_max(logε)202.0(3.65),228.5(3.44)nm；IR(KBr)ν_max 2930,2834,1464,1268,1145,1093,1026cm^-1；ECD[λ_max(Δε)]204(+22.40),236(-7.86),260(+2.64),298(-0.22)nm；¹h and¹³c NMR data, see table 2; HRESIMS [ M + Na ]]⁺m/z381.1686[M+Na]⁺(calcd for C₂₅H₂₆O₅Na,381.1678)。

TABLE 1 preparation of Compound 1¹H NMR (400MHz) and¹³c NMR (100MHz) data (Recorded in CDCl)₃,δ:ppm)

TABLE 2 preparation of Compound 2¹H NMR (400MHz) and¹³c NMR (100MHz) data (Recorded in CDCl)₃,δ:ppm)

Example 3: anti-inflammatory Activity test of Compound 1 (Compound represented by formula I) and Compound 2 (Compound represented by formula II)

Anti-inflammatory assay: samples were evaluated for anti-inflammatory activity using the Griess method. The cytotoxicity of the test compound is evaluated by MTT method using RAW264.7 mouse macrophage, and then the anti-inflammatory activity of the compound is evaluated by determining the concentration which is not toxic to the cell. Culturing RAW264.7 mouse macrophage by adopting a DMEM culture medium, inoculating the cultured mouse macrophage in a 96-well culture plate, treating the cells with samples to be detected and LPS solutions with different concentrations for 24 hours at 30000/well after the cells adhere to the wall, mixing the reacted culture medium with a Griess reagent, reacting for 10 minutes at room temperature, and measuring the absorbance at 540 nm. The test is provided with a blank control group, a modeling group and a positive control group added with indometacin, each test is repeated for 3 times, and the inhibition effect on Nitric Oxide (NO) is evaluated. The NO inhibition rate is calculated according to the following formula:

NO inhibition rate (a)_{Molding set}-A_{Drug group})/(A_{Molding set}-A_{Blank group})×100％

The results are shown in Table 3.

TABLE 3 anti-inflammatory Activity of Compound 1 (Compound of formula I), Compound 2 (Compound of formula II)

And (4) experimental conclusion: the compound 1 (the compound shown in the formula I) and the compound 2 (the compound shown in the formula II) can inhibit the release of NO in RAW264.7 macrophages stimulated by LPS, have anti-inflammatory activity and have the potential of being developed into anti-inflammatory drugs.

Although the embodiments of the present invention have been shown and described, it is understood that the above embodiments are illustrative and not restrictive, and that those skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention and that they should be included in the protection scope of the present invention.