CN115850218B

CN115850218B - Linderane type sesquiterpene dimer and preparation method and application thereof

Info

Publication number: CN115850218B
Application number: CN202310024518.8A
Authority: CN
Inventors: 文松松; 张娜; 刘永军
Original assignee: Shandong University
Current assignee: Shandong University
Priority date: 2023-01-09
Filing date: 2023-01-09
Publication date: 2023-05-05
Anticipated expiration: 2043-01-09
Also published as: CN115850218A

Abstract

The invention relates to the technical field of medicines, in particular to a linderane type sesquiterpene dimer and a preparation method and application thereof. 1 linderane sesquiterpene dimer with novel structure is separated from dry root tuber of lindera root of lindera genus of Lauraceae family, the compound is a dimer compound with C31 skeleton formed by connecting two lindera sesquiterpenes through methylene bridge, most lindera sesquiterpenes are C30 skeleton, and the structure is rare in the natural world. The invention enriches the research on the diversity of chemical components of the combined spicebush root, adds 1 new compound for the field of compounds, and also provides a new reference for the research on the chemical taxonomy of the combined spicebush root.

Description

Linderane type sesquiterpene dimer and preparation method and application thereof

Technical Field

The invention relates to the technical field of medicines, in particular to a combined spicebush root of mountain pepper genus in Lauraceae family which is a traditional Chinese medicineLindera aggregata) A linderane sesquiterpene dimer obtained by separation, and its preparation method and application are provided.

Background

Wine plays an important role in the dietary life of Chinese people. Excessive drinking not only leads to alcohol dependence, but also increases the risk of illness. Excessive drinking can cause various diseases for human body, such as alcoholic fatty liver, liver cirrhosis, alcoholic hepatitis, digestive system diseases, and neurological disorder caused by long-term drinking. Oxidative stress, fat accumulation, inflammatory reactions, apoptosis caused by alcoholic liver injury can further threaten human health. Oxidative stress, for example, can cause cell damage by exposing the cells to high concentrations of oxygen molecules or chemical derivatives of oxygen. At this time, the oxygen free radical causes the break of DNA fragments, functional defects of proteins and aggregation of various toxic substances through the interaction of proteins, fats, DNA and other biomacromolecules, and simultaneously changes the oxidation defense system of the organism, so that diseases such as diabetes, inflammation, cancer and the like are finally caused. Therefore, the lead compound with the oxidative stress protection effect is found to be of great significance for preventing or treating alcoholic injury in the natural world.

The combined spicebush root (Lindera aggregata) is a plant of the genus lindera of the family Lauraceae, is the only variety of combined spicebush root recorded in the Chinese pharmacopoeia of 2020 edition, and has long use history. This herb is pungent and warm. It has effects in promoting qi circulation, relieving pain, warming kidney, and dispelling cold, and can be used for treating stagnation of qi due to congealing cold, chest and abdominal distention, asthma, bladder deficiency cold, enuresis, frequent urination, hernia pain, and cold channel abdominal pain.

Disclosure of Invention

The invention aims to provide a preparation method of linderane sesquiterpene dimer and application thereof in the aspect of treating alcohol oxidation injury.

The technical scheme of the application is as follows:

linderane sesquiterpene dimer is named Aggreganoid G, and has a chemical structural formula shown in formula 1:

formula 1.

The invention also provides a preparation method for protecting the linderane sesquiterpene dimer, which comprises the following steps:

(1) Pulverizing dried root tuber of radix Linderae to obtain coarse powder; extracting with ethanol under reflux, filtering, mixing extractive solutions, and concentrating under reduced pressure to obtain total extract;

(2) Dispersing the total extract in the step (1) in water, and then extracting with equal volume petroleum ether to obtain petroleum ether phase extract;

(3) And (3) subjecting the extract obtained in the step (2) to silica gel column chromatography, petroleum ether-acetone gradient elution and thin-layer chromatography inspection to obtain seven components A-G, subjecting the component F to reversed-phase C-18 column chromatography and methanol-water gradient elution, and subjecting the component F to reversed-phase C-8 acetonitrile-water isocratic elution and reversed-phase C-18 methanol-water isocratic elution respectively to obtain the linderane sesquiterpene dimer, namely Aggreganoid G.

Further, in the petroleum ether-acetone gradient elution process of the step (3), the volume ratio of petroleum ether to acetone is 100:0 to 0:100.

Further, in the methanol-water gradient elution of the step (3), the volume ratio of methanol to water is 20:80 to 100:0.

Further, the volume ratio of acetonitrile to water of the acetonitrile-water isocratic elution of the step (3) is 55:45; the volume ratio of methanol to water for methanol-water isocratic elution was 65:35.

The invention also aims to protect the application of the linderane sesquiterpene dimer in preparing medicines for preventing alcohol oxidative damage.

Preferably, the use in the manufacture of a medicament for the treatment of liver fibrosis.

The invention has the beneficial effects that:

1. new compounds

According to the invention, 1 linderane sesquiterpene dimer with a novel structure is separated from dried root tuber of lindera root of lindera genus of Lauraceae family, the compound is a dimer compound with a C31 skeleton formed by connecting two lindera sesquiterpenes through a methylene bridge, most of lindera sesquiterpenes are C30 skeletons, and the structure is rare in the natural world. The invention enriches the research on the diversity of chemical components of the combined spicebush root, adds 1 new compound for the field of compounds, and also provides a new reference for the research on the chemical taxonomy of the combined spicebush root.

2. The extraction method is simple

The preparation method of the compound adopts a mature technical means in the field of natural product chemistry, and is easy to realize in actual operation.

3. Treating hepatic fibrosis

In vitro cell experiments show that Aggreganoid G has remarkable protection effect on 3% ethanol-induced HepG2 cells, the cell survival rate is 97.81%, and the 3% ethanol-induced HepG2 cells survival rate is 43.73%.

Drawings

High resolution mass spectra of compound Aggreganoid G of fig. 1;

FIG. 2 hydrogen spectrum of compound Aggreganoid G;

FIG. 3 carbon spectrum of compound Aggreganoid G;

FIG. 4 1H-1H COSY spectrum of Aggreganoid G;

FIG. 5 HSQC spectrum of Aggreganoid G;

FIG. 6 HMBC spectra of Aggreganoid G;

FIG. 7 NOESY spectrum of Aggreganoid G;

FIG. 8 ultraviolet absorbance spectra of Aggreganoid G;

FIG. 9 Infrared Spectrometry for Aggreganoid G.

Detailed Description

Example 1 preparation of Aggreganoid G

Extracting: pulverizing dried root tuber of Lindera root (5.0 kg) of Piper genus of Lauraceae family, reflux-extracting with 80% ethanol for 4 times (1-2 hr each time), filtering, mixing extractive solutions, concentrating under reduced pressure, diluting the concentrated extractive solution with water, extracting with petroleum ether of equal volume for 4 times, and recovering solvent under reduced pressure to obtain petroleum ether phase extract.

Separating: subjecting the petroleum ether phase extract (127.5G) obtained in the previous step to silica gel column chromatography, gradient eluting with petroleum ether-acetone (100:0 to 0:100) in volume ratio, performing thin layer chromatography, inspecting to obtain seven components A-G, gradient eluting with methanol-water (20:80 to 100:0) by reversed phase C18 silica gel column chromatography, separating acetonitrile-water (55:45) by semi-preparative high performance liquid chromatography C8 column chromatography, and separating methanol-water (65:35) by semi-preparative high performance liquid chromatography C18 column chromatography to obtain Aggreganoid G (5.0 mg).

Then analyzing and detecting the structure of the sample, wherein the related data of Aggreganoid G are shown in figures 1-9;

–176.3 (c 0.3, MeOH)；

ultraviolet spectrum (methanol):λ _max (log ε): 195 (4.23), 275 (3.75)；

an infrared spectrum of light is obtained,ν _max 3440, 2940, 1748, 1645, 1390, 1329, 1125, 1054, 994 cm ^-1 ；

the nuclear magnetic data are shown in Table 1;

high resolution mass spectrometrym/z 473.2683 [M + H] ⁺ (calculated value is C ₃₁ H ₃₇ O ₄ , 473.2692)。

Table 1 Nuclear magnetic data for Aggreganoid G (deuterated chloroform)

From the above analysis, the chemical structural formula is shown in formula 1:

formula 1.

Example 2 in vitro cell alcohol damage assay

Cell culture: hepG2 cells were cultured in medium containing 10% fetal bovine serum, 100U/mL penicillin and 100 EMEMU/mL streptomycin, 37 in a 5% carbon dioxide saturated incubator ^o C, culturing, and taking the cell to perform experiments in the logarithmic phase. The concentration of the cell suspension was adjusted to 2X 10 ⁵ cell/mL, inoculating cells to a 96-well plate, placing the cells in a carbon dioxide incubator for culturing for 24 hours, adding a culture medium into a blank control group, adding 3% ethanol into a model control group, adding 10 mu M Aggreganoid G solution into a sample group, culturing for 24 hours, and measuring the cell survival rate by an MTT method. The results show that the cell survival rate of the ethanol-induced group is 43.73 +/-0.95%, the cell survival rate of the sample group is 97.80+/-0.98%, and the compound has a certain protection effect on ethanol-induced cell damage.

Claims

1. The linderane sesquiterpene dimer is characterized by being named Aggreganoid G, and the chemical structural formula of the linderane sesquiterpene dimer is shown as formula 1:

formula 1.

2. A process for the preparation of the linderane sesquiterpene dimer according to claim 1, characterized by comprising the following steps:

(3) Subjecting the extract obtained in the step (2) to silica gel column chromatography, petroleum ether-acetone gradient elution and thin-layer chromatography inspection to obtain seven components A-G, subjecting the component F to reversed-phase C-18 column chromatography and methanol-water gradient elution, and subjecting the component F to reversed-phase C-8 acetonitrile-water isocratic elution and reversed-phase C-18 methanol-water isocratic elution respectively to obtain an linderane sesquiterpene dimer, namely Aggreganoid G;

in the petroleum ether-acetone gradient elution process of the step (3), the volume ratio of petroleum ether to acetone is 100:0 to 0:100;

in the methanol-water gradient elution of the step (3), the volume ratio of the methanol to the water is 20:80 to 100:0;

the volume ratio of acetonitrile and water of the acetonitrile-water isocratic elution in the step (3) is 55:45; the volume ratio of methanol to water for methanol-water isocratic elution was 65:35.

3. The use of a linderane-type sesquiterpene dimer according to claim 1 for preparing a medicament for preventing oxidative damage of alcohol.

4. The use according to claim 3, for the preparation of a medicament for the treatment of liver fibrosis.