CN115850218B - A kind of buyalane-type sesquiterpene dimer and its preparation method and application - Google Patents

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

A kind of buyalane-type sesquiterpene dimer and its preparation method and application

technical field

The invention relates to the technical field of medicine, in particular to an aggregata-type sesquiterpene dimer isolated from a traditional Chinese medicine, Lindera aggregata , and a preparation method and application thereof.

Background technique

Alcohol plays an important role in Chinese food life. Excessive drinking not only leads to alcohol dependence, but also increases the risk of people getting sick. Excessive drinking can bring many diseases to the human body, such as alcoholic fatty liver, liver cirrhosis, alcoholic hepatitis, digestive system diseases, neurological disorders and other diseases caused by long-term drinking. Oxidative stress, fat accumulation, inflammatory response, and cell apoptosis caused by alcoholic liver injury will further threaten human health. For example, oxidative stress is the cellular damage caused by exposing cells to high concentrations of oxygen molecules or chemical derivatives of oxygen. At this time, oxygen free radicals interact with biomacromolecules such as proteins, fats, and DNA, causing DNA fragment breaks, protein function defects, and various toxic substances to accumulate. and diseases such as cancer. Therefore, discovering lead compounds with oxidative stress protection from nature is of great significance for the prevention or treatment of alcohol-induced injury.

Lindera aggregata is a plant of the genus Pepper in the family Lauraceae. This plant is the only species of Lindera aggregata included in the 2020 edition of "Chinese Pharmacopoeia", and has a long history of use. This product is pungent, warm. Return lung, spleen, kidney, bladder meridian, have the effect of promoting qi and relieving pain, warming kidney and dispelling cold, used for the treatment of stagnation of qi stagnation due to cold, distending pain in the chest and abdomen, dyspnea due to inverse qi, cold bladder, enuresis, frequent urination, hernia pain, Cold abdominal pain and other diseases.

Contents of the invention

The object of the present invention is to provide a preparation method of buyalane-type sesquiterpene dimer and its application in treating alcohol oxidative damage.

The technical scheme of the application is as follows:

A kind of buyalane type sesquiterpene dimer named as Aggreganoid G, its chemical structural formula is as shown in formula 1:

Formula 1.

Another object of the present invention is to protect the preparation method of the above-mentioned buyalane-type sesquiterpene dimer, comprising the following steps:

(1) Crush the dried root tubers of black herb to obtain coarse powder; then heat and reflux with ethanol to extract, filter, combine the extracts, concentrate under reduced pressure, and obtain the total extract;

(2) Disperse the total extract of step (1) in water, then extract with an equal volume of petroleum ether to obtain a petroleum ether phase extract;

(3) The extract of step (2) was first subjected to silica gel column chromatography, petroleum ether-acetone gradient elution, and thin-layer chromatography to obtain seven components A-G, and component F was subjected to reversed-phase C-18 Column chromatography, methanol-water gradient elution, and then reversed-phase C-8 acetonitrile-water isocratic elution and reversed-phase C-18 methanol-water isocratic elution respectively, to obtain the dimerization Body, namely Aggreganoid G.

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

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

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

Another object of the present invention is to protect the application of the above-mentioned wuyalane-type sesquiterpene dimer in the preparation of drugs for preventing oxidative damage of alcohol.

Preferably, it is used in the preparation of medicines for treating liver fibrosis.

Beneficial effects of the present invention:

1. New compounds

The present invention separates and obtains a bulumane sesquiterpene dimer with a novel structure from the dry root tuber of the plant of the genus Piper genus Lauraceae. It forms a dimer compound with a C31 skeleton, while most of the bulumane sesquiterpenes have a C30 skeleton, which is relatively rare in nature. The invention enriches the research on the diversity of the chemical components of the black medicine, adds a new compound to the compound field, and also provides a new reference for the chemical taxonomy research of the black medicine.

2. The extraction method is simple

The preparation methods of the compound adopt relatively mature technical means in the field of natural product chemistry, and are easy to realize in actual operation.

3. Treatment of liver fibrosis

In vitro cell experiments showed that Aggreganoid G had a significant protective effect on HepG2 cells induced by 3% ethanol, the cell survival rate was 97.81%, and the survival rate of HepG2 cells induced by 3% ethanol was 43.73%.

Description of drawings

Figure 1 High resolution mass spectrum of compound Aggreganoid G;

The hydrogen spectrum of the compound Aggreganoid G in Fig. 2;

The carbon spectrum of Fig. 3 compound Aggreganoid G;

Figure 4 1H-1H COZY spectrum of compound Aggreganoid G;

The HSQC spectrum of Fig. 5 compound Aggreganoid G;

The HMBC spectrum of Fig. 6 compound Aggreganoid G;

Fig. 7 NOESY spectrum of compound Aggreganoid G;

The ultraviolet absorption spectrum of Fig. 8 compound Aggreganoid G;

Figure 9 is the infrared spectrum of the compound Aggreganoid G.

Detailed ways

The preparation of embodiment 1 Aggreganoid G

Extraction: Grind the dried root tuber of the plant of the genus of lauraceae Pepper (5.0 kg), reflux extraction with 80% ethanol for 4 times, each time for 1 to 2 hours, filter and combine the extracts, and decompress the extracts Concentrate, dilute the extracted concentrate with water, extract 4 times with an equal volume of petroleum ether, and recover the solvent under reduced pressure to obtain petroleum ether phase extracts.

Separation: The petroleum ether phase extract (127.5 g) obtained in the previous step was subjected to silica gel column chromatography, and was eluted with a volume ratio of petroleum ether-acetone (100:0 to 0:100) gradient, and after inspection by thin layer chromatography, A was obtained. -G seven components, component F is further eluted through reverse phase C18 silica gel column chromatography methanol-water (20:80 to 100:0) gradient elution, and then separated by semi-preparative high performance liquid chromatography C8 column acetonitrile-water ( 55:45) elution, and finally separated methanol-water (65:35) by semi-preparative high performance liquid chromatography C18 column to obtain Aggreganoid G (5.0 mg).

Then its structure was analyzed and tested, and the relevant data of Aggreganoid G are shown in Figure 1-9;

–176.3 ( c 0.3, MeOH);

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

Infrared spectrum, ν _max 3440, 2940, 1748, 1645, 1390, 1329, 1125, 1054, 994 cm ^-1 ;

See Table 1 for NMR data;

High resolution mass spectrum m/z 473.2683 [M + H] ⁺ (calcd for C ₃₁ H ₃₇ O ₄ , 473.2692).

Table 1 NMR data of compound Aggreganoid G (deuterated chloroform)

According to the above analysis, the chemical structural formula is deduced as shown in formula 1:

Formula 1.

Example 2 In vitro cell alcohol injury experiment

Cell culture: HepG2 cells were cultured in a medium containing 10% fetal bovine serum, 100 U/mL penicillin and 100 EMEMU/mL streptomycin in a 5% carbon dioxide-saturated incubator at 37 ^o C, and the logarithm of the cells was taken experiment period. Adjust the concentration of the cell suspension to 2×10 ⁵ cell/mL, inoculate the cells in a 96-well plate, and culture them in a carbon dioxide incubator for 24 hours. Add medium to the blank control group, add 3% ethanol to the model control group, and add 3% ethanol to the sample group. After adding 10 μM Aggreganoid G solution, after 24 hours of co-cultivation, the cell viability was measured by MTT method. The results showed that the cell survival rate of the ethanol-induced group was 43.73±0.95%, and the cell survival rate of the sample group was 97.80±0.98%. The compound has a certain protective effect on ethanol-induced cell damage.

Claims (4)

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:

(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) 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.

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