CN109369586B

CN109369586B - Linderane type sesquiterpene dimer and preparation method and application thereof

Info

Publication number: CN109369586B
Application number: CN201811494784.2A
Authority: CN
Inventors: 孙振亮; 邢欣; 虞贞风
Original assignee: Shanghai University of Medicine and Health Sciences
Current assignee: Shanghai University of Medicine and Health Sciences
Priority date: 2018-12-07
Filing date: 2018-12-07
Publication date: 2022-07-05
Anticipated expiration: 2038-12-07
Also published as: CN109369586A

Abstract

The invention provides an lindane sesquiterpene dimer, the structural formula of which is shown as follows,

Description

Linderane type sesquiterpene dimer and preparation method and application thereof

Technical Field

The invention belongs to the field of natural medicinal chemistry, relates to a novel compound, and particularly relates to an lindane type sesquiterpene dimer separated from a plant Dinghu Cinnamomum camphora, a preparation method and application thereof.

Background

Depression (depression) is an affective disorder characterized primarily by persistent depression, the incidence of which is rapidly rising with the exacerbation of various stressors. In addition, depression is closely related to various serious diseases such as cancer, stroke, cardiovascular diseases and the like, the pathogenesis of the depression is very complex, and mechanisms such as hypothalamus-pituitary-adrenal axis negative reaction regulation, nerve cell apoptosis, nerve cell necrosis and the like can be related to the pathogenesis of the depression from the earliest monoamine hypothesis. With the intensive research of pathogenesis, the direction is also pointed out for researching safe and effective new drugs. In recent years, especially after the research on natural medicines with neuroprotective effect has progressed to a certain extent, the method provides a wide prospect for obtaining high-efficiency medicines with low side effects clinically.

The Ting lake Zhang is small tree or shrub of plant of Linnaeus of Lauraceae. The root enlargement part is used as a medicine and mainly distributed in Guangdong, Guangxi and the like in China. The plant is often used as a substitute of the lindera strychnifolia in clinic and has a long medicinal history. Pungent and warm in nature; enters liver and stomach meridians, and has the effects of dispelling pathogenic wind, killing parasites, healing sore and stopping bleeding. The research on the plants of the genus finds that the plants are rich in various types of sesquiterpene compounds. The invention separates a new sesquiterpene dimer from the cinnamomum zeylanicum which is produced in Guangdong from Dinghu lake for the first time, and finds that the compound has better neuroprotective effect

Disclosure of Invention

Aiming at the technical problems in the prior art, the invention provides an lindane-type sesquiterpene dimer and a preparation method and application thereof, and the lindane-type sesquiterpene dimer and the preparation method and application thereof aim to solve the technical problems of limited effect and great side effect of a medicament for treating depression in the prior art.

the invention also provides a preparation method of the lindane sesquiterpene dimer, which comprises the following steps:

1) heating and refluxing dry root coarse powder of a cinnamomum camphora (Lindera chunii Merr.) in Ding lake for 1-5 times by adopting an ethanol solution, wherein each time lasts for 1-4 hours, combining extracting solutions, firstly decompressing and recovering ethanol from the extracting solutions, then adding water for dispersing, and sequentially extracting by using petroleum ether, ethyl acetate and n-butyl alcohol with the same volume to respectively obtain a petroleum ether phase, an ethyl acetate phase and an n-butyl alcohol phase;

2) subjecting the petroleum ether phase obtained in the step 1) to silica gel column chromatography, performing gradient elution through petroleum ether-acetone, inspecting through thin-layer chromatography, combining into 5 components, subjecting the third component to reversed-phase silica gel column chromatography, performing elution through a methanol-water gradient system, and performing reversed-phase C18 silica gel column chromatography to obtain lindane sesquiterpene dimer.

Further, during the petroleum ether-acetone gradient elution, the volume ratio of petroleum ether to acetone was varied from 100:0 to 0: 100.

Further, the volume ratio of methanol to water was varied from 30:70 to 100:0 during elution in a methanol-water gradient system.

The invention also provides application of the lindane sesquiterpene dimer in preparing a medicine for reducing the damage effect of corticosterone on SY5Y nerve cells.

The invention separates a new lindane type sesquiterpene dimer from a Dinghu Cinnamomum camphora (Lindera chunii Merr.) and is named as lindenanolide I.

SY5Y cell is a human neuroma blast cell line, after differentiation induced by retinoic acid. The expression of tyrosine hydroxylase, dopamine 2B2 hydroxylase, dopamine transporter and the like which are specific to catecholaminergic neurons can be used as a good cell model for researching physiological and pharmacological properties in neuroprotection research. In experimental studies, the neuroprotective effect of the samples was evaluated by observing the cell viability of SY5Y nerve cells damaged by corticosterone.

The compound is separated from the root of cinnamomum camphora (Lindera chunii Merr) in the Dinghu of the zanthoxylum genus of Lauraceae family, and the in vitro activity experiment shows that the compound can reduce the damage effect of corticosterone on SY5Y nerve cells.

Detailed Description

EXAMPLE 1 preparation of lindanalide I

Heating (95 ℃) ethanol with the mass percentage concentration of 80% to reflux and extract coarse powder of dry roots of cinnamomum hupehanum (5.0kg) for 2 times, 2 hours each time, combining extracting solutions, decompressing (150hPa) the extracting solution to recover the ethanol, adding water to disperse, and sequentially extracting with petroleum ether, ethyl acetate and n-butyl alcohol with the same volume (7L) to respectively obtain a petroleum ether phase, an ethyl acetate phase and an n-butyl alcohol phase.

Subjecting the petroleum ether phase extract to silica gel column chromatography, gradient eluting with petroleum ether-acetone (100: 0-0: 100), thin layer chromatography, and mixing to obtain 5 components (1-5). Performing reverse phase silica gel column chromatography on the third component, performing methanol-water gradient system (30: 70-100: 0), and performing reverse phase C18 silica gel column chromatography to obtain lindananolide I (2.0mg) with structural formula shown as below,

the technical indexes of the compound are as follows:

white powder;

specific rotation

(c 0.3,MeOH)；

Ultraviolet spectrum (MeOH): lambda [ alpha ]_max(logε),200(4.44),220(4.15)；

Round two chromatography (MeOH): 200(Δ ε +12.5),240(Δ ε -35.8), 255(Δ ε +8.04), nm;

infrared Spectrum, v_max 3679,2990,2362,1685,1517,1226,1375,1020,752cm^-1；

¹H and ¹³C NMR data(CDCl₃) The results are shown in Table 1,

HRESIMS m/z 597.2475[M+Na]⁺(calculation value C)₃₄H₃₈O₈ 597.2459)。

TABLE 1 Hydrogen and carbon Spectroscopy data for lindanaolide I

Example 2 experiment of neuroprotective Activity

DMEM medium containing 10% by volume of fetal calf serum, 100U/mL penicillin and 100U/mL streptomycin at 37 deg.C with 5% by volume of CO₂Culturing in an incubator with saturated humidity until SH-SY5Y cells entering logarithmic growth phase reach 70-80% fusion, digesting for 2min by pancreatin with the mass percent concentration of 0.25%, observing that the cells are circular under a mirror, suspending partial cells, immediately adding DMEM complete culture medium with the same volume as the pancreatin to stop digestion, blowing and beating the cells by a suction pipe until all the cells on the wall of a culture bottle are blown and beaten, transferring into a 15mL pointed centrifuge tube by the suction pipe, centrifuging for 5min at 1200r/min, discarding supernatant, suspending the cells by the DMEM culture medium, and passaging according to the proportion of 1: 3.

By counting cellsThe regulator regulates the SH-SY5Y cell concentration to be about 1X 10⁵At 80. mu.L/mL, the cells were inoculated into a 96-well plate and the culture was continued. After 24h, 10. mu.L of corticosterone 400. mu. mol/L lesion was added to each well, and 3 duplicate wells were set for each experimental group, with different concentrations of indenanolide I (12.5, 25, 50, 100. mu.M) being prepared. The culture was terminated after 72h, and MTT 20. mu.L was added to each well at a final mass concentration of 5g/L, 37 ℃ and 5% CO₂Continuously culturing for 4h in the incubator, after formation of bluish purple crystals, removing supernatant, adding DMSO 150 μ L into each well, shaking by shaking table for 10min to dissolve bluish purple crystals in cells completely, measuring absorbance value at 570nm wavelength with enzyme-labeling instrument, and calculating proliferation rate.

In vitro experimental study shows that the compound can reduce the damage effect of corticosterone on SY5Y cells, and the result is shown in Table 2.

TABLE 2 Effect of lindanaolide I on survival of SY5Y cells damaged by corticosterone

As can be seen from the above experiments, the lindane sesquiterpene dimer of the invention can be used as a lead compound of an antidepressant.

Claims

1. An lindane-type sesquiterpene dimer, which is characterized in that: the structural formula of the compound is shown as follows,

2. the method for preparing the lindane sesquiterpene dimer according to claim 1, which comprises the following steps:

3. The method for preparing lindane sesquiterpene dimer according to claim 2, wherein the method comprises the following steps: during the petroleum ether-acetone gradient elution, the volume ratio of petroleum ether to acetone is changed to 100:0 to 0: 100.

4. The method for preparing the lindane sesquiterpene dimer according to claim 2, wherein the method comprises the following steps: the volume ratio of methanol to water varies from 30:70 to 100:0 during elution with a methanol-water gradient system.

5. Use of the lindane sesquiterpene dimer of claim 1 in the preparation of a medicament for reducing the damaging effect of corticosterone on SY5Y nerve cells.