Cembranoid macrocyclic diterpene derivatives, preparation method therefor, and use thereof

Abstract

Cembranoid macrocyclic diterpene derivatives, a preparation method therefor, and a use thereof, belonging to the field of medical technology. Disclosed are cembranoid macrocyclic diterpene derivatives, a preparation method therefor, and a use thereof. In the present invention, a series of cembranoid macrocyclic diterpene derivatives are obtained by means of column chromatography and high-performance liquid chromatography separation of an extract from Croton oleifera. Said derivatives are capable of inhibiting LPS-induced NO release in BV-2 cells and have great potential for the preparation of a medicament for neuritis-related diseases.

Description

Cembane-type macrocyclic diterpene derivatives, preparation method and application thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and relates to cembane-type macrocyclic diterpene derivatives, a preparation method and application thereof. BACKGROUND

[0002] Cembranoid diterpenes are a class of natural products with novel, special and important biological activity of fourteen-membered ring mother nucleus, which can be classified into isopropyl type, lactone ring type, cyclic ether, open ring type and decarboxylation type according to the structure. They were first discovered in Pinus and tobacco, and a large number of cembranoid diterpene derivatives were found in marine organisms. The compounds have significant pharmacological activities, including anti-tumor, anti-inflammatory, antibacterial and anti-malaria, etc. They are a class of characteristic structures in the field of diterpene research. Among them, 4R-cembranoid (1S, 2E, 4R, 6R, 7E, 11E)-cembra-2,7,11-triene-4,6-diol) is a typical natural macrocyclic diterpene isolated from tobacco leaves, which has the characteristics of neuroprotection and anti-nicotine addiction. Therefore, 4R-cembranoid has attracted the attention of researchers as a candidate drug for neurodegenerative diseases and smoking cessation treatment.

[0003] Neurodegenerative diseases are a kind of nervous system diseases with progressive loss of neurons in central nervous system or peripheral nervous system, including pathological protein aggregation, synaptic and neuronal network dysfunction, protein homeostasis abnormalities, cytoskeleton abnormalities, energy metabolism changes, DNA and RNA defects, inflammation and other pathological characteristics, leading to diseases such as Alzheimer's disease (AD), Parkinson's disease (PD) and primary tauopathy, which seriously affect people's quality of life. Initial brain injury, trauma or genetic defects can lead to inflammatory signaling. The resident immune cells (astrocytes and microglia) in the brain can initiate the inflammatory response of cytokines such as TNF-α, IL-1β and IL-6. Therefore, regulating the inflammatory response can effectively improve and prevent nervous system lesions. The novel cembranoid diterpene involved in the present application is obtained by extraction and separation from Phyllanthus niruri, and the anti-inflammatory activity of the obtained diterpene compound is evaluated by establishing a LPS-induced BV-2 microglial inflammation model in vivo and in vitro. The compound and its anti-neuroinflammatory activity test have not been reported in patents or literatures.

[0004] SUMMARY

[0005] The purpose of the present application is to provide cembane-type macrocyclic diterpene derivatives, a preparation method and application thereof in preparing anti-neuroinflammatory drugs.

[0006] The application adopts an in vitro and in vivo LPS-induced BV-2 cell inflammation model for evaluating the anti-neuroinflammatory activity of the cembran diterpenoid compounds.

[0007] The application provides cembran macrocyclic diterpene derivatives or pharmaceutically acceptable salts thereof as shown in the following formulae I-VI:

[0008] wherein,

[0009] In formula I, the A ring is a fourteen-membered ring;

[0010] In formula II, the A ring is a fourteen-membered ring;

[0011] In formula III, the A ring is a fourteen-membered ring;

[0012] In formula IV, the A ring is a fourteen-membered ring, and the B ring is a furan ring;

[0013] In formula V, the A ring is a fourteen-membered ring, the B ring is a furan ring, and the C ring is a five-membered ether ring;

[0014] In formula VI, the A ring is a fourteen-membered ring, the B ring is a furan ring, and the C ring is a five-membered ether ring.

[0015] Further, in formula I, the A ring has two isolated double bonds, one enone conjugated double bond, and C-3, C-4 are in E configuration, C-7, C-8 are in Z configuration, C-10, C-11 are in Z configuration, and C-6 is a ketone carbonyl.

[0016] Further, in formula II, the A ring has two isolated double bonds, one enone conjugated double bond, and C-3, C-4 are in E configuration, C-7, C-8 are in Z configuration, C-10, C-11 are in Z configuration, and C-6 is a ketone carbonyl.

[0017] Further, in formula III, the A ring has two isolated double bonds, one enone conjugated double bond, and C-3, C-4 are in E configuration, C-7, C-8 are in Z configuration, C-10, C-11 are in E configuration, and C-6 is a ketone carbonyl.

[0018] Further, in formula IV, the A ring has one isolated double bond, one enone conjugated double bond, and C-3, C-4 are in E configuration, C-12, C-13 are in E configuration, and C-5 is a ketone carbonyl.

[0019] Further, in formula V, the A ring has one isolated double bond, and C-12, C-13 are in E configuration, and C-5 is a ketone carbonyl.

[0020] Further, in formula VI, the A ring has one isolated double bond, and C-11, C-12 are in E configuration, and C-5 is a ketone carbonyl.

[0021] The present application provides the cembrane diterpene derivatives, the name and structure of which are as follows:

[0022] The compounds 1-6 are respectively compound 1: croton-cembranoid A, compound 2: croton-cembranoid B, compound 3: croton-cembranoid C, compound 4: croton-cembranoid D, compound 5: croton-cembranoid E, and compound 6: croton-cembranoid F.

[0023] The present application also provides a preparation method of the cembrane diterpene derivatives, comprising the following steps:

[0024] (1) The dried branches and leaves of Phyllanthus niruri are crushed, and are extracted by heating reflux extraction method with ethanol or methanol, and the alcohol solvent is recovered under reduced pressure, and the mixture is suspended in water;

[0025] (2) The suspension is extracted with petroleum ether, dichloromethane, ethyl acetate and n-butanol in sequence, and the solvent and water are recovered respectively, and the petroleum ether layer extract, dichloromethane layer extract, ethyl acetate layer extract, n-butanol layer extract and water layer extract are obtained respectively;

[0026] (3) The petroleum ether layer extract obtained in step (2) is separated and eluted by silica gel column chromatography;

[0027] (4) The fraction containing cembrane diterpene derivative components obtained in step (3) is separated again by silica gel column chromatography or ODS column chromatography, and then the fraction containing cembrane diterpene derivative components is separated by preparative or semi-preparative HPLC chromatography, and is eluted with methanol-water or acetonitrile-water as the mobile phase, to obtain the cembrane diterpene compound;

[0028] Further, in step (1), the amount of ethanol or methanol is 3-15 times the mass of the dried branches and leaves of Phyllanthus niruri; and the ethanol or methanol is ethanol or methanol with a volume concentration of 30%-100%.

[0029] Further, in step (2), the extraction is performed 1-5 times, and the volume ratio of the extraction solvent to the suspension is (1:1)-(2:1).

[0030] Further, in the step (3), when the silica gel chromatography is a silica gel column, the elution condition is one or a mixture of two of petroleum ether, ethyl acetate, dichloromethane and methanol, specifically including, by volume ratio, petroleum ether: ethyl acetate = (100:1) to (1:1), dichloromethane: methanol = (100:0) to (15:1); when the silica gel chromatography is a GF254 silica gel plate, the elution condition is, by volume ratio, petroleum ether: ethyl acetate = (100:1) to (1:1);

[0031] Further, in the step (3), when the silica gel chromatography is a GF254 silica gel plate, the elution condition is preferably, by volume ratio, petroleum ether: ethyl acetate = 100:1, petroleum ether: ethyl acetate = 20:1, petroleum ether: ethyl acetate = 4:1, petroleum ether: ethyl acetate = 3:1, petroleum ether: ethyl acetate = 2:1 and petroleum ether: ethyl acetate = 1:1.

[0032] Further, in the step (4), the system for preparing the cembrene-type macrocyclic diterpene derivative by preparative or semi-preparative HPLC is specifically: the mobile phase is, by volume ratio, methanol: water = (1:4) to (9:1) or acetonitrile: water = (1:5) to (4:1), preferably methanol: water = (1:4) to (9:1).

[0033] The present application adopts spectroscopic means to separate the active ingredients contained in the extract obtained in the step (2). It can be seen from the results of investigating the inhibition of LPS-induced BV-2 microglial cell nitric oxide (NO) production that the active components are mainly concentrated in the petroleum ether layer extract, which shows a light yellow color with 10% sulfuric acid ethanol, confirming that the active substance is a terpenoid derivative. According to the changes of the chromophore and auxiliary chromophore, the ultraviolet full wavelength scanning shows certain regularity and characteristics, realizing the tracking separation of active diterpenes.

[0034] The cembrene diterpene skeleton isolated from the branches and leaves of Phyllanthus niruri has a conjugate feature. When an enone conjugate exists, the main characteristic of ultraviolet absorption is the existence of an absorption at about 245 nm; when a furan ring exists, the main characteristic of ultraviolet absorption is the existence of an absorption at about 280-300 nm. Accordingly, the cembrene-type macrocyclic diterpene derivative contained therein can be tracked.

[0035] The present application provides the use of the cembrene-type macrocyclic diterpene derivative or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating a neuroinflammation-related neurodegenerative disease.

[0036] The present application provides a pharmaceutical composition comprising one or more of the cembrene-type macrocyclic diterpene derivative or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0037] The application provides application of the pharmaceutical composition in preparation of a medicine for treating a neuroinflammation-related neurodegenerative disease.

[0038] The application has the following beneficial effects:

[0039] The application extracts the cembranoid diterpene derivative from dried branches and leaves of Phyllanthus niruri for the first time, and determines the structure of the cembranoid diterpene derivative by adopting separation methods such as silica gel column chromatography, ODS column chromatography and high performance liquid chromatography (HPLC) and combining with various spectroscopic techniques such as nuclear magnetic resonance spectroscopy. The cembranoid diterpene derivative prepared by the method has great application prospect in preparation of an anti-neuroinflammatory medicine. BRIEF DESCRIPTION OF DRAWINGS

[0040] Fig. 1 is an effect of the cembranoid diterpene derivative (50 μM) on the activity of BV-2 cells in the embodiment 2 of the application (Gal: galantamine);

[0041] Fig. 2 is an effect of the cembranoid diterpene derivative on the release of NO of BV-2 cells induced by LPS in the embodiment 2 of the application (Gal: galantamine). DETAILED DESCRIPTION

[0042] The following examples will further illustrate the application, but are not used to limit the application.

[0043] Example 1

[0044] The dried branches and leaves of P. officinalis (20.0 kg) were powdered and extracted with 95% ethanol by volume concentration at a ratio of 1:8 by volume to weight for 3 times, 2 h each time. The solvent was recovered under reduced pressure and the extract was dispersed in distilled water. The extract was successively extracted with petroleum ether, dichloromethane, ethyl acetate and n-butanol, each time for 3 times. The petroleum ether layer extract was obtained (300.8 g). The extract was separated by silica gel column chromatography using petroleum ether-ethyl acetate (100:1-1:1, V / V) as the mobile phase. The fractions were combined according to the results of silica gel thin layer chromatography. The petroleum ether-ethyl acetate (4:1-3:1) fractions were collected (19.4 g) and further separated. The petroleum ether-ethyl acetate (4:1) fractions were separated by ODS column chromatography using methanol-water (1:4-9:1, V / V) as the mobile phase. The yellow-brown viscous oil obtained at methanol-water (4:1) was separated by semi-preparative HPLC using methanol-water (7:3, V / V, 3 mL / min) to obtain compounds 2, 3, 5 and 6. The petroleum ether-ethyl acetate (3:1) fractions were separated by ODS column chromatography using methanol-water (1:5-8:1, V / V) as the mobile phase. The yellow-brown viscous oil obtained at methanol-water (3:1) was separated by semi-preparative HPLC using methanol-water (3:2, V / V, 3 mL / min) to obtain compound 1 and 4.

[0045] The spectral data of compounds 1-6 are as follows:

[0046] Compound 1 croton-cembranoid A

[0047] yellowish oil (methanol), 1 H-NMR (600 MHz, CDC13) δ H : 6.05 (1H, s, H-7), 5.64 (1H, dd, J = 9.7, 4.1 Hz, H-3), 4.88 (1H, m, H-10), 4.48 (1H, s, H-5), 2.33 (1H, m, H-9a), 2.31 (1H, m, H-13a), 2.20 (3H, s, H3-19), 2.14 (1H, m, H-2a), 2.09 (1H, m, H-13b), 2.08 (1H, m, H-9b), 2.06 (1H, m, H-2b), 2.05 (2H, m, H-12), 1.60 (1H, m, H-14a), 1.56 (3H, s, H3-20), 1.42 (1H, m, H-14b), 1.45 (1H, m, H-1), 1.43 (3H, s, H3-18), 1.25 (6H, d, J = 2.1 Hz, H3-16 / H3-17). 13 C-NMR (150 MHz, CDC13) δ C: 199.5 (C-6), 160.7 (C-8), 136.4 (C-11), 134.1 (C-3), 132.1 (C-4), 123.3 (C-10), 120.6 (C-7), 83.9 (C-5), 74.1 (C-15), 47.6 (C-1), 41.5 (C-9), 36.6 (C-12), 28.3 (C-14), 27.6 (C-2), 24.0 (C-13), 2 x 27.7 (C-16, C-17), 19.5 (C-19), 15.5 (C-20), 10.7 (C-18).

[0048] Compound 2 croton-cembranoid B

[0049] pale yellow oil (methanol), 1 H-NMR (600 MHz, CDC13) δ H : 6.05 (1H, s, H-7), 5.33 (1H, dd, J = 8.9, 4.6 Hz, H-3), 4.94 (1H, t, J = 6.4 Hz, H-10), 3.05 (1H, d, J = 13.0 Hz, H-5a), 2.92 (1H, d, J = 13.0 Hz, H-5b), 2.23 (1H, m, H-9a), 2.21 (2H, m, H2-13), 2.17 (1H, m, H-2a), 2.10 (1H, m, H-9b), 2.10 (1H, m, H-12a), 2.10 (3H, d, J = 1.2 Hz, H3-19), 2.04 (1H, m, H-2b), 2.00 (1H, m, H-12b), 1.63 (1H, m, H-14a), 1.60 (3H, d, J = 1.3 Hz, H3-18), 1.57 (3H, d, J = 1.3 Hz, H3-20), 1.41 (1H, tt, J = 7.1, 4.7 Hz, H-1), 1.35 (1H, m, H-14b), 1.24 (3H, s, H3-17), 1.23 (3H, s, H3-16). 13 C-NMR (150 MHz, CDC13) δ C: 200.1 (C-6), 157.1 (C-8), 136.3 (C-11), 130.3 (C-3), 129.6 (C-4), 123.8 (C-7), 123.7 (C-10), 74.1 (C-15), 48.6 (C-1), 56.8 (C-5), 41.0 (C-9), 38.0 (C-12), 28.4 (C-2), 28.2 (C-14), 24.2 (C-13), 2 x 27.8 (C-16, C-17), 18.9 (C-19), 16.3 (C-18), 15.8 (C-20).

[0050] Compound 3 croton-cembranoid C

[0051] pale yellow oil (methanol), 1 H-NMR (600 MHz, CDC13) δ H : 6.07 (1H, s, H-6), 5.28 (1H, m, H-3), 4.79 (1H, m, H-10), 3.14 (1H, m, H-9a), 3.04 (1H, d, J = 12.2 Hz, H-5a), 2.57 (1H, m, H-9b), 2.32 (1H, m, H-13a), 2.87 (1H, d, J = 12.2 Hz, H-5b), 2.21 (1H, m, H-2a), 2.13 (1H, m, H-13b), 2.01 (1H, m, H-2b), 1.98 (2H, m, H2-12), 1.78 (3H, d, J = 1.2 Hz, H3-19), 1.74 (3H, s, H3-18), 1.62 (2H, m, H2-14), 1.54 (3H, s, H3-20), 1.31 (1H, m, H-1), 1.20 (6H, d, J = 4.1 Hz, H3-16 / H3-17). 13 C-NMR (150 MHz, CDC13) δ C : 199.6 (C-6), 157.4 (C-8), 135.4 (C-11), 129.8 (C-3), 128.3 (C-4), 126.6 (C-7), 124.2 (C-10), 73.9 (C-15), 55.1 (C-5), 48.2 (C-1), 37.3 (C-12), 31.1 (C-9), 28.4 (C-14), 28.1 (C-2), 25.1 (C-13), 2 x 27.6 (C-16, C-17), 23.7 (C-19), 17.6 (C-18), 15.4 (C-20).

[0052] Compound 4 croton-cembranoid D

[0053] pale yellow oil (methanol), 1 H-NMR (600 MHz, CDC13) δ H : 7.44 (1H, s, H-19), 7.03 (1H, s, H-7), 6.37 (1H, t, J = 5.8 Hz, H-3), 5.25 (1H, t, J = 7.5 Hz, H-13), 2.57 (1H, m, H-2a), 2.54 (2H, m, H2-9), 2.39 (1H, m, H-11a), 2.28 (1H, m, H-10a), 2.24 (1H, m, H-10b), 2.11 (1H, m, H-11b), 1.96 (1H, m, H-2b), 1.91 (1H, s, H3-18), 1.86 (1H, m, H-14a), 1.58 (1H, m, H-1), 1.52 (1H, s, H3-20), 1.35 (1H, m, H-14b), 1.29 (3H, s, H3-16), 1.27 (3H, s, H3-17). 13 C-NMR (150 MHz, CDC13) δ C : 186.4 (C-5), 150.6 (C-6), 143.8 (C-19), 143.7 (C-3), 137.9 (C-12), 136.9 (C-4), 126.7 (C-8), 125.7 (C-13), 125.6 (C-7), 74.2 (C-15), 50.7 (C-1), 41.4 (C-11), 31.1 (C-2), 29.5 (C-10), 29.4 (C-14), 28.4 (C-16), 27.2 (C-17), 23.4 (C-9), 16.6 (C-20), 12.8 (C-18).

[0054] Compound 5 croton-cembranoid E

[0055] pale yellow oil (methanol), 1 H-NMR (600 MHz, CDC13) δ H7.48 (1H, s, H-19), 7.03 (1H, s, H-7), 4.97 (1H, t, J = 6.0 Hz, H-13), 3.83 (1H, m, H-3), 3.03 (1H, m, H-4), 2.72 (1H, ddd, J = 13.4, 8.3, 4.9 Hz, H-9a), 2.57 (1H, m, H-9b), 2.47 (1H, dq, J = 13.4, 6.3 Hz, H-10a), 2.29 (1H, dd, J = 14.9, 7.0 Hz, H-10b), 2.19 (1H, m, H-11a), 2.11 (1H, m, H-11b), 1.80 (1H, dd, J = 12.6, 6.0 Hz, H-2a), 1.62 (1H, dd, J = 6.0, 3.9 Hz, H-1), 1.60 (1H, m, H-14a), 1.51 (1H, m, H-2b), 1.48 (1H, s, H3-20), 1.30 (3H, s, H3-16), 1.27 (3H, d, J = 6.5 Hz, H3-18), 1.27 (1H, m, H-14b), 1.00 (3H, s, H3-17). 13 C-NMR (150 MHz, CDC13) δ C : 190.2 (C-5), 152.5 (C-6), 145.2 (C-19), 135.4 (C-12), 126.8 (C-8), 126.0 (C-13), 123.2 (C-7), 83.3 (C-15), 79.8 (C-3), 47.9 (C-4), 45.8 (C-1), 39.3 (C-11), 34.5 (C-2), 28.8 (C-16), 27.5 (C-10), 25.9 (C-14), 23.3 (C-17), 22.5 (C-9), 14.3 (C-20), 13.8 (C-18).

[0056] Compound 6 croton-cembranoid F

[0057] pale yellow oil (methanol), 1 H-NMR (600 MHz, CDC13) δ H7.39 (1H, s, H-19), 6.85 (1H, s, H-7), 4.99 (1H, d, J = 10.1 Hz, H-11), 4.11 (1H, dd, J = 9.8, 2.3 Hz, H-3), 3.21 (1H, q, J = 7.0 Hz, H-4), 2.80 (1H, m, H-9a), 2.41 (1H, m, H-9b), 2.30 (1H, m, H-10a), 1.25 (1H, m, H-10b), 2.20 (1H, d, J = 14.4 Hz, H-13a), 2.12 (1H, m, H-2a), 1.91 (1H, dd, J = 13.5, 3.8 Hz, H-14a), 1.86 (1H, m, H-2b), 1.46 (1H, tdd, J = 13.5, 4.1, 2.7 Hz, H-14b), 1.31 (3H, d, J = 7.0 Hz, H3-18), 1.30 (3H, s, H3-20), 1.01 (3H, s, H3-16), 0.90 (3H, s, H3-17), 1.03 (1H, m, H-1). 13 C-NMR (150 MHz, CDC13) δ C : 189.0 (C-5), 152.7 (C-6), 143.3 (C-19), 135.3 (C-12), 127.0 (C-8), 124.9 (C-11), 120.8 (C-7), 83.2 (C-15), 80.0 (C-3), 48.0 (C-4), 44.4 (C-1), 37.8 (C-13), 37.2 (C-2), 29.5 (C-10), 26.9 (C-16), 26.8 (C-14), 23.8 (C-9), 21.6 (C-17), 16.5 (C-18), 13.9 (C-20).

[0058] Example 2: In vitro anti-neuroinflammatory activity screening of cembrane-type macrocyclic diterpene derivatives:

[0059] (1) Cell culture

[0060] BV-2 cells were cultured in DMEM medium, 10% FBS, 37°C, 5% CO2 conditions.

[0061] (2) Activity test

[0062] MTT cytotoxicity activity test: BV-2 cells were pretreated with test monomeric compounds at a final concentration of 50 μΜ for 4 h, then LPS was added for incubation for 24 h; finally, cell survival rate was determined according to the MTT reagent instruction.

[0063] NO inhibitory activity screening: BV-2 cells were pretreated with 0-50 μM gradient concentration of the tested monomer for 4 h, then LPS was added for incubation for 24 h; finally the concentration of NO in the cell culture supernatant was determined according to the Griess kit instruction.

[0064] The experimental results of the compounds are shown in Fig. 1 and Fig. 2.

[0065] The results show that the cembane type macrocyclic diterpene derivatives can significantly inhibit the release of NO of BV-2 cells induced by LPS, and the inhibitory activities of compounds 1-3 are stronger, IC 50 which are all less than that of the positive drug (galantamine).

Claims

1. A cembrane diterpene derivative characterized in that, Specifically selected from compounds represented by the following Formulae 1 to 6: wherein, In formula I, A ring is a fourteen-membered ring; In formula II, A ring is a fourteen-membered ring; In formula III, A ring is a fourteen-membered ring; In formula IV, A ring is a fourteen-membered ring, and B ring is a furan ring; In formula V, A ring is a fourteen-membered ring, B ring is a furan ring, and C ring is a five-membered ether ring; In formula VI, A ring is a fourteen-membered ring, B ring is a furan ring, and C ring is a five-membered ether ring; The compounds shown in formula 1 to formula 6 are named as: formula 1: croton-cembranoid A, formula 2: croton-cembranoid B, formula 3: croton-cembranoid C, formula 4: croton-cembranoid D, formula 5: croton-cembranoid E, and formula 6: croton-cembranoid F.

2. Process for the preparation of cembrane diterpene derivatives according to claim 1, characterized in that, Comprising the following steps: (1) The dried branches and leaves of Phyllanthus niruri are crushed, and extracted by heating reflux extraction method with ethanol or methanol, and the alcohol solvent is recovered under reduced pressure, and the mixture is suspended with water; (2) The suspension is extracted with petroleum ether, dichloromethane, ethyl acetate and n-butanol in sequence, and the solvent and water are recovered respectively, and petroleum ether layer extract, dichloromethane layer extract, ethyl acetate layer extract, n-butanol layer extract and water layer extract are obtained respectively; (3) The petroleum ether layer extract obtained in step (2) is separated and eluted by silica gel column chromatography; (4) The fraction containing cembrane type diterpene derivatives obtained in step (3) is separated and eluted by silica gel chromatography or ODS column chromatography again, and then the fraction containing cembrane type diterpene derivatives is separated and eluted by preparative or semi-preparative HPLC chromatography with methanol-water or acetonitrile-water as the mobile phase to obtain cembrane type diterpene derivatives.

3. Process for the preparation of cembrane diterpene derivatives according to claim 2, characterized in that, In step (1), the amount of ethanol or methanol is 3-15 times of the mass of the dried branches and leaves of Phyllanthus niruri, and the volume concentration of ethanol or methanol is 30%-100%.

4. The process for the preparation of cembrane diterpene derivatives according to claim 2, characterized in that, In step (2), the extraction is performed 1-5 times, and the volume ratio of the extraction solvent to the suspension is (1:1)-(2:1).

5. The process for the preparation of cembrane diterpene derivatives according to claim 2, characterized in that, In step (3), when the silica gel chromatography is a silica gel column, the elution system used is one or a mixture of two of petroleum ether, ethyl acetate, dichloromethane and methanol; when the silica gel chromatography is a GF254 silica gel plate, the elution system used is a mixture of petroleum ether and ethyl acetate.

6. Process for the preparation of cembrane diterpene derivatives according to claim 5, characterized in that, In step (3), when the silica gel chromatography is a silica gel column, the elution system used is, by volume ratio, petroleum ether: ethyl acetate = (100:1)-(1:1), dichloromethane:methanol = (100:0)-(15:1); when the silica gel chromatography is a GF254 silica gel plate, the elution system used is, by volume ratio, petroleum ether: ethyl acetate = (100:1)-(1:1).

7. The process for the preparation of cembrane diterpene derivatives according to claim 2, characterized by, In step (4), the system for preparing cembrane type diterpene derivatives by preparative or semi-preparative HPLC is: by volume ratio, methanol: water = (1:4)-(9:1) or acetonitrile: water = (1:5)-(4:1).

8. Use of the cembrane diterpene derivative or a pharmaceutically acceptable salt thereof according to claim 1 for the manufacture of a medicament for treating neurodegenerative diseases associated with neuroinflammation.

9. A pharmaceutical composition comprising one or more of the cembrane diterpene derivatives or a pharmaceutically acceptable salt thereof according to claim 1 and a pharmaceutically acceptable carrier or excipient.

10. Use of the pharmaceutical composition according to claim 9 for the manufacture of a medicament for treating neurodegenerative diseases associated with neuroinflammation.

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