Medicine for treating neuroinflammation and application thereof

Abstract

The invention relates to a medicine for treating neuroinflammation and application thereof, and belongs to the technical field of medicines. According to the invention, a new terphenyl compound is separated and identified from a Thelephora ganbajun Zang acetone extract, and an extraction and separation method of the new compound is provided. The method comprises the following steps: extracting dried sporocarps of thelephora ganbajun Zang by using acetone to obtain an acetone extract, and sequentially carrying out normal-phase silica gel column chromatography, reverse-phase silica gel (ODS) column chromatography and high performance liquid chromatography (HPLC) to prepare the thelephora ganbajun Zang. The novel compound disclosed by the invention can be used as a potential lead compound and is used for preparing medicines for treating neuroinflammation related diseases.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a medicine for treating neuroinflammation and application thereof. Background Art

[0002] Ganbajun Zang (Thelephora ganbajun Zang) is a precious and edible higher fungus with multiple functions such as nourishing kidney essence, strengthening tendons and bones, nourishing spleen and stomach, nourishing liver and kidney, diuresis and blood pressure, anti-fatigue and anti-cancer. It is mainly produced in Yunnan and other places in my country. Modern research shows that Ganbajun Zang extract contains multiple components. Pharmacological studies have shown that the chemical components in Ganbajun Zang mainly have antibacterial, anti-tumor and antioxidant effects. Therefore, the development of the types of compounds and pharmacological uses of Ganbajun Zang extracts is a technical problem that needs to be solved in this field. Summary of the invention

[0003] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a drug for treating neuroinflammation and its application. The present invention separates and identifies new terphenyl compounds from the acetone extract of Thelephora ganbajun Zang, and experiments confirm that these compounds have the efficacy of treating diseases related to neuroinflammation.

[0004] To achieve the above object, the technical solution adopted by the present invention is:

[0005] In a first aspect, the present invention provides a terphenyl derivative, wherein the terphenyl derivative has a chemical structural formula shown in any one of the following:

[0006]

[0007] Wherein R is hydrogen, hydroxy, methoxy, benzoyloxy, phenylacetoxy, isopentenyl, alkyl, alkenyl, alkynyl, or substituted or unsubstituted aryl.

[0008] The present invention separates terphenyl derivative compounds from the dried fruiting body extract of Bacon. Studies have shown that the compounds can inhibit inflammatory factors, improve the behavior of depression mouse models, and have the effect of treating neuritis.

[0009] As a preferred embodiment of the present invention, the molecular structure of the terphenyl derivative having the chemical structure shown in formula (II) is as follows:

[0010]

[0011] Wherein, R1 is hydroxyl or methoxy; R2 is hydroxyl or methoxy; R3 is hydrogen or phenylacetyl.

[0012] As a preferred embodiment of the 001 structural formula of the present invention, R1 is a hydroxyl group; R2 is a hydroxyl group; R3 is a phenylacetyl group.

[0013] As a preferred embodiment of the 001 structural formula of the present invention, R1 is a hydroxyl group; R2 is a hydroxyl group; R3 is hydrogen.

[0014] As a preferred embodiment of the 001 structural formula of the present invention, R1 is a methoxy group; R2 is a hydroxyl group; R3 is hydrogen.

[0015] As a preferred embodiment of the 001 structural formula of the present invention, R1 is a hydroxyl group; R2 is a methoxy group; R3 is hydrogen.

[0016] As a preferred embodiment of the present invention, the molecular structure of the terphenyl derivative having the chemical structural formula shown in formula (III) is as follows:

[0017]

[0018] Among them, R is hydrogen or phenylacetyl group.

[0019] As a preferred embodiment of the present invention, the molecular structure of the terphenyl derivative having the chemical structural formula shown in formula (IV) is as follows:

[0020]

[0021] As a preferred embodiment of the present invention, the molecular structure of the terphenyl derivative having the chemical structural formula shown in formula (V) is as follows:

[0022]

[0023] Among them, R1 is hydrogen or phenylacetyl group; R2 is hydrogen or phenylacetyl group.

[0024] As a preferred embodiment of the 004 structural formula of the present invention, R1 is hydrogen; R2 is phenylacetyl group.

[0025] As a preferred embodiment of the 004 structural formula of the present invention, R1 is phenylacetyl group; R2 is hydrogen.

[0026] As a preferred embodiment of the 004 structural formula of the present invention, R1 is phenylacetyl group; R2 is phenylacetyl group.

[0027] In a second aspect, the present invention provides the use of the terphenyl derivative described in the first aspect in the preparation of a medicament for treating neuroinflammation.

[0028] It has been experimentally proven that the compound of the present invention can inhibit inflammatory factors, improve the behavior of a mouse model of depression, and has the efficacy of treating neuritis.

[0029] Thirdly, the present invention provides a method for preparing the terphenyl derivative described in the first aspect, comprising the following steps:

[0030] S1 Prepare a dried mushroom extract using an organic solvent;

[0031] S2 Perform normal-phase chromatographic gradient elution separation on the extract obtained in S1 to obtain components separated at different gradients; the eluent used for the elution is a mixed solution of dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is: dichloromethane: methanol = (100 - 0): (0 - 100);

[0032] S3 Perform reverse-phase chromatographic gradient elution separation on the components obtained in S2 to obtain components separated at different gradients; the eluent used for the elution is a mixed solution of methanol and water, and the volume ratio of methanol to water is: methanol: water = (40 - 100): (60 - 0);

[0033] S4 Perform reverse-phase high-performance liquid chromatography isocratic elution separation on the components obtained in S3, and collect the components eluted at 13.5 min or 29.8 min to obtain a natural furan lactone-type terphenylquinone compound.

[0034] As a preferred embodiment of the third aspect, the elution steps in step S2 are as follows:

[0035] (1) Elute with an eluent having a volume ratio of dichloromethane to methanol of 100:0 to obtain fraction Fr.1;

[0036] (2) Elute with an eluent having a volume ratio of dichloromethane to methanol of 98:2 to obtain fraction Fr.2;

[0037] (3) Elute with an eluent having a volume ratio of dichloromethane to methanol of 95:5 to obtain fraction Fr.3;

[0038] (4) Elute with an eluent having a volume ratio of dichloromethane to methanol of 9:1 to obtain fraction Fr.4;

[0039] (5) Elute with an eluent having a volume ratio of dichloromethane to methanol of 8:2 to obtain fraction Fr.5;

[0040] (6) Elute with an eluent having a volume ratio of dichloromethane to methanol of 1:1 to obtain fraction Fr.6;

[0041] (7) Elute with an eluent having a volume ratio of dichloromethane to methanol of 0:100 to obtain fraction Fr.7.

[0042] As a preferred embodiment of the third aspect, the component for performing reverse-phase chromatographic gradient elution in step S3 is Fr.3, and the elution steps are as follows:

[0043] (1) Elute with an eluent having a volume ratio of methanol to water of 40:0 to obtain fraction Fr.31;

[0044] (2) Elute with an eluent having a volume ratio of methanol to water of 60:40 to obtain fraction Fr.32;

[0045] (3) Elute with an eluent having a volume ratio of methanol to water of 80:20 to obtain fraction Fr.33;

[0046] (4) Elute with an eluent having a volume ratio of methanol to water of 100:0 to obtain fraction Fr.34.

[0047] As a preferred embodiment of the third aspect, the component for performing reverse-phase high-performance liquid separation in step S4 is Fr.33, and the elution program satisfies the following conditions:

[0048] (1) The mobile phase is a mixed system of methanol and water, and the volume ratio of methanol to water is: methanol: water = 70:30;

[0049] (2) Elution time: 0 - 35 min.

[0050] As a preferred embodiment of the third aspect,

[0051] The chemical structural formula of compound 1 of the component eluted at 29.8 min in S4 is a terphenyl derivative shown in 002, where R is phenylacetyl;

[0052] The chemical structural formula of compound 2 of the component eluted at 13.5 min in S4 is a terphenyl derivative shown in 002, where R is hydrogen;

[0053] The chemical structural formula of compound 3 of the component eluted at 20.6 min in S4 is a terphenyl derivative shown in 003;

[0054] The chemical structural formula of compound 4 of the component eluted at 11.3 min in S4 is a terphenyl derivative shown in 001, where R1 is hydroxyl; R2 is hydroxyl; R3 is phenylacetyl;

[0055] The chemical structural formula of compound 5 of the component eluted at 18.4 min in S4 is a terphenyl derivative shown in 004, R1 is hydrogen; R2 is phenylacetyl.

[0056] As a preferred embodiment of the third aspect, the organic solvent in S1 is acetone.

[0057] As a preferred embodiment of the third aspect, the preparation method of the Termitomyces albuminosus extract in S1 includes the following steps: The dried fruiting bodies of Termitomyces albuminosus are soaked and extracted with acetone by the soaking method, and the acetone extract is obtained after concentration.

[0058] As a preferred embodiment of the third aspect, it further includes a step of identifying the components collected in the step S4.

[0059] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0060] The present invention isolates a natural furan lactone type terphenylquinone compound from the dried fruiting body extract of Bacillus. It is found through research that this compound can inhibit inflammatory factors, improve the behavior of a mouse model of depression, and has the efficacy of treating neuritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] Figure 1 1H-NMR spectrum schematic diagram of Compound 1 1 ;

[0062] Figure 2 13C-NMR spectrum schematic diagram of Compound 1 13 ;

[0063] Figure 3 HSQC spectrum schematic diagram of Compound 1;

[0064] Figure 4 HMBC spectrum schematic diagram of Compound 1;

[0065] Figure 5 1H- 1 1 1H COSY spectrum schematic diagram;

[0066] Figure 6 Single crystal diffraction pattern of Compound 1;

[0067] Figure 7 1H-NMR spectrum schematic diagram of Compound 2 1 ;

[0068] Figure 8 13C-NMR spectrum schematic diagram of Compound 2 13 ;

[0069] Figure 9 HSQC spectrum schematic diagram of Compound 2;

[0070] Figure 10 HMBC spectrum schematic diagram of Compound 2;

[0071] Figure 11 1H- 1 1 1H COSY spectrum schematic diagram;

[0072] Figure 12 Schematic diagram of the single crystal diffraction pattern of Compound 2;

[0073] ​​Figure 13 Schematic diagram of the 1 H-NMR spectrum of Compound 3;

[0074] Figure 14 Schematic diagram of the 13 C-NMR spectrum of Compound 3;

[0075] Figure 15 Schematic diagram of the HSQC spectrum of Compound 3;

[0076] Figure 16 Schematic diagram of the HMBC spectrum of Compound 3;

[0077] Figure 17 Schematic diagram of the 1 H- 1 H COSY spectrum of Compound 3;

[0078] Figure 18 Single crystal diffraction pattern of Compound 3;

[0079] Figure 19 Schematic diagram of the 1 H-NMR spectrum of Compound 4;

[0080] Figure 20 Schematic diagram of the 13 C-NMR spectrum of Compound 4;

[0081] Figure 21 Schematic diagram of the HSQC spectrum of Compound 4;

[0082] Figure 22 Schematic diagram of the HMBC spectrum of Compound 4;

[0083] Figure 23 Schematic diagram of the 1 H- 1 H COSY spectrum of Compound 4;

[0084] Figure 24 Single crystal diffraction pattern of Compound 4;

[0085] Figure 25 Schematic diagram of the 1 H-NMR spectrum of Compound 5;

[0086] Figure 26 Schematic diagram of the 13 C-NMR spectrum of Compound 5;

[0087] Figure 27 Schematic diagram of the HSQC spectrum of Compound 5;

[0088] Figure 28 Schematic diagram of the HMBC spectrum of Compound 5;

[0089] Figure 29Of Compound 5 1 H- 1 Schematic diagram of the H-COSY spectrum;

[0090] Figure 30 Schematic diagram of the effects of Compounds 1-10 on the viability of BV2 microglia;

[0091] Figure 31 Schematic diagram of the effects of Compounds 1-9 on the release of IL-6, IL-1β and TNF-α from LPS-induced BV2 cells (Figure A: Effects of Compounds 1-9 at 10 μM on the release of pro-inflammatory factors IL-6, IL-1β and TNF-α from LPS-induced BV-2 cells; Figure B: Effects of Compounds 1-9 at 20 μM on the release of pro-inflammatory factors IL-6, IL-1β and TNF-α from LPS-induced BV-2 cells);

[0092] Figure 32 Schematic diagram of the effects of Compounds 1-9 on the mRNA expression of IL-6, IL-1β and TNF-α in LPS-stimulated BV-2 cells;

[0093] Figure 33 Schematic diagram of Compound 2 inhibiting LPS-mediated neuroinflammatory response in BV2 microglia by inhibiting the JAK2 / STAT3 signaling pathway (Figure A: Band diagram of the effects of Compound 2 at 20 μM on the JAK2 / STST3 pathway proteins in LPS-induced BV-2 cells; Figure B: Statistical chart of the protein expression level of p-JAK2 / JAK2; Figure C: Statistical chart of the protein expression level of p-STAT3 / STAT3; Figure D: Immunofluorescence detection of the intracellular localization of phosphorylated signal transduction and p-STAT3);

[0094] Figure 34 Schematic diagram of the animal behavior experiment of Compound 2;

[0095] Figure 35 Schematic diagram of the chemical structure of Compound 1;

[0096] Figure 36 Schematic diagram of the chemical structure of Compound 2;

[0097] Figure 37 Schematic diagram of the chemical structure of Compound 3;

[0098] Figure 38 Schematic diagram of the chemical structure of Compound 4;

[0099] Figure 39 Schematic diagram of the chemical structure of Compound 5. Detailed implementation manners

[0100] To better illustrate the purpose, technical solution and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0101] Example 1: Preparation of Compounds 1-10

[0102] S1: Preparation of acetone extract:

[0103] The dried fruiting bodies of *Thelephora ganbajun* were soaked and extracted with acetone (3 L) three times, 24 h each time. The extract was concentrated to obtain the acetone extract for standby;

[0104] S2: Preliminary separation of the acetone extract:

[0105] Elution column: normal-phase silica gel column chromatography column (200-300 mesh)

[0106] Eluent: a mixed solution of dichloromethane and methanol, and the elution gradient and elution components are as follows in the table:

[0107] Table 1

[0108] Dichloromethane (v) Methanol (v) Component 100％ 0 Fr.1 98％ 2％ Fr.2 95％ 5％ Fr.3 90％ 10％ Fr.4 80％ 20％ Fr.5 50％ 50％ Fr.6 0 100％ Fr.7

[0109] The specific operation is as follows:

[0110] The acetone extract was passed through a normal-phase silica gel column chromatography column and eluted with dichloromethane and methanol (dichloromethane: methanol) with volume ratios of 100:0, 98:2, 95:5, 9:1, 8:2, 1:1, and 0:100, respectively. The eluates were combined, concentrated under reduced pressure, the eluent was removed, and concentrated to an extract to obtain 7 components: Fr.1 (dichloromethane-methanol 100:0 fraction), Fr.2 (dichloromethane-methanol 98:2 fraction), Fr.3 (dichloromethane-methanol 95:5 fraction), Fr.4 (dichloromethane-methanol 9:1 fraction), Fr.5 (dichloromethane-methanol 8:2 fraction), Fr.6 (dichloromethane-methanol 1:1 fraction), and Fr.7 (dichloromethane-methanol 0:100 fraction) for standby;

[0111] S3: Separation of component Fr.3:

[0112] Elution column: ODS column (reverse-phase chromatography column 40-60 μm)

[0113] Eluent: methanol-water system, and the elution gradient and elution components are as follows in the table:

[0114] Table 2

[0115] Methanol (v) Water (v) Component 40％ 60％ Fr.31 60％ 40％ Fr.32 80％ 20％ Fr.33 100％ 0 Fr.34

[0116] The specific operation is as follows:

[0117] The component Fr.3 was gradient eluted with a methanol - water system of 40:60 - 100:0 using an ODS column (reversed - phase chromatography column) to obtain 4 fractions: Fr.31 (methanol - water 40:60 fraction), Fr.32 (methanol - water 60:40 fraction), Fr.33 (methanol - water 80:20 fraction), and Fr.34 (methanol - water 100:0 fraction);

[0118] S4: The component Fr.33 was separated by reversed - phase high - performance liquid chromatography:

[0119] Chromatographic conditions: ODS column (reversed - phase chromatography column 40 - 60μm)

[0120] Eluent: Methanol - water system. The elution gradient and eluted components are as follows in the table:

[0121] Table 3

[0122]

[0123] The specific operation is as follows:

[0124] The component Fr.33 was eluted with methanol - water (70:30) on a preparative reversed - phase high - performance liquid chromatography to obtain the following compounds:

[0125] Compound 1 (7.2 mg, t R 29.8 min), whose chemical structural formula is as Figure 35 shown, and whose single - crystal diffraction pattern is as Figure 6 shown;

[0126] The name of Compound 1 is: 4,7 - bis(4 - hydroxyphenyl) - 2,6 - dioxo - 3 - phenyl - 2,6 - dihydrobenzofuran - 5 - yl

[0127] 2 - phenylacetate.

[0128] Compound 2 (4.6 mg, t R 13.5 min), whose chemical structural formula is as Figure 36 shown, and whose single - crystal diffraction pattern is as Figure 12 shown;

[0129] The name of Compound 2 is:

[0130] 5 - hydroxy - 4,7 - bis(4 - hydroxyphenyl) - 3 - phenylbenzofuran - 2,6 - dione.

[0131] Compound 3 (1.1 mg, t R 20.6 min), whose chemical structural formula is asFigure 37 As shown, its single crystal diffraction pattern is as Figure 18 shown;

[0132] The name of Compound 3 is:

[0133] 2,5,6-trihydroxy-4,7-bis(4-hydroxyphenyl)-2-methylbenzofuran-3(2H)-one.

[0134] Compound 4 (6.0 mg, t R 11.3 min), and its chemical structural formula is as Figure 38 shown, where R1 is a hydroxyl group or a methoxy group; R2 is a hydroxyl group or a methoxy group; R3 is hydrogen or phenylacetyl; preferably, R1 is a hydroxyl group; R2 is a hydroxyl group; R3 is phenylacetyl, and its single crystal diffraction pattern is as Figure 24 shown;

[0135] The name of Compound 4 is:

[0136] 4,4”,5'-trihydroxy-3',6'-dioxo-3',6'-dihydro-[1,1':4',1”-terphenyl]-2'-yl

[0137] 2-phenylacetate.

[0138] Compound 5 (2.3 mg, t R 18.4 min), and its chemical structural formula is as Figure 39 shown, where R1 is hydrogen or phenylacetyl; R2 is hydrogen or phenylacetyl; preferably, R1 is hydrogen; R2 is phenylacetyl;

[0139] The name of Compound 5 is: 2,7,8-trihydroxy-3-(4-hydroxyphenyl)dibenzo[b,d]furan-1,4-diyl bis(2-phenylacetate).

[0140] S5: Use reverse-phase high-performance liquid chromatography to separate Component Fr.4:

[0141] Chromatographic conditions: ODS column (reverse-phase chromatographic column 40 - 60 μm)

[0142] Eluent: Methanol-water system, and the elution gradient and eluted components are as follows in the table:

[0143] Table 4

[0144] Methanol (v) Water (v) Component 40％ 60％ Fr.41 60％ 40％ Fr.42 80％ 20％ Fr.43 100％ 0 Fr.44

[0145] The specific operation is as follows:

[0146] The component Fr.4 was gradient eluted with a methanol - water system of 40:60 - 100:0 using an ODS column (reversed - phase chromatography column) to obtain 4 fractions: Fr.41 (methanol - water 40:60 fraction), Fr.42 (methanol - water 60:40 fraction), Fr.43 (methanol - water 80:20 fraction), and Fr.44 (methanol - water 100:0 fraction).

[0147] S6: The component Fr.42 was separated by reversed - phase high - performance liquid chromatography:

[0148] Chromatographic conditions: ODS column (reversed - phase chromatography column 40 - 60μm)

[0149] Eluent: methanol - water system. The elution gradient and eluted components are as follows in the table:

[0150] Table 5

[0151]

[0152] The specific operation is as follows:

[0153] The component Fr.42 was eluted with methanol - water (70:30) on a preparative reversed - phase high - performance liquid chromatography to obtain the following compounds:

[0154] Compound 6 (t R = 28.1 min, 3 mg)

[0155] Compound 8 (t R = 33.5 min, 7.2 mg)

[0156] Compound 9 (t R = 37.2 min, 5 mg)

[0157] Compound 7 (t R = 41.0 min, 6 mg)

[0158] Compound 10 (t R = 44.5 min, 3 mg).

[0159] S7: Compounds 1 - 5 were identified:

[0160] 1H - NMR, 13C - NMR, two - dimensional NMR spectra (HMBC, HSQC, 1 H - 1 H COSY), high - resolution mass spectrometry, infrared spectroscopy, ultraviolet spectroscopy, circular dichroism spectroscopy, single - crystal X - ray diffraction were used for determination. The results are as follows in the table, Figures 1 - 29 :

[0161] Table 6 of Compounds 1 - 5 11H NMR (150 MHz) NMR data (J in Hz).

[0162]

[0163]

[0164] Measured in DMSO-d6.

[0165] Table 7 of Compounds 1 - 5 13 13C NMR (150 MHz) NMR Data (δi n ppm).

[0166] No. Compound 1 Compound 2 Compound 3 Compound 4 Compound 5 1 119.6,C 120.0,C 123.2,C 121.3,C 122.9,C 2,6 132.1, CH 131.9, CH 131.7, CH 131.7, CH 131.1, CH 3,5 114.6, CH 114.0, CH 114.7, CH 114.4, CH 115.3, CH 4 158.1,C 157.8,C 156.8,C 157.0,C 157.2,C 1' 135.0,C ND 113.5,C ND 125.3,C 2' 178.1,C ND 130.3,C ND 133.9,C 3' 142.7,C ND 137.6,C ND 145.8,C 4' 115.3,C ND 110.9,C 114.8,C 114.5,C 5' 136.8,C ND 164.3,C ND 150.1,C 6' 154.7,C ND 115.1,C ND 157.6,C 1” 119.5,C 120.0,C 122.2,C 121.3,C 115.5,C 2” 130.0, CH 130.9, CH 131.5, CH 131.7, CH 146.7,C 3” 115.1, CH 114.9, CH 114.2, CH 114.4, CH 98.9, CH 4” 157.9,C 156.4,C 156.4,C 156.8,C 143.1,C 5” 115.1, CH 114.9, CH 114.2, CH 114.4, CH 142.0,C 6” 130.0, CH 130.9, CH 131.5, CH 131.7, CH 114.3, CH 1”' 167.1,C 167.7,C 196.4,C 169.5,C 170.6,C 2”' 131.2,C 131.9,C 103.1,C <![CDATA[40.7,CH2]]> <![CDATA[40.5,CH2]]> 3”' 128.2,C 129.8,C <![CDATA[22.3,CH3]]> 133.2,C 134.1,C 4”',8”' 128.4, CH 129.8, CH 129.6, CH 129.9, CH 5”',7”' 127.0, CH 127.0, CH 128.5, CH 128.8, CH 6”' 129.3, CH 130.9, CH 127.1, CH 127.4, CH 1”” 169.5,C 170.1,C 2”” <![CDATA[39.5,CH2]]> <![CDATA[40.5,CH2]]> 3”” 133.3,C 134.1,C 4””,8”” 129.4, CH 129.7, CH 5””,7”” 127.3, CH 128.6, CH 6”” 130.0, CH 127.2, CH

[0167] Measured in DMSO-d6.

[0168] Example 2: Investigation of the anti-neuroinflammatory activity of Compounds 1-10

[0169] BV2 cells (a microglial cell line of mice) were placed in DMEM medium containing 10% fetal bovine serum and cultured at 37 °C under 5% CO2 conditions. For all experiments, cells within 10 passages were used. The compounds were dissolved in DMSO to obtain a stock solution of 10 mM, and LPS was diluted to a concentration of 1 mg / mL with PBS. BV2 cells (2×10 6 / mL) were seeded in 96-well plates overnight, pretreated with different concentrations of the compounds for 1 hour, and then co-cultured with LPS (100 ng / mL) for 24 hours. The cell suspension or supernatant was collected for analysis. Cell viability was determined using a CCK8 assay kit (E-CK-A362, Elabscience). BV-2 cells were seeded into 96-well plates at a density of 2×10 5 cells / well. The results showed that Compounds 1-9 did not exhibit obvious cytotoxicity to BV-2 microglial cells at concentrations within 20 μM ( Figure 30 ).

[0170] Investigate whether Compounds 1-9 can inhibit the release of pro-inflammatory cytokines in LPS-induced BV2 cells. BV2 cells were treated with different concentrations (10 μM and 20 μM) of the compounds for 24 hours in the presence or absence of co-treatment with 100 ng / ml LPS, and then the levels of inflammatory factors in the culture supernatant were measured. The results showed that in the absence of Compounds 1-9, LPS caused an increase in the release of IL-6, IL-1β, and TNF-α in BV2 cells; after adding Compounds 1-9, the release of LPS-induced pro-inflammatory cytokines was significantly inhibited ( Figure 31 and Figure 32)。Further research found that compound 2 can reduce the production of pro-inflammatory cytokines by inhibiting the activation of the JAK2 / STAT3 signaling pathway. Figure 33 )。

[0171] Example 3: Ethology experiment of compound 2

[0172] Among various pathophysiological mechanisms of depression, more and more evidence indicates that neuroinflammation is a major factor leading to depression. Neuroinflammation refers to the inflammatory process within the nervous system, which is characterized by a complex multicellular immune response in the central nervous system (CNS) triggered by acute or chronic pathological stimuli. It has been revealed that microglia-mediated neuroinflammation plays a key role in the development of depressive-like behaviors.

[0173] Construction of LPS-induced depression model: An LPS-induced mouse depression model was used. Six-week-old C57BL / 6J mice were purchased. The experimental animals were housed in an environment with a constant temperature of 22 ± 1°C and a relative humidity of 55 ± 5%. Sufficient feed and water were provided, and the light cycle was 12 hours light / 12 hours dark (lighting was turned on at 8:00 every day). They were continuously acclimated for 5 days to eliminate transportation and environmental transition stress. The experimental mice were randomly divided into 3 experimental units (15 mice per unit): control group (Ctrl group), model group (LPS group), and treatment group (LPS + compound 2 group). The control group received only saline injection and gavage. The treatment group was gavaged with compound 2 (5 mg / kg) once a day for 1 week. From the 6th day, the model group and the treatment group dissolved LPS (L2880, Sigma) in saline and intraperitoneally injected it (1 mg / kg) continuously for 2 days. Behavioral tests were performed 24 hours after the last LPS injection. After the test, the mice were sacrificed, and the tissues were collected and stored at -80°C for subsequent experiments.

[0174] Experimental treatment group: The above model mice were injected with 5 mg / kg of compound 2.

[0175] Sucrose preference test (SPT): The two-bottle free-choice paradigm was adopted: Mice were fed with 1% sucrose solution for 3 days before the experiment for adaptation. After random grouping, they were fasted and water-deprived for 24 hours on the 3rd day of drug administration. The next day, two bottles of liquid (tap water and 1% sucrose solution) were provided for free drinking, and the bottle positions were exchanged every 12 hours to eliminate the influence of position preference. Sucrose preference rate calculation formula:

[0176] Sucrose preference rate (%) = sucrose solution consumption / total liquid consumption (water + sucrose) × 100

[0177] Forced swimming test (FST): Mice were placed in a transparent plexiglass cylinder (70 cm high, 30 cm in diameter) filled with clear water (30 cm deep) at 23 ± 1 °C. A 6-minute behavioral video was recorded, and the immobility time (s) in the last 4 minutes was analyzed. Immobility was defined as stationary floating or only maintaining minor movements with the head above the water surface. Horizontal paddling was considered swimming, and vertical wall pushing was considered climbing.

[0178] Tail suspension test (TST): The tip of the mouse's tail at 1 cm was suspended at a height of 50 cm from the ground using medical tape. After 2 minutes of adaptation, the immobility time (s) in the subsequent 4 minutes was recorded to quantify the degree of behavioral despair.

[0179] The results showed that compound 2 reversed the LPS-induced decrease in sucrose consumption ( Figure 34 A), and reduced the immobility time in TST and FST ( Figure 34 B-34C), further confirming that compound 2 has the potential to alleviate LPS-induced neuroinflammation and can be used to treat neuroinflammation.

[0180] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A terphenyl derivative, characterized in that, The terphenyl derivatives have chemical structural formulas shown in any of the following: wherein R is hydrogen, hydroxyl, methoxy, benzoyloxy, phenylacetoxy, isopentenyl, alkyl, alkenyl, alkynyl, or a substituted or unsubstituted aryl group.

2. The terphenyl derivative according to claim 1, characterized in that, The molecular structure of the terphenyl derivative having the chemical structural formula shown in formula (II) is as follows: wherein, R1 is hydroxyl or methoxy; R2 is hydroxyl or methoxy; R3 is hydrogen or phenylacetyl.

3. The terphenyl derivative according to claim 1, wherein The molecular structure of the terphenyl derivative having the chemical structural formula shown in formula (III) is as follows: wherein, R is hydrogen or phenylacetyl.

4. The terphenyl derivative according to claim 1, characterized in that, The molecular structure of the terphenyl derivative having the chemical structural formula shown in formula (IV) is as follows:

5. The terphenyl derivative according to claim 1, characterized in that, The molecular structure of the terphenyl derivative having the chemical structural formula shown in formula (V) is as follows: wherein, R1 is hydrogen or phenylacetyl; R2 is hydrogen or phenylacetyl.

6. Use of the terphenyl derivative according to any one of claims 1-5 in the preparation of a drug for treating neuroinflammation.

7. The preparation method of the terphenyl derivative according to any one of claims 1-5, characterized in that, Comprising the following steps: S1 Prepare a Termitomyces albuminosus extract using an organic solvent. S2 Perform normal-phase chromatographic gradient elution separation on the extract of S1 to obtain components separated at different gradients; the eluent used for elution is a mixed solution of dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is: dichloromethane:methanol = (100 - 0):(0 - 100); S3 Perform reverse-phase chromatographic gradient elution separation on the components of S2 to obtain components separated at different gradients; the eluent used for elution is a mixed solution of methanol and water, and the volume ratio of methanol to water is: methanol:water = (40 - 100):(60 - 0); S4 Perform reverse-phase high-performance liquid chromatography isocratic elution separation on the components of S3, and collect the components eluted at 13.5 min or 29.8 min to obtain a natural furan lactone-type terphenylquinone compound.

8. The preparation method according to claim 7, characterized in that, The elution steps of step S2 are as follows: (1) Elute with an eluent having a volume ratio of dichloromethane to methanol of 100:0 to obtain fraction Fr.1; (2) Elute with an eluent having a volume ratio of dichloromethane to methanol of 98:2 to obtain fraction Fr.2; (3) Elute with an eluent having a volume ratio of dichloromethane to methanol of 95:5 to obtain fraction Fr.3; (4) Elute with an eluent having a volume ratio of dichloromethane to methanol of 9:1 to obtain fraction Fr.4; (5) Elute with an eluent having a volume ratio of dichloromethane to methanol of 8:2 to obtain fraction Fr.5; (6) Elute with an eluent having a volume ratio of dichloromethane to methanol of 1:1 to obtain fraction Fr.6; (7) Elute with an eluent having a volume ratio of dichloromethane to methanol of 0:100 to obtain fraction Fr.

7.

9. The preparation method according to claim 8, characterized in that, The component for performing reverse-phase chromatographic gradient elution in step S3 is Fr.3, and the elution steps are as follows: (1) Elute with an eluent having a volume ratio of methanol to water of 40:0 to obtain fraction Fr.31; (2) Elute with an eluent having a volume ratio of methanol to water of 60:40 to obtain fraction Fr.32; (3) Elute with an eluent having a volume ratio of methanol to water of 80:20 to obtain fraction Fr.33; (4) Elute with an eluent having a volume ratio of methanol to water of 100:0 to obtain fraction Fr.

34.

10. The preparation method according to claim 9, characterized in that, The component for performing reverse-phase high performance liquid separation in step S4 is Fr.33, and the elution program satisfies the following conditions: (1) The mobile phase is a mixed system of methanol and water, and the volume ratio of methanol to water is: methanol: water = 70:30; (2) Elution time: 0 - 35 min.

11. The preparation method according to claim 10, characterized in that, The chemical structure of compound 1 in the component eluted at 29.8 min in S4 is the terphenyl derivative described in claim 3, wherein R is phenylacetyl; The chemical structure of compound 2 in the component eluted at 13.5 min in S4 is the terphenyl derivative described in claim 3, wherein R is hydrogen; The chemical structure of compound 3 in the component eluted at 20.6 min in S4 is the terphenyl derivative described in claim 4; The chemical structure of compound 4 in the component eluted at 11.3 min in S4 is the terphenyl derivative described in claim 2, wherein R1 is hydroxyl; R2 is hydroxyl; R3 is phenylacetyl; The chemical structure of compound 5 in the component eluted at 18.4 min in S4 is the terphenyl derivative described in claim 5, R1 is hydrogen; R2 is phenylacetyl.

12. The preparation method according to claim 3, characterized in that, The organic solvent in S1 is acetone.

13. The preparation method according to claim 8, characterized in that, The preparation method of the Termitomyces albuminosus extract in S1 includes the following steps: Use the immersion extraction method to soak the dried fruiting bodies of Termitomyces albuminosus with acetone, and concentrate to obtain an acetone extract.

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