Monoterpenoid indole alkaloid compound as well as preparation method and application thereof

Abstract

The invention discloses a monoterpenoid indole alkaloid compound as well as a preparation method and application thereof. The monoterpenoid indole alkaloid compound has a structure as shown in any one of formulas I to XII. The invention discloses a monoterpenoid indole alkaloid compound found from a medicinal plant gossypium hirsutum, and provides an extraction and separation method, a structure identification method and an anti-inflammatory effect application of the monoterpenoid indole alkaloid compound. The monoterpenoid indole alkaloid compounds 1, 4 and 8 provided by the invention have obvious activity of inhibiting release of LPS-induced RAW264.7 macrophage NO, which proves that the prepared monoterpenoid indole alkaloid compounds have good anti-inflammatory activity and can be used for preparing natural anti-inflammatory drugs.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to a monoterpene indole alkaloid compound and a preparation method and application thereof. BACKGROUND

[0002] Monoterpene indole alkaloids (MIAs) are a class of alkaloids condensed by tryptophan and camphorine, which have broad-spectrum biological activity and play an important role in the research and development of natural medicines.

[0003] There are about 50 species of Alstonia plants in the world, and 6 species are mainly produced in China, which have the effects of relieving fever, stopping malaria, stopping diarrhea and stopping bleeding, and are widely used in folk. Previous studies have shown that alkaloids with diverse structures are characteristic components of Alstonia plants. Among them, Alstonia mairei Levl., mainly distributed in the mountainous forests of Yunnan, Guizhou and Sichuan of China, is unique to China. Alstonia mairei Levl. has rich alkaloid components with rich structural types, but the research on its chemical components is relatively rare. SUMMARY

[0004] The main purpose of the present application is to provide a monoterpene indole alkaloid compound and a preparation method and application thereof in preparing anti-inflammatory drugs.

[0005] To achieve the above purpose, the present application provides a monoterpene indole alkaloid compound, which has the structure of any one of the following formula I to formula XII:

[0006]

[0007]

[0008] The present application also provides a preparation method of the above monoterpene indole alkaloid compound, which comprises the following steps:

[0009] Step one: dry Alstonia mairei Levl. branches and leaves are pulverized, and then extracted with a methanol aqueous solution, and the extract is concentrated under reduced pressure to obtain a crude extract;

[0010] Step two: water is added to the crude extract, and then hydrochloric acid aqueous solution is added after suspension to adjust the pH to 3, and then chloroform is used for extraction, and the chloroform layer is discarded to obtain an acid aqueous solution;

[0011] Step three: sodium carbonate aqueous solution is added to the acid aqueous solution to adjust the pH to 10 to obtain an alkali aqueous solution;

[0012] Step four: the alkali aqueous solution is extracted with chloroform to obtain a chloroform extract.

[0013] Step five: the chloroform fraction extract was subjected to normal phase silica gel column chromatography, the mobile phase was dichloromethane-methanol, eluted with a gradient of 100:1 to 0:1 by volume, and ten components A to J with polarity from small to large were obtained by combination;

[0014] Step six: component A was subjected to reversed phase medium pressure C 18 silica gel column chromatography, the mobile phase was methanol-water, eluted with a gradient of 50:50 to 100:0 by volume, and six sub-components A1 to A6 with polarity from large to small were obtained by combination;

[0015] Component A2 was subjected to Sephadex LH-20 gel column chromatography, the mobile phase was methanol, and three sub-components A2a to A2c with molecular weight from small to large were obtained by combination; component A2a was subjected to semi-preparative high performance liquid chromatography purification, C 18 chromatography column, the mobile phase was methanol-water-ammonia water with a volume ratio of 67:33:0.2, and the flow rate was 2.5 mL / min, to obtain the compound shown in formula VI;

[0016] Component A5 was subjected to Sephadex LH-20 gel column chromatography, the mobile phase was methanol, and two sub-components A5a and A5b with molecular weight from small to large were obtained by combination; component A5a was subjected to semi-preparative high performance liquid chromatography purification, C 18 chromatography column, the mobile phase was methanol-water-ammonia water with a volume ratio of 75:25:0.2, and the flow rate was 2.5 mL / min, to obtain the compounds shown in formula IV and formula V with polarity from large to small;

[0017] Component B was subjected to reversed phase medium pressure C 18 silica gel column chromatography, the mobile phase was methanol-water, eluted with a gradient of 50:50 to 100:0 by volume, and seven sub-components B1 to B7 with polarity from large to small were obtained by combination; component B2 was subjected to Sephadex LH-20 gel column chromatography, the mobile phase was methanol, to obtain two sub-components B2a and B2b with molecular weight from large to small; component B2b was subjected to semi-preparative high performance liquid chromatography purification, C 18 chromatography column, the mobile phase was acetonitrile-water-ammonia water with a volume ratio of 56:44:0.2, and the flow rate was 2.5 mL / min, to obtain the compound shown in formula XI;

[0018] Component B4 was subjected to Sephadex LH-20 gel column chromatography, the mobile phase was methanol, to obtain four sub-components B4a to B2d with molecular weight from large to small; component B4c was subjected to semi-preparative high performance liquid chromatography purification, C 18 chromatography column, the mobile phase was acetonitrile-water-ammonia water with a volume ratio of 40:60:0.2, and the flow rate was 2.5 mL / min, to obtain the compound shown in formula XII;

[0019] Component B6 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain two sub-components B6a and B6b in order of decreasing molecular weight; component B6a was subjected to semi-preparative high performance liquid chromatography with C 18 a chromatographic column with methanol-water-ammonia water (70:30:0.2 by volume ratio) as the mobile phase at a flow rate of 2.5 mL / min to obtain the compound shown in formula I;

[0020] Component C was subjected to reverse-phase medium-pressure C 18 silica gel column chromatography with methanol-water as the mobile phase eluted in gradient from 40:60 to 100:0 by volume ratio, and seven sub-components C1 to C7 were obtained in order of decreasing polarity; component C4 was subjected to semi-preparative high performance liquid chromatography with C 18 a chromatographic column with methanol-water-ammonia water (60:40:0.2 by volume ratio) as the mobile phase at a flow rate of 2.5 mL / min to obtain the compound shown in formula IX;

[0021] Component C5 was subjected to semi-preparative high performance liquid chromatography with C 18 a chromatographic column with acetonitrile-water-ammonia water (38:62:0.2 by volume ratio) as the mobile phase at a flow rate of 2.5 mL / min to obtain the compound shown in formula X;

[0022] Component I was subjected to reverse-phase medium-pressure C 18 silica gel column chromatography with methanol-water as the mobile phase eluted in gradient from 20:80 to 100:0 by volume ratio, and ten sub-components I1 to I10 were obtained in order of decreasing polarity; component I1 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain three sub-components I1a to I1c in order of decreasing molecular weight; component I1c was subjected to normal-phase silica gel column chromatography with dichloromethane-methanol as the mobile phase eluted in gradient from 50:1 to 2:1 by volume ratio, and two sub-components I1cA and I1cB were obtained in order of increasing polarity; component I1cB was subjected to semi-preparative high performance liquid chromatography with C 18 a chromatographic column with methanol-water-ammonia water (65:35:0.2 by volume ratio) as the mobile phase at a flow rate of 2.5 mL / min to obtain the compound shown in formula III;

[0023] Component I3 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain four sub-components I3a to I3d in order of decreasing molecular weight; component I3b was subjected to semi-preparative high performance liquid chromatography with C 18 a chromatographic column with methanol-water-ammonia water (43:57:0.2 by volume ratio) as the mobile phase at a flow rate of 2.5 mL / min to obtain the compound shown in formula VII and the compound shown in formula VIII in order of decreasing polarity;

[0024] Component I3d was purified by semi-preparative high-performance liquid chromatography (HPLC) using C1. 18 The chromatographic column was used with a mobile phase of methanol-water-ammonia (volume ratio 40:60:0.2) at a flow rate of 2.5 mL / min to obtain the compound represented by Formula II.

[0025] The present invention also provides the use of the above-mentioned monoterpenoid indole alkaloids in the preparation of anti-inflammatory drugs.

[0026] Furthermore, the dosage form of the drug is an oral dosage form or an injectable dosage form.

[0027] Furthermore, the oral dosage form is a tablet, capsule, granule, drop pill, powder, or oral liquid.

[0028] Furthermore, the injection dosage form is a water injection, a powder injection, or an oil injection.

[0029] This invention discovers a class of monoterpenoid indole alkaloids from the medicinal plant Alstonia mairei Levl., and provides methods for the extraction and separation of these compounds, methods for structural identification, and their anti-inflammatory uses.

[0030] The beneficial effects of this invention are reflected in:

[0031] The monoterpenoid indole alkaloid compounds 1, 4, and 8 provided by this invention have significant inhibitory activity against LPS-induced NO release from RAW264.7 macrophages, demonstrating that the monoterpenoid indole alkaloid compounds prepared by this invention have good anti-inflammatory activity and can be used to prepare natural anti-inflammatory drugs.

[0032] This invention isolates 12 monoterpenoid indole alkaloids from the branches and leaves of the medicinal plant *Cottonwood*, enriching the types of natural alkaloid compounds and providing a theoretical basis for the basic research on the active substances of *Cottonwood*. Attached Figure Description

[0033] Figure 1 Two-dimensional NMR correlation signals of compounds 1 and 2;

[0034] Figure 2 Measured and calculated ECD spectra of compounds 1 and 2;

[0035] Figure 3 The X-ray single-crystal diffraction pattern of compound 8 is shown.

[0036] Figure 4 NMR of compound 1 1 H spectrum;

[0037] Figure 5 NMR of compound 1 13 C spectrum.

[0038] Figure 6 NMR HSQC spectrum of compound 1;

[0039] Figure 7 NMR HMBC spectrum of compound 1;

[0040] Figure 8 NMR of compound 1 1 H- 1 H COSY spectrum;

[0041] Figure 9 NMR NOESY spectrum of compound 1. DETAILED DESCRIPTION

[0042] In order to make the skilled in the art more clearly understand the technical solutions described in the present application, the following examples are listed for illustration. It should be pointed out that the following examples do not constitute a limitation on the scope of protection required by the present application.

[0043] The raw materials, reagents or devices used in the following examples, if not specifically stated, can be obtained from conventional commercial channels, or can be obtained by existing known methods; if not specifically stated, the methods used in the examples of the present application are methods mastered by those skilled in the art.

[0044] The compound with anti-inflammatory effect provided by the present application is a monoterpene indole alkaloid compound, which is isolated and purified from the medicinal plant Alstonia mairei Levl. of the Alstonia genus of the Apocynaceae family. The present application repeatedly performs column chromatography separation and purification on the chloroform part of the 80% methanol extract of the branches and leaves of Alstonia mairei Levl. at pH 10 alkaline water layer, to obtain monoterpene indole alkaloid compounds 1-12, the structural formulas of which are respectively as shown below:

[0045]

[0046]

[0047]

[0048] The present application uses various modern spectral analysis techniques such as high-resolution mass spectrometry, NMR nuclear magnetic resonance spectroscopy (1H, 13C, 1H-1H COSY, 1H-13C HSQC, 1H-13C HMBC, 1H-1H NOESY), and the like. 1 H, 13 C, DEPT-135°, 1 H- 1The chemical structure of the compounds 1-12 is determined by H COSY, HSQC, HMBC and NOESY spectra, ultraviolet spectrum, infrared spectrum, quantum chemistry calculation ECD spectrum and X-ray single crystal diffraction. Among them, compounds 1 and 2 are new compounds, and compounds 3 and 6-12 are first found in the plant Alstonia mairei.

[0049] It is verified that the above-mentioned compounds 1, 4 and 8 provided by the present application can significantly inhibit the LPS-induced NO release activity of RAW264.7 macrophages, and can be used for preparing anti-inflammatory drugs.

[0050] The following is a specific embodiment:

[0051] Embodiment 1

[0052] Separation and preparation of compounds 1-12 and identification

[0053] (I) Separation and preparation of compounds 1-12

[0054] 1. Plant material information

[0055] The plant material was picked from Cuojiang County, Zhaotong City, Yunnan Province (26°32'~27°25'N, 102°52'~103°26'E) in April 2021, and was identified as Alstonia mairei Levl. (Alstonia mairei Levl.) of Apocynaceae.

[0056] 2. Separation and preparation process

[0057] The dried Alstonia mairei branches and leaves (20.0 kg) were pulverized, extracted with 80% methanol aqueous solution at 30°C for 5 times, 7 days each time, the extract was combined and concentrated under reduced pressure to obtain a crude extract (1.2 kg).

[0058] 8.0 L water was added to the crude extract to form a suspension, and 0.3 mol / mL hydrochloric acid aqueous solution was added to adjust the pH to 3, and chloroform was used for extraction, and then the chloroform layer was discarded to remove the neutral components in the extract, and an acid aqueous solution was obtained.

[0059] 10% sodium carbonate aqueous solution was added to the acid aqueous solution to adjust the pH to 10, and an alkali aqueous solution was obtained.

[0060] The alkali aqueous solution was extracted with chloroform to obtain a chloroform extract (230.0 g).

[0061] The chloroform extract was mixed with 200-300 mesh silica gel, and subjected to normal phase silica gel column chromatography, the mobile phase was dichloromethane-methanol (100:1 to 0:1, V / V) gradient elution, and TLC thin layer chromatography plate was used for sample detection, and 10 components (A-J) with different polarity from small to large were obtained by combination.

[0062] Fraction A was subjected to reversed-phase medium pressure C 18 Silica gel column chromatography with gradient elution of methanol-water (50:50 to 100:0, V / V) combined with TLC thin layer chromatography plate spotting detection, and six sub-fractions (A1-A6) were obtained by combining the fractions with polarity from large to small.

[0063] Fraction A2 was subjected to Sephadex LH-20 gel column chromatography with methanol as mobile phase combined with TLC thin layer chromatography plate spotting detection, and three sub-fractions (A2a-A2c) were obtained by combining the fractions with molecular weight from small to large. Fraction A2a was subjected to semi-preparative high performance liquid chromatography purification, C 18 chromatography column with methanol-water-25wt% ammonia water (67:33:0.2, V / V) as mobile phase at a flow rate of 2.5 mL / min, to obtain compound 6 (retention time t R = 18.5 min, weight 27.6 mg).

[0064] Fraction A5 was subjected to Sephadex LH-20 gel column chromatography with methanol as mobile phase combined with TLC thin layer chromatography plate spotting detection, and two sub-fractions (A5a and A5b) were obtained by combining the fractions with molecular weight from small to large. Fraction A5a was subjected to semi-preparative high performance liquid chromatography purification, C 18 chromatography column with methanol-water-25wt% ammonia water (75:25:0.2, V / V) as mobile phase at a flow rate of 2.5 mL / min, to obtain compounds 4 (retention time t R = 18.5 min, weight 27.6 mg) and 5 (retention time t R = 30.9 min, weight 21.7 mg) with polarity from large to small.

[0065] Fraction B was subjected to reversed-phase medium pressure C 18 Silica gel column chromatography with gradient elution of methanol-water (50:50 to 100:0, V / V) combined with TLC thin layer chromatography plate spotting detection, and seven sub-fractions (B1-B7) were obtained by combining the fractions with polarity from large to small.

[0066] Fraction B2 was subjected to Sephadex LH-20 gel column chromatography with methanol as mobile phase to obtain two sub-fractions B2a and B2b with molecular weight from large to small; Fraction B2b was subjected to semi-preparative high performance liquid chromatography purification, C 18 chromatography column with acetonitrile-water-25wt% ammonia water (56:44:0.2, V / V) as mobile phase at a flow rate of 2.5 mL / min, to obtain compound 11 (retention time t R = 26.9 min, weight 19.4 mg).

[0067] Fraction B4 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to give four sub-fractions B4a-B2d in order of decreasing molecular weight. Sub-fraction B4c was subjected to semi-preparative high performance liquid chromatography with C 18 a column and the mobile phase of acetonitrile-water-25wt% ammonia water (40:60:0.2, V / V) at a flow rate of 2.5 mL / min to give compound 12 (retention time t R = 47.3 min, weight 10.4 mg).

[0068] Fraction B6 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to give two sub-fractions B6a and B6b in order of decreasing molecular weight. Sub-fraction B6a was subjected to semi-preparative high performance liquid chromatography with C 18 a column and the mobile phase of methanol-water-25wt% ammonia water (70:30:0.2, V / V) at a flow rate of 2.5 mL / min to give compound 1 (retention time t R = 36.1 min, weight 6.8 mg).

[0069] Fraction C was subjected to reverse phase medium pressure C 18 silica gel column chromatography with gradient elution of methanol-water (40:60 to 100:0, V / V) in combination with TLC thin layer chromatography plate spotting detection to give seven sub-fractions (C1-C7) in order of decreasing polarity.

[0070] Fraction C4 was subjected to semi-preparative high performance liquid chromatography with C 18 a column and the mobile phase of methanol-water-25wt% ammonia water (60:40:0.2, V / V) at a flow rate of 2.5 mL / min to give compound 9 (retention time t R = 56.4 min, weight 14.7 mg).

[0071] Fraction C5 was subjected to semi-preparative high performance liquid chromatography with C 18 a column and the mobile phase of acetonitrile-water-25wt% ammonia water (38:62:0.2, V / V) at a flow rate of 2.5 mL / min to give compound 10 (retention time t R = 31.6 min, weight 6.5 mg).

[0072] Fraction I was subjected to reverse phase medium pressure C 18 silica gel column chromatography with gradient elution of methanol-water (20:80 to 100:0, V / V) in combination with TLC thin layer chromatography plate spotting detection to give ten sub-fractions (11-110) in order of decreasing polarity.

[0073] Component II was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain three sub-components IIa-IIc in order of decreasing molecular weight. Component IIc was subjected to normal-phase silica gel column chromatography with dichloromethane-methanol (50:1 to 2:1, V / V) as the mobile phase in gradient elution, combined with TLC thin-layer chromatography plate spotting detection to obtain two sub-components IIcA and IIcB in order of decreasing polarity. Component IIcB was subjected to semi-preparative high-performance liquid chromatography purification, C 18 chromatography column with methanol-water-25wt% ammonia water (65:35:0.2, V / V) as the mobile phase at a flow rate of 2.5 mL / min to obtain compound 3 (retention time t R = 40.3 min, weight 4.9 mg).

[0074] Component I3 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain four sub-components I3a-I3d in order of decreasing molecular weight. Component I3b was subjected to semi-preparative high-performance liquid chromatography purification, C 18 chromatography column with methanol-water-25wt% ammonia water (43:57:0.2, V / V) as the mobile phase at a flow rate of 2.5 mL / min to obtain compounds 7 (retention time t R = 29.5 min, weight 6.2 mg) and 8 (retention time t R = 32.8 min, weight 4.5 mg) in order of decreasing polarity.

[0075] Component I3d was subjected to semi-preparative high-performance liquid chromatography purification, C 18 chromatography column with methanol-water-25wt% ammonia water (40:60:0.2, V / V) as the mobile phase at a flow rate of 2.5 mL / min to obtain compound 2 (retention time t R = 17.9 min, weight 16.1 mg).

[0076] (B) Structure identification of compounds 1-12

[0077] The structures of compounds 1-12 were determined by combining various spectroscopic analysis methods including high-resolution mass spectrometry, optical rotation, ultraviolet spectroscopy, infrared spectroscopy, nuclear magnetic resonance, and X-ray single crystal diffraction. Compounds 1 and 2 are new compounds, and compounds 3 and 6-12 are first discovered in the plant Gossypium australe. See Figures 1 to 9 :

[0078] Compound 1 (mareine C): yellow oil; [α] D 25 +18 (c 0.1, methanol); ultraviolet (methanol) λ max(log ε) 229 (2.9), 282 (2.4), 288 (2.3) nm; infrared (potassium bromide) v max 3439, 2925, 2854, 1958, 1633, 1458, 1195, 1076 cm -1 ; ECD (methanol) 210 (Δε, +0.2), 215 (Δε, +5.7), 245 (Δε, -15.3), 278 (Δε, +2.6) nm; 1 H and 13 C NMR data are listed in Table 1; high resolution mass spectrum HRESIMS m / z 383.1976 [M+H] + (C 22 H 27 N2O4, 383.1971).

[0079] Combining the high resolution mass spectrum of compound 1 and 13 C NMR nuclear magnetic data analysis its molecular formula is C 22 H 26 N2O4. The NMR data of compound 1 (Table 1) contains a set of characteristic peak signals of indole ring (δ H 7.49, d, J = 7.8 Hz, H-9; 7.40, d, J = 8.2 Hz, H-12; 7.18, dd, J = 8.2, 7.1 Hz, H-11; 7.11, dd, J = 7.8, 7.1 Hz, H-10; δ C 136.0, C-13; 133.0, C-2; 127.6, C-8; 122.0, C-11; 119.6, C-10; 118.2, C-9; 111.5, C-12; 107.3, C-7), a carbonyl carbon signal (δ C 168.0, C-22), a set of tri-substituted olefinic carbon signals (δ H 7.53, s, H-17; δ C 155.7, C-17; 109.9, C-16), an oxymethine signal (δ H 4.58, dq, J = 10.4, 6.2 Hz; δ C 71.4, CH-19), an oxymethylene signal (δ H 3.84, d, J = 10.8 Hz; 3.36, d, J = 10.8 Hz; δ C 68.5, CH2-23), a methoxy signal (δ H 3.72, s; δ C 51.2, OCH3) and a methyl signal (δ H 1.41, d, J = 6.2 Hz; δC 18.8, CH3-18). By careful comparison of the NMR data of compound 1 and compound 4, it was found that the structures of these two compounds are similar, the main difference is that there is an additional methylene oxide signal (δ 68.5, CH2-23) and a quaternary carbon signal (δ 58.8, C-3) in compound 1 instead of an aminomethyl signal (δ 58.8, CH2-3) in compound 4. H 3.84, d, J = 10.8 Hz; 3.36, d, J = 10.8 Hz; δ C 68.5, CH2-23) and one quaternary carbon signal (δ C 58.8, C-3), instead of an aminomethyl signal (δ H 3.32, br d, J = 11.7, δ C 60.0, CH-3). Combined with the HMBC correlation of H2-23 with C-2, C-3 and C-14 in the 2D NMR spectrum and the molecular formula of compound 1 C 22 H 26 N2O4, it was speculated that C-23 is connected with C-3 by methylene oxide. Therefore, compound 1 is a hydroxymethylated derivative of compound 4. The relative configuration of compound 1 was determined by NOESY correlation signals and coupling constants. First, it was assumed that H-20 is in the α configuration, and in its NOESY spectrum, the correlation signal of H-20 (δ 4.57, d, J = 10.4 Hz) with H-15 (δ 2.45, ddd, J = 12.8, 4.4, 4.1 Hz) was observed, which determined that H-15 is in the α configuration. In addition, H-15 and H-14a (δ 2.38, dd, J = 14.7, 4.1 Hz), H2-23 and H-14b (δ 2.33, d, J = 14.7 Hz) and H-19 (δ 1.52, m) were correlated, which determined that H-19 is in the β configuration. The absolute configuration of compound 1 was determined by calculation of ECD, and the results showed that the measured ECD spectrum of compound 1 is consistent with the calculated (3S, 15S, 19S, 20S)-1 spectrum. Therefore, the absolute configuration of compound 1 is 3S, 15S, 19S, 20S. H 1.52, m) with H-15 (δ H 2.45, ddd, J = 12.8, 4.4, 4.1 Hz) was observed, which determined that H-15 is in the α configuration. In addition, H-15 and H-14a (δ H 2.38, dd, J = 14.7, 4.1 Hz), H2-23 and H-14b (δ H 4.57, d, J = 10.4, 6.2 Hz) and H-19 (δ 1.52, m) were correlated, which determined that 23-CH2OH is in the α configuration and H-19 is in the β configuration. The absolute configuration of compound 1 was determined by calculation of ECD, and the results showed that the measured ECD spectrum of compound 1 is consistent with the calculated (3S, 15S, 19S, 20S)-1 spectrum. Therefore, the absolute configuration of compound 1 is 3S, 15S, 19S, 20S.

[0080] Table 1.1H and 1 H and 13 C NMR data (deuterated chloroform)

[0081] No. H (J in Hz) ​ C ]]> ​ No. H (J in Hz) ​ delta C ]]> 2 133.0 14β 2.14, dd (14.7, 12.8) 3 58.8 15 2.45, ddd (12.8, 4.4, 4.1) 27.7 5α 2.90, dd (14.2, 6.7) 45.2 16 109.9 5β 3.51, ddd (14.2, 11.6, 6.1) 17 7.53,s 155.7 6α 3.08, ddd (16.2, 11.6, 6.7) 16.5 18 1.41,d(6.2) 18.8 6β 2.59, dd (16.2, 6.1) 19 4.57, dq (10.4, 6.2) 71.4 7 107.3 20 1.52,m 37.7 8 127.6 21α 2.83, dd (12.4, 2.1) 47.6 9 7.49,d(7.8) 118.2 21β 3.15, dd (12.4, 4.1) 10 7.11, dd (7.8, 7.1) 119.6 22 168.0 11 7.18, dd (8.2, 7.1) 122.0 23α 3.36,d(10.8) 68.5 12 7.40,d(8.2) 111.5 23β 3.84,d(10.8) 13 136.0 OCH3 3.72,s 51.2 14α 2.38, dd (14.7, 4.1) 33.7

[0082] Compound 2 (mareine D): colorless oil; [α] D 25 +56 (c 0.1, methanol); UV (methanol) λ max(log ε) 209 (3.0), 228 (2.9), 260 (2.8), 278 (2.5) nm; IR (KBr) v max 3431, 2926, 1958, 1632, 1353, 1196, 1076 cm -1 ; ECD (methanol) 215 (Δε, -7.5), 228 (Δε, +7.8), 243 (Δε, -17.9) nm; 1 H and 13 C NMR data are listed in Table 2; high resolution mass spectrum HRESIMS m / z 389.1627 [M + (C 21 H 27 ClN2O3. The calculated value for C19H20CIN2O3is 389.1626).

[0083] Combining the high resolution mass spectrum of compound 2 with 13 C NMR nuclear magnetic data analysis its molecular formula is C 21 H 27 ClN2O3. The NMR data of compound 2 (Table 2) is similar to that of known compound 12-methoxytubotaiwine, the main difference is that there is an extra set of gem-dihydrogen signals (δ H 5.75, d, J = 10.0 Hz; 5.61, d, J = 10.0 Hz; δ C 66.7, N4-CH2Cl), and one methoxy signal is missing. The nuclear magnetic data of CH2-3, CH2-5 and CH-21 in compound 2 move to the deshielded region compared with 12-methoxytubotaiwine, combined with the molecular formula of compound 2, it can be inferred that the chloromethyl (CH2Cl) in the structure is connected to the N4 atom. From the two-dimensional nuclear magnetic spectrum, the HMBC correlation signals of N4-CH2Cl with C-3, C-5 and C-21 can be observed, and the HMBC correlation signals of H-9 / H-10 / H-11 with C-3, C-5 and C-21 can be observed, which further confirms that the chloromethyl (CH2Cl) is connected to the N4 atom. 1 H- 1H COSY correlation signals thus proved the structure of compound 2 as N4-chloromethyl-12-methoxytubotaiwine. The chloromethyl CH2C1 group correlated with H-5a (3.80, overlap), H-20 (2.22, overlap) and H-21 (4.61, br s) indicated the a configuration of N4-CH2C1. The experimental ECD spectrum of compound 2 was consistent with the calculated (4S,7R,15R,20R,21S)-2 spectrum trend. Therefore, the absolute configuration of compound 2 was 4S,7R,15R,20R,21S. Up to now, five tubotaiwine-type alkaloids have been reported from medicinal plants of Alstonia genus, including tubotaiwine, tubotaiwine N-oxide, lagunamine, 12-methoxytubotaiwine and alstoscholarinine G. Compound 2 (mareine D) represents the first 12-hydroxytubotaiwine-type alkaloid and the sixth tubotaiwine-type alkaloid reported from medicinal plants of Alstonia genus.

[0084] Table 2.1H NMR data of compound 2 in DMSO-d6 1 H and 13 C NMR data (deuterated methanol)

[0085]

[0086] Compound 3 (4R-tetrahydroalstonine N-oxide): 1 H NMR (600 MHz, deuterated chloroform) δ H : 3.30 (m, H-3), 7.44 (dd, J = 7.7, 1.2 Hz, H-9), 7.09 (ddd, J = 7.7, 7.7, 1.0 Hz, H-10), 7.13 (ddd, J = 7.7, 7.7, 1.2 Hz, H-11), 7.24 (dd, J = 7.7, 1.0 Hz, H-12), 7.59 (s, H-17), 1.44 (d, J = 6.0 Hz, H3-18), 4.40 (dq, J = 10.0, 6.1 Hz, H-19), 1.54 (q, J = 12.2 Hz, H-20), 3.76 (s, H3-23); 13 C NMR (150 MHz, deuterated chloroform) δ C: 136.84 (C-2), 61.65 (C-3), 49.49 (C-5), 19.13 (C-6), 112.11 (C-7), 126.42 (C-8), 117.93 (C-9), 119.39 (C-10), 121.80 (C-11), 111.03 (C-12), 136.13 (C-13), 30.28 (C-14), 33.07 (C-15), 109.01 (C-16), 154.94 (C-17), 18.66 (C-18), 74.15 (C-19), 39.15 (C-20), 45.00 (C-21), 167.98 (C-22), 51.45 (C-23).

[0087] Compound 4 (tetrahydroalstonine): 1 H NMR (600 MHz, Deuterated chloroform) δ H : 3.32 (dd, J = 11.7, 2.3 Hz, H-3), 2.92 (overlap, H-5a), 3.09 (dd, J = 12.3, 2.0 Hz, H-5b), 7.45 (dd, J = 7.7, 1.2 Hz, H-9), 7.07 (ddd, J = 7.7, 7.7, 1.0 Hz, H-10), 7.12 (ddd, J = 7.7, 7.7, 1.2 Hz, H-11), 7.25 (dd, J = 7.7, 1.0 Hz, H-12), 7.57 (s, H-17), 1.39 (d, J = 6.2 Hz, H3-18), 4.49 (dq, J = 10.1, 6.2 Hz, H-19), 1.53 (q, J = 12.2 Hz, H-20), 3.75 (s, H3-23), 7.92 (s, NH); 13 C NMR (150 MHz, Deuterated chloroform) δ C : 134.71 (C-2), 38.52 (C-3), 53.67 (C-5), 34.37 (C-6), 109.65 (C-7), 127.29 (C-8), 121.48 (C-9), 110.94 (C-10), 119.48 (C-11), 118.18 (C-12), 136.10 (C-13), 21.88 (C-14), 31.43 (C-15), 108.13 (C-16), 155.92 (C-17), 18.65 (C-18), 72.62 (C-19), 59.94 (C-20), 56.41 (C-21), 168.18 (C-22), 51.31 (C-23).

[0088] Compound 5 (tabersonine):1 H NMR (600 MHz, deuteriochloroform) δ H : 3.46 (ddd, J = 15.8, 4.8, 1.5 Hz, H-3a), 3.19 (dt, J = 15.8, 1.5 Hz, H-3b), 3.04 (m, H-5a), 2.71 (m, H-5b), 2.07 (dt, J = 11.3, 6.5 Hz, H-6a), 1.80 (ddd, J = 11.3, 6.5, 1.5 Hz, H-6b), 7.23 (dd, J = 7.7, 1.2 Hz, H-9), 6.87 (ddd, J = 7.7, 7.7, 1.0 Hz, H-10), 7.14 (ddd, J = 7.7, 7.7, 1.2 Hz, H-11), 6.82 (dd, J = 7.7, 1.0 Hz, H-12), 5.80 (ddd, J = 9.9, 4.8, 1.5 Hz, H-14), 5.71 (d, J = 9.9, 1.5, 1.5 Hz, H-15), 2.55 (dd, J = 15.0, 1.9 Hz, H-17a), 2.44 (d, J = 15.0 Hz, H-17b), 0.64 (t, J = 7.5 Hz, H3-18), 1.00 (dq, J = 14.5, 7.5 Hz, H-19a), 0.86 (dq, J = 14.5, 7.5 Hz, H-19b), 3.77 (s, H3-23); 1 H NMR (600 MHz, deuteriochloroform) δ C : 166.80 (C-2), 51.02 (C-3), 50.97 (C-5), 44.54 (C-6), 55.14 (C-7), 138.09 (C-8), 121.49 (C-9), 120.59 (C-10), 127.67 (C-11), 109.32 (C-12), 143.22 (C-13), 124.88 (C-14), 133.10 (C-15), 92.15 (C-16), 26.94 (C-17), 7.49 (C-18), 28.49 (C-19), 41.35 (C-20), 70.08 (C-21), 169.03 (C-22), 50.97 (C-23).

[0089] Compound 6 (melodinine X3): 1 H NMR (600 MHz, deuteriochloroform) δ H3.50 (m, H-3), 3.26 (ddd, J = 8.7, 8.7, 3.0 Hz, H-5a), 3.15 (ddd, J = 8.7, 8.7, 8.7 Hz, H-5b), 2.27 (m, H-6a), 2.19 (m, H-6b), 7.40 (d, J = 7.7 Hz, H-9), 6.94 (dd, J = 7.7, 1.3 Hz, H-10), 7.18 (dd, J = 7.7, 1.3 Hz, H-11), 6.85 (d, J = 7.7 Hz, H-12), 5.74 (dd, J = 10.5, 4.1 Hz, H-14), 5.88 (d, J = 10.5 Hz, H-15), 2.69 (dd, J = 15.4, 1.6 Hz, H-17a), 2.06 (d, J = 15.4 Hz, H-17b), 0.67 (t, J = 7.5 Hz, H3-18), 1.07 (m, H-19a), 0.97 (m, H-19b), 3.62 (dd, J = 10.6, 4.5 Hz, H-22a), 3.36 (dd, J = 10.6, 10.6 Hz, H-22b), 3.78 (s, H3-24); 13 H NMR (600 MHz, Methanol-d4) δ C : 165.71 (C-2), 57.77 (C-3), 54.14 (C-5), 41.95 (C-6), 56.47 (C-7), 139.15 (C-8), 122.30 (C-9), 121.52 (C-10), 121.86 (C-11), 109.12 (C-12), 143.11 (C-13), 122.30 (C-14), 134.09 (C-15), 89.92 (C-16), 33.40 (C-17), 8.0 (C-18), 31.70 (C-19), 35.61 (C-20), 65.15 (C-21), 63.12 (C-22), 169.82 (C-23), 50.96 (C-24).

[0090] Compound 7 (peraksine): 1 H NMR (600 MHz, Methanol-d4) δ H: 2.59 (d, J = 15.5 Hz, H-6β), 7.37 (dd, J = 7.6, 1.1 Hz, H-9), 6.96 (ddd, J = 7.6, 7.6, 1.1 Hz, H-10), 7.04 (ddd, J = 7.6, 7.6, 1.1 Hz, H-11), 7.27 (dd, J = 7.6, 1.1 Hz, H-12), 2.25 (br s, H-15), 5.00 (d, J = 1.8 Hz, H-17), 1.40 (d, J = 7.0 Hz, H-18); 13 H NMR (600 MHz, Methanol-d4) δ C : 139.49 (C-2), 53.13 (C-3), 45.80 (C-5), 29.59 (C-6), 104.59 (C-7), 128.91 (C-8), 118.83 (C-9), 119.86 (C-10), 122.09 (C-11), 112.05 (C-12), 138.39 (C-13), 34.31 (C-14), 31.30 (C-15), 43.11 (C-16), 98.66 (C-17), 14.02 (C-18), 56.84 (C-19), 34.79 (C-20), 66.40 (C-21).

[0091] Compound 8 (17-epi-peraksine): 1 H NMR (600 MHz, Methanol-d4) δ H : 2.59 (d, J = 15.5 Hz, H-6β), 7.37 (dd, J = 7.6, 1.1 Hz, H-9), 6.96 (ddd, J = 7.6, 7.6, 1.1 Hz, H-10), 7.04 (ddd, J = 7.6, 7.6, 1.1 Hz, H-11), 7.27 (dd, J = 7.6, 1.1 Hz, H-12), 2.25 (br s, H-15), 5.00 (d, J = 1.8 Hz, H-17), 1.40 (d, J = 7.0 Hz, H-18); 13 H NMR (600 MHz, Methanol-d4) δ C: 139.68 (C-2), 53.04 (C-3), 48.96 (C-5), 28.17 (C-6), 104.04 (C-7), 128.83 (C-8), 118.78 (C-9), 119.90 (C-10), 122.14 (C-11), 112.09 (C-12), 138.36 (C-13), 34.07 (C-14), 23.95 (C-15), 42.03 (C-16), 95.24 (C-17), 14.12 (C-18), 56.75 (C-19), 35.85 (C-20), 61.15 (C-21).

[0092] Compound 9 (vincamine): 1 H NMR (600 MHz, deuterated chloroform) δ H : 7.49 (m, H-9), 7.13 (overlap, H-10), 7.13 (overlap, H-11), 7.09 (m, H-12) 2.22 (d, J = 14.2 Hz, H-17a), 2.13 (d, J = 14.2 Hz, H-17b), 0.91 (t, J = 7.6 Hz, H3-19), 4.59 (s, H-21), 3.83 (s, H3-23); 13 C NMR (150 MHz, deuterated chloroform) δ C : 131.48 (C-2), 44.40 (C-3), 50.95 (C-5), 16.83 (C-6), 105.93 (C-7), 128.98 (C-8), 118.50 (C-9), 121.67 (C-10), 120.26 (C-11), 110.30 (C-12), 134.08 (C-13), 20.78 (C-14), 25.09 (C-15), 81.88 (C-16), 44.58 (C-17), 28.89 (C-18), 7.6 (C-19), 35.10 (C-20), 59.14 (C-21), 174.51 (C-22), 54.33 (C-23).

[0093] Compound 10 (vincapusine): 1 H NMR (600 MHz, deuterated chloroform) δ H: 1.78 (m, H-3a), 2.10 (m, H-3p), 3.34 (ddd, J = 14.0, 10.2, 7.9 Hz, H-5a), 3.17 (dd, J = 14.0, 7.9 Hz, H-5p), 2.67 (ddd, J = 16.6, 7.9, 2.0 Hz, H-6a), 2.89 (m, H-6p), 7.44 (m, H-9), 7.12 (overlap, H-10), 7.12 (overlap, H-11), 6.95 (m, H-12), 3.75 (m, H-14), 3.97 (d, J = 2.9 Hz, H-15), 2.77 (d, J = 12.0 Hz, H-17a), 2.42 (d, J = 12.0 Hz, H-17p), 1.07 (t, J = 7.5 Hz, H3-19), 4.19 (s, H-21), 4.19 (s, H3-23), 1.55 (s, -OH); 13 C NMR (150 MHz, chloroform-d) δ C : 134.41 (C-2), 46.49 (C-3), 50.50 (C-5), 18.72 (C-6), 111.08 (C-7), 130.50 (C-8), 118.62 (C-9), 121.05 (C-10), 122.66 (C-11), 111.22 (C-12), 137.13 (C-13), 66.62 (C-14), 82.20 (C-15), 90.65 (C-16), 45.91 (C-17), 26.19 (C-18), 9.5 (C-19), 44.25 (C-20), 56.83 (C-21), 168.73 (C-22), 53.46 (C-23).

[0094] Compound 11 (uncarine E): 1 H NMR (600 MHz, chloroform-d) δ H: 2.57 (dd, J = 11.6, 2.9 Hz, H-3), 3.22 (ddd, J = 8.6, 8.6, 2.3 Hz, H-5p), 7.27 (dd, J = 7.7, 1.0 Hz, H-9), 7.02 (ddd, J = 7.7, 7.7, 1.0 Hz, H-10), 7.19 (ddd, J = 7.7, 7.7, 1.0 Hz, H-11), 6.92 (dd, J = 7.7, 1.0 Hz, H-12), 0.88 (ddd, J = 13.1, 11.6, 11.6 Hz, H-14p), 7.42 (s, H-17), 1.42 (d, J = 6.1 Hz, H3-18), 4.36 (qd, J = 10.2, 6.2 Hz, H-19), 3.29 (dd, J = 12.0, 2.0 Hz, H-21p), 3.60 (s, H3-23), 8.94 (br s, NH); 13 C NMR (150 MHz, Chloroform-d) δ C : 181.77 (C-2), 71.31 (C-3), 54.22 (C-5), 34.95 (C-6), 57.11 (C-7), 133.88 (C-8), 124.58 (C-9), 122.57 (C-10), 127.78 (C-11), 109.94 (C-12), 140.49 (C-13), 30.28 (C-14), 30.54 (C-15), 109.92 (C-16), 155.08 (C-17), 18.75 (C-18), 72.25 (C-19), 37.97 (C-20), 53.61 (C-21), 167.75 (C-22), 51.11 (C-23).

[0095] Compound 12 (uncarine C): 1 H NMR (600 MHz, Chloroform-d) δ H : 1.99 (m, H-6a), 7.19 (dd, J = 7.5, 1.0 Hz, H-9), 7.04 (ddd, J = 7.5, 7.5, 1.3 Hz, H-10), 7.17 (ddd, J = 7.5, 7.5, 1.0 Hz, H-11), 6.85 (dd, J = 7.5, 1.3 Hz, H-12), 1.71 (ddd, J = 12.0, 4.6, 2.8 Hz, H-14a), 1.49 (ddd, J = 12.0, 12.0, 12.0 Hz, H-14p), 7.48 (s, H-17), 1.40 (d, J = 6.1 Hz, H3-18), 4.55 (qd, J = 10.4, 6.1 Hz, H-19), 3.60 (s, H3-23), 8.39 (br s, NH).13 C NMR (150 MHz, Deuterated chloroform) δ C : 181.26 (C-2), 74.58 (C-3), 55.36 (C-5), 34.77 (C-6), 56.23 (C-7), 133.58 (C-8), 123.26 (C-9), 122.79 (C-10), 128.10 (C-11), 109.65 (C-12), 140.81 (C-13), 29.68 (C-14), 31.09 (C-15), 109.30 (C-16), 155.41 (C-17), 19.17 (C-18), 72.34 (C-19), 37.95 (C-20), 53.84 (C-21), 167.90 (C-22), 51.09 (C-23).

[0096] Single crystal data: Compound 8, C 19 H 21 N2O2·H2O, molecular weight M = 327.39, cell parameters α = 90.00°, β = 99.39°, γ = 90.00° and The diffraction experiment temperature was 273 K, and the diffraction wavelength was Monoclinic, space group C121, Z = 4, diffraction experiment crystal size was 0.22 x 0.20 x 0.18 mm 3 , absorption coefficient was 0.726 mm -1 , F(000) = 700.0, diffraction collection angle was 4.343° to 66.621°, the minimum and maximum diffraction indexes (h, k and l) were -23 ≤ h ≤ 9, -9 ≤ k ≤ 8, -12 ≤ l ≤ 12. The total number of diffraction points was 25459, the number of independent diffraction points was 2552 [R (int) = 0.1544] and [R (sigma) = 0.1387], the degree of completion was 87%, the residual factor R1 and the weighted residual factor wR2 of the observable diffraction points [I > = 2σ (I)] were 0.0674 and 0.2024, respectively, the fitting factor F 2 was 1.164, and the Flack parameter was 0.02 (6).

[0097] Example 2

[0098] Test of anti-inflammatory performance of compound 1-12

[0099] I. Experimental method

[0100] NO is a key factor in mediating inflammatory response and plays an important role in endotoxin-induced body injury. Excessive NO can promote the release of various cytokines from cells and aggravate inflammatory response. In this study, LPS was used to induce RAW264.7 cells in vitro to establish an inflammatory model, and monoterpenoid indole alkaloids 1-12 were used for intervention. The release of NO in the cell supernatant was measured to determine the level of inflammatory regulation of RAW264.7 cells and evaluate their anti-inflammatory activity.

[0101] RAW264.7 cells in the logarithmic growth phase were inoculated in a 96-well cell culture plate, and the cell concentration was adjusted to 1×10 6 对照组 实验组 对照组 空白组 50 50

[0102] II. Results

[0103] The results of the inhibitory activity of compounds 1-12 on LPS-induced NO release from RAW264.7 macrophages are shown in Table 3.

[0104] Table 3. Inhibitory activity of compounds 1-12 on LPS-induced NO release from RAW264.7 cells

[0105]

[0106]

[0107] Conclusion: Compounds 1, 4, and 8 have significant inhibitory activity on LPS-induced NO release from RAW264.7 macrophages, with IC 50 values of 4.4±1.8, 4.6±0.9, and 9.2±0.3 μΜ, respectively. Compounds 7, 11, and 12 have better inhibitory activity on LPS-induced NO release from RAW264.7 macrophages, with IC​​​​​50 The values were 17.3 ± 1.1, 20.4 ± 2.1 and 11.3 ± 0.6 μΜ, respectively. In addition, at a concentration of 50 μΜ, none of the compounds 1-12 showed significant cytotoxicity to RAW264.7 cells, with or without the addition of LPS (1 μg / mL).

[0108] The above description is only the preferred embodiment of the application, not to limit the application, any modification, equivalent replacement, improvement, etc. within the spirit and principle of the application, should be included in the protection scope of the application.

Claims

1. A class of monoterpenoid indole alkaloids, characterized in that, has the structure of any one of the following formula I to formula XII:

2. The method for preparing monoterpenoid indole alkaloids as described in claim 1, characterized in that, The method comprises the following steps: Step one: dry cotton branches and leaves are powdered and extracted with a methanol aqueous solution, and the extract is concentrated under reduced pressure to obtain a crude extract; Step two: water is added to the crude extract, and after being suspended, hydrochloric acid aqueous solution is added to adjust the pH to 3, and then chloroform is used for extraction, and the chloroform layer is discarded to obtain an acid aqueous solution; Step three: sodium carbonate aqueous solution is added to the acid aqueous solution to adjust the pH to 10 to obtain an alkali aqueous solution; Step four: the alkali aqueous solution is extracted with chloroform to obtain a chloroform fraction extract; Step five: the chloroform fraction extract is subjected to normal phase silica gel column chromatography, the mobile phase is dichloromethane-methanol, and elution is performed in a gradient of 100:1 to 0:1 by volume, and ten components A to J with decreasing polarity are obtained by combination; Step six: component A was subjected to reverse phase medium pressure C 18 Silica gel column chromatography, the mobile phase was methanol-water, eluted with gradient of 50:50 to 100:0 by volume ratio, combined to get six sub-components A1 to A6 from large to small polarity; Fraction A2 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase, and three sub-fractions A2a to A2c with increasing molecular weight were obtained by combining. Fraction A2a was subjected to semi-preparative high performance liquid chromatography with C 18 chromatography column with methanol-water-ammonia water (67:33:0.2 by volume ratio) as the mobile phase at a flow rate of 2.5 mL / min to obtain a compound shown as formula VI. Component A5 was chromatographed on Sephadex LH-20 gel column with methanol as the mobile phase to give two sub-components A5a and A5b with molecular weight from small to large. Component A5a was purified by semi-preparative high performance liquid chromatography with C 18 chromatography column with methanol-water-ammonia water (75:25:0.2 by volume) as the mobile phase at a flow rate of 2.5 mL / min to give compounds of formula IV and formula V in order of polarity from large to small. Component B was subjected to reversed-phase medium-pressure C 18 The component B2 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain two sub-components B2a and B2b in the order of decreasing molecular weight. The component B2b was subjected to semi-preparative high-performance liquid chromatography with C 18 The component B2 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain two sub-components B2a and B2b in the order of decreasing molecular weight. The component B2b was subjected to semi-preparative high-performance liquid chromatography with C Fraction B4 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain four sub-fractions B4a to B2d in the order of decreasing molecular weight; Fraction B4c was subjected to semi-preparative high performance liquid chromatography with C 18 chromatography column with acetonitrile-water-ammonia water (40:60:0.2 by volume) as the mobile phase at a flow rate of 2.5 mL / min to obtain the compound shown in formula Xl; Fraction B6 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain two sub-fractions B6a and B6b in the order of decreasing molecular weight; fraction B6a was subjected to semi-preparative high performance liquid chromatography with C 18 chromatography column with the mobile phase of methanol-water-ammonia water at a volume ratio of 70:30:0.2 at a flow rate of 2.5 mL / min to obtain the compound shown in formula I; Fraction C was purified by reversed-phase medium pressure C 18 Silica gel column chromatography was performed using methanol-water as mobile phase with a gradient elution from 40:60 to 100:0 (v / v) to give seven sub-fractions C1 to C7 with decreasing polarity. Fraction C4 was further purified by semi-preparative high performance liquid chromatography using C 18 Chromatography column was used with methanol-water-ammonia water (60:40:0.2, v / v) as mobile phase at a flow rate of 2.5 mL / min to give the compound shown as formula IX. Component C5 was purified by semi-preparative high performance liquid chromatography, using C 18 chromatography column with a mobile phase of acetonitrile-water-ammonia water (38:62:0.2 by volume) at a flow rate of 2.5 mL / min to obtain the compound shown as Formula X; Component I was subjected to reversed-phase medium-pressure C 18 The silica gel column chromatography was performed with methanol-water as the mobile phase, and elution was performed by gradient elution with a volume ratio of 20:80 to 100:0, and ten sub-components I1 to I10 with decreasing polarity were obtained by combination. Component I1 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase, and three sub-components I1a to I1c with decreasing molecular weight were obtained. Component I1c was subjected to normal-phase silica gel column chromatography with dichloromethane-methanol as the mobile phase, and elution was performed by gradient elution with a volume ratio of 50:1 to 2:1, and two sub-components I1cA and I1cB with increasing polarity were obtained by combination. Component I1cB was subjected to semi-preparative high-performance liquid chromatography purification, and C 18 The chromatographic column was subjected to high-performance liquid chromatography purification with methanol-water-ammonia water as the mobile phase at a volume ratio of 65:35:0.2, and a flow rate of 2.5 mL / min, and a compound represented by formula III was obtained. Component I3 was subjected to Sephadex LH-20 gel column chromatography with methanol as the mobile phase to obtain four sub-components I3a to I3d in descending order of molecular weight; component I3b was subjected to semi-preparative high performance liquid chromatography with C 18 chromatography column with the mobile phase of methanol-water-ammonia water at a volume ratio of 43:57:0.2 at a flow rate of 2.5 mL / min to obtain compounds of formula VII and compounds of formula VIII in descending order of polarity. Component I3d was purified by semi-preparative HPLC using a C 18 The column was eluted with a mobile phase of methanol-water-ammonia (40:60:0.2 by volume) at a flow rate of 2.5 mL / min to give the compound of formula II.

3. The use of the monoterpene indole alkaloid compound in claim 1 in the preparation of an anti-inflammatory drug.

4. Use according to claim 3, characterized in that, The dosage form of the drug is an oral dosage form or an injection dosage form.

5. Use according to claim 4, characterized in that, The oral dosage form is a tablet, a capsule, a granule, a dripping pill, a powder or an oral liquid.

6. Use according to claim 4, characterized in that, The injection dosage form is an aqueous injection, a powder injection or an oil injection.

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