Losane alkaloid and dimer compound thereof, and preparation method and application of losane alkaloid and dimer compound thereof

Abstract

The invention relates to lotus alkane alkaloids and dimer compounds thereof as well as a preparation method and application of the dimer compounds, in particular to four lotus alkane alkaloid dimer compounds and eight lotus alkane alkaloid compounds extracted and separated from stephania elegans, as well as a preparation method and application of the four lotus alkane alkaloid dimer compounds and the eight lotus alkane alkaloid compounds, and belongs to the technical field of medicines. The structural general formula of the compound is shown in the specification, in the formula, R1 is hydrogen, and R2 and R3 are both methoxyl; or R1 and R2 are-OCH2O-so as to constitute a five-membered ring, and R3 is hydrogen. According to the invention, an LPS-induced BV-2 microglial cell overactivity model is used for evaluating the neuroinflammation resistance of the prepared lotus alkane alkaloid and the dimer compound thereof, and the result shows that the new compound can inhibit the release of LPS-induced overactivity BV-2 microglial cell NO, shows obvious neuroinflammation resistance activity, and has a good application prospect. The application of the novel compound in development of drugs for treating neurodegenerative diseases is clarified. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to liensinine-type alkaloids and their dimer compounds, as well as their preparation methods and applications, and specifically relates to liensinine-type alkaloids and liensinine-type alkaloid dimer compounds in Stephania venosa (Willd.) Hook. f. var. hainana Lo, and their preparation methods and applications, belonging to the field of pharmaceutical technology. Background Art

[0002] Stephania venosa (Willd.) Hook. f. var. hainana Lo is a plant of the genus Stephania in the family Menispermaceae. There are about 60 species of the genus Stephania globally, distributed in tropical and subtropical regions of Asia and Africa, and a few are produced in Oceania. There are 39 species and 1 variety in China, produced in provinces and regions south of the Yangtze River, with the most species in Yunnan and Guangxi. Stephania venosa (Willd.) Hook. f. var. hainana Lo is a vine, distributed in Yunnan, Guangxi, Guizhou, Sichuan and other places in China. It is recorded in "New Outline of Chinese Materia Medica" that it has the effects of clearing heat and detoxifying, dispelling wind and removing dampness, regulating qi and relieving pain, etc., and can be used to treat rheumatoid arthritis, carbuncles and sores, heat stroke dysentery, mumps, pharyngitis and stomatitis, etc. Modern pharmacological studies have shown that Stephania venosa (Willd.) Hook. f. var. hainana Lo has anti-inflammatory and anti-tumor effects, and its main chemical components are isoquinoline alkaloids, flavonoids, etc.

[0003] In recent years, certain progress has been made in the research on the chemical components and biological activities of Stephania venosa (Willd.) Hook. f. var. hainana Lo, but there are still many structurally complex unknown compounds that have not been discovered, which limits the further application of Stephania venosa (Willd.) Hook. f. var. hainana Lo in terms of biological activities. The present invention uses the dried whole herb of Stephania venosa (Willd.) Hook. f. var. hainana Lo as the raw material, isolates and identifies 12 new liensinine-type alkaloids and their dimer compounds, and systematically evaluates their existing biological activities, providing an important basis for in-depth research on the pharmacological activities of Stephania venosa (Willd.) Hook. f. var. hainana Lo and the development of new drugs. Summary of the Invention

[0004] The purpose of the present invention is to provide liensinine-type alkaloids and their dimer compounds, as well as their preparation methods and applications, and in particular to provide the structures, preparation methods of 12 liensinine-type alkaloids and their dimer compounds isolated and identified from Stephania venosa (Willd.) Hook. f. var. hainana Lo, and their applications in the preparation of drugs for preventing or treating neurodegenerative diseases.

[0005] The liensinine-type alkaloids and their dimer compounds, and their pharmaceutically acceptable salts and isomers have the following general structural formulas (I) - (VIII):

[0006]

[0007] Among them, R1 is hydrogen, and both R2 and R3 are methoxy; or R1 and R2 are -OCH2O- to form a five-membered ring, and R3 is hydrogen.

[0008] The Huashi - type alkaloids, their dimer - like compounds, pharmaceutically acceptable salts and isomers thereof in the present invention, and the compounds are one of the following structural formulas:

[0009]

[0010] Among them, R1 is hydrogen, and both R2 and R3 are methoxy groups; or R1 and R2 are -OCH2O- to form a five - membered ring, and R3 is hydrogen.

[0011] The Lianhuashi - type alkaloids, their dimer - like compounds, pharmaceutically acceptable salts and isomers thereof in the present invention specifically disclose the following 12 specific compounds:

[0012]

[0013] Compound 1: (-)(10R,13R,14S,16S,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine A

[0014] Compound 2: (-)(10S,13R,14S,16S,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine B

[0015] Compound 3: (-)(10S,13R,14S,16S,6"S,7"S,8"R,10"S,13"R,14"S)-bishernsubanine C

[0016] Compound 4: (-)(10S,13R,14S,16R,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine D

[0017] Compound 5: (-)(10S,13R,14S)-hernsubanine G

[0018] Compound 6: (+)(6S,7S,8R,10S,13R,14S,2'R)-hernsubanine H

[0019] Compound 7: (-)(6S,7S,8R,10S,13R,14S)-hernsubanine I

[0020] Compound 8: (-)(6S,7S,8R,10S,13S,14S)-hernsubanine J

[0021] Compound 9: (+)(6S,7S,8R,10S,13R,14S,2'R)-hernsubanine K

[0022] Compound 10: (-)(6S,7S,8R,10S,13R,14S)-hernsubanine L

[0023] Compound 11: (-)(6S,7S,8R,10S,13S,14S)-hernsubanine M

[0024] Compound 12: (-)(7R,8R,10S,13S,14S,16R)-hernsubanine N.

[0025] Another object of the present invention is to provide a method for preparing the above-mentioned liensinine-type alkaloids and their dimer compounds 1-12, comprising the following steps:

[0026] (1) Extract the dried whole herb of Stephania hernandifolia with ethanol or methanol, and recover the extract to obtain a crude extract;

[0027] (2) Disperse the crude extract obtained in step (1) in water, adjust the pH value to 2, extract with petroleum ether or cyclohexane, adjust the pH value of the aqueous layer to 7, and extract with dichloromethane, chloroform or ethyl acetate to obtain total alkaloids;

[0028] (3) Separate the total alkaloids obtained in step (2) by silica gel column chromatography, and perform gradient elution with mixed solvent A, wherein the mixed solvent A is a mixed solvent of petroleum ether and ethyl acetate, a mixed solvent of petroleum ether and acetone, a mixed solvent of chloroform and acetone, a mixed solvent of dichloromethane and acetone, a mixed solvent of chloroform and methanol or a mixed solvent of dichloromethane and methanol;

[0029] (4) Separate the fractions obtained in step (3) by ODS column chromatography, and perform gradient elution with mixed solvent B as the mobile phase, wherein the mixed solvent B is a mixed solvent of methanol and water or a mixed solvent of acetonitrile and water;

[0030] (5) Further separate the fractions obtained in step (4) by HPLC, and perform gradient elution with mixed solvent C as the mobile phase to obtain compounds 1-12, wherein the mixed solvent C is a mixed solvent of methanol and water or a mixed solvent of acetonitrile and water.

[0031] In the above technical solution, in step (1), heating under reflux with ethanol extraction or heating under reflux with methanol extraction is carried out 2-5 times, and the above organic solvents are recovered under reduced pressure to obtain a crude extract, wherein the volume concentration of ethanol is 70%-95%, or the volume concentration of methanol is 60%-90%, and the mass-volume ratio of the dried whole herb of Stephania hernandifolia to ethanol or methanol is 1:8-1:20 g / mL.

[0032] Preferably, the volume concentration of the ethanol is 75% - 90%.

[0033] Preferably, the volume concentration of the methanol is 70% - 85%.

[0034] Preferably, the mass - volume ratio of the dried whole herb of Stephania hernandifolia to ethanol or methanol is 1:10 - 1:15 g / mL.

[0035] In the above - mentioned technical solution, in step (2), the crude extract obtained in step (1) is dispersed in water, the pH value is adjusted to 2 by adding HCl, extracted with petroleum ether or cyclohexane, the pH value of the aqueous layer is adjusted to 7 by adding sodium carbonate solution, and then extracted with dichloromethane, chloroform or ethyl acetate to obtain total alkaloids. Among them, each organic solvent is extracted 2 - 4 times, and the volume ratio of the aqueous phase to the organic phase is 1:1 - 1:5.

[0036] Preferably, the extraction times of each organic solvent are 3 times.

[0037] Preferably, the volume ratio of the aqueous phase to the organic phase is 1:1 - 1:3.

[0038] The present invention adopts the "acid - base extraction" method for the extraction and separation of compounds. Among them, the acid is hydrochloric acid (HCl), and the base is sodium carbonate.

[0039] Furthermore, the volume fraction of the HCl solution is 3% - 6%, and the volume fraction of the sodium carbonate solution is 2% - 10%.

[0040] Further preferably, the volume fraction of the HCl solution is 4% - 5%, and the volume fraction of the sodium carbonate solution is 5%.

[0041] In the above - mentioned technical solution, in step (3), the mixed solvent A is a mixed solvent composed of petroleum ether and ethyl acetate in a volume ratio of 15:1 - 0:1, or a mixed solvent composed of petroleum ether and acetone in a volume ratio of 15:1 - 0:1, or a mixed solvent composed of dichloromethane and acetone in a volume ratio of 100:1 - 5:1, or a mixed solvent composed of chloroform and acetone in a volume ratio of 100:1 - 5:1, or a mixed solvent composed of dichloromethane and methanol in a volume ratio of 100:0 - 10:1, or a mixed solvent composed of chloroform and methanol in a volume ratio of 100:0 - 10:1.

[0042] Furthermore, when the mixed solvent A is a mixed solvent of petroleum ether and ethyl acetate or a mixed solvent of petroleum ether and acetone, the volume ratio is preferably 10:1 - 0:1.

[0043] Further, when the mixed solvent A is a mixed solvent of dichloromethane and acetone or a mixed solvent of chloroform and acetone, the volume ratio is preferably 100:1 to 10:1.

[0044] Further, when the mixed solvent A is a mixed solvent of dichloromethane and methanol or a mixed solvent of chloroform and methanol, the volume ratio is preferably 100:1 to 15:1.

[0045] In the above technical solution, in the step (4), the mixed solvent B is a mixed solvent composed of methanol and water in a volume ratio of 2:8 to 9:1, or a mixed solvent composed of acetonitrile and water in a volume ratio of 2:8 to 9:1.

[0046] Further, when the mixed solvent B is a mixed solvent of methanol and water, the volume ratio is preferably 3:7 to 8:2.

[0047] Further, when the mixed solvent B is a mixed solvent of acetonitrile and water, the volume ratio is preferably 2:8 to 8:2.

[0048] In the above technical solution, in the step (5), the mixed solvent C is a mixed solvent composed of methanol and water in a volume ratio of 3:7 to 8:2, or a mixed solvent composed of acetonitrile and water in a volume ratio of 2:8 to 6:4.

[0049] Further, when the mixed solvent C is a mixed solvent of methanol and water, the volume ratio is preferably 3:7 to 8:2.

[0050] Further, when the mixed solvent C is a mixed solvent of acetonitrile and water, the volume ratio is preferably 2:8 to 6:4.

[0051] In the above technical solution, in the step (5), according to the separation situation, 1‰ to 2‰ of diethylamine can be added to the mixed solvent C to improve the separation degree.

[0052] Another object of the present invention is to provide a pharmaceutical composition, comprising the above-mentioned liensinine-type alkaloids and their dimer compounds and their pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier.

[0053] Another object of the present invention is to provide the use of the above-mentioned liensinine-type alkaloids and their dimer compounds and their pharmaceutically acceptable salts or the above-mentioned pharmaceutical composition in the preparation of drugs for preventing or treating neurodegenerative diseases.

[0054] The present invention established an LPS-induced BV-2 microglia over-activation model to evaluate the anti-neuroinflammatory effects of the prepared lotusan-type alkaloids and their dimer derivatives 1-12. The results showed that the new compounds 1-12 could inhibit the release of NO from LPS-induced over-activated BV-2 microglia, demonstrating significant anti-neuroinflammatory activity. Therefore, the newly prepared lotusan-type alkaloid compounds in the present invention can be applied in the development of drugs for treating neurodegenerative diseases.

[0055] Advantages of the present invention: The present invention for the first time provides 12 lotusan-type alkaloids and their dimer derivatives isolated and identified from the dried whole herb of Stephania hernandifolia (Willd.) Walp., as well as the preparation method, and systematically evaluated their activities in neuroprotection and analgesia, and clarified their applications in the development and treatment of drugs for neurodegenerative diseases. Detailed implementation modes

[0056] The following non-limiting examples can enable those of ordinary skill in the art to more comprehensively understand the present invention, but do not limit the present invention in any way.

[0057] In the following examples, the test methods are conventional methods unless otherwise specified; the reagents and materials can be obtained from commercial sources unless otherwise specified.

[0058] Example 1

[0059] (1) 5000 g of the dried whole herb of Stephania hernandifolia (Willd.) Walp. was refluxed and extracted 3 times with 80% ethanol (dosage: 50 L), and the extract was recovered under reduced pressure to obtain a crude extract;

[0060] (2) The 80% ethanol crude extract obtained in step (1) was dissolved in water, adjusted to pH 2 with 4.5% HCl, and extracted 3 times with petroleum ether (volume ratio of aqueous phase to petroleum ether 1:1); 5% sodium carbonate solution was added to the aqueous layer to adjust the pH to 7, and then extracted 3 times with dichloromethane (volume ratio of aqueous phase to dichloromethane 1:1) to obtain total alkaloids;

[0061] (3) The total alkaloids obtained in step (2) were separated by silica gel column chromatography, and eluted successively with dichloromethane and methanol mixed solvents at ratios of 100:1, 50:1, 30:1, 20:1, and 15:1;

[0062] (4) The fractions obtained with dichloromethane:methanol mixed solvents at ratios of 50:1 - 15:1 in step (3) were subjected to ODS chromatography and gradient eluted with methanol and water mixed solvents at ratios of 3:7, 4:6, 5:5, 6:4, and 8:2.

[0063] (5) The fractions obtained from the methanol:water mixed solvent of 4:6 to 8:2 in the above step (4) were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min and a mobile phase of methanol:water = 40:60, to obtain the lianhuashane-type alkaloid compound 5 (t R = 10 min) (yield 0.00049‰) and 10 (t R = 51 min) (yield 0.00006‰);

[0064] (6) The fractions obtained from the methanol:water mixed solvent of 4:6 to 8:2 in the above step (4) were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min and a mobile phase of methanol:water = 65:35, to obtain the lianhuashane-type alkaloid dimer compound 3 (t R = 15 min) (yield 0.00007‰), the lianhuashane-type alkaloid compound 6 (t R = 10 min) (yield 0.00039‰), compound 7 (t R = 26 min) (yield 0.00006‰) and compound 9 (t R = 29 min) (yield 0.00024‰);

[0065] (7) The fractions obtained from the methanol:water mixed solvent of 4:6 to 8:2 in the above step (4) were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min and a mobile phase of methanol:water = 70:30 (plus 1‰ diethylamine), to obtain the lianhuashane-type alkaloid compound 2 (t R = 33 min) (yield 0.0010‰), compound 8 (t R = 13 min) (yield 0.00035‰), compound 11 (t R = 16 min) (yield 0.00029‰) and compound 12 (t R = 24 min) (yield 0.00006‰);

[0066] (8) The fractions obtained from the methanol:water mixed solvent of 4:6 to 8:2 in the above step (4) were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min and a mobile phase of acetonitrile:water = 45:55 (plus 1‰ diethylamine), to obtain the lianhuashane-type alkaloid dimer compound 1 (t R = 18 min) (yield 0.00043‰) and compound 4 (t R = 21 min) (yield 0.00045‰).

[0067] The structures of compounds 1-12 were identified based on their physicochemical properties and spectral data.

[0068] The structure identification data of compound 1 are as follows:

[0069] White amorphous powder (methanol), positive reaction with modified bismuth potassium iodide, . HR-ESI-MS gave the quasi-molecular ion peak m / z 759.3058 [M+H] + (calcd. 759.3091 for C 41 H 47 N2O 12 ), indicating its molecular formula is C 41 H 46 N2O 12 , and the degree of unsaturation is 20. 1 1H NMR (600 MHz, CD3OD) gave the characteristic signals of an 8,10-oxy-bridged lianasane-type alkaloid nucleus at δ H 4.95 (1H, d, J = 6.3 Hz, H-10") and a characteristic signal of a 10-hydroxy lianasane-type alkaloid nucleus at δ H 4.26 (1H, dd, J = 11.0, 5.0 Hz, H-10). Four isolated aromatic hydrogen proton signals were given in the low field region at δ H 6.93 (1H, d, J = 0.6 Hz, H-1), 6.88 (1H, s, H-4), 6.71 (1H, s, H-1"), 6.69 (1H, s, H-4"); an olefinic hydrogen signal at δ H 5.76 (1H, dd, J = 6.8, 3.0 Hz, H-6); two sets of methylenedioxy hydrogen signals at δ H 5.91 (2H, s), 5.89 (1H, d, J = 1.0 Hz), 5.80 (1H, d, J = 1.0 Hz). Three sets of methoxy hydrogen signals were visible in the high field region at δ H 3.53 (3H, s, 8"-OCH H 3), 3.50 (3H, s, 7-OCH H 3), 3.35 (3H, s, 7"-OCH H 3); two sets of N-methyl hydrogen signals at δ H 2.58 (3H, s, 17"-NCH H 3), 2.20 (3H, s, 17-NCH H 3). 13 13C NMR (150 MHz, CD3OD) gave 41 carbon signals, including the characteristic carbon signals of the 8,10-oxy-bridged lianasane-type alkaloid oxygen bridge at δ C104.6 (C-8"), 78.3 (C-10"); two carbonyl carbon signals δ C 192.8 (C-8), 173.2 (C-2'); two sets of benzene ring carbon signals δ C 149.3 (C-3), 149.2 (C-3"), 148.5 (C-2), 146.2 (C-2"), 139.1 (C-12"), 134.9 (C-11"), 134.7 (C-12), 132.9 (C-11), 108.2 (C-1"), 107.3 (C-4), 107.0 (C-4"), 106.7 (C-1) and one set of double bond carbon signals δ C 152.0 (C-7), 115.4 (C-6); two methylenedioxy carbon signals δ C 102.3×2 (O C H2O), three methoxy carbon signals δ C 58.1 (7"-O C H3), 55.4 (7-O C H3), 51.9 (8"-O C H3); two N-methyl carbon signals δ C 39.3 (17"-N C H3), 33.0 (17-N C H3); Combining with the HSQC spectrum, there are also seven sets of methylene signals, and the data attribution is shown in Table 1 and Table 3. It is speculated that compound 1 is a dimer of liensinine-type alkaloids.

[0070] According to 1 H- 1 H COSY spectrum, the correlation between δ H 5.76 (H-6) and δ H 2.96 (H-5a), 2.90 (H-5b), the correlation between δ H 4.26 (H-10) and δ H 1.48 (H-9b), the correlation between δ H 2.85 (H-16) and δ H 2.07 (H-15a), 1.94 (H-1'), the existence of the fragments C-5-C-6, C-9-C-10, C-15-C-16-C-1' is determined; According to δ H 5.28 (H-6") and δ H 3.75 (H-7"), 2.30 (H-5"a), 2.28 (H-5"b), the correlation between δ H 4.95 (H-10") and δ H 2.71 (H-9"a), the correlation between δ H3.38 (H-16") is related to δ H 1.99, 1.91 (H-15"), which determines the existence of fragments C-5"-C-6"-C-7", C-9"-C-10", and C-15"-C-16". Combining with the HMBC spectrum δ H 1.94 (H-1') is remotely related to δ C 173.2 (C-2'), 61.9 (C-16), 46.6 (C-15), and 1 Based on the chemical shift of H-6" in the H NMR spectrum, it is speculated that two molecules of lianhuashinine-type alkaloids in the structure are connected by an acetyl bond at C-16 and C-6". According to δ H 3.50 (7-OC H 3) is remotely related to δ C 152.0 (C-7), δ H 3.53 (8"-OC H 3) is remotely related to δ C 104.7 (C-8"), δ H 3.35 (7"-OC H 3) is remotely related to δ C 82.9 (C-7"), which determines the substitution positions of the three methoxy groups. In addition, δ H 2.58 (17"-NC H 3) is remotely related to δ C 77.0 (C-14"), 54.9 (C-16"), δ H 2.20 (17-NC H 3) is remotely related to δ C 72.3 (C-14), 61.9 (C-16), which determines the substitution positions of the two nitrogen methyl groups. δ H 5.92, 5.89, 5.76 (OC H 2O×2) is remotely related to δ C 149.2 (C-3"), 146.2 (C-2"), δ C 148.8 (C-3), 147.8 (C-2), indicating that the methylenedioxy substitution is at C-2 / C-3 and C-2" / C-3" positions. In summary, the planar structure of compound 1 is determined.

[0071] The relative configuration of compound 1 is determined by 1 the coupling constants of key hydrogens in the H NMR spectrum and the NOESY spectrum. In the NOESY spectrum, according to δ H 4.26 (H-10) is related to δ H 2.77 (H-9a), δ H 2.85 (H-16) is related to δ H2.20(17-NC H 3), 2.09(H-15a) are correlated with δ H 2.09(H-15a) and δ H 1.48(H-9b) are correlated, suggesting that the spatial orientations of 10-OH, H-16 and the ethylamine bridge at the 13,14 positions are the same. The relative configuration of 1-i is 10R*, 13R*, 14S*, 16S*. According to J H-6",H-7" = 4.1 Hz, it is determined that H-6" and H-7" are cis. In the NOESY spectrum, δ H 4.95(H-10") is correlated with 3.53(8"-OC H 3), δ H 3.75(H-7") and δ H 2.58(17"-NC H 3), 2.26(H-5"b) are correlated, δ H 3.38(H-16a") and δ H 2.26(H-5"b) are correlated, suggesting that the spatial orientations of H-6", H-7" and the ethylamine bridge at the 13",14" positions are the same. The relative configuration of 1-ii is 6"S*, 7"S*, 8"R*, 10"S*, 13"S*, 14"S*. Further, the configurations of the two molecules of lianhuashanine-type alkaloid fragments in the structure are determined by base hydrolysis experiments. After base hydrolysis, two lianhuashanine-type alkaloid compounds 1-i and 1-ii are prepared. After identification, compound 1-i is a new compound; compound 1-ii is the same as compound N,O-dimethylstephine, but its absolute configuration has not been determined. The absolute configuration of 1-i is determined to be 10R, 13R, 14S, 16S by using the TDDFT method for ECD calculation, and the absolute configuration of compound 1-ii is 6S, 7S, 8R, 10S, 13S, 14S. Finally, the absolute configuration of compound 1 is determined to be 10R, 13R, 14S, 16S, 6"S, 7"S, 8"R, 10"S, 13"S, 14"S by comparing the measured and calculated ECD data.

[0072] In summary, compound 1 is identified as (-)(10R,13R,14S,16S,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine A, a new compound not reported in the literature after retrieval.

[0073] Table 1 1H NMR data of compound 1 and its base hydrolysis products 1-i and 1-ii

[0074]

[0075]

[0076] a : The test solvent was CD3OD, b : The test solvent was CDCl3.

[0077] The structural identification data of Compound 2 are as follows:

[0078] White amorphous powder (methanol), positive reaction to modified bismuth potassium iodide, . HR-ESI-MS gave the quasi-molecular ion peak m / z 759.3120 [M+H] + (calcd. 759.3091 for C 41 H 47 N2O 12 ), indicating its molecular formula is C 41 H 46 N2O 12 , and the degree of unsaturation is 20. 1 1H NMR (600 MHz, CD3OD) gave the characteristic signals of an 8,10-oxo-bridged nelumbine-type alkaloid nucleus at δ H 4.83 (1H, d, J = 6.2 Hz, H-10") and the characteristic signal of a 10-hydroxy nelumbine-type alkaloid nucleus at δ H 4.62 (1H, dd, J = 8.7, 4.8 Hz, H-10). Four isolated aromatic hydrogen proton signals were given in the low-field region at δ H 6.93 (1H, s, H-1), 6.89 (1H, s, H-4), 6.61 (1H, s, H-4"), 6.53 (1H, s, H-1"); an olefinic hydrogen signal at δ H 5.83 (1H, m, H-6); two sets of methylenedioxy hydrogen signals at δ H 5.93 (1H, J = 1.1 Hz), 5.92 (1H, J = 1.1 Hz), 5.76 (1H, s), 5.63 (1H, s). Three sets of methoxy hydrogen signals were visible in the high-field region at δ H 3.62 (3H, s, 7-OC H 3), 3.51 (3H, s, 8"-OC H 3), 3.33 (3H, s, 7"-OC H 3); two sets of N-methyl hydrogen signals at δ H 2.56 (3H, s, 17"-NC H 3), 2.45 (3H, s, 17-NC H 3). 13 13C NMR (150 MHz, CD3OD) gave 41 carbon signals, including the characteristic carbon signal of the 8,10-oxo bridge of nelumbine-type alkaloid at δ C104.5 (C-8"), 78.2 (C-10"); two carbonyl carbon signals δ C 195.8 (C-8), 173.4 (C-2'); two sets of benzene ring carbon signals δ C 149.1 (C-3"), 149.0 (C-3), 147.6 (C-2), 146.1 (C-2"), 138.9 (C-12"), 137.5 (C-12), 134.7 (C-11"), 133.0 (C-11), 108.1 (C-4"), 107.4 (C-4), 107.3 (C-1), 106.9 (C-1") and a set of double bond carbon signals δ C 152.1 (C-7), 113.9 (C-6); two methylenedioxy carbon signals δ C 102.3, 102.2 (O C H2O), three methoxy carbon signals δ C 58.0 (7"-O C H3), 55.5 (7-O C H3), 51.9 (8"-O C H3); two nitrogen-methyl carbon signals δ C 39.3 (17"-N C H3), 32.2 (17-N C H3); Combining with the HSQC spectrum, there are also seven sets of methylene signals. The data attribution is shown in Table 2 and Table 3. The NMR data of compound 2 is similar to that of 1, and the main differences are in the H-9 / 10 and C-13 / 14 positions.

[0079] According to the δ in the HMBC spectrum H 1.98 (H-1'a), 1.54 (H-1'b) and δ C 173.4 (C-2'), 58.3 (C-16), 44.7 (C-15) long-range correlations, combined with 1 the chemical shift value of H-6" in 1H NMR, it is speculated that the two molecules of lianhuashane-type alkaloids in the structure are connected by an acetyl bond at the C-16 and C-6" positions. According to δ H 3.63 (7-OC H 3) and δ C 152.1 (C-7), δ H 3.51 (8"-OC H 3) and δ C 104.5 (C-8"), δ H 3.33 (7"-OC H 3) and δ C 82.8 (C-7") long-range correlations, the substitution positions of the three methoxy groups are determined. In addition, δH 2.56(17"-NC H 3) is related to δ C 76.9(C-14"), 54.9(C-16"), δ H 2.45(17-NC H 3) is related to δ C It is remotely related to 70.2(C-14), 58.3(C-16), and the substitution positions of two N-methyl groups are determined. δ H 5.93, 5.92, 5.76, 5.63(OC H 2O×2) is related to δ C It is remotely related to 149.1(C-3"), 149.0(C-3), 147.6(C-2), 146.1(C-2"), suggesting that the substitution of methylenedioxy is at the C-2, C-3 and C-2", C-3" positions. In summary, it is determined that Compound 2 has the same planar structure as Compound 1.

[0080] Similarly, according to the NOESY spectrum, δ H 2.45(17-NC H 3) is related to δ H 4.62(H-10), 2.25(H-9a) are related, δ H 2.66(H-16) is related to δ H 2.17(H-15a) is related, δ H 1.89(H-15b) is related to δ H 1.51(H-1') is related, suggesting that the spatial orientations of H-10, H-16 and the ethylamine bridge at the 13, 14 positions are the same; according to J H-6",H-7" = 4.1Hz, it is determined that H-6" and H-7" are cis. Combining with δ in the NOESY spectrum H 4.83(H-10") is related to δ H 3.51(8"-OCH3) is related, δ H 3.71(H-7") is related to δ H 3.36(H-16a"), 2.56(17"-NC H 3), 2.23(H-5"b) are related, suggesting that the spatial orientations of H-6", H-7" and the ethylamine bridge at the 13", 14" positions are the same. The relative configuration of 2 is determined to be 10S*, 13R*, 14S*, 16S*, 6"S*, 7"S*, 8"R*, 10"S*, 13"S*, 14"S*. Further, the absolute configuration of Compound 2 is determined to be 10S, 13R, 14S, 16S, 6"S, 7"S, 8"R, 10"S, 13"S, 14"S by comparing the experimental and calculated ECD data.

[0081] In summary, compound 2 was identified as (-)(10S,13R,14S,16S,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine B, which is a new compound not reported in the literature after retrieval.

[0082] The structural identification data of compound 3 are as follows:

[0083] White amorphous powder (methanol), positive reaction to modified bismuth potassium iodide . HR-ESI-MS gave the quasi-molecular ion peak m / z 773.2894 [M+H] + (calcd. 773.2916 for C 41 H 45 N2O 13 ), suggesting its molecular formula is C 41 H 44 N2O 13 , and the degree of unsaturation is 21. 1 The 1H NMR (600 MHz, CD3OD) spectrum gave the characteristic signals of an 8,10-oxy-bridged nelumbine-type alkaloid nucleus at δ H 4.88 (1H, d, J = 6.2 Hz, H-10") and the characteristic signals of a 10-hydroxy nelumbine-type alkaloid nucleus at δ H 4.62 (1H, dd, J = 8.8, 4.8 Hz, H-10). Four isolated aromatic hydrogen proton signals were given in the low field region at δ H 6.94 (1H, s, H-1), 6.89 (1H, s, H-4), 6.72 (1H, s, H-4"), 6.56 (1H, s, H-1"); an olefinic hydrogen signal at δ H 5.84 (1H, dd, J = 5.9, 3.9 Hz, H-6); two sets of methylenedioxy hydrogen signals at δ H 5.93 (1H, J = 1.1 Hz), 5.93 (1H, J = 1.1 Hz), 5.80 (1H, d, J = 1.1 Hz), 5.67 (1H, d, J = 1.1 Hz). Three sets of methoxy hydrogen signals were visible in the high field region at δ H 3.63 (3H, s, 7-OC H 3), 3.59 (3H, s, 8"-OC H 3), 3.34 (3H, s, 7"-OC H 3); two sets of nitrogen-methyl hydrogen signals at δ H 3.02 (3H, s, 17"-NC H 3), 2.45 (3H, s, 17-NC H 3). 1313C NMR (150 MHz, CD3OD) gave 41 carbon signals, including the characteristic carbon signals of the oxygen bridge of 8,10-epoxylotusine-type alkaloids, δ C 102.7 (C-8"), 77.2 (C-10"); three carbonyl carbon signals, δ C 195.7 (C-8), 176.1 (C-16"), 173.1 (C-2'); two sets of benzene ring carbon signals, δ C 149.5 (C-3"), 149.0 (C-3), 147.6 (C-2), 146.8 (C-2"), 137.5 (C-12), 135.5 (C-12"), 135.1 (C-11"), 133.1 (C-11), 108.2 (C-4"), 107.4 (C-4), 107.3 (C-1), 107.1 (C-1") and a set of double bond carbon signals, δ C 152.1 (C-7), 113.9 (C-6); two methylenedioxy carbon signals, δ C 102.5, 102.3 (O C H2O), three methoxy carbon signals, δ C 58.1 (7"-O C H3), 55.6 (7-O C H3), 52.3 (8"-O C H3); two N-methyl carbon signals, δ C 32.1 (17-N C H3), 28.7 (17"-N C H3); Combining with the HSQC spectrum, there are also six sets of methylene signals, and the data attribution is shown in Table 2 and Table 3. Comparing with the NMR data of compound 2, the main difference is that the 16th position is oxidized to a carbonyl group.

[0084] According to the HMBC spectrum, the correlations between δ H 3.02 (17"-NC H 3) and δ C 176.1 (C-16"), 75.4 (C-14"), and the correlations between δ H 2.45 (17-NC H 3) and δ C 70.2 (C-14), 58.3 (C-16) determined the substitution positions of the two N-methyl groups and the carbonyl group. Similar to the above compound, the planar structure of compound 3 was determined based on the key HMBC correlations. Through 1The coupling constants of the key hydrogens in the 1H NMR spectrum and the NOESY spectrum determined the relative configuration of Compound 3 as 10S*,13R*,14S*,16S*,6"S*,7"S*,8"R*,10"S*,13"R*,14"S*. Further comparison of the experimental and calculated ECD data determined the absolute configuration of Compound 3 as 10S,13R,14S,16S,6"S,7"S,8"R,10"S,13"R,14"S.

[0085] In summary, Compound 3 was identified as (-)(10S,13R,14S,16S,6"S,7"S,8"R,10"S,13"R,14"S)-bishernsubanine C, which was a new compound not reported in the literature after retrieval.

[0086] The structure identification data of Compound 4 are as follows:

[0087] White needle crystals (methanol), positive reaction to modified bismuth potassium iodide . The HR-ESI-MS spectrum gave the quasi-molecular ion peak m / z 759.3082[M+H] + (calcd. 759.3091 for C 41 H 47 N2O 12 ), suggesting its molecular formula was C 41 H 46 N2O 12 , and the degree of unsaturation was 20. 1 1H NMR (600 MHz, CD3OD) gave the characteristic signals of an 8,10-oxo-bridged nelumbianine-type alkaloid nucleus at δ H 4.77 (1H, d, J = 6.2 Hz, H-10") and the characteristic signals of a 10-hydroxy-nelumbianine-type alkaloid nucleus at δ H 4.56 (1H, dd, J = 11.0, 5.6 Hz, H-10). Four isolated aromatic hydrogen proton signals were given in the low field region at δ H 6.99 (1H, s, H-1), 6.71 (1H, s, H-4), 6.61 (1H, s, H-4"), 6.59 (1H, s, H-1"); an olefinic hydrogen signal at δ H 5.92 (1H, m, H-6); two sets of methylenedioxy hydrogen signals at δ H 5.93 (1H, J = 1.2 Hz), 5.92 (1H, J = 1.2 Hz), 5.82 (1H, J = 0.9 Hz), 5.61 (1H, J = 0.9 Hz). Three sets of methoxy hydrogen signals were visible in the high field region at δ H 3.67 (3H, s, 7-OC H3), 3.48 (3H, s, 8"-OC H 3), 3.35 (3H, s, 7"-OC H 3); Two sets of nitrogen-methyl hydrogen signals δ H 2.56 (3H, s, 17"-NC H 3), 2.47 (3H, s, 17-NC H 3). 13 13C NMR (150 MHz, CD3OD) gave 41 carbon signals, including the characteristic carbon signals of the 8,10-oxo bridge in lotusan-type alkaloids with an oxo bridge δ C 104.4 (C-8"), 78.1 (C-10"); Two carbonyl carbon signals δ C 198.9 (C-8), 173.4 (C-2'); Two sets of benzene ring carbon signals δ C 149.0 (C-3"), 148.4 (C-3), 147.4 (C-2), 146.0 (C-2"), 139.2 (C-12), 138.8 (C-12"), 134.6 (C-11"), 132.7 (C-11), 108.1 (C-4"), 107.4 (C-4), 107.3 (C-1"), 106.3 (C-1) and a set of double bond carbon signals δ C 152.3 (C-7), 115.3 (C-6); Two methylenedioxy carbon signals δ C 102.2, 102.1 (O C H2O), Three methoxy carbon signals δ C 57.9 (7"-O C H3), 55.6 (7-O C H3), 51.8 (8"-O C H3); Two nitrogen-methyl carbon signals δ C 39.2 (17"-N C H3), 34.3 (17-N C H3); Combining with the HSQC spectrum, it can be seen that there are also seven sets of methylene signals, and the data attribution is shown in Table 2 and Table 3. The NMR data of compound 4 is similar to that of compound 2, and the main difference is in the H-15 and H-16 positions. It is speculated that the configuration at the 16th position is different.

[0088] Similarly, the planar structure of compound 4 was determined based on the key correlations in the HMBC spectrum. Through 1The coupling constants of the key hydrogens in the \(^1H\) NMR spectrum and the NOESY spectrum determined the relative configuration of compound 4 as 10S*,13R*,14S*,16R*,6"S*,7"S*,8"R*,10"S*,13"S*,14"S*. Further comparison of experimental and calculated ECD data determined the absolute configuration of compound 4 as 10S,13R,14S,16R,6"S,7"S,8"R,10"S,13"S,14"S.

[0089] In summary, compound 4 was identified as (-)(10S,13R,14S,16R,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine D, a new compound not reported in the literature after retrieval.

[0090] Table 2 \(^1H\) NMR data of compounds 2 - 4

[0091]

[0092] a : The test solvent was CD3OD, b : The test solvent was CDCl3.

[0093] Table 3 \(^{13}\)C NMR data of compounds 1 - 4

[0094]

[0095]

[0096] a : The test solvent was CD3OD, b : The test solvent was CDCl3.

[0097] The structural identification data of compound 5 are as follows:

[0098] Pale yellow amorphous powder (methanol), positive reaction to modified bismuth potassium iodide, [α] 2 D 0 -186.6 (c 0.50, CH3OH). HR-ESI-MS gave the quasi-molecular ion peak m / z 358.1291 [M + H] + (calcd. 358.1285 for C 19 H 20 NO6), suggesting its molecular formula is C 19 H 19 NO6, and the degree of unsaturation is 11. 1 The \(^1H\) NMR (600 MHz, CD3OD) gave the characteristic signals of the lotusane-type alkaloid nucleus δ H4.58 (1H, dd, J = 10.8, 5.3 Hz, H-10). Two isolated aromatic hydrogen proton signals are given in the low field region at δ H 7.03 (1H, s, H-1), 6.90 (1H, s, H-4); one vinylic hydrogen signal at δ H 6.08 (1H, dd, J = 6.8, 2.9 Hz, H-6); a set of methylenedioxy hydrogen signals at δ H 5.97 (1H, d, J = 1.0 Hz), 5.95 (1H, d, J = 1.0 Hz). A set of methoxy hydrogen signals can be seen in the high field region at δ H 3.68 (3H, s, 7-OC H 3); a set of N-methyl hydrogen signals at δ H 3.05 (3H, s, 17-NC H 3). The chemical shift of this N-methyl indicates that the C-16 position of the compound is oxidized to a carbonyl group. 13 13C NMR (150 MHz, CD3OD) gives 19 carbon signals, including two carbonyl carbon signals at δ C 192.9 (C-8), 176.0 (C-16); a set of benzene ring carbon signals at δ C 149.3 (C-3), 148.5 (C-2), 134.7 (C-12), 132.9 (C-11), 107.3 (C-4), 106.7 (C-1) and a set of double bond carbon signals at δ C 152.7 (C-7), 116.4 (C-6); one methylenedioxy carbon signal at δ C 102.6 (O C H2O), one methoxy carbon signal at δ C 55.9 (7-O C H3); one N-methyl carbon signal at δ C 28.4 (17-N C H3); The NMR data were assigned by combining with the HSQC spectrum, as shown in Table 4 and Table 6.

[0099] According to the HMBC spectrum, the long-range correlation between δ H 3.68 (7-OC H 3) and δ C 152.7 (C-7) determined the substitution position of the methoxy group. According to the long-range correlations between δ H 3.05 (17-NC H 3) and δ C 176.0 (C-16), 71.7 (C-14), and the long-range correlations between δ H 2.97 (H-15a), 2.57 (H-15b) and δ C176.0 (C-16) long-range correlation determined the substitution positions of the nitrogen-methyl and carbonyl groups. δ H 5.97, 5.95 (-OC H 2O-) showed long-range correlation with δ C 149.3 (C-3), 148.5 (C-2), suggesting the substitution of methylenedioxy group at C-2 and C-3 positions. In summary, the planar structure of compound 5 was determined. According to the NOESY spectrum, δ H 4.58 (H-10) showed long-range correlation with δ H 3.05 (17-NC H 3), 2.57 (H-15b), suggesting that the spatial orientations of H-10 and the ethylamine bridge at positions 13 and 14 are the same. The absolute configuration of compound 5 was determined to be 10S, 13R, 14S by X-ray single crystal diffraction method.

[0100] In summary, compound 5 was identified as (-)(10S, 13R, 14S)-hernsubanine G, which is a new compound not reported in the literature after retrieval.

[0101] The structure identification data of compound 6 are as follows:

[0102] White amorphous powder (methanol), positive reaction to modified bismuth potassium iodide test, . HR-ESI-MS gave the quasi-molecular ion peak m / z 474.2105 [M + H] + (calcd. 474.2123 for C 25 H 32 NO8), suggesting the molecular formula C 25 H 31 NO8, and the degree of unsaturation is 11. 1 1H NMR (600 MHz, CD3OD) gave the characteristic signals of the 8,10-oxy-bridged lianhuashanine alkaloid nucleus δ H 5.03 (1H, d, J = 6.1 Hz, H-10). Two isolated aromatic hydrogen proton signals were given in the low field region δ H 6.80 (1H, s, H-4), 6.73 (1H, s, H-1); a set of methylenedioxy hydrogen signals δ H 5.95 (1H, d, J = 1.1 Hz, H-18a), 5.92 (1H, d, J = 1.1 Hz, H-18b); a hydrogen signal on the carbon substituted by an acyloxy group δ H 5.32 (1H, m, H-6). Two sets of methoxy hydrogen signals were visible in the high field region δ H 3.59 (3H, s, 8-OC H 3), 3.38 (3H, s, 7-OC H3); A set of nitrogen-methyl hydrogen signals δ H 3.05(3H, s, 17-NC H 3), and the chemical shift of this nitrogen-methyl indicates that the C-16 position of the compound is oxidized to a carbonyl group; a hydrogen signal on an oxygen-substituted carbon δ H 3.64(1H, d, J = 4.1Hz, H-7); Two sets of methyl hydrogen signals δ H 0.80(3H, d, J = 7.4Hz, C H 3-4'), 0.78(3H, t, J = 7.0Hz, C H 3-5'). 13 C NMR(150MHz, CD3OD) gives 25 carbon signals, including the characteristic carbon signals of the 8,10-oxo bridge in lotusan-type alkaloids with an oxo bridge δ C 102.7(C-8), 77.2(C-10); Two carbonyl carbon signals δ C 178.0(C-1'), 176.1(C-16); A set of benzene ring carbon signals δ C 149.6(C-3), 147.0(C-2), 135.6(C-11), 135.3(C-12), 108.5(C-4), 107.0(C-1); Two methoxy carbon signals δ C 58.0(7-O C H3), 52.2(8-O C H3); A nitrogen-methyl carbon signal δ C 28.7(17-N C H3). The NMR data were assigned by combining with the HSQC spectrum, as shown in Table 4 and Table 6.

[0103] According to the HMBC spectrum, δ H 5.32(H-6) and δ C 178.0(C-1') long-range correlation, δ H 0.78(H-5') and 178.0(C-1') long-range correlation indicate the presence of a 2-methylbutyrate group in the structure and substitution at the C-6 position. At the same time, δ H 3.59(8-OC H 3), 3.38(7-OC H 3) are respectively correlated with δ C 102.7(C-8), 81.8(C-7) long-range correlation to determine the substitution positions of the methoxy groups. In addition, δ H 3.05(17-NC H 3) is correlated with δ C 176.1(C-16), 75.4(C-14) long-range correlation, δ H2.65 (H-15a), 2.56 (H-15b) are remotely correlated with δ C 176.1 (C-16), determining the substitution positions of the nitrogen-methyl and carbonyl groups. δ H 5.95, 5.92 (-OC H2 O-) is remotely correlated with δ C 149.6 (C-3), 147.0 (C-2), suggesting the presence of a methylenedioxy group at C-2 and C-3 positions. In summary, the planar structure of compound 6 was determined. Through 1 the coupling constants of the key hydrogens in the 1H NMR spectrum and the NOESY spectrum, the relative configuration of compound 6 was determined. Based on J H-6,H-7 = 4.1 Hz, H-6 and H-7 were determined to be cis. In the NOESY spectrum, δ H 3.64 (H-7) is correlated with 2.10 (H-5b), 3.05 (17-NC H 3), δ H 2.65 (H-5a) is correlated with δ H 6.73 (H-1), 2.65 (H-15a), δ H 2.56 (H-15b) is correlated with δ H 1.80 (H-9a), suggesting that the spatial orientations of H-6, H-7 and the ethylamine bridge at positions 13 and 14 are the same, and the relative configuration of compound 6 was determined to be 6S*, 7S*, 8R*, 10S*, 13R*, 14S*. Further, the side chain was determined to be (2'R)-methylbutyric acid compound through base hydrolysis experiment. Further, the absolute configuration of compound 6 was determined to be 6S, 7S, 8R, 10S, 13R, 14S, 2'R by comparing the experimental and calculated ECD data.

[0104] In summary, compound 6 was identified as (+)(6S,7S,8R,10S,13R,14S,2'R)-hernsubanine H, a new compound not reported in the literature.

[0105] The structure identification data of compound 7 are as follows:

[0106] White amorphous powder (methanol), positive reaction to modified bismuth potassium iodide test, HR-ESI-MS gave a quasi-molecular ion peak at m / z 554.1996 [M + H] + (calcd. 554.2024 for C 29 H 32 NO 10 ), suggesting a molecular formula of C 29 H 31 NO 10 with an unsaturation degree of 14.1 1H NMR (600 MHz, CD3OD) gave the characteristic signals of the 8,10-oxo-lotusane alkaloid nucleus at δ H 5.03 (1H, d-like, J = 6.1 Hz, H-10). Two isolated aromatic hydrogen proton signals appeared in the low field region at δ H 6.65 (1H, s, H-1), 6.62 (1H, s, H-4); a set of ABX coupled system hydrogen signals at δ H 7.29 (1H, d, J = 2.0 Hz, H-2'), 7.10 (1H, dd, J = 8.2, 2.0 Hz, H-6′), 6.86 (1H, d, J = 8.2 Hz, H-5'); a set of methylenedioxy hydrogen signals at δ H 5.77 (1H, d, J = 1.4 Hz, H-18a), 5.28 (1H, d, J = 1.4 Hz, H-18b). Four methoxy hydrogen signals were visible in the high field region at δ H 3.91 (3H, s, 4'-OCH H 3), 3.89 (3H, s, 3'-OCH H 3), 3.64 (3H, s, 8-OCH H 3), 3.48 (3H, s, 7-OCH H 3); a nitrogen-methyl hydrogen signal at δ H 3.08 (3H, s, 17-NCH H 3), and the chemical shift of this nitrogen-methyl group indicated that the C-16 position of the compound was oxidized to a carbonyl group; a carbon on the acyloxy-substituted carbon 13 13C-NMR (150 MHz, CD3OD) gave 29 carbon signals, including the characteristic carbon signals of the 8,10-oxo-lotusane alkaloid oxo bridge at δ C 102.9 (C-8), 77.5 (C-10); two carbonyl carbon signals at δ C 175.1 (C-16), 167.6 (C-7'); 12 aromatic carbon signals at δ C 154.7 (C-4'), 149.5 (C-3'), 149.4 (C-3), 146.7 (C-2), 134.8 (C-12), 134.6 (C-11), 125.5 (C-6'), 123.2 (C-1'), 113.9 (C-2'), 111.2 (C-5'), 108.1 (C-4), 107.6 (C-1); a methylenedioxy carbon signal at δ C 102.2 (C-18); four methoxy carbon signals at δ C 58.3 (7-OCH C 3), 56.6 (3′-OCH CH3), 56.5 (4'-O C H3), 52.3 (8-O C H3); a signal of a nitrogen-methyl carbon, δ C 28.6 (17-N C H3); The NMR data were assigned by combining with the HSQC spectrum, as shown in Table 4 and Table 6.

[0107] In the HMBC spectrum, δ H 7.29 (H-2′), 7.10 (H-6′) showed long-range correlations with 167.6 (C-7′), δ H 5.43 (H-6) and δ C 167.6 (C-7') showed long-range correlations, indicating the presence of 3,4-dimethoxycinnamoyl group substituted at C-6. In addition, δ H 3.91 (4′-OC H 3), 3.89 (3′-OC H 3), 3.64 (8-OC H 3), 3.48 (7-OC H 3) showed long-range correlations with δ C 154.7 (C-4'), 149.5 (C-3'), 102.9 (C-8), 83.1 (C-7) respectively to determine the substitution positions of the methoxy groups. δ H 3.08 (17-NC H 3) showed long-range correlations with δ C 175.1 (C-16), 75.5 (C-14); δ H 2.61 (H-15a), 2.52 (H-15b) showed long-range correlations with δ C 175.1 (C-16) to determine the substitution positions of the nitrogen-methyl and carbonyl groups. δ H 5.77 (H-18a), 5.28 (H-18b) showed long-range correlations with δ C 149.4 (C-3), 146.7 (C-2), indicating the substitution of methylenedioxy group at C-2 and C-3. The relative configuration of compound 7 was determined by 1 the coupling constants of key hydrogens in the H-NMR spectrum and the NOESY spectrum. J H-6,H-7 = 4.1 Hz, determining that H-6 and H-7 are cis. In the NOESY spectrum, δ H 3.79 (H-7) was correlated with δ H 2.13 (H-5b), 3.08 (17-NC H 3); δ H 2.68 (H-5a) was correlated with δ H 6.65 (H-1), 2.61 (H-15a); δH 2.52(H - 15b) is related to δ H 1.84(H - 9a), suggesting that the hydrogen atoms at positions 6 and 7 and the ethylamine bridge at positions 13 and 14 have the same spatial orientation, indicating that the relative configuration of compound 7 is 6S*, 7S*, 8R*, 10R*, 13R*, 14S*. Further, the absolute configuration of compound 7 was determined to be 6S, 7S, 8R, 10S, 13R, 14S by comparing experimental and calculated ECD data.

[0108] In summary, compound 7 was identified as (-)(6S,7S,8R,10S,13R,14S)-hernsubanine I, a new compound not reported in the literature after retrieval.

[0109] The structure identification data of compound 8 are as follows:

[0110] Pale yellow amorphous powder (methanol), positive reaction to modified bismuth potassium iodide reaction, . HR - ESI - MS gave the quasi - molecular ion peak m / z 552.2216 [M + H] + (calcd. 552.2228 for C 30 H 34 NO9), suggesting the molecular formula C 30 H 33 NO9, with an unsaturation degree of 15. 1 1H NMR (600 MHz, CDCl3) gave the characteristic signals of the 8,10 - oxy - bridge lotusanine - type alkaloid nucleus δ H 4.90 (1H, d, J = 6.1 Hz, H - 10). A set of trans - double - bond hydrogen signals were given in the low - field region δ H 7.16 (1H, d, J = 16.0 Hz, H - 7'), 5.55 (1H, d, J = 16.0 Hz, H - 8'); A set of ABX - coupled system hydrogen signals δ H 6.98 (1H, d, J = 2.0 Hz, H - 2'), 6.93 (1H, dd, J = 8.3, 2.0 Hz, H - 6'), 6.82 (1H, d, J = 8.3 Hz, H - 5'); A set of methylenedioxy hydrogen signals δ H 5.64 (1H, d, J = 1.5 Hz), 5.00 (1H, d, J = 1.5 Hz); Two isolated aromatic hydrogen proton signals δ H 6.65 (1H, s, H - 1), 6.47 (1H, s, H - 4); A hydrogen signal on the carbon substituted by an acyloxy group δ H 5.33 (1H, m, H - 6). Three sets of methoxy hydrogen signals were visible in the high - field region δ H 3.92 (3H, s, 4'-OCH 3), 3.55 (3H, s, 8 - OC H 3), 3.39 (3H, s, 7 - OC H 3); A set of signals for the hydrogen of the nitrogen - methyl group, δ H 2.57 (3H, s, 17 - NC H 3); A signal for the hydrogen of the methine - oxygen group, δ H 3.74 (1H, d, J = 4.3 Hz, H - 7). 13 C NMR (150 MHz, CDCl3) gives 30 carbon signals, including the characteristic carbon signals of the oxygen - bridge in 8,10 - epoxy - lianasane - type alkaloids, δ C 103.4 (C - 8), 77.0 (C - 10); A signal for the carbonyl carbon, δ C 167.3 (C - 9'); Two sets of signals for the benzene - ring carbons, δ C 148.3 (C - 4'), 147.8 (C - 3), 145.8 (C - 3'), 144.9 (C - 2), 137.5 (C - 12), 133.4 (C - 11), 128.4 (C - 1'), 121.3 (C - 6'), 113.5 (C - 2'), 110.6 (C - 5'), 106.9 (C - 4), 106.1 (C - 1) and a set of signals for the double - bond carbons 143.3 (C - 7'), 116.6 (C - 8'); A signal for the methylenedioxy carbon, δ C 101.0 (O C H2O); Three signals for the methoxy carbons, δ C 57.4 (7 - O C H3), 56.1 (4' - O C H3), 51.8 (8 - O C H3); A signal for the nitrogen - methyl carbon, δ C 38.8 (17 - N C H3); Combining with the HSQC spectrum, there are also four sets of methylene signals. The data attribution is shown in Table 4 and Table 6.

[0111] According to the HMBC spectrum, δ H 6.93 (H - 6') has a long - range correlation with δ C 148.3 (C - 4'), 113.5 (C - 2'), δ H 7.16 (H - 7') has a long - range correlation with δ C 167.3 (C - 9'), 121.3 (C - 6'), 113.5 (C - 2'), δ H 5.33 (H - 6) has a correlation with δ C 167.3 (C - 9'), combined with 1The chemical shift of H-6 in ¹H NMR suggests the presence of 3,4-dimethoxycinnamoyl group substituted at C-6. According to δ H 6.65 (H-1) shows long-range correlations with δ C 145.8 (C-3), 137.5 (C-12), 77.0 (C-10), and δ H 6.47 (H-4) shows long-range correlations with δ C 144.9 (C-2), 133.4 (C-11), 49.8 (C-13). δ H 5.64, 5.00 (-OCH H2 O-) shows long-range correlations with δ C 147.8 (C-3), 144.9 (C-2), suggesting the substitution of methylenedioxy group at C-2 and C-3. In the HMBC spectrum, δ H 3.92 (4'-OCH H 3), 3.55 (8-OCH H 3), 3.39 (7-OCH H 3) show long-range correlations with δ C 148.3 (C-4'), 103.4 (C-8), 81.6 (C-7) respectively, determining the substitution positions of the three methoxy groups. δ H 2.57 (17-NCH H 3) shows long-range correlations with δ C 75.7 (C-14), 54.1 (C-16), determining the substitution position of the N-methyl group. In summary, the planar structure of compound 8 is determined. The relative configuration of compound 8 is determined by the coupling constants of the key hydrogens in the 1 ¹H NMR spectrum and the NOESY spectrum. According to J H-6,H-7 = 4.3 Hz, it is determined that H-6 and H-7 are cis. In the NOESY spectrum, δ H 3.74 (H-7) is correlated with δ H 2.31 (H-5b), 2.57 (17-NCH H 3), δ H 2.31 (H-5b) is correlated with δ H 1.87 (H-15a), δ H 1.87 (H-15b) is correlated with δ HRelated to 1.54(H-9b), indicating that the spatial orientations of the ethylamine bridges at H-6, H-7 and positions 13 and 14 are the same, and the relative configuration of compound 8 is determined to be 6S*,7S*,8R*,10S*,13S*,14S*. The side chain was determined to be the (2'R)-methylbutyric acid compound through base hydrolysis experiments. Further comparison of experimental and calculated ECD data determined the absolute configuration of compound 8 to be 6S,7S,8R,10S,13S,14S.

[0112] In summary, compound 8 was identified as (-)(6S,7S,8R,10S,13S,14S)-hernsubanine J, a new compound not reported in the literature after retrieval.

[0113] Table 4 1H NMR data of compounds 5 - 8

[0114]

[0115]

[0116] a : The test solvent was CD3OD, b : The test solvent was CDCl3.

[0117] The structure identification data of compound 9 are as follows:

[0118] Pale yellow amorphous powder (methanol), positive reaction with modified bismuth potassium iodide, [α]93.2 (c 0.50, CH3OH). HR-ESI-MS gave the quasi-molecular ion peak m / z 490.2439 [M+H] + (calcd. 490.2441 for C 26 H 36 NO8), indicating the molecular formula C 26 H 35 NO8, with an unsaturation degree of 10. 1 1H NMR (600 MHz, CD3OD) gave the characteristic signals of the 8,10-oxy-bridge lotanocarpine alkaloid nucleus at δ H 4.99 (1H, d, J = 6.2 Hz, H-10). A set of hydrogen signals of the 1,2,3,4-tetrasubstituted benzene ring appeared in the low field region at δ H 6.91 (1H, d, J = 8.2 Hz, H-1), 6.87 (1H, d, J = 8.2 Hz, H-2); the hydrogen signal on the carbon substituted by an acyloxy group at δ H 5.32 (1H, m, H-6). Four sets of methoxy hydrogen signals were visible in the high field region at δ H 3.88 (3H, s, 4-OC H 3), 3.86 (3H, s, 3-OCH 3), 3.59 (3H, s, 8 - OC H 3), 3.38 (3H, s, 7 - OC H 3); A set of nitrogen - methyl hydrogen signals δ H 3.06 (3H, s, 17 - NC H 3), and the chemical shift of this nitrogen - methyl indicates that the C - 16 position of the compound is oxidized to a carbonyl group; A methine hydrogen signal δ H 3.63 (1H, d, J = 4.2 Hz, H - 7); Two sets of methyl - carbon signals δ H 0.76 (3H, t, J = 7.5 Hz, C H 3 - 4'), 0.71 (3H, d, J = 6.9 Hz, C H 3 - 5'). 13 C NMR (150 MHz, CD3OD) gives 26 carbon signals, including the characteristic carbon signals of the 8,10 - oxygen - bridge in the nelumbine - type alkaloid oxygen - bridge δ C 102.6 (C - 8), 77.4 (C - 10); Two carbonyl - carbon signals δ C 178.1 (C - 1'), 176.5 (C - 16); A set of benzene - ring carbon signals δ C 154.8 (C - 3), 149.1 (C - 4), 135.2 (C - 11), 133.5 (C - 12), 121.4 (C - 1), 111.4 (C - 2); Four methoxy - carbon signals δ C 60.9 (4 - O C H3), 57.9 (3 - O C H3), 56.1 (7 - O C H3), 52.2 (8 - O C H3); A nitrogen - methyl carbon signal δ C 28.7 (17 - N C H3). Combining with the HSQC spectrum, there are also four sets of methylene signals. The data attribution is shown in Table 5 and Table 6.

[0119] In the HMBC spectrum, δ H 5.32 (H - 6) has a long - range correlation with 178.1 (C - 1'), δ H 1.58 (H - 2'), 1.29 (H - 3'a), 1.15 (H - 3'b), 0.71 (H - 5') have long - range correlations with 178.1 (C - 1'), indicating the presence of a 2 - methylbutyrate group in the structure and it is substituted at the C - 6 position. δ H 3.88 (4 - OC H 3), 3.86 (3 - OC H 3), 3.59 (8 - OC H3), 3.38 (7-OC H 3) were respectively remotely correlated with δ C 149.1 (C-4), 154.8 (C-3), 102.6 (C-8), 83.1 (C-7), and the substitution positions of the four methoxy groups were determined. δ H 3.06 (17-NC H 3) was remotely correlated with δ C 176.5 (C-16), 76.3 (C-14), δ H 2.90 (H-15a), 2.62 (H-15b) were remotely correlated with δ C 176.5 (C-16), and the substitution positions of the nitrogen-methyl and carbonyl groups were determined. In summary, the planar structure of Compound 9 was determined. Through 1 the coupling constants of the key hydrogens in the 1H NMR spectrum and the NOESY spectrum, the relative configuration of Compound 9 was determined. According to J H-6,H-7 = 4.2 Hz, it was determined that H-6 and H-7 were cis. According to the NOESY spectrum, δ H 3.63 (H-7) was correlated with δ H 1.96 (H-5b), 3.06 (17-NC H 3), δ H 1.96 (H-5b) was correlated with δ H 2.90 (H-15a), δ H 2.62 (H-15b) was correlated with δ H 1.76 (H-9b), suggesting that the spatial orientations of H-6, H-7 and the ethylamine bridge at the 13, 14 positions were the same, and the relative configuration of Compound 9 was determined to be 6S*, 7S*, 8R*, 10S*, 13R*, 14S*. Through base hydrolysis experiments, the side chain was determined to be the (2'R)-methylbutyric acid compound. Further, by comparing the experimental and calculated ECD data, the absolute configuration of Compound 9 was determined to be 6S, 7S, 8R, 10S, 13R, 14S, 2'R.

[0120] In summary, Compound 9 was identified as (+)(6S,7S,8R,10S,13R,14S,2'R)-hernsubanine K, which is a new compound not reported in the literature after retrieval.

[0121] The structure identification data of Compound 10 are as follows:

[0122] White amorphous powder (methanol), positive reaction to modified bismuth potassium iodide reaction, . HR-ESI-MS gave the quasi-molecular ion peak m / z 582.2365 [M + H] + (calcd. 582.2339 for C31 H 36 NO 10 ), suggesting the molecular formula is C 31 H 35 NO 10 , with an unsaturation degree of 14. 1 1H NMR (600 MHz, CD3OD) gave the characteristic signals of the 8,10-oxy-bridge nelumbine-type alkaloid nucleus at δ H 5.08 (1H, d-like, J = 6.1 Hz, H-10). A set of trans double bond hydrogen signals were given in the low field region at δ H 7.06 (1H, d, J = 16.0 Hz, H-7'), 5.46 (1H, d, J = 16.0 Hz, H-8'); A set of ortho-coupled hydrogen signals on the tetrasubstituted benzene ring at δ H 6.98 (1H, d, J = 8.2 Hz, H-1), 6.68 (1H, d, J = 8.2 Hz, H-2); A set of ABX coupling system hydrogen signals at δ H 6.97 (1H, d, J = 8.2 Hz, H-5'), 6.95 (1H, dd, J = 8.2, 1.8 Hz, H-6'), 6.93 (1H, d, J = 1.8 Hz, H-2'). Five methoxy hydrogen signals were visible in the high field region at δ H 3.94 (3H, s, 4'-OC H 3), 3.82 (3H, s, 4-OC H 3), 3.63 (3H, s, 8-OC H 3), 3.46 (3H, s, 3-OC H 3), 3.44 (3H, s, 7-OC H 3); A nitrogen-methyl hydrogen signal at δ H 3.08 (3H, s, 17-NC H 3), and the chemical shift of this nitrogen-methyl group indicates that the C-16 position of the compound is oxidized to a carbonyl group; A hydrogen signal on the carbon substituted by an acyloxy group at δ H 5.38 (1H, m, H-6); A hydrogen signal on the oxygen-linked carbon at δ H 3.75 (1H, d, J = 4.1 Hz, H-7). 13 13C-NMR (150 MHz, CD3OD) gave 31 carbon signals, including the characteristic carbon signals of the 8,10-oxy-bridge nelumbine-type alkaloid oxygen bridge at δ C 101.4 (C-8), 76.2 (C-10); Two carbonyl carbon signals at δ C 175.1 (C-16), 167.1 (C-9′); 12 aromatic carbon signals at δ C153.4 (C-3), 149.9 (C-4'), 147.6 (C-4), 146.5 (C-3'), 133.3 (C-11), 131.8 (C-12), 127.5 (C-1'), 121.5 (C-6'), 120.1 (C-1), 113.3 (C-2'), 111.0 (C-5'), 110.2 (C-2); a set of double-bond carbon signals δ C 144.2 (C-7'), 114.5 (C-8'); five methoxy carbon signals δ C 59.5 (4-O C H3), 56.5 (7-O C H3), 54.9 (4'-O C H3), 54.3 (3-O C H3), 50.8 (8-O C H3); a nitrogen-methyl carbon signal δ C 27.3 (17-N C H3). The NMR data were assigned by combining with the HSQC spectrum, as shown in Table 4 and Table 6.

[0123] In the HMBC spectrum, δ H 6.97 (H-5') was remotely correlated with δ C 146.5 (C-3'), δ H 6.95 (H-6'), 6.93 (H-2') and δ C 149.9 (C-4′), 144.2 (C-7') were remotely correlated. Combining with 1 the chemical shift of δ H-6 5.38 in 1H NMR, it was speculated that there was a 3,4-dimethoxycinnamoyl group in the structure and it was substituted at C-6. Meanwhile, in the HMBC spectrum, δ H 3.94 (4'-OC H 3), 3.82 (3'-OC H 3), 3.63 (8-OC H 3), 3.46 (3-OC H 3), 3.44 (7-OC H 3) were remotely correlated with δ C 149.9 (C-4'), 146.5 (C-3'), 101.4 (C-8), 153.4 (C-3), 81.6 (C-7) respectively to determine the substitution positions of the methoxy groups. δ H 3.08 (17-NC H 3) was correlated with δ C175.1 (C-16), 75.0 (C-14) are remotely correlated, determining the substitution position of the nitrogen-methyl. In summary, the planar structure of compound 10 is determined. According to J H-6,H-7 = 4.1 Hz, it is determined that H-6 and H-7 are cis. In the NOESY spectrum, δ H 3.75 (H-7) is correlated with 1.95 (H-5b), 3.08 (17-NC H 3); δ H 1.95 (H-5b) is correlated with 2.88 (H-15a); δ H 2.61 (H-15b) is correlated with 1.82 (H-9b), indicating that the hydrogens at the 6,7 positions and the ethylamine bridge at the 13,14 positions have the same spatial orientation, suggesting that the relative configuration of compound 10 is 6S*, 7S*, 8R*, 10R*, 13R*, 14S*. Further comparison of experimental and calculated ECD data determined the absolute configuration of compound 10 to be 6S, 7S, 8R, 10S, 13R, 14S.

[0124] In summary, compound 10 was identified as (-)(6S,7S,8R,10S,13R,14S)-hernsubanine L, which is a new compound not reported in the literature after retrieval.

[0125] The structure identification data of compound 11 are as follows:

[0126] White amorphous powder (methanol), positive reaction with modified bismuth potassium iodide, . HR-ESI-MS gave the quasi-molecular ion peak m / z 568.2529 [M+H] + (calcd. 568.2541 for C 31 H 38 NO9), suggesting the molecular formula C 31 H 37 NO9, and the degree of unsaturation is 14. 1 HNMR (600 MHz, CD3OD) gave the characteristic signals of the 8,10-oxy-bridge nelumbine-type alkaloid nucleus δ H 4.99 (1H, d, J = 6.2 Hz, H-10). A set of trans double bond hydrogen signals δ H 7.00 (1H, d, J = 16.0 Hz, H-7'), 5.41 (1H, d, J = 16.0 Hz, H-8'); a set of ABX coupling system hydrogen signals δ H6.97 (1H, d, J = 8.0 Hz, H-5'), 6.93 (1H, dd, J = 8.0, 2.0 Hz, H-6'), 6.91 (1H, d, J = 2.0 Hz, H-2'); a set of hydrogen signals of 1,2,3,4-tetrasubstituted benzene ring δ H 6.90 (1H, d, J = 8.1 Hz, H-1), 6.60 (1H, d, J = 8.1 Hz, H-4); a hydrogen signal on the carbon substituted by an acyloxy group δ H 5.36 (1H, m, H-6). Five sets of hydrogen signals of methoxy groups can be seen in the high-field region δ H 3.91 (3H, s, 4'-OC H 3), 3.74 (3H, s, 4-OC H 3), 3.53 (3H, s, 8-OC H 3), 3.41 (3H, s, 3-OC H 3), 3.40 (3H, s, 7-OC H 3); a hydrogen signal on the carbon of the methine group connected to oxygen δ H 3.81 (1H, d, J = 4.3 Hz, H-7); a set of hydrogen signals of N-methyl δ H 2.60 (3H, s, 17-NC H 3). 13 C NMR (150 MHz, CD3OD) gives 31 carbon signals, including the characteristic carbon signals of the oxygen bridge of 8,10-oxoaporphine alkaloid oxygen bridge δ C 104.7 (C-8), 78.7 (C-10); a carbonyl carbon signal δ C 168.6 (C-9'); two sets of benzene ring carbon signals δ C 154.8 (C-3), 151.3 (C-4'), 149.0 (C-4), 147.9 (C-3'), 136.8 (C-12), 134.7 (C-11), 129.0 (C-1'), 122.8 (C-6'), 121.2 (C-1), 114.7 (C-2'), 112.4 (C-5'), 110.9 (C-2) and a set of double bond carbon signals δ C 145.3 (C-7'), 116.3 (C-8'); five methoxy carbon signals δ C 60.8 (4-O C H3), 57.8 (7-O C H3), 56.4 (4'-O C H3), 55.7 (3-O C H3), 51.8 (8-O C H3); a N-methyl carbon signal δ C39.3(17-N C H3); The NMR data were assigned by combining with the HSQC spectrum, as shown in Table 5 and Table 6.

[0127] In the HMBC spectrum, δ H 6.97 (H-5') was remotely correlated with 147.9 (C-3'), δ H 6.93 (H-6') was remotely correlated with δ C 151.3 (C-4'), 114.7 (C-2'), δ H 7.00 (H-7') was remotely correlated with δ C 168.6 (C-9'), 122.8 (C-6'), 114.7 (C-2'); Combining with 1 the chemical shift of H-6 in 1H NMR, it was suggested that there was a 3,4-dimethoxycinnamoyl group substituted at C-6. δ H 3.91 (4'-OC H 3), 3.74 (4-OC H 3), 3.53 (8-OC H 3), 3.41 (3-OC H 3), 3.40 (7-OC H 3) were remotely correlated with δ C 151.3 (C-4'), 149.0 (C-4), 104.7 (C-8), 154.8 (C-3), 82.9 (C-7) respectively, determining the substitution positions of the five methoxy groups. δ H 2.60 (17-NC H 3) was remotely correlated with δ C 77.9 (C-14), 55.2 (C-16), determining the substitution position of the nitrogen-methyl group. In summary, the planar structure of compound 11 was determined. By 1 the coupling constants of the key hydrogens in the 1H NMR spectrum and the NOESY spectrum, the relative configuration of compound 11 was determined. According to J H-6,H-7 = 4.3 Hz, it was determined that H-6 and H-7 were cis. In the NOESY spectrum, δ H 3.81 (H-7) was correlated with δ H 2.14 (H-5b), 2.60 (17-NC H 3), δ H 2.87 (H-5a) was correlated with δ H 2.36 (H-15a), δ H 1.91 (H-15b) was correlated with δ HIt is related to 1.62(H-9b), indicating that the spatial orientations of the ethylamine bridges at H-6, H-7 and positions 13 and 14 are the same. The relative configuration of compound 11 was determined to be 6S*,7S*,8R*,10S*,13S*,14S*. Further comparison of experimental and calculated ECD data confirmed the absolute configuration of compound 11 as 6S,7S,8R,10S,13S,14S.

[0128] In summary, compound 11 was identified as (-)(6S,7S,8R,10S,13S,14S)-hernsubanine M, a new compound not reported in the literature after retrieval.

[0129] The structure identification data of compound 12 are as follows:

[0130] White amorphous powder (methanol), positive reaction to modified bismuth potassium iodide . HR-ESI-MS gave the quasi-molecular ion peak m / z 406.1860 [M+H] + (calcd. 406.1867 for C 21 H 28 NO7), indicating the molecular formula C 21 H 27 NO7, with an unsaturation degree of 9. 1 1H NMR (600 MHz, CD3OD) gave the characteristic signals of the 8,10-oxabicyclo[3.2.1]octane-type alkaloid nucleus δ H 4.83 (1H, d, J = 6.3 Hz, H-10). A set of hydrogen signals of the 1,2,3,4-tetrasubstituted benzene ring were given in the low field region δ H 6.81 (1H, d, J = 8.2 Hz, H-2), 6.77 (1H, d, J = 8.2 Hz, H-1). Four sets of methoxy hydrogen signals were visible in the high field region δ H 3.89 (3H, s, 4-OC H 3), 3.84 (3H, s, 3-OC H 3), 3.51 (3H, s, 8-OC H 3), 3.43 (3H, s, 7-OC H 3); a set of nitrogen-methyl hydrogen signal δ H 2.85 (3H, s, 17-NC H 3); hydrogen signals on two oxygen-bearing carbons δ H 4.43 (1H, s, H-7), 4.18 (1H, s, H-16). 13 13C NMR (150 MHz, CD3OD3) spectrum gave 21 carbon signals, including the characteristic signal of the oxygen bridge of the 8,10-oxabicyclo[3.2.1]octane-type alkaloid oxygen bridge δ C108.5 (C-8), 78.1 (C-10); one keto carbonyl carbon signal δ C 206.6 (C-6); a set of benzene ring carbon signals δ C 154.9 (C-3), 149.5 (C-4), 135.2 (C-12), 132.8 (C-11), 121.2 (C-1), 112.0 (C-2); four methoxy carbon signals δ C 60.9 (4-O C H3), 59.8 (7-O C H3), 56.2 (3-O C H3), 52.2 (8-O C H3); one nitrogen-methyl carbon signal δ C 37.7 (17-N C H3); The NMR data were assigned by combining with the HSQC spectrum, as shown in Table 5 and Table 6.

[0131] In the HMBC spectrum, δ H 6.81 (H-2) has long-range correlations with δ C 149.5 (C-4), 135.2 (C-11), δ H 6.77 (H-1) has long-range correlations with δ C 154.9 (C-3), 132.8 (C-12), 78.1 (C-10), δ H 3.84 (3-OC H 3), 3.89 (4-OC H 3), 3.43 (7-OC H 3), 3.51 (8-OC H 3) has long-range correlations with δ C 154.9 (C-3), 149.5 (C-4), 89.7 (C-7), 108.5 (C-8) respectively, indicating that the four methoxy groups are substituted at positions C-3, C-4, C-7 and C-8; According to δ H 2.85 (17-NC H 3) has long-range correlations with δ C 101.0 (C-16), 78.8 (C-14), indicating the presence of the nitrogen-methyl group; δ H 4.18 (H-16) has long-range correlations with δ C 52.0 (C-13), 37.7 (N C H3), indicating that the hydroxyl group is substituted at position C-16. In summary, the planar structure of compound 12 was determined. In the NOESY spectrum, δ H 3.61 (H-5a) is correlated with 4.43 (H-7), 2.60 (H-15a), δ H3.51 (8-OC H 3) and δ H 2.85 (-NC H 3), δ H 4.18 (H-16) and δ H 2.85 (-NC H 3) are related, suggesting that the spatial orientations of H-7 and H-16 are the same as those of the ethylamine bridge at the 13th and 14th positions, and the relative configuration of compound 12 is determined to be 7R*, 8R*, 10S*, 13S*, 14S*, 16R*. The absolute configuration of compound 12 is further determined to be 7R, 8R, 10S, 13S, 14S, 16R by comparing experimental and calculated ECD data.

[0132] In summary, compound 12 was identified as (-)(7R,8R,10S,13S,14S,16R)-hernsubanine N, which is a new compound not reported in the literature after retrieval.

[0133] Table 5 1H NMR data of compounds 9 - 12

[0134]

[0135]

[0136] a : The test solvent was CD3OD, b : The test solvent was CDCl3

[0137] Table 6 13C NMR data of compounds 5 - 12

[0138]

[0139]

[0140] a : The test solvent was CD3OD, b : The test solvent was CDCl3

[0141] Example 2

[0142] (1) 3000 g of the dried whole herb of Stephania hernandifolia was refluxed with 95% ethanol for 2 times (dosage: 30 L), and the extract was recovered under reduced pressure to obtain a crude extract;

[0143] (2) The 95% ethanol crude extract obtained in step (1) above was dissolved in water, adjusted to pH 2 with 5% HCl, and extracted 3 times with petroleum ether (the volume ratio of the aqueous phase to petroleum ether was 1:2); 5% sodium carbonate solution was added to the aqueous layer to adjust the pH to 7, and it was extracted 3 times with ethyl acetate (the volume ratio of the aqueous phase to ethyl acetate was 1:2) to obtain total alkaloids;

[0144] (3) The total alkaloids obtained in the above step (2) are separated by silica gel column chromatography and eluted successively with dichloromethane and acetone mixed solvents at ratios of 100:1, 100:2, 100:3, 100:5, 100:8, and 10:1.

[0145] (4) The fractions obtained from the dichloromethane:acetone mixed solvent at ratios of 100:3 to 10:1 in the above step (3) are subjected to ODS chromatography and gradient eluted with methanol and water mixed solvents at ratios of 3:7, 5:5, 6:4, 7:3, and 8:2.

[0146] (5) The fractions obtained from the methanol:water mixed solvent at ratios of 5:5 to 8:2 in the above step (4) are separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of methanol:water = 32:68, to obtain the lianhuashane-type alkaloid compound 5 (t R = 9 min) (yield 0.00042‰) and compound 10 (t R = 45 min) (yield 0.00005‰);

[0147] (6) The fractions obtained from the methanol:water mixed solvent at ratios of 5:5 to 8:2 in the above step (4) are separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of methanol:water = 62:38, to obtain the lianhuashane-type alkaloid dimer compound 3 (t R = 19 min) (yield 0.00007‰), the lianhuashane-type alkaloid compound 6 (t R = 12 min) (yield 0.00040‰), compound 7 (t R = 34 min) (yield 0.00006‰), and compound 9 (t R = 41 min) (yield 0.00025‰);

[0148] (7) The fractions obtained from the methanol:water mixed solvent at ratios of 5:5 to 8:2 in the above step (4) are separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 50:50 (plus 1‰ diethylamine), to obtain the lianhuashane-type alkaloid compound 2 (t R = 27 min) (yield 0.0011‰), compound 8 (t R = 12 min) (yield 0.00032‰), compound 11 (t R = 14 min) (yield 0.00028‰), and compound 12 (t R = 20 min) (yield 0.00006‰);

[0149] (8) The fraction obtained from the methanol:water mixed solvent at 5:5 - 8:2 in step (4) above was separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase being acetonitrile:water = 42:58 (plus 1‰ diethylamine), to obtain the dimer-type compound 1 of lianhuashane alkaloids (t R = 23 min) (yield 0.00044‰) and compound 4 (t R = 28 min) (yield 0.00045‰).

[0150] The structural identification methods of lianhuashane alkaloids and their dimer-type compounds 1 - 12 are shown in Example 1.

[0151] Example 3

[0152] (1) 2000 g of the dried whole herb of Stephania hernandifolia was refluxed and extracted 3 times with 75% methanol (dosage: 30 L), and the extract was recovered under reduced pressure to obtain the crude extract;

[0153] (2) The 75% methanol crude extract obtained in the above step (1) was dissolved in water, adjusted to pH 2 with 4% HCl, and extracted 3 times with cyclohexane (volume ratio of aqueous phase to cyclohexane is 1:1); 5% sodium carbonate solution was added to the aqueous layer to adjust the pH to 7, and extracted 3 times with chloroform (volume ratio of aqueous phase to chloroform is 1:2) to obtain the total alkaloids;

[0154] (3) The total alkaloids obtained in the above step (2) were separated by silica gel column chromatography, and eluted successively with petroleum ether and ethyl acetate mixed solvents of 10:1, 8:1, 5:1, 3:1, 2:1, 1:1, 0:1;

[0155] (4) The fractions obtained from the petroleum ether:ethyl acetate mixed solvent at 5:1 - 0:1 in the above step (3) were subjected to ODS chromatography and gradient eluted with acetonitrile and water mixed solvents of 2:8, 3:7, 5:5, 6:4, 7:3, 8:2;

[0156] (5) The fractions obtained from the acetonitrile:water mixed solvent at 2:8 - 7:3 in the above step (4) were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase being acetonitrile:water = 25:75, to obtain the lianhuashane alkaloid compound 5 (t R = 13 min) (yield 0.00040‰) and compound 10 (t R = 42 min) (yield 0.00005‰);

[0157] (6) The fractions obtained from the acetonitrile: water mixed solvent at 2:8 to 7:3 in the above step (4) were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase was acetonitrile: water = 48:52, to obtain the dimer compounds of liensinine-type alkaloids 3 (t R = 17 min) (yield 0.00006‰), liensinine-type alkaloid compounds 6 (t R = 13 min) (yield 0.00038‰), compound 7 (t R = 24 min) (yield 0.00006‰) and compound 9 (t R = 39 min) (yield 0.00023‰);

[0158] (7) The fractions obtained from the acetonitrile: water mixed solvent at 2:8 to 7:3 in the above step (4) were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase was methanol: water = 72:28 (plus 1.5‰ diethylamine), to obtain the liensinine-type alkaloid compounds 2 (t R = 29 min) (yield 0.0009‰), compound 8 (t R = 11 min) (yield 0.00031‰), compound 11 (t R = 12 min) (yield 0.00027‰) and compound 12 (t R = 20 min) (yield 0.00006‰);

[0159] (8) The fractions obtained from the acetonitrile: water mixed solvent at 2:8 to 7:3 in the above step (4) were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase was acetonitrile: water = 48:52 (plus 1‰ diethylamine), to obtain the dimer compounds of liensinine-type alkaloids 1 (t R = 15 min) (yield 0.00040‰) and compound 4 (t R = 18 min) (yield 0.00041‰).

[0160] The structure identification methods of liensinine-type alkaloids and their dimer compounds 1 - 12 are shown in Example 1.

[0161] Example 4

[0162] (1) 4000 g of the dried whole herb of Stephania hernandifolia was refluxed with 80% methanol 3 times (dosage: 50 L), and the extract was recovered under reduced pressure to obtain the crude extract;

[0163] (2) Dissolve the 85% methanol extract obtained in the above step (1) in water, add 5% HCl to adjust the pH value to 2, and extract with cyclohexane (the volume ratio of the aqueous phase to cyclohexane is 1:2) for 3 times; then add 5% sodium carbonate solution to the aqueous layer to adjust the pH value to 7, and extract with ethyl acetate (the volume ratio of the aqueous phase to ethyl acetate is 1:1) for 3 times to obtain total alkaloids;

[0164] (3) Subject the total alkaloids obtained in the above step (2) to silica gel column chromatography and elute successively with a mixed solvent of chloroform and methanol at ratios of 100:1, 50:1, 30:1, 20:1, and 15:1;

[0165] (4) Subject the fractions obtained from chloroform:methanol 50:1 to 15:1 in the above step (3) to ODS chromatography and perform gradient elution with a mixed solvent of acetonitrile and water at ratios of 2:8, 3:7, 5:5, 6:4, 7:3, and 8:2;

[0166] (5) Subject the fractions obtained from acetonitrile:water mixed solvent 2:8 to 7:3 in the above step (4) to HPLC-UV chromatographic separation and preparation, detect at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of methanol:water = 36:64 to obtain lianhuashane-type alkaloid compounds 5 (t R = 6 min) (yield 0.00042‰) and compound 10 (t R = 36 min) (yield 0.00005‰);

[0167] (6) Subject the fractions obtained from acetonitrile:water mixed solvent 2:8 to 7:3 in the above step (4) to HPLC-UV chromatographic separation and preparation, detect at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 50:50 to obtain lianhuashane-type alkaloid dimer compounds 3 (t R = 14 min) (yield 0.00007‰), lianhuashane-type alkaloid compounds 6 (t R = 11 min) (yield 0.00038‰), compound 7 (t R = 19 min) (yield 0.00006‰) and compound 9 (t R = 28 min) (yield 0.00022‰);

[0168] (7) Subject the fractions obtained from methanol:water mixed solvent 5:5 to 8:2 in the above step (4) to HPLC-UV chromatographic separation and preparation, detect at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 52:48 (plus 1‰ diethylamine) to obtain lianhuashane-type alkaloid compounds 2 (t R = 25 min) (yield 0.0010‰), compound 8 (t R= 11 min) (yield 0.00030 ‰), compound 11 (t R = 12 min) (yield 0.00028 ‰) and compound 12 (t R = 17 min) (yield 0.00006 ‰);

[0169] (8) The fraction obtained from the acetonitrile: water mixed solvent 2:8 - 7:3 in step (4) above was separated and prepared by HPLC - UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase was methanol: water = 68:32 (plus 1‰ diethylamine), to obtain the lotusan - type alkaloid dimer compounds 1 (t R = 23 min) (yield 0.00041 ‰) and compound 4 (t R = 40 min) (yield 0.00040 ‰).

[0170] The structural identification methods of lotusan - type alkaloids and their dimer compounds 1 - 12 are shown in Example 1.

[0171] Example 5

[0172] (1) 4000 g of the dried whole herb of Stephania hernandifolia was refluxed with 70% ethanol 3 times (dosage: 40 L), and the extract was recovered under reduced pressure to obtain a crude extract;

[0173] (2) The 70% ethanol extract obtained in step (1) above was dissolved in water, adjusted to pH 2 with 4.5% HCl, and extracted 3 times with petroleum ether (volume ratio of aqueous phase to petroleum ether is 1:1); 5% sodium carbonate solution was added to the aqueous layer to adjust the pH to 7, and extracted 3 times with dichloromethane (volume ratio of aqueous phase to dichloromethane is 1:1) to obtain total alkaloids;

[0174] (3) The total alkaloids obtained in step (2) above were separated by silica gel column chromatography, and eluted successively with petroleum ether and acetone mixed solvents 10:1, 8:1, 5:1, 3:1, 2:1, 1:1, 0:1;

[0175] (4) The fraction obtained from the petroleum ether: acetone mixed solvent 8:1 - 1:1 in step (3) above was subjected to ODS chromatography, and gradient elution was carried out with methanol and water mixed solvents 3:7, 4:6, 5:5, 6:4, 8:2;

[0176] (5) The fraction obtained from the methanol: water mixed solvent 4:6 - 8:2 in step (4) above was separated and prepared by HPLC - UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase was acetonitrile: water = 28:72, to obtain the lotusan - type alkaloid compounds 5 (t R = 9 min) (yield 0.00042 ‰) and compound 10 (tR = 37 min) (yield 0.00006‰);

[0177] (6) The fractions obtained from the methanol:water mixed solvent 4:6 - 8:2 in step (4) above were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase was methanol:water = 70:30, to obtain the lianhuashane-type alkaloid dimer compound 3 (t R = 12 min) (yield 0.00007‰), the lianhuashane-type alkaloid compound 6 (t R = 8 min) (yield 0.00040‰), compound 7 (t R = 19 min) (yield 0.00006‰), and compound 9 (t R = 22 min) (yield 0.00025‰);

[0178] (7) The fractions obtained from the methanol:water mixed solvent 4:6 - 8:2 in step (4) above were separated and prepared by HPLC-UV chromatography, detected at 254 nm, with a flow rate of 3 mL / min, and the mobile phase was acetonitrile:water = 45:55 (plus 1‰ diethylamine), to obtain the lianhuashane-type alkaloid dimer compound 1 (t R = 19 min) (yield 0.00040‰), compound 2 (t R = 44 min) (yield 0.0010‰), compound 4 (t R = 24 min) (yield 0.00039‰), and the lianhuashane-type alkaloid compound 8 (t R = 16 min) (yield 0.00032‰), compound 11 (t R = 21 min) (yield 0.00028‰), and compound 12 (t R = 39 min) (yield 0.00006‰).

[0179] The structure identification methods of the lianhuashane-type alkaloids and their dimer compounds 1 - 12 are shown in Example 1.

[0180] Example 6

[0181] (1) 2000 g of the dried whole herb of Stephania hernandifolia was refluxed and extracted 3 times with 85% methanol (dosage: 30 L), and the extract was recovered under reduced pressure to obtain a crude extract;

[0182] (2) Dissolve the 70% ethanol extract obtained in the above step (1) in water, adjust the pH value to 2 by adding 4% HCl, and extract 3 times with cyclohexane (the volume ratio of the aqueous phase to cyclohexane is 1:2); add 5% sodium carbonate solution to the aqueous layer to adjust the pH value to 7, and extract 3 times with chloroform (the volume ratio of the aqueous phase to chloroform is 1:2) to obtain total alkaloids;

[0183] (3) Subject the total alkaloids obtained in the above step (2) to silica gel column chromatography and elute successively with chloroform and acetone mixed solvents at ratios of 100:1, 100:2, 100:3, 100:5, 100:8, 10:1;

[0184] (4) Subject the fractions obtained from the chloroform:acetone mixed solvent at ratios of 100:3 to 10:1 in the above step (3) to ODS chromatography and perform gradient elution with acetonitrile and water mixed solvents at ratios of 2:8, 3:7, 5:5, 6:4, 7:3, 8:2;

[0185] (5) Subject the fractions obtained from the acetonitrile:water mixed solvent at ratios of 2:8 to 7:3 in the above step (4) to HPLC-UV chromatographic separation and preparation, detect at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 23:77, to obtain the lianhuashane-type alkaloid compound 5 (t R = 14 min) (yield 0.00044‰) and compound 10 (t R = 49 min) (yield 0.00006‰);

[0186] (6) Subject the fractions obtained from the acetonitrile:water mixed solvent at ratios of 2:8 to 7:3 in the above step (4) to HPLC-UV chromatographic separation and preparation, detect at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 45:55, to obtain the lianhuashane-type alkaloid dimer compound 3 (t R = 20 min) (yield 0.00006‰), the lianhuashane-type alkaloid compound 6 (t R = 16 min) (yield 0.00039‰), compound 7 (t R = 34 min) (yield 0.00007‰) and compound 9 (t R = 56 min) (yield 0.00019‰);

[0187] (7) Subject the fractions obtained from the acetonitrile:water mixed solvent at ratios of 2:8 to 7:3 in the above step (4) to HPLC-UV chromatographic separation and preparation, detect at 254 nm, with a flow rate of 3 mL / min, and a mobile phase of methanol:water = 72:28 (plus 1‰ diethylamine), to obtain the lianhuashane-type alkaloid dimer compound 1 (t R = 21 min) (yield 0.00041‰), compound 2 (tR = 29 min) (yield 0.0009‰), compound 4 (t R = 28 min) (yield 0.00043‰), and lotuscane-type alkaloid compound 8 (t R = 11 min) (yield 0.00030‰), compound 11 (t R = 12 min) (yield 0.00028‰), and compound 12 (t R = 20 min) (yield 0.00006‰).

[0188] The structural identification methods of lotuscane-type alkaloids and their dimer compounds 1-12 are shown in Example 1.

[0189] Anti-neuroinflammatory activity test of lotuscane-type alkaloids and their dimer compounds 1-12 prepared in Examples 1-6:

[0190] (1) Experimental principle: Neuroinflammation caused by overactivation of microglia plays an important role in mediating the occurrence of neurodegenerative diseases. Inhibiting overactivated microglia may become a new target for drug discovery. By establishing an in vitro screening model of LPS-induced overactivated BV-2 microglia, with the NO release amount as the detection index, the anti-neuroinflammatory activities of novel lotuscane-type alkaloids and their dimer compounds 1-12 were evaluated.

[0191] (2) Experimental method:

[0192] ① Culture of mouse microglial cell line BV-2

[0193] All glassware and metal instruments (culture flasks, pipettes, solution bottles, etc.) used in cell culture and model establishment were autoclaved at 121 °C for 30 min to thoroughly remove contaminated LPS. The cell culture medium was prepared based on DMEM medium containing 10% fetal bovine serum. Microglia were passaged at a concentration of about 2.0×10 5 cells / mL in a 5% CO2, 37 °C culture flask. By the third day, the adherent cells accounted for about 70-80% of the bottom area of the culture flask. The adherent cells were digested with trypsin and passaged to another culture flask. The BV-2 cells after cryopreservation and resuscitation in an -80 °C ultra-low temperature freezer were used as the first generation, and the 3rd-8th generation BV-2 cells were selected for the experiment.

[0194] ② Drug preparation method

[0195] The test compounds were all dissolved in DMSO to prepare a stock solution (100 mM) and stored at -20 °C. Before use, they were diluted with DMEM culture medium to 100 μM, 30 μM, 10 μM, and 1 μM in sequence. The final concentration of DMSO < 1‰.

[0196] ③ Detection of the inhibitory effect of compounds on LPS-activated microglia by the Griess method

[0197] BV-2 microglial cells in the logarithmic growth phase were taken, and the cell density was adjusted to 2.0×10 5 cells / mL with fresh DMEM culture medium containing 10% fetal bovine serum. They were inoculated into 96-well plates, 100 μL / well, and cultured in an incubator at 37°C and 5% CO2. After 24 h of adherent cell culture, the fresh culture medium without serum was replaced, and drug treatment was carried out simultaneously. The test concentrations of the compounds were set at 100 μM, 30 μM, 10 μM, 1 μM and co-treated with LPS. At the same time, a blank control was set. The final concentration of LPS in each drug administration group was 100 ng / mL. After the cells were treated with drugs and continued to be cultured for 24 h, the supernatant was collected, and the NO 2- content in the supernatant was detected by the Griess colorimetric method.

[0198] ④ Detection of the effect of compounds on the cell viability of microglia by the MTT method

[0199] BV-2 microglial cells cultured in the logarithmic growth phase were taken, and the cell density was adjusted to 2.0×10 5 cells / mL with fresh DMEM culture medium containing 10% fetal bovine serum. They were inoculated into 96-well plates, 100 μL / well, and cultured in an incubator at 37°C and 5% CO2. After 24 h of adherent cell culture, the fresh culture medium was replaced, and drug treatment was carried out simultaneously. The doses of the compounds were set at 100 μM, 30 μM, 10 μM, 1 μM and co-treated with LPS. At the same time, a blank control was set. The final concentration of LPS in each drug administration group was 100 ng / mL. After the cells were treated with drugs and continued to be cultured for 24 h, then MTT solution, 10 μL / well, was added to the cell solution, and the cells and 0.25 mg / mL MTT were co-incubated at 37°C for 3 h. The culture medium was aspirated, and then 150 μL of DMSO solution was added to measure its optical density OD value. The data was processed using the microplate reader software, and the average value of the OD values of 3 wells for each sample was calculated. The cell viability (CV%) was calculated using the average value according to the following formula.

[0200] Cell viability % = (average value of OD values of the sample group / average value of OD values of the blank control group) × 100%

[0201] ⑤ Statistical method

[0202] All data were analyzed using the SPSS (19.0) statistical software package. Results were expressed as mean ± standard error. To evaluate the overall differences, one-way ANOVA was used to analyze the homogeneity of variance among group means, and Dunnett's test was combined for inter-group comparison. Levene's test was used for the homogeneity test of variances among multiple samples. When p > 0.05, the variances were homogeneous, and Dunnett's two-sided T test was used to analyze the differences in means among multiple groups. When p < 0.05, the variances were heterogeneous, and Dunnett T3 test was used to analyze the differences in means among multiple groups.

[0203] ⑥IC 50 Calculation method

[0204] Nonlinear regression fitting was used to calculate IC with parameters such as each dose and inhibition rate. 50 。

[0205] (3) Experimental results:

[0206] The experimental results are shown in Table 7.

[0207] Table 7 Experimental results of the effects of Compounds 1 - 12 on the release of NO from LPS-activated BV-2 microglia

[0208]

[0209] Note: *P < 0.05, **P < 0.01, ***P < 0.001 compared with the LPS-induced group; ### P < 0.001 compared with the control group.

[0210] As can be seen from the results in Table 7, the novel lotusanane-type alkaloid compounds 1 (1 μM, 10 μM, 30 μM, 100 μM), 2 (30 μM, 100 μM), 3 (30 μM, 100 μM), 4 (1 μM, 10 μM, 30 μM, 100 μM), 5 (100 μM), 6 (30 μM, 100 μM), 7 (30 μM, 100 μM), 8 (10 μM, 30 μM, 100 μM), 9 (30 μM, 100 μM), 10 (30 μM, 100 μM), 11 (30 μM, 100 μM), and 12 (30 μM, 100 μM) prepared in Examples 1 - 6 could significantly inhibit the release of NO from LPS-induced over-activated BV-2 microglia.

Claims

1. Lotusane-type alkaloids, their dimer compounds, pharmaceutically acceptable salts and isomers thereof, characterized in that: It has the following structural general formulas (I) to (VIII): Wherein, R1 is hydrogen, and both R2 and R3 are methoxy groups; or R1 and R2 are -OCH2O- to form a five-membered ring, and R3 is hydrogen.

2. The lotusan type alkaloids, their dimer compounds, pharmaceutically acceptable salts and isomers according to claim 1, characterized in that: The compound is one of the following structural formulas: Wherein, R1 is hydrogen, and both R2 and R3 are methoxy groups; or R1 and R2 are -OCH2O- to form a five-membered ring, and R3 is hydrogen.

3. The liensinine-type alkaloids, their dimer compounds, pharmaceutically acceptable salts and isomers according to claim 1 or 2, characterized in that: The compound is specifically one of the following structural formulas: Compound 1: (-)(10R,13R,14S,16S,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine A Compound 2: (-)(10S,13R,14S,16S,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine B Compound 3: (-)(10S,13R,14S,16S,6"S,7"S,8"R,10"S,13"R,14"S)-bishernsubanine C Compound 4: (-)(10S,13R,14S,16R,6"S,7"S,8"R,10"S,13"S,14"S)-bishernsubanine D Compound 5: (-)(10S,13R,14S)-hernsubanine G Compound 6: (+)(6S,7S,8R,10S,13R,14S,2'R)-hernsubanine H Compound 7: (-)(6S,7S,8R,10S,13R,14S)-hernsubanine I Compound 8: (-)(6S,7S,8R,10S,13S,14S)-hernsubanine J Compound 9: (+)(6S,7S,8R,10S,13R,14S,2'R)-hernsubanine K Compound 10: (-)(6S,7S,8R,10S,13R,14S)-hernsubanine L Compound 11: (-)(6S,7S,8R,10S,13S,14S)-hernsubanine M Compound 12: (-)(7R,8R,10S,13S,14S,16R)-hernsubanine N.

4. The preparation method of the liensinine-type alkaloid and its dimer compounds according to claim 3, characterized in that: It includes the following steps: (1) Extract the dried whole herb of Stephania hernandifolia with ethanol or methanol, and recover the extract to obtain a crude extract; (2) Disperse the crude extract obtained in step (1) with water, adjust the pH value to 2, extract with petroleum ether or cyclohexane, adjust the pH value of the aqueous layer to 7, and extract with dichloromethane, chloroform or ethyl acetate to obtain total alkaloids; (3) The total alkaloids obtained in step (2) are separated by silica gel column chromatography and gradient elution is carried out with mixed solvent A, wherein the mixed solvent A is a mixed solvent of petroleum ether and ethyl acetate, a mixed solvent of petroleum ether and acetone, a mixed solvent of chloroform and acetone, a mixed solvent of dichloromethane and acetone, a mixed solvent of chloroform and methanol or a mixed solvent of dichloromethane and methanol; (4) The fractions obtained in step (3) are separated by ODS column chromatography and gradient elution is carried out with mixed solvent B as the mobile phase, wherein the mixed solvent B is a mixed solvent of methanol and water or a mixed solvent of acetonitrile and water; (5) The fractions obtained in step (4) are further separated by HPLC and gradient elution is carried out with mixed solvent C as the mobile phase to obtain Compounds 1-12, wherein the mixed solvent C is a mixed solvent of methanol and water or a mixed solvent of acetonitrile and water.

5. The preparation method according to claim 4, characterized in that: In step (1), heating under reflux with ethanol extraction or heating under reflux with methanol extraction is carried out 2-5 times, and the above organic solvents are recovered under reduced pressure to obtain a crude extract, wherein the volume concentration of ethanol is 70%-95%, the volume concentration of methanol is 60%-90%, and the mass-volume ratio of the dried whole herb of Stephania hernandifolia to ethanol or methanol is 1:8-1:20 g / mL.

6. The preparation method according to claim 4, characterized in that: In step (2), the crude extract obtained in step (1) is dispersed in water, HCl is added to adjust the pH value to 2, extracted with petroleum ether or cyclohexane, sodium carbonate solution is added to the aqueous layer to adjust the pH value to 7, and extracted with dichloromethane, chloroform or ethyl acetate to obtain total alkaloids, wherein each organic solvent is extracted 2-4 times, and the volume ratio of the aqueous phase to the organic phase is 1:1-1:

5.

7. The preparation method according to claim 4, wherein: In step (3), the mixed solvent A is a mixed solvent composed of petroleum ether and ethyl acetate in a volume ratio of 15:1-0:1, or a mixed solvent composed of petroleum ether and acetone in a volume ratio of 15:1-0:1, or a mixed solvent composed of dichloromethane and acetone in a volume ratio of 100:1-5:1, or a mixed solvent composed of chloroform and acetone in a volume ratio of 100:1-5:1, or a mixed solvent composed of dichloromethane and methanol in a volume ratio of 100:0-10:1, or a mixed solvent composed of chloroform and methanol in a volume ratio of 100:0-10:1; In step (4), the mixed solvent B is a mixed solvent composed of methanol and water in a volume ratio of 2:8-9:1, or a mixed solvent composed of acetonitrile and water in a volume ratio of 2:8-9:1; In step (5), the mixed solvent C is a mixed solvent composed of methanol and water in a volume ratio of 3:7-8:2, or a mixed solvent composed of acetonitrile and water in a volume ratio of 2:8-6:

4.

8. The preparation method according to claim 4 or 7, characterized in that: In step (5), according to the separation situation, 1‰-2‰ of diethylamine can be added to the mixed solvent C to improve the separation degree.

9. A pharmaceutical composition, characterized in that: Comprising the liensinine-type alkaloids and their dimer compounds and pharmaceutically acceptable salts and pharmaceutically acceptable carriers according to any one of claims 1-3.

10. Use of the lianhuashane-type alkaloid, its dimer compounds, and pharmaceutically acceptable salts thereof according to any one of claims 1 to 3, or the pharmaceutical composition according to claim 9, in the preparation of a medicament for preventing or treating neurodegenerative diseases.

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