Piperidine alkaloid compound in purslane as well as extraction and separation method and application of piperidine alkaloid compound

Abstract

The invention belongs to the technical field of traditional Chinese medicine extraction and separation, and particularly relates to a piperidine alkaloid compound extracted, separated and identified from purslane and an extraction and separation method of the piperidine alkaloid compound. The molecular formula of the novel alkaloid compound is C10H14N6O3, and the chemical name of the novel alkaloid compound is 2-(6-amino-9H-purin-9-yl) piperidine-3, 4, 5-triol. The invention further provides an extraction and separation method of the novel alkaloid compound. The extraction and separation method sequentially adopts water decoction extraction, macroporous resin column chromatography, ODS column chromatography, Sephadex LH-20 chromatography and a high performance liquid chromatograph for separation, purification and preparation. The structure of the alkaloid compound is determined as a novel alkaloid compound by mass spectrum, hydrogen spectrum, carbon spectrum and two-dimensional nuclear magnetic spectrum analysis methods. The novel alkaloid compound has anti-inflammatory activity, anti-cholinesterase activity and anti-oxidation activity. The novel alkaloid compound and the salt or derivative thereof can be used as raw materials for drug development and pharmacological activity research.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine extraction and separation, and in particular relates to a piperidine alkaloid compound in purslane and an extraction and separation method and application thereof. Background Art

[0002] Purslane is an annual herbaceous plant belonging to the family Portulacaceae and the order Portulacales in plant chemical taxonomy. It is widely distributed in tropical and subtropical regions of the world. Because its leaves resemble horse teeth, it is called "Purslane"; and because it is cold-resistant, has strong vitality and a long life, it is called "longevity vegetable".

[0003] Purslane contains a variety of chemical components, mainly including: alkaloids, flavonoids, organic acids, terpenes, coumarins, polysaccharides, amino acids and minerals. Among them, there are many types of alkaloid compounds in Purslane, such as uracil, adenine, norepinephrine, dopamine, adenosine, N, N-dicyclohexylurea, purslanamide, allantoin, N-trans-feruloyltyramine, iseluxine, oleracrylimide A, oleracrylimide B, oleracrylimideE, oleraisoquinoline A, etc. The chemical components contained in Purslane are closely related to its diverse pharmacological effects. Modern pharmacological studies have shown that Purslane has antioxidant, neuroprotective, anti-inflammatory, antibacterial, anti-tumor, hypoglycemic, liver-protecting and immune-enhancing effects. There are many kinds of chemical components in Purslane and their pharmacological activities are diverse. The development and separation of compounds in Purslane can provide a basis for in-depth research on Purslane. Summary of the invention

[0004] In view of the above problems, the present invention provides a piperidine alkaloid compound extracted from Portulaca oleracea. Studies have shown that the piperidine alkaloid compound of the present invention has anti-inflammatory activity, anticholinesterase activity and antioxidant activity. At the same time, a simple, rapid, environmentally friendly and high-purity extraction and separation method for the compound of the present invention is provided.

[0005] To achieve the above-mentioned object of the present invention, the present invention provides a piperidine alkaloid compound having a molecular formula of C 10 H 14 N6O3, chemical name is 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol, chemical structure is:

[0006]

[0007] To achieve the above-mentioned object of the present invention, the present invention also provides a method for extracting and separating a piperidine alkaloid compound in Portulaca oleracea, which specifically comprises the following steps:

[0008] Step 1: Take dried purslane, decoct in water to extract, concentrate the extract, cool to room temperature, and set aside the liquid medicine.

[0009] Step 2: The concentrated solution in step 1 is subjected to a macroporous resin, and eluted with water and ethanol of different concentrations, and the 30% ethanol portion is recovered under reduced pressure to obtain an extract, and a concentrate is obtained for use.

[0010] Step 3: Separate the concentrate in step 2 through an ODS column (Octadecylsilyl, octadecylsilane bonded silica gel filler) using a methanol-water gradient elution, detect by thin layer chromatography, develop color, combine the 30% methanol elution portion and evaporate to dryness to obtain a concentrate for use.

[0011] Step 4: The concentrate obtained in step 3 is separated by ODS column chromatography again, using methanol-water gradient elution, detected by thin layer chromatography, color development, and the 30% methanol elution portion is combined and evaporated to dryness to obtain a concentrate for use.

[0012] Step 5: The product obtained in step 4 is further separated by chromatography on a pretreated Sephadex LH-20 column, and isocratically eluted with 10% methanol to obtain several elution fractions, which are detected by thin layer chromatography, developed, and the elution fractions with developed color are combined, and the combined elution fractions are concentrated to dryness under reduced pressure for later use;

[0013] Step 6: The concentrate obtained in step 5 is separated and prepared by HPLC (high performance liquid chromatography), and isocratic elution is performed using methanol-0.1% formic acid (volume percentage) as a mobile phase to finally obtain the piperidine alkaloid compound of the present invention.

[0014] Furthermore, in step 1, the water decocting and extraction are performed twice, each time for 2 hours, and the amount of water used is 8 to 16 times the amount of the medicinal material.

[0015] Furthermore, in step 2, the macroporous resin is AB-8 macroporous resin, and the volume ratio of ethanol to water is 30:70, 50:50, 70:30 and 100:0 for gradient elution.

[0016] Furthermore, in step 3, the volume ratio of methanol to water is 10:90, 30:70, 50:50, 70:30 and 100:0 for gradient elution; and the particle size of ODS is 40 to 70 μm.

[0017] Furthermore, in step 4, the volume ratio of methanol to water is 10:90, 30:70, 50:50, 70:30 and 100:0 for gradient elution; and the particle size of ODS is 40 to 70 μm.

[0018] Furthermore, in step 5, the methanol elution procedure is 10% methanol isocratic elution.

[0019] Furthermore, the pretreatment process of the ODS and dextran gel is: soaking in methanol for 24 hours, loading on the column, washing with methanol until there is no turbidity when dripped into water, and then balancing with the initial mobile phase.

[0020] Furthermore, in step 6, the volume ratio of methanol to 0.1% formic acid is 10:90, and the retention time of the compound is 3.506 min.

[0021] The present invention also provides a use of the 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol isolated from the purslane medicinal material in the preparation of anti-inflammatory drugs, anticholinesterase drugs and antioxidant drugs.

[0022] Compared with the prior art, the invention has the following beneficial effects.

[0023] The separation and pharmacological activity research of the compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol in purslane described in the present invention have not been reported in plants; the present invention provides a new alkaloid compound derived from purslane and an extraction and separation method for the compound of the present invention, which sequentially adopts water decoction extraction, macroporous resin column chromatography, ODS column chromatography, dextran gel column (Sephadex LH-20) chromatography and high performance liquid chromatography for separation, purification and preparation, and successfully extracts and separates a new alkaloid compound. The method has only six operating steps, the operating method is simple and fast, water is used for extraction, the process method is environmentally friendly, and the purity of the compound separated by the method is high, greater than 90%; in addition, studies have shown that the above compounds have anti-inflammatory activity, anticholinesterase activity and antioxidant activity, so the new compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol and its salts and derivatives can be used as raw materials for drug development and pharmacological activity research. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The novel alkaloid compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention 1 H-NMR spectrum.

[0025] Figure 2 The novel alkaloid compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention 13 C-NMR spectrum.

[0026] Figure 3 The DEPT spectrum of the new alkaloid compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention is shown in FIG.

[0027] Figure 4 The HSQC spectrum of the new alkaloid compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention is shown in FIG.

[0028] Figure 5 This is the HMBC spectrum of the new alkaloid compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention.

[0029] Figure 6 The figure is the COESY spectrum of the new alkaloid compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention.

[0030] Figure 7 The ROESY spectrum of the new alkaloid compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention is shown in FIG.

[0031] Figure 8 It is a high-resolution mass spectrum of the new alkaloid compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention. DETAILED DESCRIPTION

[0032] The following examples will help to understand the present invention, but these examples are only for illustrating the present invention, and the present invention is not limited to these contents. The operating methods in the examples are all conventional operating methods in the art.

[0033] Example 1 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol.

[0034] The present invention provides a piperidine alkaloid compound having a molecular formula of C 10 H 14 N6O3, chemical name is 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol, chemical structure is:

[0035]

[0036] Table 1 shows the NMR data of 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol. The solvent used in NMR is DMSO-d6.

[0037] Table 1 NMR data of 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention.

[0038]

[0039] 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol: yellow-brown oily substance, easily soluble in water. After being spotted on a silica gel thin layer plate, the spots are sprayed with dilute potassium bismuth iodide test solution and appear orange-yellow, indicating that the compound is an alkaloid component. 1 H-NMR, 13 C-NMR and HR-ESI-TOF-MS signals indicate that the possible molecular formula of the compound is C 10 H 14 N6O3, unsaturation is 7. HR-ESI-TOF-MS gives m / z [MH] - The quasi-molecular ion peak is 265.1050, and the calculated value (C 10 H 13 N6O3 - =265.1054). According to 13 C NMR and DEPT spectra show that there are 10 carbon resonance peaks in the compound, including 1 methylene (δ C 61.26), 6 methines (δ C 88.30,73.56,70.54,85.71,151.97,140.49), 3 quaternary carbon resonance peaks (δ C The NMR data of the compound are listed in Table 1. 1 The H NMR spectrum shows two isolated heteroaromatic methine resonances (δ H 8.11,8.24) according to 1 In the H-NMR spectrum and HMBC spectrum, H-2'(δ H 8.11) respectively with C-4'(δ C 148.32) and C-5'(δ C 118.90) has a strong correlation; H-8'(δ H 8.24) have strong correlations with C-4' and C-5', respectively, and according to 13The chemical shifts of C-2', C-4', and C-8' in the C-NMR spectrum are in the low field, proving that they are connected to N atoms respectively; and the chemical shift of C-6' is in a lower field compared with the above, inferring that it is connected to -NH2; the chemical shift of C-5' is in a higher field relative to the above three C atoms, proving that its other end is connected to a C atom, thus proving the existence of a purine structure. 1 H-NMR spectrum and HMBC spectrum, H-4(δ H 4.35) and C-2(δ C 88.30), C-6(δ C 61.36) related, H-3(δ H 4.69) and C-5(δ C 85.71) related, H-6 (δ C 3.76,3.83) and C-2(δ C 88.30), indicating the existence of a six-membered ring. 1 H- 1 The correlation between H-2 and H-3, the correlation between H-3 and H-4, the correlation between H-4 and H-5, and the correlation between H-5 and H-6 in the HCOSY spectrum can support the above-mentioned six-membered ring. 13 The chemical shifts of C-3, C-4 and C-5 in the C-NMR spectrum are in the low field, which indicates that they are connected to hydroxyl groups respectively; the chemical shift of C-6 is also in the low field, which indicates that it may be connected to the N atom, and it is inferred that there is a piperidine structure. According to the strong correlation between H-2 and C-4' and C-8' in the HMBC spectrum, it is shown that the piperidine ring is connected to the purine ring through C-2-N-9'. Based on the above information, combined with the molecular weight of the HR-ESI-TOF-MS spectrum, it can be determined that this new alkaloid has the above structure.

[0040] The present invention also provides a method for extracting and separating 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol, which comprises the following specific steps:

[0041] Step 1: Weigh 250 kg of dried purslane medicinal materials, use water decoction to extract, the amount used is 10 times the medicinal material, extract twice, each time for 2 hours, combine the extracts, heat and concentrate, cool to room temperature, and obtain the medicinal solution for use.

[0042] Step 2: evaporate the drug solution obtained in step 1 to dryness, separate it through AB-8 macroporous resin column chromatography, use ethanol-water (30:70, 50:50, 70:30 and 100:0, v:v) gradient elution, collect the 30% ethanol portion, recover it to the extract under reduced pressure, and obtain the concentrate for use.

[0043] Step 3: Separate the concentrate in step 2 through a pretreated ODS column with a filler particle size of 40 to 70 μm, and use methanol-water (10:0, 30:70, 50:50, 70:30 and 100:0, v:v) gradient elution, detect by thin layer chromatography, develop color, combine the 30% methanol elution fractions, concentrate to dryness under reduced pressure, and set aside.

[0044] Step 4: The obtained product in step 3 is separated by pre-treated ODS column chromatography, the filler particle size is 40-70 μm, and gradient elution is performed with methanol-water (10:0, 30:70, 50:50, 70:30 and 100:0, v / v), and detected by thin layer chromatography, color development, and the 30% methanol elution part is combined, concentrated to dryness under reduced pressure, and used for standby. The ODS pretreatment process is to soak in methanol for 24 hours, after loading the column, wash with methanol until there is no turbidity when dripped into water, and then balance with the initial mobile phase.

[0045] Step 5: The obtained product in step 4 is further chromatographed and separated on a pretreated Sephadex LH-20 column, and isocratically eluted with 10% methanol to obtain several elution parts, which are detected by thin layer chromatography, color developed, and the elution parts with color developed are combined, and the combined elution parts are concentrated to dryness under reduced pressure for standby use. The pretreatment process of the dextran gel is to soak it in methanol for 24 hours, load it onto the column, wash it with methanol until there is no turbidity when it is dropped into water, and then balance it with the initial mobile phase.

[0046] Step 6: The fraction obtained in step 5 was separated and prepared by HPLC, with methanol and 0.1% formic acid in a volume ratio of 10:90 as the mobile phase and detection wavelengths of 210 and 254 nm to separate and prepare the new alkaloid compound of the present invention, and the purity determined by normalization method was 96%.

[0047] Example 2 Anti-inflammatory effect of 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention.

[0048] 1Main materials.

[0049] 1.1 Drugs and reagents: The new alkaloids used in the experiment were prepared by the above method with a purity of 96%. DMEM high-glucose medium, fetal bovine serum (Hyclone, USA); penicillin and streptomycin (Hangzhou Sijiqing Company); LPS (Sigma, USA); ELISA kits for IL-1β, TNF-α and inflammatory mediator PGE2 (Cayman, USA); cell lysate.

[0050] 1.2 Cell line: RAW264.7 macrophages (ATCC cell bank, USA).

[0051] 1.3 Grouping: Divided into control group, LPS group and experimental group.

[0052] 2 Experimental methods.

[0053] 2.1 Cell culture: Add 10% fetal bovine serum and 1% antibiotics (100 U / mL penicillin and 100 μg / mL streptomycin) to DMEM high-glucose medium and store at 4°C.

[0054] 2.2 CCK-8 method to determine cell viability: The three groups of RAW264.7 macrophages in logarithmic growth phase were inoculated in 96-well culture plates at a cell density of 1×10 4 / mL, 100μL per well, 37℃, 5% CO2, after overnight culture, different concentrations of the compound of the present invention 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol (5μM~50μM) were added to the experimental group, and LPS with a concentration of 1μg / mL was added to the LPS group and the experimental group after incubation for 1h. A zero adjustment group (culture solution containing DMSO solvent) was set up, and 3 duplicate wells were set up in each group to investigate the effect of the addition of drugs on the cells. After the cells in the above groups were cultured for 24h, 10μL of CCK-8 was added to the cells in each well, and the cells were incubated for another 4h at 37℃ and 5% CO2, and the absorbance of each well was measured at a wavelength of 450nm by an enzyme marker.

[0055] 2.3ELISA method for determination of inflammatory factors IL-1β, TNF-α and inflammatory mediator PGE2: RAW264.7 macrophages in logarithmic growth phase were inoculated in 24-well culture plates at a cell density of 1×10 5 / mL, 1mL per well, cultured overnight at 37°C and 5% CO2, the experimental group added the compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol (1μM~20μM) of the present invention and cultured for 1h, then LPS (final concentration of 1μg / mL) was added to each well, and incubated for 24h, with 3 wells in each group. ELISA method was used to determine the content of IL-1β, TNF-α and inflammatory mediator PGE2.

[0056] 3 Experimental results.

[0057] The experimental results show that the compound of the present invention has no effect on the proliferation of LPS-induced macrophages RAW264.7; it can effectively inhibit the secretion of excessive inflammatory cytokines IL-1β, TNF-α and inflammatory mediator PGE2 produced by LPS-induced macrophages RAW264.7 in a concentration-dependent manner.

[0058] The relative cell viability test results of the inventive compounds are shown in Table 2.

[0059] Table 2 Effects of the compounds of the present invention on the relative survival rate of RAW264.7 macrophages.

[0060]

[0061] The results of ELISA determination of inflammatory factors IL-1β, TNF-α and inflammatory mediator PGE2 are shown in Table 3.

[0062] Table 3 Effects of the compounds of the present invention on the levels of IL-1β, TNF-α and inflammatory mediator PGE2 secreted by RAW264.7 cells induced by LPS (mean±standard deviation, n=3).

[0063]

[0064]

[0065] Note: * P<0.05 compared with the control group. # P<0.05 compared with LPS group.

[0066] Example 3 Anticholinesterase effect of 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention.

[0067] 1Main materials.

[0068] 1.1 Drugs and reagents: The new alkaloids used in the experiment were prepared by the above method with a purity of 96%. Physostigmine (Shanghai Hanxiang Biotechnology Co., Ltd.), acetylcholine iodide (ATCI) and acetylcholinesterase (AChE) (Dalian Meilun Biotechnology Co., Ltd.), dithiodinitroformic acid (DTNB) (Shanghai Jinshui Biotechnology Co., Ltd.), disodium hydrogen phosphate and sodium dihydrogen phosphate (Shanghai Sinopharm Reagent Co., Ltd.).

[0069] 1.2 Experimental instruments and equipment: HBS-1096A96-well microplate reader (Nanjing Detie Experimental Equipment Co., Ltd.), 1 / 100,000 balance (METTLER, Switzerland), HH-4 digital display constant temperature water bath (Jiangsu Jintan Ronghua Instrument Manufacturing Co., Ltd.).

[0070] 2 Experimental methods.

[0071] In this experiment, the anticholinesterase activity of each compound was determined according to the modified Ellman method. 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol and physostigmine were accurately weighed and prepared into five series of sample solutions with concentrations of 31.25μM, 62.5μM, 125μM, 250μM and 500μM using methanol. The specific operation is as follows: add 140 μL of phosphate buffered saline (0.1M, pH=8.0, containing 0.1 mol / L disodium hydrogen phosphate and sodium dihydrogen phosphate), 20 μL of sample solution, 15 μL of AChE (0.2U / mL) to a 96-well ELISA plate, incubate at 37°C for 10 minutes, add 10 μL of ATCI (4mmol / L) and 10 μL of DTNB (15mmol / L), incubate at 37°C for 20 minutes, place the 96-well plate in an ELISA instrument, and measure the absorbance of each group of samples at a wavelength of 405nm. Among them, methanol was used instead of the sample solution as the blank group, and physostigmine was used instead of the sample solution as the positive control group. The cholinesterase inhibition rate of each compound was calculated according to the following formula (A represents absorbance):

[0072] Inhibition rate (%) = (A 空白 -A 样品 ) / A 空白 ×100%

[0073] 3 Experimental results.

[0074] The experimental results show that the novel alkaloid compound of the present invention exhibits certain anticholinesterase activity, and the inhibitory effect on cholinesterase increases with the increase of the compound concentration, showing a dose-dependent trend.

[0075] The anticholinesterase activities of the novel alkaloids of the present invention are shown in Table 4.

[0076] Table 4 Anticholinesterase activities of the compounds of the present invention.

[0077]

[0078] Example 4 Antioxidant effect of 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol of the present invention.

[0079] 1Main materials.

[0080] 1.1 Drugs and reagents: The new alkaloid used in the experiment was prepared by the above method with a purity of 96%. 1,1-Diphenyl-2-picrylhydrazyl free radical (DPPH) (Sigma, USA), butylated hydroxyanisole (BHA) (Shanghai Xiangrui Biological Co., Ltd.), methanol (chromatographic grade, Tianjin Kaixin Chemical Industry Co., Ltd.).

[0081] 1.2 Experimental instruments and equipment: Hitachi UV-3010 ultraviolet-visible spectrophotometer (Hitachi, Japan), 1 / 100,000 balance (METTLER, Switzerland).

[0082] 2 Experimental methods.

[0083] In this experiment, the DPPH free radical scavenging method was used to determine the antioxidant activity of each compound. 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol and BHA were accurately weighed, and five series of sample solutions with concentrations of 12.5μM, 25μM, 50μM, 100μM and 200μM were prepared with methanol. In addition, the DPPH solution was prepared as needed, and an appropriate amount of DPPH was accurately weighed and prepared with methanol to a solution with a concentration of 80μM, and the whole process was protected from light. The specific operation is as follows: 1mL of the sample solution was fully mixed with 1mL of the DPPH solution, and placed at room temperature and in a light-proof environment for 10min. After setting the detection wavelength of the ultraviolet spectrophotometer to 517nm, the absorbance value of the sample was measured. Among them, methanol was used instead of the sample solution as the blank group, BHA was used instead of the sample solution as the positive control group, and a mixed solution of 1mL of methanol and 1mL of the sample solution was used as the control group. The DPPH scavenging rate of each compound was calculated according to the following formula (A represents absorbance):

[0084] DPPH clearance rate (%) = (1-(A 样品 -A 对照 ) / A 空白 )×100%

[0085] 3 Experimental results.

[0086] The experimental results show that the novel alkaloid compound of the present invention exhibits certain antioxidant activity, and the antioxidant effect increases with the increase of the compound concentration, showing a dose-dependent trend.

[0087] The antioxidant activities of the novel alkaloid compounds of the present invention are shown in Table 5.

[0088] Table 5 Antioxidant activity of the compounds of the present invention.

[0089]

[0090] In summary, the present invention provides a compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol and an extraction and separation method thereof, which uses water decoction extraction, macroporous resin column chromatography, ODS column chromatography, Sephadex LH-20 and high performance liquid chromatography for separation, purification and preparation, and successfully extracts and separates a new alkaloid compound. The operation method is simple, rapid and environmentally friendly, and the compound separated by the method has a high purity. Since the obtained compound is extracted from the commonly used traditional Chinese medicine Portulaca oleracea, it has anti-inflammatory activity, anticholinesterase activity and antioxidant activity; therefore, the new compound 2-(6-amino-9H-purin-9-yl)piperidine-3,4,5-triol and its salts and derivatives of the present invention can provide ideas for drug development and have broad development prospects.

Claims

1. A piperidine alkaloid compound isolated from Portulaca oleracea, characterized in that: The molecular formula is: C 10 H 14 N6O3, chemical name is 2-(6-amino-9 H -purin-9-yl)piperidine-3,4,5-triol, the chemical structure is: 。 2. A method for extracting and separating a piperidine alkaloid compound from Portulaca oleracea, characterized in that: The specific steps include: Step 1: Take dried purslane medicinal material, decoct it in water, concentrate the extract, cool it to room temperature, and obtain the medicinal solution for later use; Step 2: The concentrated solution in step 1 is subjected to a macroporous resin, and eluted with water and ethanol of different concentrations, and the 30% ethanol portion is recovered under reduced pressure to obtain an extract, and a concentrate is obtained for standby use; Step 3: Separate the concentrate in step 2 through an ODS column, use a methanol-water gradient elution, detect by thin layer chromatography, develop color, combine the 30% methanol elution portion and evaporate to dryness to obtain a concentrate for use; Step 4: The concentrate obtained in step 3 is separated by ODS column chromatography, eluted by methanol-water gradient, detected by thin layer chromatography, color developed, and the 30% methanol elution portion is combined and evaporated to dryness to obtain a concentrate for use; Step 5: The product obtained in step 4 is further separated by pre-treated dextran gel column chromatography, isocratically eluted with 10% methanol to obtain several elution fractions, which are detected by thin layer chromatography, developed, and the developed elution fractions are combined, and the combined elution fractions are concentrated to dryness under reduced pressure for later use; Step 6: The concentrate obtained in step 5 is separated and prepared by HPLC, and isocratic elution is performed using methanol-0.1% formic acid (volume percentage) as a mobile phase to finally obtain the new alkaloid compound of the present invention.

3. The extraction and separation method according to claim 2, characterized in that: In the step 1, the extraction is performed by decoction twice, each time for 2 hours, and the amount of water used is 8 to 16 times the amount of the medicinal material.

4. The extraction and separation method according to claim 2, characterized in that: In the step 2, the macroporous resin is AB-8 macroporous resin, and the volume ratio of ethanol to water is 30:70, 50:50, 70:30 and 100:0 for gradient elution.

5. The extraction and separation method according to claim 2, characterized in that: In the step 3, the volume ratio of methanol to water is 10:90, 30:70, 50:50, 70:30 and 100:0 for gradient elution; and the particle size of ODS is 40-70 μm.

6. The extraction and separation method according to claim 2, characterized in that: In step 4, the volume ratio of methanol to water is 10:90, 30:70, 50:50, 70:30 and 100:0 for gradient elution; and the particle size of ODS is 40-70 μm.

7. The extraction and separation method according to claim 2, characterized in that: In step 5, the methanol elution procedure is 10% methanol isocratic elution.

8. The extraction and separation method according to claim 2, characterized in that: The pretreatment process of the ODS and dextran gel is as follows: soaking in methanol for 24 hours, loading onto the column, washing with methanol until there is no turbidity when dripped into water, and then balancing with the initial mobile phase.

9. The extraction and separation method according to claim 2, characterized in that: In step 6, the volume ratio of methanol to 0.1% formic acid is 10:90, and the retention time of the compound is 3.506 min.

10. A use of the piperidine alkaloid compound as claimed in claim 1, characterized in that: The compound is used for preparing anti-inflammatory drugs, anticholinesterase drugs and antioxidant drugs or health products.

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