Beta-carboline alkaloid coumaroyl glucoside compound, extraction method and application thereof, and monoamine oxidase inhibitor

Abstract

The present application provides a beta-coclinic alkaloid coumaroyl glucoside compound, an extraction method and application thereof, and a monoamine oxidase inhibitor. The beta-coclinic alkaloid coumaroyl glucoside compound provided by the present application has a structure shown in formula 1 or formula 2, is novel in structure, has good monoamine oxidase inhibitory activity, can slow down the consumption of monoamine neurotransmitters in the brain, slow down oxidative damage, realize the treatment of neurodegenerative diseases, and can be used for preparing a monoamine oxidase inhibitor or an anti-neurodegenerative disease drug, and has a wide application prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of traditional Chinese medicine, and particularly relates to a beta-carboline alkaloid feruloyl glucoside compound, an extraction method and application thereof, and a monoamine oxidase inhibitor. BACKGROUND

[0002] Monoamine oxidase (MAO) is a class of vital flavoproteins, which exist on the outer membrane of mitochondria and are subdivided into two subtypes, MAO-A and MAO-B. Monoamine oxidase plays an important role in regulating the metabolism of monoamine neurotransmitters in the human body, and is closely related to a series of neurodegenerative diseases such as depression, Parkinson's disease and Alzheimer's disease.

[0003] Specifically, MAO-A has strong catalytic activity on substances such as serotonin and adrenaline, while MAO-B shows higher preference for the metabolism of substances such as benzylamine and beta-phenylethylamine. Serotonin, adrenaline, benzylamine and beta-phenylethylamine play a vital role in regulating movement, emotion, perception and cognitive function, and their imbalance is often closely related to the occurrence of diseases such as depression, Parkinson's disease and Alzheimer's disease.

[0004] When the activity of monoamine oxidase in the brain is abnormally enhanced, the metabolism of monoamine neurotransmitters is accelerated, and a large amount of hydrogen peroxide is generated. This process not only leads to functional disorders of the nervous system, but also may cause oxidative stress damage, which is an important predisposing factor for various neurodegenerative diseases. In fact, scientists have found a significant increase in the expression level of monoamine oxidase in brain tissue samples of patients with depression, Parkinson's disease and Alzheimer's disease.

[0005] In view of the core role of monoamine oxidase in neurotransmitter metabolism and disease pathogenesis, the development of effective monoamine oxidase inhibitors has become a research hotspot. Such inhibitors are expected to slow down the consumption rate of monoamine neurotransmitters by inhibiting the activity of monoamine oxidase, thereby reducing oxidative stress damage and providing new hope for the intervention and treatment of neurodegenerative diseases such as depression and Parkinson's disease. At present, there are few studies on monoamine oxidase inhibitors, and new types are urgently needed. SUMMARY

[0006] The present application aims to provide a beta-carboline alkaloid feruloyl glucoside compound, an extraction method and application thereof, and a monoamine oxidase inhibitor. The beta-carboline alkaloid feruloyl glucoside compound provided by the present application has good monoamine oxidase inhibitory activity and can be used for preparing a novel monoamine oxidase inhibitor or a novel anti-neurodegenerative disease drug.

[0007] To achieve the above-mentioned object of the application, the application provides the following technical solutions.

[0008] The application provides a β-carboline alkaloid feruloyl glucoside compound having a structure shown in formula 1 or formula 2.

[0009]

[0010] The application provides an extraction method of the β-carboline alkaloid feruloyl glucoside compound described in the above technical solution, comprising the following steps.

[0011] (1) Alcohol precipitation is performed on fruit juice of Nitraria tangutorum Bob, to obtain an extract;

[0012] (2) The extract is dissolved in water, and macroporous resin column chromatography is performed on the extract solution by using pure water and 40% ethanol aqueous solution as eluents in sequence, to obtain an elution part of 40% ethanol aqueous solution;

[0013] (3) Column chromatography separation is performed on the elution part of 40% ethanol aqueous solution under gradient elution conditions by using MCI resin as a stationary phase and methanol-water as an eluent, to obtain four components, denoted as Fr 40-1 ~Fr 40-4 ; the volume fraction of methanol in the eluent of step (3) is 0% to 80%;

[0014] (4) Column chromatography separation is performed on the Fr 40-1 component under gradient elution conditions by using MCI resin as a stationary phase and methanol-water as an eluent, to obtain eight components, denoted as Fr 40-1-1 ~Fr 40-1-8 ; the volume fraction of methanol in the eluent of step (4) is 0% to 50%;

[0015] (5) Column chromatography separation is performed on the Fr 40-1-8 component under gradient elution conditions by using C18 bonded silica gel as a stationary phase and acetonitrile-water as an eluent, to obtain five components, denoted as Fr 40-1-8-1 ~Fr 40-1-8-5 ; the volume fraction of acetonitrile in the eluent of step (5) is 15% to 35%.

[0016] (6) Column chromatography separation is performed on the Fr 40-1-8-3 component under isocratic elution conditions by using C18 bonded silica gel as a stationary phase and acetonitrile-water as an eluent, to obtain a β-carboline alkaloid feruloyl glucoside compound having a structure shown in formula 1; the volume fraction of acetonitrile in the eluent of step (6) is 15%;

[0017] (7) performing column chromatography separation on the Fr 40-1-8-5 components under isocratic elution condition with C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent to obtain a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 2; the volume fraction of acetonitrile in the eluent of the step (7) is 20%;

[0018] The step (6) and the step (7) are not limited in time sequence.

[0019] Preferably, in the step (1), the reagent used for the alcohol precipitation is an ethanol aqueous solution; the volume fraction of the ethanol aqueous solution is 50% to 75%.

[0020] Preferably, the alcohol precipitation is performed for 3 to 4 times; the time for each alcohol precipitation is independently 10 to 12 hours, and the volume ratio of the ethanol aqueous solution to the Nitraria tangutorum Bob. fruit juice is independently 2 to 4:1.

[0021] Preferably, in the step (3), the volume fraction of methanol in the eluent used for the column chromatography separation is 0%, 20%, 40% and 80% in sequence.

[0022] Preferably, in the step (4), the volume fraction of methanol in the eluent used for the column chromatography separation is 0%, 10%, 20%, 30%, 40% and 50% in sequence.

[0023] Preferably, in the step (2), the chromatographic column used for the macroporous resin column chromatography is an AB-8 macroporous resin chromatographic column.

[0024] Preferably, in the step (6), the Fr 40-1-8-3 components are subjected to the column chromatography separation by using a preparative liquid XCharge C18 chromatographic column;

[0025] In the step (7), the Fr 40-1-8-5 components are subjected to the column chromatography separation by using a preparative liquid XCharge C18 chromatographic column.

[0026] The application provides application of the β-carboline alkaloid coumaroyl glucoside compound in the above technical solution in preparation of a monoamine oxidase inhibitor or an anti-neurodegenerative disease drug.

[0027] The application provides a monoamine oxidase inhibitor containing the β-carboline alkaloid coumaroyl glucoside compound in the above technical solution.

[0028] The present application provides a beta-carboline alkaloid coumaroyl glucoside compound with a structure shown in formula 1 or formula 2, which is novel in structure, has good monoamine oxidase inhibitory activity, can slow down the consumption of monoamine neurotransmitters in the brain, slow down oxidative damage, achieve the treatment of neurodegenerative diseases, and can be used for preparing a monoamine oxidase inhibitor or an anti-neurodegenerative disease drug, and has a wide application prospect. The examples show that the beta-carboline alkaloid coumaroyl glucoside compound has good MAO-A inhibitory activity and MAO-B inhibitory activity; the IC 50 value of the beta-carboline alkaloid coumaroyl glucoside compound with the structure shown in formula 1 for MAO-A inhibitory activity is 121.7 μmol / L, the IC 50 value for MAO-B inhibitory activity is 113.2 μmol / L; the IC 50 value of the beta-carboline alkaloid coumaroyl glucoside compound with the structure shown in formula 2 for MAO-A inhibitory activity is 101.2 μmol / L, and the IC 50 value for MAO-B inhibitory activity is 115.9 μmol / L.

[0029] The present application also provides an extraction method of the beta-carboline alkaloid coumaroyl glucoside compound, which uses the fruits of Tangutian white thorn as raw materials, and separates the beta-carboline alkaloid coumaroyl glucoside compound by using ethanol extraction combined with a series of column chromatography separation. The extraction method is simple and fast, and the purity of the beta-carboline alkaloid coumaroyl glucoside compound obtained by the method is higher than 95% BRIEF DESCRIPTION OF DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0031] Figure 1 The H-NMR spectrum of the beta-carboline alkaloid coumaroyl glucoside compound Tangutoid LI is shown in the following figure. 1

[0032] Figure 2 The C-NMR spectrum of the beta-carboline alkaloid coumaroyl glucoside compound Tangutoid LI is shown in the following figure. 13

[0033] Figure 3 The HSQC spectrum of the beta-carboline alkaloid coumaroyl glucoside compound Tangutoid LI is shown in the following figure.​​

[0034] Figure 4 HMBC spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LI;

[0035] Figure 5 HSQC spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LI; 1 H- 1 H COSY spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LI;

[0036] Figure 6 NOESY spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LI;

[0037] Figure 7 TOCSY spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LI;

[0038] Figure 8 (-)-HRESIMS spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LI;

[0039] Figure 9 IR spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LI;

[0040] Figure 10 UV spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LI;

[0041] Figure 1 HMBC spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LII; 1 H-NMR spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;

[0042] Figure 2 HSQC spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LII; 13 C-NMR spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;

[0043] Figure 3 HSQC spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;

[0044] Figure 4 HMBC spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;

[0045] Figure 5 HSQC spectrum of β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;1 H- 1 H COSY spectrum;

[0046] Figure 6 NOESY spectrum of the β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;

[0047] Figure 7 TOCSY spectrum of the β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;

[0048] Figure 8 (-)-HRESIMS spectrum of the β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;

[0049] Figure 9 IR spectrum of the β-carboline alkaloid feruloyl glucoside compound Tangutoid LII;

[0050] Figure 10 UV spectrum of the β-carboline alkaloid feruloyl glucoside compound Tangutoid LII. DETAILED DESCRIPTION

[0051] The present application provides a β-carboline alkaloid feruloyl glucoside compound, having or the structure shown in formula 2:

[0052]

[0053] The molecular formula of the β-carboline alkaloid feruloyl glucoside compound is C 32 H 36 N2O 13 The structure is novel, has good monoamine oxidase inhibitory activity, can slow down the consumption of monoamine neurotransmitters in the brain, slow down oxidative damage, achieve the treatment of neurodegenerative diseases, and can be used for preparing a monoamine oxidase inhibitor or an anti-neurodegenerative disease drug, and has a wide application prospect.

[0054] The present application also provides an extraction method of the β-carboline alkaloid feruloyl glucoside compound in the above technical solution, comprising the following steps:

[0055] (1) alcohol precipitation is carried out on the fruit juice of Nitraria tangutorum Bob, to obtain an extract;

[0056] (2) the extract is dissolved with water, and macroporous resin column chromatography is carried out on the extract liquid by using pure water and 40% (volume fraction) ethanol aqueous solution as eluent in sequence, to obtain the elution part of 40% (volume fraction) ethanol aqueous solution;

[0057] (3) column chromatography separation of the elution part of the 40% volume fraction of the ethanol aqueous solution under gradient elution conditions with MCI resin as the stationary phase and methanol-water as the eluent to obtain four components, denoted as Fr 40-1 ~Fr 40-4 ; the volume fraction of methanol in the eluent of step (3) is 0%-80%;

[0058] (4) column chromatography separation of the Fr 40-1 components under gradient elution conditions with MCI resin as the stationary phase and methanol-water as the eluent to obtain eight components, denoted as Fr 40-1-1 ~Fr 40-1-8 ; the volume fraction of methanol in the eluent of step (4) is 0%-50%;

[0059] (5) column chromatography separation of the Fr 40-1-8 components under gradient elution conditions with C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent to obtain five components, denoted as Fr 40-1-8-1 ~Fr 40-1-8-5 ; the volume fraction of acetonitrile in the eluent of step (5) is 15%-35%.

[0060] (6) column chromatography separation of the Fr 40-1-8-3 components under isocratic elution conditions with C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent to obtain a β-carboline alkaloid coumaroyl glucoside compound with the structure shown in formula 1; the volume fraction of acetonitrile in the eluent of step (6) is 15%;

[0061] (7) column chromatography separation of the Fr 40-1-8-5 components under isocratic elution conditions with C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent to obtain a β-carboline alkaloid coumaroyl glucoside compound with the structure shown in formula 2; the volume fraction of acetonitrile in the eluent of step (7) is 20%;

[0062] The steps (6) and (7) are not limited in time sequence.

[0063] In the present application, if not specifically stated, the raw materials used are all commercially available goods well known to those skilled in the art or are prepared by methods well known to those skilled in the art.

[0064] The present application obtains an extract by alcohol precipitation of the Nitraria tangutorum fruit juice. The present application obtains Nitraria tangutorum fruit juice by crushing and concentrating the Nitraria tangutorum fruit successively. In the specific embodiments of the present application, the crushing method is juicing. The present application obtains crushed material by crushing the Nitraria tangutorum fruit, and obtains Nitraria tangutorum fruit juice by concentrating the crushed material.

[0065] After obtaining the juice of *Nitraria tangutorum*, the present invention performs alcohol precipitation on the juice to obtain an extract. The reagent used for alcohol precipitation in the present invention can be an aqueous ethanol solution. In the present invention, the volume fraction of the aqueous ethanol solution can be 50%–75%; in specific embodiments of the present invention, the volume fraction of the aqueous ethanol solution can be 50%, 55%, 60%, 65%, 70%, or 75%. The number of alcohol precipitation cycles in the present invention can be 3–4 times; in specific embodiments of the present invention, the number of alcohol precipitation cycles can be 3 or 4 times. The present invention does not have special requirements for the temperature of alcohol precipitation; conventional operating temperatures in the art are acceptable. In the embodiments of the present invention, alcohol precipitation is carried out at room temperature (20–35°C). In this invention, the alcohol precipitation time for each precipitation can be independently set to 10–12 hours, and the volume ratio of the ethanol-water solution to the Tangut thorn juice can be independently set to 2–4:1. In specific embodiments of this invention, the alcohol precipitation time for each precipitation is independently set to 10, 11, or 12 hours, and the volume ratio of the ethanol-water solution to the Tangut thorn juice is independently set to 2:1, 3:1, or 4:1. After each alcohol precipitation, the resulting liquid can be subjected to solid-liquid separation, and the liquid material can be collected and used as the material to be processed for the next extraction. After the final alcohol precipitation, the resulting liquid material is concentrated to obtain the extract. In this invention, the solid-liquid separation can be performed by filtration; the concentration can be performed by vacuum distillation. This invention uses alcohol precipitation to remove proteins, pectin, and polysaccharides from the Tangut thorn juice; simultaneously, ethanol can be recovered by distillation to obtain a crude extract containing β-carboline alkaloids, coumaroyl glucoside compounds.

[0066] After obtaining the extract, the present invention dissolves the extract in water, and sequentially uses pure water and a 40% (v / v) ethanol aqueous solution as eluents to perform macroporous resin column chromatography, obtaining a 40% (v / v) ethanol aqueous solution eluent. In the present invention, the macroporous adsorption resin used for macroporous adsorption column chromatography can be AB-8 macroporous resin. In a specific embodiment of the present invention, the extract is loaded onto an AB-8 macroporous resin chromatographic column, and sequentially uses pure water, a 40% (v / v) ethanol aqueous solution, and a 95% (v / v) ethanol aqueous solution as eluents for column chromatography separation, obtaining pure water eluent, a 40% (v / v) ethanol aqueous solution eluent, and a 95% (v / v) ethanol aqueous solution eluent, respectively. Subsequent further separation processes do not involve the pure water eluent and the 95% (v / v) ethanol aqueous solution eluent, and will not be described in detail here.

[0067] After obtaining the elution fraction of 40% ethanol aqueous solution, the present application carries out column chromatography separation of the elution fraction of 40% ethanol aqueous solution with MCI resin as the stationary phase and methanol-water as the eluent under gradient elution condition, to obtain 4 components, denoted as Fr 40-1 ~Fr 40-4 In the specific embodiment of the present application, the elution fraction of 40% ethanol aqueous solution is dissolved with water and then loaded on the MCI gel chromatography column for column chromatography separation. In the present application, the volume fraction of methanol in the eluent used in the column chromatography separation is 0% to 80%; in the specific embodiment of the present application, the volume fraction of methanol in the eluent used in the column chromatography separation is 0%, 20%, 40% and 80% in turn, i.e. the eluent used in the column chromatography separation is pure water, 20% methanol-water, 40% methanol-water and 80% methanol-water in turn. After elution with 80% methanol-water as the eluent is completed, the present application can use pure methanol as the eluent to wash the chromatography column used in the column chromatography separation. In the specific embodiment of the present application, the elution fraction of 40% ethanol aqueous solution is loaded on the MCI gel chromatography column, and pure water, 20% methanol-water, 40% methanol-water, 80% methanol-water and pure methanol are used as the eluent in turn for gradient elution.

[0068] After obtaining the Fr 40-1 component, the present application carries out column chromatography separation of the Fr 40-1 component with MCI resin as the stationary phase and methanol-water as the eluent under gradient elution condition, to obtain 8 components, denoted as Fr 40-1-1 ~Fr 40-1-8 In the present application, the volume fraction of methanol in the eluent used in the column chromatography separation is 0% to 50%; in the specific embodiment of the present application, the volume fraction of methanol in the eluent used in the column chromatography separation is 0%, 10%, 20%, 30%, 40% and 50% in turn, i.e. the eluent used in the column chromatography separation is pure water, 10% methanol-water, 20% methanol-water, 30% methanol-water, 40% methanol-water and 50% methanol-water in turn. After elution with 50% methanol-water as the eluent is completed, the present application can use pure methanol as the eluent to wash the chromatography column used in the column chromatography separation. In the specific embodiment of the present application, the Fr 40-1 component is loaded on the MCI gel chromatography column, and pure water, 10% methanol-water, 20% methanol-water, 30% methanol-water, 40% methanol-water, 50% methanol-water and pure methanol are used as the eluent in turn for gradient elution.

[0069] After obtaining the Fr 40-1-8 component, the present application carries out column chromatography separation of the Fr 40-1-7The components are subjected to column chromatography separation to obtain five components, denoted as Fr 40-1-8-1 ~ Fr 40-1-8-5 In the present application, the volume fraction of acetonitrile in the eluent is 15% to 35%. In the specific embodiment of the present application, Fr 40-1-8 The components are subjected to column chromatography separation to obtain five components, denoted as Fr

[0070] After obtaining Fr 40-1-8-3 The components are subjected to column chromatography separation to obtain five components, denoted as Fr 40-1-8-3 The components are subjected to column chromatography separation to obtain the β-carboline alkaloid coumaroyl glucoside compound with the structure shown in Formula 1. The present application is characterized in that the Fr 40-1-8-3 The components are subjected to column chromatography separation to obtain five components, denoted as Fr 40-1-8-3 The components are subjected to column chromatography separation to obtain five components, denoted as Fr

[0071] After obtaining Fr 40-1-8-5 The components are subjected to column chromatography separation to obtain five components, denoted as Fr 40-1-8-5 The components are subjected to column chromatography separation to obtain the β-carboline alkaloid coumaroyl glucoside compound with the structure shown in Formula 2. The present application is characterized in that the Fr 40-1-8-5 The components are subjected to column chromatography separation to obtain five components, denoted as Fr 40-1-8-5 The components are subjected to column chromatography separation to obtain five components, denoted as Fr

[0072] The present application uses the fruit of Nitraria tangutorum Bobr as raw material, adopts ethanol extraction, and combines a series of column chromatography separation to obtain the β-carboline alkaloid coumaroyl glucoside compound. The extraction method is simple and fast, and the purity of the β-carboline alkaloid coumaroyl glucoside compound obtained is above 95%.

[0073] The present application also provides the application of the β-carboline alkaloid coumaroyl glucoside compound in the preparation of monoamine oxidase inhibitors or anti-neurodegenerative disease drugs.

[0074] The beta-carboline alkaloid feruloyl glucoside compound has good MAO-A inhibitory activity and MAO-B inhibitory activity; the beta-carboline alkaloid feruloyl glucoside compound having the structure shown in formula 1 has an IC 50 value of 121.7 μmol / L, and an IC 50 value of 113.2 μmol / L; the beta-carboline alkaloid feruloyl glucoside compound having the structure shown in formula 2 has an IC 50 value of 101.2 μmol / L, and an IC 50 value of 115.9 μmol / L. The beta-carboline alkaloid feruloyl glucoside compound can slow down the consumption of monamine neurotransmitters in the brain, slow down oxidative damage, achieve the treatment of depression, Parkinson's disease and other neurodegenerative diseases, and thus can be used for preparing a monoamine oxidase inhibitor or an anti-neurodegenerative disease drug,

[0075] The application further provides a monoamine oxidase inhibitor containing the beta-carboline alkaloid feruloyl glucoside compound.

[0076] The technical solutions in the application will be clearly and completely described below with reference to the embodiments in the application. Obviously, the described embodiments are only some of the embodiments of the application, but not all the embodiments. Based on the embodiments in the application, all the other embodiments obtained by a person of ordinary skill in the art without creative work fall within the protection scope of the application.

[0077] The room temperature mentioned in the embodiments is 20-35 DEG C.

[0078] Embodiment 1

[0079] (1) 110 kg of Nitraria tangutorum fruits are juiced to obtain broken materials; the broken materials are concentrated under reduced pressure to obtain 25 L of Nitraria tangutorum fruit juice. 50 L of an ethanol aqueous solution (75% by volume) is added to the Nitraria tangutorum fruit juice, and alcohol precipitation is carried out at room temperature for 12 h, after which filtration is carried out, and the supernatant is collected; the above alcohol precipitation operation is repeated for three times, and the supernatant obtained by the last alcohol precipitation is distilled under reduced pressure to recover ethanol to obtain an extract.

[0080] (2) The extract obtained in step (1) is dissolved in water, and the obtained extract solution is loaded onto an AB-8 macroporous resin chromatographic column. Gradient elution is performed using pure water, 40% ethanol-water and 95% ethanol-water as eluents in sequence. After the end, pure water elution fractions (denoted as Fr0), 40% ethanol-water elution fractions (denoted as Fr 40 ) and 95% ethanol-water elution fractions (denoted as Fr 95 ) are obtained respectively.

[0081] (3) The Fr 40 component obtained in step (2) is dissolved in water, and then the obtained solution is loaded onto an MCI gel chromatographic column. Gradient elution is performed using pure water, 20% methanol-water, 40% methanol-water, 80% methanol-water and pure methanol in sequence. After the end, four components are obtained, denoted as Fr 40-1 ~ Fr 40-4 .

[0082] (4) The Fr 40-1 component obtained in step (3) is loaded again onto the MCI gel chromatographic column in step (3). Gradient elution is performed using pure water, 10% methanol-water, 20% methanol-water, 30% methanol-water, 40% methanol-water, 50% methanol-water and pure methanol in sequence. After the end, eight components are obtained, denoted as Fr 40-1-1 ~ Fr 40-1-8 .

[0083] (5) The Fr 40-1-8 component obtained in step (4) is loaded onto a preparative liquid phase Megress C18 chromatographic column. Gradient elution is performed using an aqueous acetonitrile solution with a volume fraction of 15% to 35%. After the end, five components are obtained in sequence, denoted as Fr 40-1-8-1 ~ Fr 40-1-8-5 .

[0084] (6) The Fr 40-1-8-3 component obtained in step (5) is loaded onto a preparative liquid phase XCharge C18 chromatographic column. Isocratic elution is performed using an aqueous acetonitrile solution with a volume fraction of 15%. A β-carboline alkaloid feruloyl glucoside compound (395 mg) in Nitraria tangutorum Bobr is obtained, and the β-carboline alkaloid feruloyl glucoside compound is named as compound Tangutoid LI.

[0085] (7) The Fr 40-1-8-5 component obtained in step (5) is loaded onto a preparative liquid phase XCharge C18 chromatographic column. Isocratic elution is performed using an aqueous acetonitrile solution with a volume fraction of 20%. A β-carboline alkaloid feruloyl glucoside compound (30.5 mg) in Nitraria tangutorum Bobr is obtained, and the β-carboline alkaloid feruloyl glucoside compound is named as compound Tangutoid LII

[0086] Test Example 1

[0087] Using nuclear magnetic resonance hydrogen spectroscopy (NMR) 1 H-NMR), carbon nuclear magnetic resonance (NMR) 13 C-NMR, heteronuclear single quantum correlation spectroscopy (HSQC), heteronuclear multi-bond correlation spectroscopy (HMBC), hydrogen-hydrogen correlation spectroscopy (H-NMR), ... 1 H- 1 The structures of the compounds Tangutoid LI and Tangutoid LII prepared in Example 1 were analyzed using H COSY, NOESY, TOCSY, HRESIMS, IR, and UV spectroscopy. The results are as follows: Figures 1 to 20 As shown, compounds Tangutoid LI and Tangutoid LII... 1 H-NMR and 13 C-NMR data were obtained in DMSO-d6, and the NMR data are shown in Table 1.

[0088] Table 1. Compounds Tangutoid LI and Tangutoid LII 1 H-NMR (600MHz) and 13 C-NMR (150MHz) data

[0089]

[0090]

[0091] The specific spectral data of compounds Tangutoid LI and Tangutoid LII are as follows:

[0092] Compound Tangutoid LI: white powder,[α] D 20.2 -24.25(c 0.24,MeOH); UV(MeOH)λ max nm(logε):314.50(0.2335); 291.00(0.2163); 222.00(0.4843); IR(KBr)ν max :3407,3068,2910,1686,1629,1513,1393,1170,1065cm-1; NMR spectroscopic date see table 1; HRESIMS m / z:655.2180[MH] -(calculated for C 32 H 35 N2O 13 655.2145).

[0093] Compound Tangutoid LII: white powder, [α] D 20.0 -23.70 (c 0.20, MeOH); UV (MeOH) λ max nm (log ε): 315.00 (0.2393), 291.00 (0.2251), 223.00 (0.5140), 202.50 (0.4535); IR (KBr) v max : 3402, 2926, 1692, 1604, 1514, 1451, 1384, 1261, 1169, 1071 cm-1; NMR spectroscopic data see table 1; HRESIMS m / z 655.2177 [M-H] - (calculated for C 32 H 35 N2O 13 655.2145).

[0094] In addition, the spectra of compounds Tangutoid LI and Tangutoid LII can clearly show the structures of the two β-carboline alkaloid coumaroyl glucoside compounds, and can also prove that the two β-carboline alkaloid coumaroyl glucoside compounds have high purity, wherein the purity of compound Tangutoid LI is 95%, and the purity of Tangutoid LII is 96%. The structural formulas of compounds Tangutoid LI and Tangutoid LII are as follows:

[0095]

[0096] Test Example 2

[0097] The compounds Tangutoid LI and Tangutoid LII obtained in Example 1 were subjected to monoamine oxidase inhibitory activity test, and the specific operation was as follows:

[0098] (1) Monoamine oxidase extraction

[0099] Take 10 g of pig liver (purchased) and put it in a juicer cup, add 400 g of 0.3 mol / L sucrose solution pre-cooled to 4°C to the juicer cup, turn on the juicer, and obtain pig liver serum. Then centrifuge the pig liver serum at a centrifugal force of 1000 g (rotational speed of 2100 r / min) at 4°C for 10 min, remove the surface scum, and collect the supernatant. Then centrifuge the supernatant at a centrifugal force of 10000 g (rotational speed of 6800 r / min) at 4°C for 30 min, and collect the precipitate. Add 25 mL of phosphate buffer solution (PBS, concentration of 0.1 mol / L, pH value = 7.4) pre-cooled to 25°C to the precipitate, divide and store at -80°C, and obtain a monoamine oxidase enzyme solution.

[0100] (2) Monoamine oxidase inhibitor activity evaluation

[0101] Step 1: Add the compound Tangutoid LI obtained in Example 1 to dimethyl sulfoxide (DMSO) to obtain a gradient concentration of the compound solution, which is ready for use. Take 2 μL of the compound solution respectively, and add 100 μL of PBS aqueous solution (concentration of 0.1 mol / L) to obtain a compound-PBS complex solution.

[0102] Step 2: Take two identical culture dishes, respectively labeled as reaction system A and reaction system B, and add 8 μL of monoamine oxidase enzyme solution and 102 μL of compound-PBS complex solution to each of reaction system A and reaction system B, and incubate at 37°C for 15 min; add 40 μL of Amplex Red solution (0.25 mg / mL), 10 μL of horseradish peroxidase solution (0.3 U / mL), and 40 μL of 5-hydroxytryptamine hydrochloride solution (as a MAO-A substrate) with a concentration of 0.1 mmol / L to reaction system A; add 40 μL of Amplex Red solution (0.25 mg / mL), 10 μL of horseradish peroxidase solution (0.3 U / mL), and 40 μL of benzylamine hydrochloride solution (as a MAO-B substrate) with a concentration of 0.1 mmol / L to reaction system B; react reaction system A and reaction system B at 37°C for 24 min, and then measure the optical density difference before and after the reaction at 570 nm to calculate the inhibition rate I of compound Tangutoid LI on MAO-A or MAO-B.

[0103] Step 3: Repeat steps 1 and 2, except that Tangutoid LI is replaced by Tangutoid LII, and calculate the inhibition rate I of compound Tangutoid LII on MAO-A or MAO-B.

[0104] The inhibition rate I is calculated using the following formula: I = 1 - ΔD1 / ΔD2, where ΔD1 is the difference in optical density before and after the reaction in the experimental group, and ΔD2 is the difference in optical density before and after the reaction in the blank group without monoamine oxidase solution.

[0105] The half-maximal inhibitory concentrations (IC50) of compounds Tangutoid LI and Tangutoid LII against MAO-A and MAO-B were calculated using Graphpad 8.0.2. 50 IC 50 The test results are shown in Table 2.

[0106] Table 2. MAO-A and MAO-B inhibitory activities of compounds Tangutoid LI and Tangutoid LII.

[0107] Compound MAO-AIC 50 (μmol / L) MAO-B IC 50 (μmol / L) Tangutoid LI 121.7±4.7 113.2±5.3 Tangutoid LII 101.2±3.6 115.9±6.2

[0108] Note: IC in Table 2 50 The test results are the average value ± SD (n = 3).

[0109] As shown in Table 2, compounds Tangutoid LI and Tangutoid LII exhibit excellent inhibitory activities against MAO-A and MAO-B. Among them, the inhibitory activity IC50 of compound Tangutoid LI against MAO-A is [missing value]. 50 The value was 121.7 μmol / L, and the IC50 value for the inhibitory activity against MAO-B was 121.7 μmol / L. 50 The value was 113.2 μmol / L; the inhibitory activity IC50 of compound Tangutoid LII against MAO-A was 113.2 μmol / L. 50 The value was 101.2 μmol / L, and the IC50 value for the inhibitory activity against MAO-B was 101.2 μmol / L. 50 The value was 115.9 μmol / L.

[0110] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A β-carboline alkaloid coumaroyl glucoside compound characterized by, having a structure represented by Formula 1 or Formula 2:

2. The extraction method of the β-carboline alkaloid coumaroyl glucoside compound according to claim 1, characterized by, comprising the following steps: (1) alcohol precipitation of Nitraria tangutorum Bob. fruit juice to obtain an extract; (2) dissolving the extract with water, and sequentially using pure water and an ethanol aqueous solution with a volume fraction of 40% as eluents to perform macroporous resin column chromatography on the extract to obtain an ethanol aqueous solution with a volume fraction of 40% elution fraction; (3) column chromatography separation of the elution part of the 40% volume fraction of ethanol aqueous solution under gradient elution condition with MCI resin as the stationary phase and methanol-water as the eluent, to obtain 4 components, recorded as Fr 40-1 ~Fr 40-4 ; the volume fraction of methanol in the eluent of step (3) is 0% to 80%. (4) the MCI resin is used as the stationary phase, and methanol-water is used as the eluent to perform gradient elution on the Fr 40-1 fractions under the condition of step (4) to obtain 8 fractions, denoted as Fr 40-1-1 40-1-8 ; the volume fraction of methanol in the eluent of step (4) is 0% to 50%.​ (5) column chromatography is carried out on C18 bonded silica gel with acetonitrile-water as eluent under gradient elution conditions to separate Fr 40-1-8 ; the column chromatography is carried out on C18 bonded silica gel with acetonitrile-water as eluent under gradient elution conditions to separate Fr 40-1-8-1 ~Fr 40-1-8-5 ; the volume fraction of acetonitrile in the eluent of step (5) is 15% to 35%. (6) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr was removed under isogradient elution conditions. 40-1-8-3 The components were separated by column chromatography to obtain β-carbamoline alkaloid coumaroyl glucoside compounds with the structure shown in Formula 1; the volume fraction of acetonitrile in the eluent of step (6) was 15%; (7) the C18 bonded silica gel as the stationary phase, acetonitrile-water as the eluent, under the condition of isocratic elution, Fr 40-1-8-5 is separated by column chromatography to obtain the β-carboline alkaloid coumaroyl glucoside compound with the structure shown in formula 2; the volume fraction of acetonitrile in the eluent of step (7) is 20%. The steps (6) and (7) are not limited in time sequence.

3. The extraction method according to claim 2, characterized in that, In step (1), the reagent used for alcohol precipitation is an ethanol aqueous solution; the volume fraction of the ethanol aqueous solution is 50%-75%.

4. The extraction method according to claim 2 or 3, characterized in that, The number of alcohol precipitation is 3-4 times; the time of each alcohol precipitation is independently 10-12 h, and the volume ratio of the ethanol aqueous solution to the Nitraria tangutorum Bob. fruit juice is independently 2-4:

1.

5. The extraction method of claim 2, wherein, In step (3), the volume fraction of methanol in the eluent is 0%, 20%, 40% and 80% in sequence.

6. The extraction method of claim 2, wherein, In step (4), the volume fraction of methanol in the eluent is 0%, 10%, 20%, 30%, 40% and 50% in sequence.

7. The extraction method of claim 2, wherein, In step (2), the chromatographic column used for macroporous resin column chromatography is an AB-8 macroporous resin chromatographic column.

8. The extraction method of claim 2, wherein, In the step (6), the Fr 40-1-8-3 The chromatographic column used for column chromatographic separation of the components is a preparative liquid XCharge C18 chromatographic column. In the step (7), the Fr 40-1-8-5 The chromatographic column used for column chromatographic separation of the components is a preparative liquid XCharge C18 chromatographic column.

9. Use of the β-carboline alkaloid coumaroyl glucoside compound in claim 1 in the preparation of a monoamine oxidase inhibitor or an anti-neurodegenerative disease drug.

10. A monoamine oxidase inhibitor comprising the β-carboline alkaloid coumaroyl glucoside compound in claim 1.

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