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

Abstract

The invention provides a beta-carboline alkaloid coumaroyl glucoside compound, an extraction method and application thereof, and a monoamine oxidase inhibitor. The beta-carboline alkaloid coumaroyl glucoside compound provided by the invention has a structure as shown in a formula 1 or a formula 2, is novel in structure, has good monoamine oxidase inhibitory activity, can slow down monoamine neurotransmitter consumption in the brain, slow down oxidative damage and realize treatment of neurodegenerative diseases, and has good application prospects. Further, the compound can be used for preparing a monoamine oxidase inhibitor or preparing a medicine for resisting neurodegenerative diseases, can also be used for developing health-care food, and is wide in application prospect. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine, and specifically relates to a β-carboline alkaloid coumaroyl glucoside compound, an extraction method and application thereof, and a monoamine oxidase inhibitor. Background Art

[0002] Monoamine oxidase (MAO) is a vital flavin protein that exists on the outer membrane of mitochondria and is divided into two subtypes: monoamine oxidase A (MAO-A) and monoamine oxidase B (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 a strong catalytic activity on substances such as serotonin and adrenaline, while MAO-B shows a higher preference for the metabolism of substances such as benzylamine and β-phenylethylamine. Substances such as serotonin, adrenaline, benzylamine, and β-phenylethylamine play a vital role in regulating movement, emotions, perception, and cognitive functions. 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 metabolic process of monoamine neurotransmitters is accelerated, accompanied by the generation of a large amount of hydrogen peroxide. This process not only leads to dysfunction of the nervous system, but also may cause oxidative stress damage, becoming an important cause of 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] Given the central role of monoamine oxidase in neurotransmitter metabolism and disease pathogenesis, the development of effective monoamine oxidase inhibitors has become a hot topic in current research. Such inhibitors are expected to reduce oxidative stress damage by inhibiting the activity of monoamine oxidase and slowing down the consumption of monoamine neurotransmitters, thus providing new hope for the intervention and treatment of neurodegenerative diseases such as depression and Parkinson's disease. Currently, there are few studies on monoamine oxidase inhibitors, and new types are urgently needed. Summary of the invention

[0006] The object of the present invention is to provide a β-carboline alkaloid coumaroyl glucoside compound, an extraction method and application thereof, and a monoamine oxidase inhibitor. The β-carboline alkaloid coumaroyl glucoside compound provided by the present invention has good monoamine oxidase inhibitory activity and can be used to prepare a novel monoamine oxidase inhibitor or a novel drug for preventing neurodegenerative diseases.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 1 or Formula 2:

[0009]

[0010] The present invention provides a method for extracting the β-carboline alkaloid coumaroyl glucoside compound described in the above technical solution, comprising the following steps:

[0011] (1) subjecting the Nitraria tanguticus fruit juice to alcohol precipitation to obtain an extract;

[0012] (2) dissolving the extract in water, using pure water and 40% by volume ethanol aqueous solution as eluents in sequence, subjecting the extract to macroporous resin column chromatography to obtain a 40% by volume ethanol aqueous solution elution fraction;

[0013] (3) Using MCI resin as the stationary phase and methanol-water as the eluent, the elution portion of the 40% ethanol aqueous solution was subjected to column chromatography under gradient elution conditions to obtain four components, which were recorded as Fr 40-1 ~Fr 40-4 ; The volume fraction of methanol in the eluent of step (3) is 0% to 80%;

[0014] (4) Using MCI resin as the stationary phase and methanol-water as the eluent, Fr 40-1 The components were separated by column chromatography to obtain 8 components, which were recorded 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) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8 The components were separated by column chromatography to obtain 5 components, which were recorded 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) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8-3 The components are separated by column chromatography to obtain a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 1; the volume fraction of acetonitrile in the eluent of step (6) is 15%;

[0017] (7) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8-5 The components are separated by column chromatography 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 step (7) is 20%;

[0018] There is no time sequence limitation for step (6) and step (7).

[0019] Preferably, in 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 3 to 4 times; the time of each alcohol precipitation is independently 10 to 12 hours, and the volume ratio of the ethanol aqueous solution to the Nitraria tanguticus 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% respectively.

[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% respectively.

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

[0024] Preferably, in step (6), the Fr 40-1-8-3 The chromatographic column used for column chromatography separation of components was a preparative liquid phase XCharge C18 chromatographic column;

[0025] In the step (7), the Fr 40-1-8-5 The chromatographic column used for column chromatography separation of the components is a preparative liquid phase XCharge C18 chromatographic column.

[0026] The present invention provides the use of the β-carboline alkaloid coumaroyl glucoside compound described in the above technical solution in the preparation of monoamine oxidase inhibitors or anti-neurodegenerative disease drugs or health foods.

[0027] The present invention provides a monoamine oxidase inhibitor, comprising the β-carboline alkaloid coumaroyl glucoside compound described in the above technical solution.

[0028] The present invention provides a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 1 or Formula 2, which has a novel structure and good monoamine oxidase inhibitory activity, can slow down the consumption of monoamine neurotransmitters in the brain, slow down oxidative damage, and achieve the treatment of neurodegenerative diseases, and can be used to prepare monoamine oxidase inhibitors or prepare drugs for anti-neurodegenerative diseases. It can also be used for the development of health foods, and has broad application prospects. The embodiments show that the β-carboline alkaloid coumaroyl glucoside compound has good MAO-A inhibitory activity and MAO-B inhibitory activity; wherein, the IC value of the β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 1 on the MAO-A inhibitory activity is 50 The value was 121.7 μmol / L, and the IC value of MAO-B inhibitory activity was 50 The value is 113.2 μmol / L; the IC value of the inhibitory activity of the β-carboline alkaloid coumaroyl glucoside compound with the structure shown in Formula 2 on MAO-A 50 The value was 101.2 μmol / L, and the IC value of MAO-B inhibitory activity was 50 The value is 115.9μmol / L.

[0029] The present invention also provides a method for extracting a β-carboline alkaloid coumaroyl glucoside compound. The present invention uses the fruit of Nitraria tanguticus as a raw material, and uses ethanol extraction combined with a series of column chromatography separations to separate and obtain the β-carboline alkaloid coumaroyl glucoside compound. The extraction method of the present invention is simple and fast, and the purity of the β-carboline alkaloid coumaroyl glucoside compound obtained thereby is relatively high, reaching more than 95%. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0031] Figure 1 It is a β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI 1 H-NMR spectrum;

[0032] Figure 2 It is a β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI 13 C-NMR spectrum;

[0033] Figure 3This is the HSQC spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI;

[0034] Figure 4 This is the HMBC spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI;

[0035] Figure 5 It is a β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI 1 H- 1 H COSY spectrum;

[0036] Figure 6 This is the NOESY spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI;

[0037] Figure 7 This is the TOCSY spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI;

[0038] Figure 8 This is the (-)-HRESIMS spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI;

[0039] Fig. 9 This is the IR spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI;

[0040] Fig.10 This is the UV spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LI;

[0041] Fig.11 It is a β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII 1 H-NMR spectrum;

[0042] Fig.12 It is a β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII 13 C-NMR spectrum;

[0043] Fig.13 This is the HSQC spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII;

[0044] Fig.14 This is the HMBC spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII;

[0045] Fig.15It is a β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII 1 H- 1 H COSY spectrum;

[0046] Fig.16 This is the NOESY spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII;

[0047] Fig.17 This is the TOCSY spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII;

[0048] Fig.18 This is the (-)-HRESIMS spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII;

[0049] Fig.19 This is the IR spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII;

[0050] Fig. 20 This is the UV spectrum of the β-carboline alkaloid coumaroyl glucoside compound Tangutoid LII. DETAILED DESCRIPTION

[0051] The present invention provides a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 2:

[0052]

[0053] The molecular formula of the β-carboline alkaloid coumaroyl glucoside compound of the present invention is C 32 H 36 N2O 13 It has a novel structure and good monoamine oxidase inhibitory activity. It can slow down the consumption of monoamine neurotransmitters in the brain, slow down oxidative damage, and achieve the treatment of neurodegenerative diseases. It can then be used to prepare monoamine oxidase inhibitors or drugs against neurodegenerative diseases. It can also be used in the development of health foods and has broad application prospects.

[0054] The present invention also provides a method for extracting the β-carboline alkaloid coumaroyl glucoside compound described in the above technical solution, comprising the following steps:

[0055] (1) subjecting the Nitraria tanguticus fruit juice to alcohol precipitation to obtain an extract;

[0056] (2) dissolving the extract in water, using pure water and 40% by volume ethanol aqueous solution as eluents in sequence, subjecting the extract to macroporous resin column chromatography to obtain a 40% by volume ethanol aqueous solution elution fraction;

[0057] (3) Using MCI resin as the stationary phase and methanol-water as the eluent, the elution portion of the 40% ethanol aqueous solution was subjected to column chromatography under gradient elution conditions to obtain four components, which were recorded as Fr 40-1 ~Fr 40-4 ; The volume fraction of methanol in the eluent of step (3) is 0% to 80%;

[0058] (4) Using MCI resin as the stationary phase and methanol-water as the eluent, Fr 40-1 The components were separated by column chromatography to obtain 8 components, which were recorded as Fr 40-1-1 ~Fr 40-1-8 ; The volume fraction of methanol in the eluent of step (4) is 0% to 50%;

[0059] (5) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8 The components were separated by column chromatography to obtain 5 components, which were recorded 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%.

[0060] (6) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8-3 The components are separated by column chromatography to obtain a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 1; the volume fraction of acetonitrile in the eluent of step (6) is 15%;

[0061] (7) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8-5 The components are separated by column chromatography 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 step (7) is 20%;

[0062] There is no time sequence limitation for step (6) and step (7).

[0063] In the present invention, unless otherwise specified, the raw materials used are commercially available products well known to those skilled in the art or are prepared by methods well known to those skilled in the art.

[0064] The present invention performs alcohol precipitation on the Nitraria tanguticum fruit juice to obtain an extract. The present invention can sequentially crush and concentrate the Nitraria tanguticum fruit to obtain the Nitraria tanguticum fruit juice. In a specific embodiment of the present invention, the crushing method is juicing. The present invention crushes the Nitraria tanguticum fruit to obtain a crushed product; and concentrates the crushed product to obtain the Nitraria tanguticum fruit juice.

[0065] After obtaining the Nitraria tanguticus fruit juice, the present invention performs alcohol precipitation on the Nitraria tanguticus fruit juice to obtain an extract. The reagent used for the alcohol precipitation of the present invention can be an ethanol aqueous solution. In the present invention, the volume fraction of the ethanol aqueous solution can be 50% to 75%; in a specific embodiment of the present invention, the volume fraction of the ethanol aqueous solution can be 50%, 55%, 60%, 65%, 70% or 75%. The number of alcohol precipitations of the present invention can be 3 to 4 times; in a specific embodiment of the present invention, the number of alcohol precipitations can be 3 or 4 times. The present invention has no special requirements for the temperature of the alcohol precipitation, and the conventional operating temperature in the field can be used. In the embodiment of the present invention, the alcohol precipitation is carried out at room temperature (20 to 35°C). In the present invention, the time of each alcohol precipitation can be independently 10 to 12 hours, and the volume ratio of ethanol aqueous solution to Tangut Nitraria tangutica juice can be independently 2 to 4:1; in a specific embodiment of the present invention, the time of each alcohol precipitation is independently 10 hours, 11 hours or 12 hours, and the volume ratio of ethanol aqueous solution to Tangut Nitraria tangutica juice is independently 2:1, 3:1 or 4:1. After each alcohol precipitation, the present invention can separate the obtained liquid into solid and liquid, collect the liquid material, and use the liquid material as the material to be processed for the next extraction; after the last alcohol precipitation, the obtained liquid material is concentrated to obtain an extract. In the present invention, the solid-liquid separation method can be filtration; the concentration can be reduced pressure distillation. The present invention adopts the extraction method of alcohol precipitation to remove protein, pectin and polysaccharide in Tangut Nitraria tangutica juice; at the same time, ethanol can be recovered by distillation to obtain a crude extract containing β-carboline alkaloid coumaroyl glucoside compounds.

[0066] After obtaining the extract, the present invention dissolves the extract in water, uses pure water and 40% ethanol aqueous solution as eluents in sequence, and performs macroporous resin column chromatography on the extract to obtain an ethanol aqueous solution elution portion with a volume fraction of 40%. In the present invention, the macroporous adsorption resin used for the macroporous adsorption resin column chromatography can be AB-8 macroporous resin. In a specific embodiment of the present invention, the extract is loaded on an AB-8 macroporous resin chromatographic column, and pure water, 40% ethanol aqueous solution and 95% ethanol aqueous solution are used as eluents in sequence to perform column chromatography separation, and the pure water elution portion, the 40% ethanol aqueous solution elution portion and the 95% ethanol aqueous solution elution portion are respectively obtained. The pure water elution portion and the 95% ethanol aqueous solution elution portion are not involved in the subsequent further separation process, and no further introduction is made here.

[0067] After obtaining the 40% volume fraction ethanol aqueous solution elution portion, the present invention uses MCI resin as the stationary phase and methanol-water as the eluent to separate the 40% volume fraction ethanol aqueous solution elution portion by column chromatography under gradient elution conditions to obtain four components, which are recorded as Fr 40-1 ~Fr 40-4 . In a specific embodiment of the present invention, the elution portion of the ethanol aqueous solution with a volume fraction of 40% is dissolved with water and then loaded on an MCI gel chromatography column for column chromatography separation. In the present invention, the volume fraction of methanol in the eluent used for column chromatography separation is 0% to 80%; in a specific embodiment of the present invention, the volume fraction of methanol in the eluent used for column chromatography separation is 0%, 20%, 40% and 80% in sequence, that is, the eluents used for column chromatography separation are pure water, 20% methanol-water, 40% methanol-water and 80% methanol-water in sequence. After elution using 80% methanol-water as the eluent is completed, the present invention can use pure methanol as the eluent to wash the chromatographic column used for column chromatography separation. In a specific embodiment of the present invention, the elution portion of the ethanol aqueous solution with a volume fraction of 40% is loaded on an MCI gel chromatography column, and pure water, 20% methanol-water, 40% methanol-water, 80% methanol-water and pure methanol are used as eluents in sequence to perform gradient elution.

[0068] Get Fr 40-1 After the components are separated, the present invention uses MCI resin as the stationary phase and methanol-water as the eluent to separate Fr 40-1 The components were separated by column chromatography to obtain 8 components, which were recorded as Fr 40-1-1 ~Fr 40-1-8. In the present invention, the volume fraction of methanol in the eluent used for column chromatography separation is 0% to 50%; in a specific embodiment of the present invention, the volume fraction of methanol in the eluent used for column chromatography separation is 0%, 10%, 20%, 30%, 40% and 50% in sequence, that is, the eluents used for column chromatography separation are pure water, 10% methanol-water, 20% methanol-water, 30% methanol-water, 40% methanol-water and 50% methanol-water in sequence. After elution with 50% methanol-water as the eluent is completed, the present invention can use pure methanol as the eluent to wash the chromatographic column used for column chromatography separation. In a specific embodiment of the present invention, Fr 40-1 The components were loaded onto an MCI gel chromatography column, and gradient elution was performed using pure water, 10% methanol-water, 20% methanol-water, 30% methanol-water, 40% methanol-water, 50% methanol-water and pure methanol as eluents in sequence.

[0069] Get Fr 40-1-8 After the components are separated, the present invention uses C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent to separate Fr 40-1-7 The components were separated by column chromatography to obtain 5 components, which were recorded as Fr 40-1-8-1 ~Fr 40-1-8-5 In the present invention, the volume fraction of acetonitrile in the eluent is 15% to 35%. In a specific embodiment of the present invention, Fr 40-1-8 The components were loaded onto a preparative liquid phase Megress C18 chromatographic column and gradient elution was performed using 15% to 35% acetonitrile-water as an eluent.

[0070] Get Fr 40-1-8-3 After the components are separated, the present invention uses C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent to separate Fr 40-1-8-3 The components are separated by column chromatography to obtain a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 1. 40-1-8-3 The chromatographic column used for column chromatography separation of the components can be a preparative liquid phase XCharge C18 chromatographic column. In the present invention, the volume fraction of acetonitrile in the eluent is 15%. In a specific embodiment of the present invention, Fr 40-1-8-3 The components were loaded onto a preparative liquid phase XCharge C18 chromatographic column and eluted with 15% acetonitrile-water for isocratic elution.

[0071] Get Fr 40-1-8-5 After the components are separated, the present invention uses C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent to separate Fr 40-1-8-5 The components are separated by column chromatography to obtain a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 2.40-1-8-5 The chromatographic column used for column chromatography separation of the components can be a preparative liquid phase XCharge C18 chromatographic column. In the present invention, the volume fraction of acetonitrile in the eluent is 20%. In a specific embodiment of the present invention, Fr 40-1-8-5 The components were loaded onto a preparative liquid phase XCharge C18 chromatographic column and eluted with 15% acetonitrile-water for isocratic elution.

[0072] The present invention uses the fruit of Nitraria tanguticus as a raw material, adopts ethanol extraction, and combines a series of column chromatography separation methods to separate and obtain the β-carboline alkaloid coumaroyl glucoside compound. The extraction method of the present invention is simple and fast, and the purity of the obtained β-carboline alkaloid coumaroyl glucoside compound is high, reaching more than 95%.

[0073] The present invention also provides the use of the β-carboline alkaloid coumaroyl glucoside compound described in the above technical solution in the preparation of monoamine oxidase inhibitors or anti-neurodegenerative disease drugs or health foods.

[0074] The β-carboline alkaloid coumaroyl glucoside compound of the present invention has good MAO-A inhibitory activity and MAO-B inhibitory activity; wherein the IC value of the MAO-A inhibitory activity of the β-carboline alkaloid coumaroyl glucoside compound having the structure shown in Formula 1 is 50 The value was 121.7 μmol / L, and the IC value of MAO-B inhibitory activity was 50 The value is 113.2 μmol / L; the IC value of the inhibitory activity of the β-carboline alkaloid coumaroyl glucoside compound with the structure shown in Formula 2 on MAO-A 50 The value was 101.2 μmol / L, and the IC value of MAO-B inhibitory activity was 50 The value is 115.9 μmol / L. The β-carboline alkaloid coumaroyl glucoside compound of the present invention can slow down the consumption of monoamine neurotransmitters in the brain, slow down oxidative damage, and achieve the treatment of neurodegenerative diseases such as depression and Parkinson's disease, and can be used to prepare monoamine oxidase inhibitors or drugs against neurodegenerative diseases.

[0075] The present invention also provides a monoamine oxidase inhibitor, comprising the β-carboline alkaloid coumaroyl glucoside compound described in the above technical solution. In the present invention, the monoamine oxidase inhibitor may include an active ingredient and a pharmaceutical excipient; the active ingredient is the β-carboline alkaloid coumaroyl glucoside compound described in the above technical solution.

[0076] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0077] The room temperature mentioned in the examples is 20-35°C.

[0078] Example 1

[0079] (1) 110 kg of Nitraria tanguticum fruit was taken and juiced to obtain a crushed product; the crushed product was concentrated under reduced pressure to obtain 25 L of Nitraria tanguticum juice. 50 L of ethanol aqueous solution (75% by volume) was added to the Nitraria tanguticum juice, and alcohol precipitation was performed at room temperature for 12 h. After the precipitation, the supernatant was filtered and collected; the above alcohol precipitation operation was repeated 3 times, and the supernatant obtained from the last alcohol precipitation was subjected to reduced pressure distillation to recover ethanol to obtain an extract.

[0080] (2) The extract obtained in step (1) was dissolved in water, and the obtained extract was loaded onto an AB-8 macroporous resin column, and gradient elution was performed using pure water, 40% ethanol-water, and 95% ethanol-water as eluents in sequence; after the elution, the pure water elution portion (denoted as Fr0), the 40% ethanol-water elution portion (denoted as Fr1), and the 95% ethanol-water elution portion (denoted as Fr2) were obtained. 40 ) and the 95% ethanol-water elution portion (denoted as Fr 95 ).

[0081] (3) The Fr obtained in step (2) 40 The components were dissolved in water, and then the obtained solution was loaded on an MCI gel chromatography column, and gradient elution was performed using pure water, 20% methanol-water, 40% methanol-water, 80% methanol-water and pure methanol in sequence; after the elution, 4 components were obtained, which were recorded as Fr 40-1 ~Fr 40-4 .

[0082] (4) Take the Fr obtained in step (3) 40-1 The fraction was loaded onto the MCI gel chromatography column in step (3) again, and gradient elution was 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 elution, 8 fractions were obtained, which were recorded as Fr 40-1-1 ~Fr 40-1-8 .

[0083] (5) Take the Fr obtained in step (4) 40-1-8The components were loaded onto a preparative liquid phase Megress C18 chromatographic column and gradient eluted with an acetonitrile aqueous solution having a volume fraction of 15% to 35%. After the elution, five components were obtained in sequence, which were recorded as Fr 40-1-8-1 ~Fr 40-1-8-5 .

[0084] (6) The Fr obtained in step (5) 40-1-8-3 The components were loaded onto a preparative liquid phase XCharge C18 chromatographic column and isocratically eluted with an acetonitrile aqueous solution having a volume fraction of 15% to obtain a β-carboline alkaloid coumaroyl glucoside compound (395 mg) from Nitraria tanguticus. The β-carboline alkaloid coumaroyl glucoside compound was named Tangutoid LI.

[0085] (7) The Fr obtained in step (5) 40-1-8-5 The components were loaded onto a preparative liquid phase XCharge C18 column and eluted isocratically with a 20% volume fraction of acetonitrile aqueous solution to obtain a β-carboline alkaloid coumaroyl glucoside compound (30.5 mg) from Nitraria tanguticus. The β-carboline alkaloid coumaroyl glucoside compound was named TangutoidLII.

[0086] Test Example 1

[0087] The H NMR spectroscopy ( 1 H-NMR), carbon nuclear magnetic resonance spectroscopy ( 13 C-NMR), heteronuclear single quantum correlation spectroscopy (HSQC), heteronuclear multiple bond correlation spectroscopy (HMBC), hydrogen-hydrogen correlation spectroscopy ( 1 H- 1 The structures of the compounds Tangutoid LI and Tangutoid LII prepared in Example 1 were tested by H COSY, nuclear Overhauser effect spectroscopy (NOESY), total correlation spectroscopy (TOCSY), high resolution electrospray ionization mass spectrometry (HRESIMS), infrared spectroscopy (IR) and ultraviolet spectroscopy (UV). The results are as follows: Figures 1 to 20 As shown, the compounds Tangutoid LI and Tangutoid LII 1 H-NMR and 13 The C-NMR data were obtained by measuring in DMSO-d6. The C-NMR data are shown in Table 1.

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

[0089]

[0090]

[0091] The spectroscopic data of 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(logε):315.00(0.2393),291.00(0.2251),223.00(0.5140),202.50(0.4535); IR(KBr)ν max :3402,2926,1692,1604,1514,1451,1384,1261,1169,1071cm-1; NMRspectroscopic date see table 1; HRESIMS m / z 655.2177[MH] - (calculated for 32 H 35 N2O 13 655.2145).

[0094] In addition, the spectra of the 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 purity of the two β-carboline alkaloid coumaroyl glucoside compounds is high, among which the purity of the compound Tangutoid LI is 95% and the purity of Tangutoid LII is 96%. The structural formulas of the compounds Tangutoid LI and Tangutoid LII are shown below:

[0095]

[0096] Test Example 2

[0097] The monoamine oxidase inhibitory activity test of the compounds Tangutoid LI and Tangutoid LII obtained in Example 1 was performed as follows:

[0098] (1) Monoamine oxidase extraction

[0099] Take 10g of pig liver (commercially available) and place it in a juicer cup, add 400g of 0.3mol / L sucrose aqueous solution precooled to 4°C to the juicer cup, start squeezing, and obtain pig liver slurry. Then centrifuge the pig liver slurry for 10min at a centrifugal force of 1000g (rotation speed of 2100r / min) and 4°C, remove the surface foam, and collect the supernatant. Then centrifuge the supernatant for 30min at a centrifugal force of 10000g (rotation speed of 6800r / min) and 4°C to collect the precipitate. Add 25mL of phosphate buffer (PBS, concentration of 0.1mol / L, pH value = 7.4) precooled to 25°C to the precipitate, store it at -80°C after aliquoting, and obtain monoamine oxidase enzyme solution.

[0100] (2) Evaluation of monoamine oxidase inhibitor activity

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

[0102] Step 2: Take two culture dishes of the same specifications, mark them as reaction system A and reaction system B, add 8 μL of monoamine oxidase solution and 102 μL of compound-PBS complex solution to reaction system A and reaction system B respectively, 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 0.1 mmol / L 5-hydroxytryptamine hydrochloride solution (as MAO-A substrate) 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 0.1 mmol / L 5-hydroxytryptamine hydrochloride solution (as MAO-A substrate) to reaction system B; Red solution (0.25 mg / mL), 10 μL horseradish peroxidase solution (0.3 U / mL) and 40 μL benzylamine hydrochloride solution with a concentration of 0.1 mmol / L (as MAO-B substrate); reaction system A and reaction system B were reacted at 37°C for 24 minutes, and the optical density difference before and after the reaction was measured 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 calculation formula of inhibition rate I is as follows: I=1-ΔD1 / ΔD2, ΔD1 is the difference in optical density before and after the reaction of the experimental group, and ΔD2 is the difference in optical density before and after the reaction of the blank group without adding monoamine oxidase enzyme solution.

[0105] Graphpad 8.0.2 was used to calculate the half-inhibitory concentration (IC) of Tangutoid LI and Tangutoid LII on MAO-A and MAO-B. 50 ), IC 50 The test results are shown in Table 2.

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

[0107] Compound <![CDATA[MAO-AIC 50 (μmol / L)]]> <![CDATA[MAO-BIC 50 (μmol / L)]]> TangutoidLI 121.7±4.7 113.2±5.3 TangutoidLII 101.2±3.6 115.9±6.2

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

[0109] As shown in Table 2, Tangutoid LI and Tangutoid LII have excellent MAO-A and MAO-B inhibitory activities. The inhibitory activity IC of Tangutoid LI on MAO-A is 50 The value is 121.7μmol / L, and the inhibitory activity IC of MAO-B 50 The value is 113.2μmol / L; the inhibitory activity IC of compound Tangutoid LII on MAO-A 50 The value is 101.2μmol / L, and the inhibitory activity IC of MAO-B 50 The value is 115.9μmol / L.

[0110] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A β-carboline alkaloid coumaroyl glucoside compound, characterized in that: It has a structure shown in Formula 1 or Formula 2:

2. The method for extracting the β-carboline alkaloid coumaroyl glucoside compound according to claim 1, characterized in that: The following steps are involved: (1) subjecting the Nitraria tanguticus fruit juice to alcohol precipitation to obtain an extract; (2) dissolving the extract in water, using pure water and 40% by volume ethanol aqueous solution as eluents in sequence, subjecting the extract to macroporous resin column chromatography to obtain a 40% by volume ethanol aqueous solution elution fraction; (3) Using MCI resin as the stationary phase and methanol-water as the eluent, the elution portion of the 40% ethanol aqueous solution was subjected to column chromatography under gradient elution conditions to obtain four components, which were recorded as Fr 40-1 ~Fr 40-4 ; The volume fraction of methanol in the eluent of step (3) is 0% to 80%; (4) Using MCI resin as the stationary phase and methanol-water as the eluent, Fr 40-1 The components were separated by column chromatography to obtain 8 components, which were recorded as Fr 40-1-1 ~Fr 40-1-8 ; The volume fraction of methanol in the eluent of step (4) is 0% to 50%; (5) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8 The components were separated by column chromatography to obtain 5 components, which were recorded 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%. (6) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8-3 The components are separated by column chromatography to obtain a β-carboline alkaloid coumaroyl glucoside compound having a structure shown in Formula 1; the volume fraction of acetonitrile in the eluent of step (6) is 15%; (7) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-8-5 The components are separated by column chromatography 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 step (7) is 20%; There is no time sequence limitation for step (6) and step (7).

3. The extraction method according to claim 2, characterized in that In 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%.

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

1.

5. The extraction method according to claim 2, characterized in that In the step (3), the volume fraction of methanol in the eluent is 0%, 20%, 40% and 80% respectively.

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

7. The extraction method according to claim 2, characterized in that In step (2), the chromatographic column used for the macroporous resin column chromatography is an AB-8 macroporous resin chromatographic column.

8. The extraction method according to claim 2, characterized in that In the step (6), the Fr 40-1-8-3 The chromatographic column used for column chromatography separation of components was a preparative liquid phase XCharge C18 chromatographic column; In the step (7), the Fr 40-1-8-5 The chromatographic column used for column chromatography separation of the components is a preparative liquid phase XChargeC18 chromatographic column.

9. Use of the β-carboline alkaloid coumaroyl glucoside compound according to claim 1 in the preparation of monoamine oxidase inhibitors or anti-neurodegenerative disease drugs or health foods.

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

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