Beta-carboline alkaloid derivative, extraction method and application thereof, and monoamine oxidase inhibitor

Abstract

The invention provides a beta-carboline alkaloid derivative, an extraction method and application thereof, and a monoamine oxidase inhibitor. The beta-carboline alkaloid derivative provided by the invention has a structure as shown in formula 1, is novel in structure, has good monoamine oxidase inhibitory activity, can slow down consumption of monoamine neurotransmitters 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 beta-carboline alkaloid derivative, an extraction method and application thereof, and a monoamine oxidase inhibitor. Background Art

[0002] Monoamine oxidase (MAO) is a type of flavin protein present on the outer membrane of mitochondria, which is divided into two subtypes: monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B). The preferred reaction substrates of MAO-A are 5-hydroxytryptamine, adrenaline, etc., and the preferred reaction substrates of MAO-B are benzylamine and β-phenylethylamine. The control of movement, mood regulation, perception and cognitive function, as well as the occurrence of related diseases are closely related to the monoamine signaling mechanism. In vivo, MAO-A is mainly related to depression, and MAO-B is related to Parkinson's disease. If the activity of monoamine oxidase in the brain is too strong, it will over-metabolize monoamine neurotransmitters in the body and generate excessive hydrogen peroxide, inducing nervous system disorders and oxidative damage. Oxidative damage is one of the causes of many neurodegenerative diseases, such as depression, Parkinson's disease, Alzheimer's disease, etc. In particular, in the brain slices of patients with depression and Parkinson's disease after death, monoamine oxidase with excessive expression levels was detected. Therefore, by inhibiting the activity of monoamine oxidase, the consumption of monoamine neurotransmitters in the brain can be slowed down, oxidative damage can be slowed down, and the purpose of treating neurodegenerative diseases such as depression and Parkinson's disease can be achieved.

[0003] Nitraria tangutorum Bobr. is a wild plant with both medicinal and edible properties. Its fruit is an important plant resource, rich in polysaccharides, β-carboline alkaloids, flavonoids and phenolic acid components, and has high economic, ecological and medicinal value. The fruit of Nitraria tangutorum Bobr. is often used by the public to treat spleen and stomach weakness, indigestion, neurasthenia, hyperglycemia, dizziness, colds, milk retention, etc. At present, there are few studies on the extract of Nitraria tangutorum fruit, and there is no report on the extraction and isolation of substances that inhibit monoamine oxidase activity from the fruit of Nitraria tangutorum Bobr. Summary of the invention

[0004] The object of the present invention is to provide a β-carboline alkaloid derivative and an extraction method and application thereof, and a monoamine oxidase inhibitor. The β-carboline alkaloid derivative provided by the present invention has good monoamine oxidase inhibitory activity and can be used to prepare a monoamine oxidase inhibitor or a drug for treating neurodegenerative diseases.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a β-carboline alkaloid derivative having a structure shown in Formula 1:

[0007]

[0008] The present invention provides a method for extracting β-carboline alkaloid derivatives according to the above technical solution, comprising the following steps:

[0009] (1) subjecting the Nitraria tanguticus fruit juice to alcohol precipitation to obtain an extract;

[0010] (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;

[0011] (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%;

[0012] (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%;

[0013] (5) Using LH-20 gel as the stationary phase and methanol-water as the eluent, Fr 40-1-7 The components were separated by column chromatography to obtain 10 components, which were recorded as Fr 40-1-7-1 ~Fr 40-1-7-10 ; The volume fraction of methanol in the eluent of step (5) is 0%, 20%, 40% and 60% respectively;

[0014] (6) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-7-9 The components are separated by column chromatography to obtain a β-carboline alkaloid derivative having a structure shown in Formula 1; the volume fraction of acetonitrile in the eluent is 15%.

[0015] 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%.

[0016] Preferably, the alcohol precipitation is performed 3 to 4 times.

[0017] Preferably, 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.

[0018] Preferably, in step (3), the volume fraction of methanol in the eluent used for column chromatography separation is 0%, 20%, 40% and 80% respectively.

[0019] Preferably, in step (4), the volume fraction of methanol in the eluent used for column chromatography separation is 0%, 10%, 20%, 30%, 40% and 50% respectively.

[0020] Preferably, in step (2), the chromatographic column used in the macroporous resin column chromatography is an AB-8 macroporous resin chromatographic column.

[0021] The present invention provides the use of the beta-carboline alkaloid derivatives described in the above technical solution in the preparation of monoamine oxidase inhibitors or anti-neurodegenerative disease drugs or health foods.

[0022] The present invention provides a monoamine oxidase inhibitor, comprising the β-carboline alkaloid derivative described in the above technical solution.

[0023] The present invention provides a β-carboline alkaloid derivative having a structure shown in Formula 1, 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. It can be used to prepare monoamine oxidase inhibitors or drugs for treating neurodegenerative diseases, and can also be used for the development of health foods, with broad application prospects. The embodiments show that the β-carboline alkaloid derivative has good MAO-A inhibitory activity, IC 50 The value is 151.4μmol / L; it also has good MAO-B inhibitory activity, IC 50 The value is 122.3μmol / L.

[0024] The present invention also provides a method for extracting β-carboline alkaloid derivatives. The present invention uses the fruit of Nitraria tanguticus as a raw material, and separates the β-carboline alkaloid derivatives by ethanol extraction combined with a series of column chromatography separations. The extraction method of the present invention is simple and fast, and the purity of the obtained β-carboline alkaloid derivatives is high, reaching 95% or more. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] 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.

[0026] Figure 1 It is a β-carboline alkaloid derivative Tangutoid LIII 1 H-NMR spectrum;

[0027] Figure 2 It is a β-carboline alkaloid derivative Tangutoid LIII 13 C-NMR spectrum;

[0028] Figure 3 This is the HSQC spectrum of Tangutoid LIII, a β-carboline alkaloid derivative;

[0029] Figure 4 This is the HMBC spectrum of Tangutoid LIII, a β-carboline alkaloid derivative;

[0030] Figure 5 It is a β-carboline alkaloid derivative Tangutoid LIII 1 H- 1 H COSY spectrum;

[0031] Figure 6 This is the NOESY spectrum of Tangutoid LIII, a β-carboline alkaloid derivative;

[0032] Figure 7 The TOCSY spectrum of Tangutoid LIII, a β-carboline alkaloid derivative;

[0033] Figure 8 This is the (-)-HRESIMS spectrum of Tangutoid LIII, a β-carboline alkaloid derivative;

[0034] Fig. 9 It is the IR spectrum of Tangutoid LIII, a β-carboline alkaloid derivative;

[0035] Fig.10 This is the UV spectrum of Tangutoid LIII, a β-carboline alkaloid derivative. DETAILED DESCRIPTION

[0036] The present invention provides a β-carboline alkaloid derivative having a structure shown in Formula 1:

[0037]

[0038] The molecular formula of the β-carboline alkaloid derivatives of the present invention is C 32 H 38 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.

[0039] The present invention also provides a method for extracting β-carboline alkaloid derivatives according to the above technical solution, comprising the following steps:

[0040] (1) subjecting the Nitraria tanguticus fruit juice to alcohol precipitation to obtain an extract;

[0041] (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;

[0042] (3) Using MCI resin as the stationary phase and methanol-water as the eluent, the 40% ethanol aqueous solution elution portion 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%;

[0043] (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%;

[0044] (5) Using LH-20 gel as the stationary phase and methanol-water as the eluent, Fr 40-1-7 The components were separated by column chromatography to obtain 10 components, which were recorded as Fr 40-1-7-1 ~Fr 40-1-7-10 ; The volume fraction of methanol in the eluent of step (5) is 0%, 20%, 40% and 60% respectively;

[0045] (6) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-7-9The components are separated by column chromatography to obtain a β-carboline alkaloid derivative having a structure shown in Formula 1; the volume fraction of acetonitrile in the eluent is 15%.

[0046] 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.

[0047] 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.

[0048] 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 derivatives.

[0049] 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 a 40% ethanol aqueous solution elution portion. In the present invention, the resin column used for the macroporous resin column chromatography can be an AB-8 macroporous resin chromatographic column. 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 a pure water elution portion, a 40% ethanol aqueous solution elution portion and a 95% ethanol aqueous solution elution portion are obtained respectively. In the present invention, the column chromatography separation is performed under gradient elution conditions. 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.

[0050] 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.

[0051] 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.

[0052] Get Fr 40-1-7 After the components are separated, the present invention uses LH-20 gel as the stationary phase and methanol-water as the eluent to separate Fr 40-1-7 The components were separated by column chromatography to obtain 10 components, which were recorded as Fr 40-1-7-1 ~Fr 40-1-7-10 . In the present invention, the volume fraction of methanol in the eluent is 0%, 20%, 40% and 60% in sequence. In a specific embodiment of the present invention, the eluents used for column chromatography separation are pure water, 20% methanol-water, 40% methanol-water and 60% methanol-water in sequence. After elution with 60% 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 40-1-7 The components were loaded onto an LH-20 gel chromatography column, and gradient elution was performed using pure water, 20% methanol-water, 40% methanol-water, 60% methanol-water and pure methanol as eluents in sequence.

[0053] Get Fr 40-1-7-9 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 40-1-7-9 The components are separated by column chromatography to obtain a β-carboline alkaloid derivative having a structure shown in Formula 1. In the present invention, the volume fraction of acetonitrile in the eluent is 15%. In a specific embodiment of the present invention, Fr 40-1-7-9 The components were loaded onto a preparative liquid phase Megress C18 chromatographic column and eluted with 15% acetonitrile-water for isocratic elution.

[0054] The invention uses the fruit of Nitraria tanguticus as raw material, adopts ethanol extraction, and combines a series of column chromatography separation methods to separate and obtain the β-carboline alkaloid derivatives. The extraction method of the invention is simple and fast, and the purity of the obtained β-carboline alkaloid derivatives is high, which is 95%.

[0055] The present invention also provides the use of the β-carboline alkaloid derivatives described in the above technical solution in the preparation of monoamine oxidase inhibitors or anti-neurodegenerative disease drugs or health foods.

[0056] The β-carboline alkaloid derivatives of the present invention have good MAO-A inhibitory activity, IC 50 The value is 151.4μmol / L; it also has good MAO-B inhibitory activity, IC 50 The value is 122.3μmol / L, which 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. It can also be used to prepare monoamine oxidase inhibitors or drugs against neurodegenerative diseases.

[0057] The present invention also provides a monoamine oxidase inhibitor, comprising the β-carboline alkaloid derivatives 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 derivatives described in the above technical solution.

[0058] 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.

[0059] The room temperature condition described in the embodiment is 20-35°C.

[0060] Example 1

[0061] (1) 110 kg of Nitraria tanguticum fruit were 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.

[0062] (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 ).

[0063] (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 .

[0064] (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 .

[0065] (5) Take the Fr obtained in step (4) 40-1-7 The components were loaded onto an LH-20 gel chromatography column and gradient eluted with pure water, 20% methanol-water, 40% methanol-water, 60% methanol-water and pure methanol. Ten components were obtained, recorded as Fr 40-1-7-1 ~Fr 40-1-7-10 .

[0066] (6) The Fr obtained in step (5) 40-1-7-9 The components were loaded onto a preparative liquid phase Megress C18 chromatographic column and isocratically eluted with a 15% by volume acetonitrile aqueous solution to obtain a β-carboline alkaloid derivative (125.8 mg) from Nitraria tanguticus. The β-carboline alkaloid derivative was named Tangutoid LIII.

[0067] Test Example 1

[0068] 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- 1The structure of the compound Tangutoid LIII prepared in Example 1 was 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 10 As shown, the compound Tangutoid LIII 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.

[0069] Table 1 Compound Tangutoid LIII 1 H-NMR (600 MHz) and 13 C-NMR (150 MHz) data

[0070]

[0071]

[0072] The spectroscopic data of compound Tangutoid LIII are as follows: white powder, [α] D 20.1 -35.70(c0.118,MeOH); UV(MeOH)λ max nm(logε):315.00(0.2059),291.50(0.1895),223.50(0.3970),201.50(0.2937); IR(KBr)ν max :3385,2933,1692,1630,1604,1514,1395,1262,1170cm -1 ;NMR spectroscopic date see table 1; HRESIMS m / z 673.2256[MH]-(calculated for C 32 H 37 N2O 14 673.2250).

[0073] In addition, the purity of the compound Tangutoid LIII is relatively high, at 95%. The structural formula of the compound Tangutoid LIII is shown below:

[0074]

[0075] Test Example 2

[0076] The monoamine oxidase inhibitory activity test of the compound Tangutoid LIII obtained in Example 1 was performed as follows:

[0077] (1) Monoamine oxidase extraction

[0078] 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.

[0079] (2) Evaluation of monoamine oxidase inhibitor activity

[0080] The compound Tangutoid LIII obtained in Example 1 was added to dimethyl sulfoxide (DMSO) to obtain a compound solution with gradient concentrations for later use. 2 μL of the compound solution was mixed with 100 μL of PBS aqueous solution to obtain a compound-PBS composite solution.

[0081] Take two culture dishes of the same specifications and mark them as reaction system A and reaction system B, respectively. 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 0.1 mmol / L benzylamine hydrochloride solution (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 LIII on MAO-A or MAO-B.

[0082] 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.

[0083] Graphpad 8.0.2 was used to calculate the half-inhibitory concentration (IC) of Tangutoid LIII on MAO-A and MAO-B. 50 ), IC 50 The test results are shown in Table 2.

[0084] Table 2 MAO-A and MAO-B inhibitory activity of compound Tangutoid LIII

[0085] Compound <![CDATA[MAO-AIC 50 (μmol / L)]]> <![CDATA[MAO-BIC 50 (μmol / L)]]> TangutoidLIII 151.4±7.7 122.3±6.3

[0086] Note: IC in Table 2 50 The test results are mean ± SD (n = 3).

[0087] As shown in Table 2, the compound Tangutoid LIII has excellent MAO-A and MAO-B inhibitory activity. The inhibitory activity IC of the compound Tangutoid LIII on MAO-A is 50 The value is 151.4μmol / L, and the inhibitory activity IC of MAO-B 50 The value is 122.3μmol / L.

[0088] 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 derivative, characterized in that: It has the structure shown in formula 1:

2. The method for extracting β-carboline alkaloid derivatives 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 LH-20 gel as the stationary phase and methanol-water as the eluent, Fr 40-1-7 The components were separated by column chromatography to obtain 10 components, which were recorded as Fr 40-1-7-1 ~Fr 40-1-7-10 ; The volume fraction of methanol in the eluent of step (5) is 0%, 20%, 40% and 60% respectively; (6) Using C18 bonded silica gel as the stationary phase and acetonitrile-water as the eluent, Fr 40-1-7-9 The components are separated by column chromatography to obtain a β-carboline alkaloid derivative having a structure shown in Formula 1; the volume fraction of acetonitrile in the eluent is 15%.

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 alcohol precipitation is performed 3 to 4 times.

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

1.

6. The extraction method according to claim 2, characterized in that In step (3), the volume fractions of methanol in the eluent used for column chromatography separation are 0%, 20%, 40% and 80% respectively.

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

8. 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.

9. Use of the β-carboline alkaloid derivative 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 derivative according to claim 1.

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