Spermidine glycoside alkaloid compound as well as preparation and application thereof and medicine

Abstract

The invention relates to a novel spermidine glycoalkaloid compound as well as an application and a preparation method of the novel spermidine glycoalkaloid compound. The novel compound provided by the invention comprises three compounds (capparisane D, capparisane E and capparisane F), is obtained by extracting and separating from a cauliflower (capparaceae) caper vine plant caper spinosa, and is spermidine alkaloid containing a glycoside structure. Wherein the cappissane F is a glucosinolate-containing spermidine alkaloid which is separated from a natural product for the first time, and a biological activity experiment shows that the cappissane F has beta2-adrenergic receptor antagonistic activity and can be applied to medicines for preventing and treating diseases related to the receptor, such as rheumatoid arthritis, arrhythmia and the like.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of natural medicinal chemistry, and relates to a new spermidine glycoalkaloid compound. More particularly, it relates to three new spermidine glycoalkaloid compounds, capparissane D, capparissane E and capparissane F, a preparation method thereof, and the use of capparissane F as a β2-adrenergic receptor active ingredient for treating diseases related thereto, such as rheumatoid arthritis, arrhythmia, etc. In vitro activity shows that capparissane F has significant β2-adrenergic receptor antagonistic activity, and can be used as a lead compound for developing new drugs for treating or inhibiting the above-mentioned diseases. BACKGROUND

[0002] Natural products are a class of compounds with outstanding pharmacological activity. According to statistics, more than 60% of small molecule drugs are directly or indirectly related to natural products [1] . Alkaloids are a class of compounds of natural products, which are structurally diverse, rich in types, and often have good activity. According to relevant statistics, the number of alkaloids in natural products accounts for about 20%, but the activity accounts for as high as 50% [2] . Systematic separation and purification of natural products rich in alkaloids is conducive to obtaining more potential high-activity alkaloid compounds, and has great significance for new drug discovery.

[0003] Capparis spinosa L. is a plant of Capparaceae, and is mainly distributed in the Middle East and Mediterranean countries. In China, Capparis spinosa L. mainly grows in Xinjiang, Tibet, Gansu, etc. As a kind of local traditional medicine in Xinjiang Uygur Autonomous Region, the flowers, fruits and roots of Capparis spinosa L. can be used as medicine. Traditional pharmacological studies have shown that Capparis spinosa L. has the effects of dispelling wind and dampness, relieving pain and swelling, etc. In particular, Capparis spinosa L. is often used externally to treat rheumatoid arthritis, periarthritis of shoulder and other diseases in the Uygur folk. Modern pharmacological studies have shown that Capparis spinosa L. has a series of activities such as anti-inflammatory and analgesic, anti-diabetic, anti-tumor, immune regulation, etc.

[0004] Adrenergic receptors (ADRs) are a class of tissue receptors that mediate the action of catecholamines. Based on their differences in response to norepinephrine, they can be divided into two main classes: α and β. Further, α receptors include α1 and α2, mainly distributed in the smooth muscle of the skin, kidneys, and gastrointestinal tract; β receptors include β1, β2, and β3, mainly distributed in cardiomyocytes and smooth muscle. Among them, β2-adrenergic receptors (β2-ARs) have the most significant regulatory effect on immune cells. Rheumatoid arthritis (RA) is a systemic autoimmune disease. Many factors, including humoral immunity, imbalance of immune cells, lack of immune tolerance, and the production of pro-inflammatory cytokines, are closely related to the pathogenesis of RA. [3] Studies have shown that the β2-AR antagonist propranolol can reduce inflammatory activity and pain levels in patients with rheumatoid arthritis (RA). [4] The β2-AR agonist salbutamol can also effectively inhibit joint inflammation. [5] Therefore, it can be seen that β2-AR is closely related to RA. Combined with the pharmacological effect of raphana in treating RA, the discovery of β2-AR receptor active compounds in raphana has important potential and research value. [1] Newman DJ, Cragg G M. Natural products as sources of new drugs over the nearly four decades from 01 / 1981 to 09 / 2019. Journal of natural products[J],2020,83(3):770-803.

[0005] [2] Shi Q, Hui S, Zhang AH, et al. Natural alkaloids: basic aspects, biological roles, and future perspectives. Chinese Journal of Natural Medicines[J], 2014, 12(6): 401-406.

[0006] [3] Liu Yan. Study on the effect and mechanism of adrenaline β2 receptor signaling on Treg cells in a mouse model of rheumatoid arthritis. Doctoral dissertation, 2019.

[0007] [4]Levine J D, Dardick S J, Roizen M F, et al. Contribution of sensory afferents and sympathetic efferents to joint injury in experimental arthritis. Journal of Neuroscience [J], 1986, 6(12): 3423-3429.

[0008] [5]Malfait A-M, Malik A S, Marinova-Mutafchieva L, et al. The β2-adrenergic agonist salbutamol is a potent suppressor of established collagen-induced arthritis: mechanisms of action. The Journal of Immunology [J], 1999, 162(10): 6278-6283. SUMMARY

[0009] The application provides a class of spermidine glycoside alkaloids compounds with novel structure, or crystal form thereof, or chiral isomer thereof, or glycoside thereof, or pharmaceutically acceptable salt thereof, or solvate thereof, or prodrug thereof, or metabolite thereof, which includes three new compounds, and the structural formula (I) is as follows:

[0010] The application also provides a method for preparing the three compounds (I), and the steps are as follows: the fruits of Capparis spinosa are extracted with 1% HAc / H2O (v:v) as the extraction solvent at room temperature for 3 hours, and then the solvent is removed by rotary evaporation. The extract is filtered by a 10KD ultrafiltration membrane, and the obtained filtrate is treated by anion exchange resin PA408. 1% HAc / H2O (v:v) is used as the loading and elution liquid, and the elution is performed for 3BV, and then the loading liquid and the elution liquid are combined and concentrated, so that the alkaloid component is obtained. The component is separated by multi-step high-performance preparative liquid chromatography, so that three compounds capparissane D, capparissane E and capparissane F are obtained.

[0011] The obtained three compounds are subjected to beta2-adrenergic receptor (beta2-AR) activity test, and human epidermal carcinoma A431 cells are used for the activity test, and results show that capparissane F shows micromolar level antagonistic activity to beta2-AR receptor, and can become a potential lead compound for treating rheumatoid arthritis and other diseases.

[0012] The compound can be obtained by separation and purification from plants, or can be obtained by synthesis through chemical methods well known to those skilled in the art.

[0013] The compound can be used alone or in combination, and can be combined with a pharmaceutically suitable carrier or excipient to form an oral or non-oral dosage form according to a conventional method.

[0014] Obviously, according to the above content of the present application, according to the ordinary technical knowledge and common means in the art, other various forms of modification, replacement or change can be made without departing from the above basic technical idea of the present application.

[0015] The three novel compounds (capparissane D, capparissane E and capparissane F) provided by the present application are spermidine alkaloids containing glycoside structure, which are extracted and separated from capparidaceae capparis liana plant capparis divaricata.

[0016] The present application has the following advantages: the target compound is a novel spermidine alkaloid; capparissane F is the first spermidine alkaloid containing thioglycoside separated from natural products, and the compound has micromolar level beta2-AR receptor antagonistic activity, and has application prospect for the development of rheumatoid arthritis drugs which are currently highly concerned. The biological activity experiment shows that it has beta2-adrenergic receptor antagonistic activity, and can be used in drugs for preventing and treating diseases related to the receptor, such as rheumatoid arthritis, arrhythmia and the like. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 H NMR spectrum of capparissane D 1 H NMR spectrum of capparissane D

[0018] Figure 2 C NMR spectrum of capparissane D 13 C NMR spectrum of capparissane D

[0019] Figure 3 H NMR spectrum of capparissane E 1 H NMR spectrum of capparissane E

[0020] H NMR spectrum of capparissane E 1 H NMR spectrum of capparissane E

[0020] H NMR spectrum of capparissane E 1 H NMR spectrum of capparissane E

[0020] Figure 4 H NMR spectrum of capparissane E 13 H NMR spectrum of capparissane E

[0021] Figure 5 H NMR spectrum of capparissane E 1 H NMR spectrum of capparissane E

[0022] Figure 6 H NMR spectrum of capparissane E 13 H NMR spectrum of capparissane E

[0023] Figure 7 H,H-COSY and key HMBC of capparissane D, capparissane E and capparissane F

[0024] Figure 8 Dose response of capparissane F on β2-AR DETAILED DESCRIPTION

[0025] The following examples are intended to illustrate the present application and not to further limit it, which can be implemented in any way described in the summary of the invention.

[0026] Preparation example of compound of formula (II) of the present application:

[0027] Preparation of compounds and structure identification:

[0028] In the following preparation examples, the preparation system includes Alliance (Waters Corporation), including e2695 separation unit, 2998 PDA detector, data processing by empow er 3; Auto-Purification (Waters) automatic purification system, including 2545 separation unit, 2767 sample manager, 2489 dual wavelength detector; novasep HPLC industrial high pressure liquid preparation chromatography (novasep separation technology Co., Ltd.) and HIPERSEP software. Reagents include chromatographic grade methanol and acetonitrile purchased from Thermo Fisher Scientific, chromatographic grade formic acid (FA), acetic acid (HAc), ammonium formate (NH4FA) purchased from Bailingwei Technology Co., Ltd. (Hebei, China), laboratory water from Milli-Q ultrapure water purification system (Billerica, MA, USA), and preparation grade methanol purchased from Shanghai Xingke High Purity Solvent Co., Ltd. (Shanghai, China).

[0029] NMR spectra were recorded on a Bruker AVIII-600 NMR spectrometer (Bruker, German) and the compounds were dissolved in deuterated methanol (MeOD). Mass spectrometry was performed on an Agilent 1290 Infinity LC / 6540 Q-TOF MS liquid chromatography-mass spectrometry system, including a high-pressure binary pump, an autosampler, a column oven system, a diode array detector, an electrospray ion source, and Agilent MassHunter software. Mass spectrometry detection was performed in positive ion scanning mode, with a scan range of m / z MS: 100-1000 and MS / MS: 50-1000. Other conditions were as follows: nitrogen flow rate, 8 L / min; nitrogen temperature, 350 °C; nebulizer gas pressure, 35 psi; capillary voltage, 3500 V; and collision voltage, 75 V.

[0030] The preparation and compound identification steps were as follows:

[0031] (1) Extraction of medicinal materials: 25 kg of fruits of Capparis miclana Hook.f.et Thoms were extracted with 250 L of 1% acetic acid / water at room temperature for 3 hours, and then rotary evaporated to a volume of about 30 L. Then, the filtrate (membrane permeate) was obtained by filtration using a 10 KD ultrafiltration membrane, and then treated with anion exchange resin PA408 (Mitsubishi Chemical Corporation, Japan). The treatment process mainly included three steps of equilibration, sample loading, and elution. The equilibration step used 1% HAc / H2O (v:v) to equilibrate 3 BV (column volume), the sample loading step was to directly load the filtrate of the ultrafiltration membrane, and the elution step used 1% acetic acid (HAc) / water to elute 3 BV after sample loading. The sample loading liquid and the elution liquid were combined as the Capparis miclana alkaloid component.

[0032] (2) The alkaloid component obtained in step (1) was subjected to first-dimensional separation and purification using an FC8 SCX (inner diameter x length = 200 mm x 250 mm, filler particle size 40 μm) chromatographic column, with a flow rate of 1.2 L / min, mobile phase A being 0.1% FA / MeOH (v / v), B being 0.1% FA / H2O (v / v), and C being 80 mM NH4FA / MeOH:H2O (50:50, v:v), with an elution gradient of 0-20 min, 5% B1; 20-40 min, 50% B; 40-60 min, 100% C. The sample collected at 40-60 min was concentrated and freeze-dried to obtain a solid sample F3.

[0033] (3) The CSGS-F3 obtained in step (2) was subjected to second dimensional separation and purification using a FC8 SCX (inner diameter x length 100 mm x 250 mm, packing particle size 5 μm) column at a flow rate of 320 mL / min, with mobile phase A being 10 mM NH4FA / MeOH:H2O (95:5, v / v) and B being 10 mM NH4FA / H2O, and gradient elution conditions being 0-5 min, 5% A; 5-8 min, 5-32% A (linear, same below); 8-48 min, 32-63% A; 48-53 min, 63-100% A; 53-65 min, 100% A, to obtain 7 sub-fractions F3-1 to F3-7 by chromatographic peak collection.

[0034] (4) The sub-fraction F3-2 (collection time 10-20 min) obtained in step (3) was subjected to third dimensional separation and purification using a CYS (inner diameter x length 50 mm x 250 mm, packing particle size 7 μm) column at a flow rate of 80 mL / min, with mobile phase A being ACN and B being 0.1% FA / H2O (v / v), and gradient elution conditions being 0-50 min, 95-60% A (linear, same below); 50-52 min, 60-10% A; 52-65 min, 10% A, to obtain 10 sub-fractions F3-2-1 to F3-2-10 by chromatographic peak collection.

[0035] (5) The sub-fraction F3-2-7 (collection time 30-38 min) obtained in step (4) was subjected to fourth dimensional separation and purification using a C18 HCE (inner diameter x length 50 mm x 250 mm, packing particle size 7 μm) column, with mobile phase A being 0.1% FA / MeOH (v / v) and B being 0.1% FA / H2O (v / v), and gradient elution conditions being 0-40 min, 0-25% A (linear, same below); 40-42 min, 25-90% A; 42-50 min, 90% A. 7 sub-fractions F3-2-7-1 to F3-2-7-7 were obtained by chromatographic peak collection.

[0036] (6) The fractions F3-2-7-2 (collection time 20-22 min), F3-2-7-5 (collection time 27-28 min), and F3-2-7-6 (collection time 30-32 min) obtained in step (5) were separated and purified in the fifth dimension by passing them through a PHPCE (inner diameter × length 10 mm × 250 mm, packing particle size 7 μm) chromatographic column. The flow rate was 3.3 mL / min, the mobile phase A was 0.1% FA / ACN (v / v), and the mobile phase B was 0.1% FA / H2O (v / v). The gradient elution (linear) conditions were 0-25 min, 20% A (F3-2-7-2); 0-25 min, 21% A (F3-2-7-5); 0-25 min, 26% A (F3-2-7-6). The main peaks collected were F3-2-7-2-1 (collection time 11-14 min), F3-2-7-5-1 (collection time 10-14 min), and F3-2-7-6-1 (collection time 15-18 min), corresponding to the compounds capparissane E, capparissane D, and capparissane F.

[0037] (7) The structure identification of the compound and related information are as follows:

[0038] capparissane D:C 31 H 39 N3O9, white solid, HR-ESI-MS (m / z): 598.2763 [M+H] + (calcdfor C 31 H 40 N3O9,598.2759); 1H NMR (600MHz, MeOD) δ7.20(2H,d,J=8.2Hz,H-4,H-28),7.73(2H,d,J=8.2Hz,H-5,H-29),7.57(1H,d,J=16.1Hz,H-7),6.61(1H,d,J=16.1Hz,H-8),3.2 8(2H,m,H-11),1.52(2H,m,H-12),1.69(2H,m,H-13),2.98(4H,m,H-14,H- 16),1.95(2H,m,H-17),3.51(2H,m,H-18),5.92(1H,d,J=15.5Hz,H-21),7. 34(1H,d,J=15.5Hz,H-22),6.99(1H,dd,J=8.5,1.6Hz,H-24),7.25(1H,d, J=8.5Hz,H-25),6.47(1H,d,J=1.6Hz,H-27),5.11(1H,d,J=7.6Hz,H-1′),3 .58(1H,m,H-2′),3.51(1H,m,H-3′),3.44(1H,m,H-4′),3.49(1H,m,H-5′) ,3.91(1H,dd,J=12.1,1.7Hz,H-6a′),3.72(1H,dd,J=12.1,5.9Hz,H-6b′); 13 C NMR(150MHz,MeOD)δ151.9(C-1),158.3(C-3),124.2(C-4,C-28),130.9(C-5,C-29),134.3(C-6),141.1( C-7),123.7(C-8),168.7(C-9),40.2(C-11),26.7(C-12),24.6(C-13),49.6(C-14),46.2(C-16),27.3(C- 17),37.5(C-18),168.5(C-20),120.5(C-21),140.7(C-22),130.7(C-23),126.4(C-24),118.1(C-25),14 9.5(C-26),112.5(C-27),102.2(C-1′),75.0(C-2′),78.0(C-3′),71.3(C-4′),78.4(C-5′),62.5(C-6′). 1 H-NMR and 13 The C-NMR spectrum is as follows Figure 1 and Figure 2 As shown, the key two-dimensional NMR information of the compound is as follows: Figure 7 As shown.

[0039] capparissane E:C 31 H 39 N3O9, a white solid, HR-ESI-MS (m / z): 598.2764 [M+H] + (calcd for C 31 H 40 N3O9, 598.2759); 1 1H NMR (600 MHz, MeOD) δ 7.13 (2H, d, J = 8.2 Hz, H-4, H-28), 7.69 (2H, d, J = 8.2 Hz, H-5, H-29), 7.60 (1H, d, J = 16.1 Hz, H-7), 6.66 (1H, d, J = 16.1 Hz, H-8), 3.49 (2H, m, H-11), 1.71 (2H, m, H-12), 1.73 (2H, m, H-13), 2.96 (4H, m, H-14, H-16), 1.61 (2H, m, H-17), 2.58 (2H, m, H-18), 5.81 (1H, d, J = 12.9 Hz, H-21), 6.49 (1H, d, J = 12.9 Hz, H-22), 6.93 (1H, dd, J = 8.5, 1.8 Hz, H-24), 7.23 (1H, d, J = 8.5 Hz, H-25), 6.94 (1H, d, J = 1.6 Hz, H-27), 5.07 (1H, d, J = 7.6 Hz, H-1′), 3.54 (1H, m, H-2′), 3.47 (1H, m, H-3′), 3.41 (1H, m, H-4′), 3.46 (1H, m, H-5′), 3.89 (1H, dd, J = 12.1, 1.8 Hz, H-6a′), 3.71 (1H, dd, J = 12.1, 5.5 Hz, H-6b′); 13C NMR(150MHz,MeOD)δ149.9(C-1),159.2(C-3),123.2(C-4,C-28),131.1(C-5,C-29),132.4(C-6),141.9( C-7),121.4(C-8),170.0(C-9),37.9(C-11),21.8(C-12),25.6(C-13),47.4(C-14),45.8(C-16),27.2(C- 17),37.7(C-18),170.2(C-20),122.7(C-21),135.7(C-22),130.9(C-23),127.3(C-24),118.0(C-25),14 9.2(C-26),117.4(C-27),102.4(C-1′),74.9(C-2′),78.0(C-3′),71.3(C-4′),78.4(C-5′),62.5(C-6′). 1 H-NMR and 13 The C-NMR spectrum is as follows Figure 3 and Figure 4 As shown, the key two-dimensional NMR information of the compound is as follows: Figure 7 As shown.

[0040] capparissane F:C 31 H 40 N3O9S, white solid, HR-ESI-MS (m / z): 630.2484 [M+H]+ (calcd for C 31 H 40 N3O9S, 630.2480); 1H NMR (600MHz, MeOD) δ7.12(2H,d,J=8.3Hz,H-4,H-28),7.69(2H,d,J=8.3Hz,H-5,H-29),7.67(1H,d,J=16.0Hz,H-7),6.67(1H,d,J=16.0Hz ,H-8),3.53(2H,m,H-11),1.97(2H,m,H-12),3.02(2H,m,H-13),2.90 (2H,m,H-15),1.41(2H,m,H-16),1.35(2H,m,H-17),2.49(2H,m,H-18 ),6.90(1H,s,H-22),6.86(1H,dd,J=8.5,1.2Hz,H-24),6.88(1H,d,J=8.5Hz,H-25),6.76(1H,d,J=1.2Hz,H-27),4.40(1H,d,J=9.6Hz,H- 1′),3.23(1H,m,H-2′),3.30(1H,m,H-3′),3.21(1H,m,H-4′),3.21(1H,m,H-5′),3.82(1H,dd,J=11.0,1.0Hz,H-6a′),3.55(1H,m,H-6b′); 13 C NMR(150MHz,MeOD)δ148.3(C-1),159.5(C-3),122.8(C-4,C-28),131.3(C-5,C-29),132.1(C-6),143.1( C-7),120.6(C-8),169.9(C-9),36.6(C-11),26.6(C-12),43.7(C-13),46.3(C-14),24.8(C-16),27.4(C- 17),40.7(C-18),170.6(C-20),124.5(C-21),139.8(C-22),127.8(C-23),128.5(C-24),117.5(C-25),1 49.9(C-26),116.0(C-27),88.5(C-1′),73.9(C-2′),79.6(C-3′),71.5(C-4′),82.4(C-5′),63.1(C-6′). 1 H-NMR and 13 The C-NMR spectrum is as follows Figure 5 and Figure 6 As shown, the key two-dimensional NMR information of the compound is as follows: Figure 7 As shown.

[0041] Activity test examples:

[0042] Experimental methods:

[0043] β2-AR activity assays were performed using human epidermal cancer cells (A431) cultured in DMEM medium containing 10% fetal bovine serum (FBS). Cells were seeded in 384-well plates at a density of approximately 30,000 cells / well. Activity screening was performed using dynamic mass resetting (DMR) assays on an Epic system.

[0044] The first step in the dose-response curve assay involved adding a serially diluted compound in DMSO, resulting in 14 concentration points. The final concentrations in the wells were 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.563 μM, 0.781 μM, 0.391 μM, 0.196 μM, 0.098 μM, 0.049 μM, 0.024 μM, and 0.012 μM. After establishing a baseline for 2 minutes, the DMR signal induced by the analyte compound was recorded over 1 hour. The highest signal value within 1 hour for each concentration was recorded and denoted as S1. The second step involved adding a probe molecule (the β2-AR agonist isoprenaline, in DMSO, with a final concentration of 1.25 nM in the wells), and recording the DMR response signal for another hour. Similarly, the highest signal value within 1 hour for each concentration was recorded and denoted as S2. Finally, the S2 data are plotted as a concentration-response value curve, which is the dose-response curve.

[0045] For the compound Capparissane F, its DMR signal in S1 showed no significant change with increasing concentration, but in S2 it dose-dependently reduced the DMR signal produced by the β2-AR agonist isoprenaline in A431 cells, indicating that it does indeed possess β2-AR antagonistic activity. The β2-AR receptor dose-response curve of Capparissane F is shown below. Figure 8 As shown, IC is calculated 50 The concentration was 40.07 μM. Therefore, this compound is a lead compound for treating or improving β2-AR-related activities such as rheumatoid arthritis.

Claims

1. An agmatine glycoside alkaloid, characterized by: It is one or two or three of the following compounds of spermidine glycoside alkaloids as shown in the following structure as formula (I); The spermidine glycoside alkaloids include one or two or more of the following compounds of formula (I), or crystal form thereof, or pharmaceutically acceptable salt thereof.

2. A process for the preparation of agmatine glycoside alkaloid as claimed in claim 1, wherein the process comprises: Comprising the following steps: (1) medicinal material extraction: taking the fruits of Capparis spinosa L., extracting with 0.1-2% acetic acid aqueous solution at room temperature or heating (temperature range 50-100℃) for 2-24 hours, then filtering with 10-50KD filter membrane, obtaining the filtrate, and then treating with anion exchange resin; collecting the sample solution and water eluent with flow-through mode, and then concentrating after combining, as Capparis spinosa alkaloid component; (2) purifying the alkaloid component obtained in step (1) with FC8SCX column in the first dimension separation, mobile phase A is 0.1-1% FA / MeOH (v / v), B is 0.1-1% FA / H2O (v / v), C is 10-200mM NH4FA / MeOH:H2O (50:50, v:v), column specification is 100mm×250mm (inner diameter×length), flow rate is 320mL / min, elution gradient: 0-20min, 5% B1; 20-40min, 50% B; 40-60min, 100% C; collecting the sample of 40-60min, and then concentrating after combining, as F3; (3) purifying F3 obtained in step (2) with FC8SCX column in the second dimension separation, mobile phase A is 10-100mM NH4FA / MeOH:H2O (95:5, v / v), B is 10-100mM NH4FA / H2O, column specification is 100mm×250mm (inner diameter×length), flow rate is 320mL / min, gradient elution conditions are 0-5min, 5% A; 5-8min, 5-32% A; 8-48min, 32-63% A; 48-53min, 63-100% A; 53-65min, 100% A, collecting 7 sub-fractions F3-1-F3-7 according to chromatographic peaks; (4) purifying the sub-fraction F3-2 (collection time 10-20min) obtained in step (3) with CYS in the third dimension separation, mobile phase A is ACN, B is 0.1-1% FA / H2O (v / v), column specification is 50mm×250mm (inner diameter×length), flow rate is 80mL / min, gradient elution conditions are 0-50min, 95-60% A; 50-52min, 60-10% A; 52-65min, 10% A, collecting 10 sub-fractions F3-2-1-F3-2-10 according to chromatographic peaks; (5) The sub-fraction F3-2-7 (collection time 30-38 min) obtained in step (4) was subjected to fourth dimensional separation and purification using a C18 HCE chromatographic column, the mobile phase A was 0.1%-1% FA / MeOH (v / v), B was 0.1%-1% FA / H2O (v / v), the column size was 50 mm x 250 mm (inner diameter x length), the flow rate was 80 mL / min, and the gradient elution conditions were 0-40 min, 0-25% A; 40-42 min, 25-90% A; 42-50 min, 90% A. Seven sub-fractions F3-2-7-1 to F3-2-7-7 were collected according to the chromatographic peaks; (6) The sub-fractions F3-2-7-2 (collection time 20-22 min), F3-2-7-5 (collection time 27-28 min), and F3-2-7-6 (collection time 30-32 min) obtained in step (5) were subjected to fifth dimensional separation and purification using a PHPCE chromatographic column, the column size was 10 mm x 250 mm (inner diameter x length), the flow rate was 3.3 mL / min, the mobile phase A was 0.1%-1% FA / ACN (v / v), B was 0.1%-1% FA / H2O (v / v), and the gradient elution conditions were 0-25 min, 20% A (F3-2-7-2); 0-25 min, 21% A (F3-2-7-5); 0-25 min, 26% A (F3-2-7-6). The main peaks were collected as F3-2-7-2-1 (collection time 11-14 min), F3-2-7-5-1 (collection time 10-14 min), and F3-2-7-6-1 (collection time 15-18 min), corresponding to compounds capparissane E, capparissane D, and capparissane F.

3. Use of spermidine glycoside alkaloids in the preparation of a medicament for preventing and / or treating one or more than two β2-adrenergic receptor related diseases.

4. The use according to claim 3, wherein: the spermidine glycoside alkaloids comprise one or more than two of the three spermidine glycoside alkaloid compounds, or crystal forms thereof, or pharmaceutically acceptable salts thereof; and the β2-adrenergic receptor related diseases comprise one or more than two of rheumatoid arthritis or arrhythmia.

5. A medicament comprising one or more than two of the three spermidine glycoside alkaloid compounds, or crystal forms thereof, or isomers thereof, or pharmaceutically acceptable salts thereof. Also included is a pharmaceutical composition comprising one or more than two of any other pharmaceutically acceptable adjuvant, carrier, diluent, or other active ingredient. ​ 6. The medicament according to claim 5, characterized in that: ​

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