阅读说明：本技术 从香蕉花中提取的生物碱类化合物及其提取方法 (Alkaloid compound extracted from banana flower and extraction method thereof ) 是由 何雪梅 孙健 唐雅园 刘国明 叶冬青 李丽 李昌宝 李志春 杨莹 李杰民 郑凤锦 于 2021-01-15 设计创作，主要内容包括：本发明属于生物碱类化合物提取的技术领域,具体涉及一种从香蕉花中提取的生物碱类化合物,它由以下步骤提取得到：(1)将新鲜香蕉花晒干,浸提,过滤,减压浓缩,获得乙醇提取物；(2)将乙醇提取物分散至水中,依次采用石油醚和乙酸乙酯萃取,得到乙酸乙酯浸膏；(3)将乙醇乙酯浸膏溶解于甲醇中,分别通过洗脱液为甲醇和氯仿-甲醇的硅胶色谱柱进行洗脱；(4)再通过高效液相色谱和硅胶色谱柱进行洗脱；(5)将依次通过高效液相色谱、硅胶色谱柱、高效液相色谱恒冲,用氯仿-甲醇洗脱,收集洗脱液,获得生物碱类化合物。通过本发明方法制备的生物碱类化合物收率高,且具有较佳的抑制血小板聚集活性。(The invention belongs to the technical field of alkaloid compound extraction, and particularly relates to an alkaloid compound extracted from banana flowers, which is extracted by the following steps: (1) sun drying fresh banana flower, extracting, filtering, and concentrating under reduced pressure to obtain ethanol extract; (2) dispersing the ethanol extract into water, and sequentially extracting with petroleum ether and ethyl acetate to obtain ethyl acetate extract; (3) dissolving the ethyl acetate extract in methanol, and eluting with silica gel chromatographic column containing methanol and chloroform-methanol as eluents; (4) then eluting by high performance liquid chromatography and silica gel chromatographic column; (5) sequentially subjecting to high performance liquid chromatography, silica gel chromatographic column, and high performance liquid chromatography under constant pressure, eluting with chloroform-methanol, and collecting eluate to obtain alkaloid compounds. The alkaloid compound prepared by the method has high yield and better activity of inhibiting platelet aggregation.)

1. The alkaloid compound extracted from banana flower is characterized by having the following chemical structural formula:

2. the method for extracting alkaloid compounds according to claim 1, wherein: the method comprises the following steps:

(1) sun drying fresh banana flower, extracting with ethanol for 3 times, each time for 1 day, mixing extractive solutions, filtering, and concentrating under reduced pressure to obtain ethanol extract;

(2) dispersing the ethanol extract into water, and sequentially extracting with petroleum ether and ethyl acetate to obtain ethyl acetate extract;

(3) dissolving the ethyl alcohol extract in methanol, eluting by a silica gel chromatographic column with methanol as eluent, and then performing gradient elution by a silica gel chromatographic column with chloroform-methanol as gradient eluent, wherein the gradient elution is sequentially performed by the chloroform-methanol volume ratio of 100:1 → 60:1 → 30:1 → 10:1 → 5: 1;

(4) concentrating the receiving solution of the gradient elution part of 60:1 in the step (3), performing gradient elution by using high performance liquid chromatography with methanol with the volume percentage of 20% and the volume percentage of 26% in sequence, and performing gradient elution by using a silica gel chromatographic column, wherein the gradient elution is chloroform-methanol, and the gradient elution is performed by using chloroform-methanol with the volume ratio of 100:1 → 60:1 → 30:1 → 10:1 in sequence;

(5) and (3) combining and concentrating the receiving solution of the gradient elution part with the ratio of 30:1 in the step (4), performing constant flushing by using methanol with the volume percentage of 43% through high performance liquid chromatography, performing constant flushing by using a silica gel chromatographic column with the volume ratio of chloroform to methanol of 15:1, continuously performing constant flushing by using high performance liquid chromatography with the volume percentage of 40% and using chloroform to methanol of 4:1 as an eluent, and collecting and concentrating the eluent with the Rf value of 0.5 to obtain the required product.

3. The method for extracting alkaloid compounds according to claim 2, wherein: in the step (1), the temperature of the reduced pressure concentration is 25-40 ℃, and the pressure is (-0.06) — (-0.1) MPa.

4. The method for extracting alkaloid compounds according to claim 2, wherein: in the step (2), petroleum ether and ethyl acetate are extracted for 2-4 times respectively.

5. The method for extracting alkaloid compounds according to claim 2, wherein: in the step (1), before leaching, the banana flowers are treated as follows: adding ethanol into the sun-dried banana flower, pulping, homogenizing, and performing dynamic ultrahigh pressure micro-jet treatment.

6. The method for extracting alkaloid compounds according to claim 5, wherein: the ratio of banana flower to ethanol is 1g to 40-50 ml.

7. The method for extracting alkaloid compounds according to claim 5, wherein: the homogenization conditions are as follows: the pressure is 15-20MPa, and the time is 15-20 min.

8. The method for extracting alkaloid compounds according to claim 5, wherein: the dynamic ultrahigh pressure microjet treatment conditions are as follows: the pressure is 120-125MPa, the temperature is 50-60 ℃, and the time is 50-80 min.

Technical Field

The invention belongs to the technical field of alkaloid compound extraction, and particularly relates to an alkaloid compound extracted from banana flowers and an extraction method thereof.

Background

The banana flower is waste which is almost equal to fruit in quantity and is generated after the bananas are ripe and picked, each banana plant generates male flower with the weight of about 2kg, and the annual banana flower yield in China is about 500 ten thousand tons or more according to the planting density of 180 bananas per mu and the area of bananas in China. The banana flower is sweet in taste, slightly pungent and cool in nature, has the effects of reducing phlegm and relieving distension and fullness, calming the liver and removing blood stasis and the like, and is suitable for symptoms such as chest and diaphragm fullness, abdominal distension and pain, acid regurgitation, dizziness, rheumatic pain and the like. Banana flowers are used as vegetables in many asian countries, such as srilanca, malaysia, indonesia, etc., and they are often consumed in a cooked and fried form. In India, banana flowers have been consumed for thousands of years as a medicinal material for improving female breast milk and alleviating dysmenorrhea. A large amount of banana flowers in banana producing areas in China are treated as waste materials, and due to the fact that the banana flowers are high in water content and long in rotting time, insect pests such as weevils are easily caused, secondary pollution to banana garden environment is not caused, a large amount of plant resources are wasted, and the banana flowers are not fully utilized at the present stage.

In the production process of bananas, wastes such as banana flowers, stems and leaves, which are almost equal to fruits, are generated, and the maximum value of bananas is not facilitated, so how to improve the comprehensive utilization of banana wastes is one of the problems to be solved by the banana industry at present. The banana flower has higher nutritive value and good development prospect, and domestic scholars research the chemical components of the banana flower, namely the banana flower contains substances such as saponin, flavone and the like with high content; the research of foreign scholars shows that the water, ethanol and chloroform extracts of banana flowers have strong blood sugar reducing efficacy, and the fresh and tender parts of the flower diameter and the flowers can also be eaten as vegetables. At present, the previous research on active substances mainly focuses on flavones in banana flowers, and the research on other components in banana flowers is less. At present, researches on banana peanut alkaloid compounds and extraction methods thereof are not found, and researches on the properties and the application of the novel alkaloid compounds in banana flowers are not reported.

Disclosure of Invention

The invention aims to solve the technical problems and provides an alkaloid compound extracted from banana flowers and an extraction method thereof.

The technical scheme of the invention is as follows:

the invention provides an alkaloid compound extracted from banana flowers, which has the following chemical structural formula:

the invention also provides an extraction method of the alkaloid compounds extracted from the banana flowers, which comprises the following extraction steps:

(1) sun drying fresh banana flower, extracting with ethanol for 3 times, each time for 1 day, mixing extractive solutions, filtering, and concentrating under reduced pressure to obtain ethanol extract;

(2) dispersing the ethanol extract into water, and sequentially extracting with petroleum ether and ethyl acetate to obtain ethyl acetate extract;

(3) dissolving the ethyl alcohol extract in methanol, eluting by a silica gel chromatographic column with methanol as eluent, and then performing gradient elution by a silica gel chromatographic column with chloroform-methanol as gradient eluent, wherein the gradient elution is sequentially performed by the chloroform-methanol volume ratio of 100:1 → 60:1 → 30:1 → 10:1 → 5: 1;

(4) concentrating the receiving solution of the gradient elution part of 60:1 in the step (3), performing gradient elution by high performance liquid chromatography with 20% and 26% methanol in sequence, and performing gradient elution by a silica gel chromatographic column, wherein the gradient elution is chloroform-methanol, and the gradient elution is performed in sequence with the chloroform-methanol volume ratio of 100:1 → 60:1 → 30:1 → 10: 1;

(5) and (3) combining and concentrating the receiving solution at the gradient elution part of 30:1 in the step (4), performing constant flushing by using high performance liquid chromatography with 43% methanol, performing constant flushing by using a silica gel chromatographic column at a chloroform-methanol volume ratio of 15:1, continuously performing constant flushing by using high performance liquid chromatography with 40% methanol, using chloroform-methanol volume ratio of 4:1 as an eluent, and collecting and concentrating the eluent with the Rf value of 0.5 to obtain the required product.

The percentages of methanol in the invention are volume percentages.

In the extraction of the alkaloid compounds, the obtained ethanol extract is relatively dilute, and for the convenience of subsequent purification, preferably, in the step (1) of the invention, the temperature for reduced pressure concentration is 25-40 ℃, and the pressure is (-0.06) — (-0.1) MPa.

In order to increase the extraction rate and sufficiently extract the alkaloid compounds, preferably, in the step (2) of the present invention, petroleum ether and ethyl acetate are extracted 2 to 4 times respectively.

Before leaching, in order to improve the extraction rate, preferably, in the step (1) of the invention, before leaching, the banana flowers are treated as follows: adding ethanol into the sun-dried banana flower, homogenizing, and carrying out dynamic ultrahigh pressure micro-jet treatment. The banana flower is subjected to comprehensive effects of strong shearing, high-speed impact, pressure instant release, vortex and the like in a reaction cavity of the micro-jet homogenizer, so that banana flower raw material particles are more uniformly dispersed, mixed and micronized in water; the crushing degree of the banana flower raw material cells is increased, and the solvent can easily enter the cells by utilizing the permeability or penetrability of the cells, so that the solvent can quickly reach the dissolving balance with the effective components in the cells, and the extraction is more favorable.

In the dynamic ultrahigh pressure micro-jet treatment, the ratio of material to liquid is also one of important elements for improving the yield, if the liquid amount is too low, the extraction cannot be fully carried out, if the liquid amount is too large, the ethanol extract is relatively more dispersed, in a reaction cavity, under the violent action, the structure of the alkaloid compound can be damaged, and the yield is reduced, so in order to improve the yield, the ratio of material to liquid of the banana flower and the ethanol is preferably 1g:40-50 ml.

Homogenization is also called homogenizing, and is a process of micronizing and homogenizing a dispersion in a suspension (or emulsion) system, and the process simultaneously plays roles of reducing the size of the dispersion and improving the distribution uniformity of the dispersion, and for better performing subsequent dynamic ultrahigh pressure micro-jet, the homogenization is performed first, preferably, the homogenization conditions are as follows: the pressure is 15-20MPa, and the time is 15-20 min.

When the banana flower raw material is treated by the dynamic ultrahigh pressure microjet, the subsequent purification yield is influenced by the treatment pressure, temperature and treatment time, if the purification time is not enough, the highest yield cannot be reached, and under the action of instantaneous high pressure in a certain time, the alkaloid compound can fully permeate into the solvent in a very short time, and can be dissolved in a certain time to quickly reach the balance, if the treatment time is continuously prolonged, the extraction rate cannot be improved, the extraction period is also prolonged, the cost is improved, in addition, the increase of the treatment temperature is favorable for the dissolution of the alkaloid compound, the extraction rate is increased, but the structure of the alkaloid compound is damaged too high, the extraction of the alkaloid compound is not favorable, and in a certain pressure range, the pressure is increased, the ethanol extract particles are favorably refined, the dissolution of effective substances is favorable, but the pressure is too high, the ethanol extract is lost, which is not beneficial to extraction. Therefore, in order to improve the extraction rate, the dynamic ultrahigh pressure micro-jet treatment conditions of the present invention are preferably as follows: the pressure is 120-125MPa, the temperature is 50-60 ℃, and the time is 50-80 min.

The inventor finds that the alkaloid compound has certain activity of inhibiting platelet aggregation, so that various thrombotic and embolic diseases can be prevented and treated.

Due to the adoption of the technical scheme, the invention has the beneficial effects that:

1. the extraction method can be used for extracting the novel alkaloid compound from the banana flower, the yield is high, the quality is stable, and the test of the applicant shows that the alkaloid compound has the activity of inhibiting the platelet aggregation and is expected to be used for preparing the medicament for inhibiting the platelet aggregation.

2. Before leaching, the banana flower is pulped and then is subjected to homogenization treatment, and then is subjected to dynamic ultrahigh-pressure micro-jet treatment, so that solid-phase particles of the banana flower can be refined, cell walls of raw materials of the banana flower are broken, the mass transfer speed is accelerated, the follow-up purification is facilitated, and the yield is improved.

3. The method takes the banana flowers as the raw material to extract the alkaloid, can change waste into valuable, reduces the production cost and improves the economic benefit of the banana planting industry.

Drawings

FIG. 1 is an IR (KBr) spectrum of a compound obtained in example 1 of the present invention;

FIG. 2 is an ESI-MS spectrum of a compound obtained in example 1 of the present invention;

FIG. 3 is a HR-ESI-MS spectrum of the compound obtained in example 1 of the present invention;

FIG. 4 shows the obtention of example 1 of the present inventionProcess for preparing compounds¹An H-NMR spectrum;

FIG. 5 shows the preparation of the compound obtained in example 1 of the present invention¹³A C-NMR spectrum;

FIG. 6 shows the preparation of the compound obtained in example 1 of the present invention¹H-¹H COSY map;

FIG. 7 is an HSQC spectrum of the compound obtained in example 1 of the present invention;

FIG. 8 is an HMBC profile of the compound obtained in example 1 of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The percentages in the present invention are all by weight unless otherwise specified.

Example 1

(1) Sun drying 7kg fresh banana flower, extracting with ethanol for 3 times, each time for 1 day, mixing extractive solutions, filtering, and concentrating under reduced pressure at 25 deg.C and-0.06 MPa until the liquid has no taste to obtain ethanol extract;

(2) dispersing the ethanol extract into water, and extracting with petroleum ether and ethyl acetate for 3 times to obtain 965g of ethyl acetate extract;

(3) dissolving the ethyl alcohol extract in methanol, adding silica gel 2 times of the weight of the 100-mesh ethyl alcohol extract, mixing the samples, after the solvent is volatilized, eluting by using a silica gel chromatographic column with methanol as eluent, and then performing gradient elution by using a silica gel chromatographic column with chloroform-methanol as gradient eluent, wherein the gradient elution is sequentially performed by using chloroform-methanol in a volume ratio of 100:1 → 60:1 → 30:1 → 10:1 → 5: 1;

(4) concentrating the receiving solution of the gradient elution part of 60:1 in the step (3), performing gradient elution by using high performance liquid chromatography with 20% by volume of methanol and 26% by volume of methanol sequentially, and performing gradient elution by using a silica gel chromatographic column, wherein the gradient elution is chloroform-methanol, and the gradient elution by using the silica gel chromatographic column is performed sequentially with the chloroform-methanol volume ratio of 100:1 → 60:1 → 30:1 → 10: 1;

(5) combining and concentrating the receiving solution of the gradient elution part of 30:1 in the step (4), performing constant flushing by using methanol with the volume percentage of 43% through high performance liquid chromatography, performing constant flushing by using a silica gel chromatographic column with the volume ratio of chloroform to methanol of 15:1, continuously performing constant flushing by using methanol with the volume percentage of 40% through high performance liquid chromatography, dissolving by using chloroform to methanol with the volume ratio of 4:1 as a solvent, spotting the dissolved solution onto a Thin Layer Chromatography (TLC) silica gel plate by using a capillary tube, collecting and concentrating the eluent with the Rf value of 0.5, displaying the TLC plate in an orange red color, and recrystallizing the obtained crystals by using methanol to obtain 24mg of a target product which is white powder.

The white powder was analyzed for its bopp properties as follows:

UV(MeOH)λ_max: 221(sh),298 nm; IR (KBr) spectrum (see FIG. 1) showing aromatic aldehyde group-containing (1639 cm)^-1) And benzene rings (1526,1491 and 1452 cm)^-1) Characteristic absorption peak of (a); ESI-MS M/z 284[ M + Na ]]⁺(see FIG. 2); HR-ESI-MS M/z 284.0894[ M + Na ]]⁺(calculation value C)₁₄H₁₅NO₄Na,284.0893) (see FIG. 3), binding¹H-NMR (see FIG. 4) and¹³C-NMR (see FIG. 5) spectrum confirmed the molecular formula to be C₁₄H₁₅NO₄The unsaturation degree is 8. Of the present Compound¹H-NMR and¹³the C-NMR signals are fully assigned in Table 1, where:

¹H-NMR Spectroscopy (acetone-d)₆500MHz) shows 15 proton signals, of which delta_H2.86(2H, br.t, J ═ 7.8Hz) and 4.47(2H, br.t, J ═ 7.8Hz) are a set of methylene proton signals coupled to each other, and the structures are presumed to contain-CH₂-CH₂And (3) fragment. Aldehyde proton signal delta can be observed in the low field region_H9.53(1H, s); in addition, 2 groups of intercoupled aromatic proton signals were also observed: one group is ABX coupled proton signals delta on benzene ring_H6.52(1H, dd, J ═ 8.0,2.0Hz),6.71(1H, d, J ═ 8.0Hz), and 6.72(1H, d, J ═ 2.0); one group is proton signal delta on pyrrole ring_H6.94(1H, d, J ═ 4.0Hz) and 6.16(1H, d, J ═ 4.0 Hz).

¹³C-NMR(acetone-d₆125MHz) shows 14 carbon signals, respectively: 1 aldehyde carbon; group 1 benzene ring carbon signals; combined with a carbon signal from group 1 pyrrole ring and 3 saturated methylene carbons¹H-¹H COSY (see FIG. 6) and HSQC (see FIG. 7) can obtain C-3/C-4 linkage, C-5 '/C-6' linkage and C-1 '/C-2' linkage. The above NMR characteristics are very similar to the compound pyrrolezanthine (see Y.P.Yang, M.J.Chen, C.M.Teng, Y.L.Chang, I.L.Tsai, I.S.Chen, Phytochemistry,2002,61(5), 567-572.), with the main differences: the benzene ring in pyrrolezanthine is 1 ', 4 ' -para-disubstituted, while the benzene ring in the compound is 1 ', 3 ', 4 ' -trisubstituted. The above-mentioned position of the benzene ring substituent was further confirmed by HMBC (see fig. 8): h-2 "is associated with C-4", H-5 "is associated with C-3" and H-6 "is associated with C-2" and C-4 ".

In summary, the compound of the white powder of the above example is named 3 ″ -hydroxypyrolizanthine, which is a novel alkaloid, and the chemical structural formula of the obtained white powder can be determined as follows:

importance of the above-mentioned Compounds¹H-¹H COSYAnd HMBCThe relationship is as follows:

of the above compounds¹H-NMR and¹³the C-NMR signals are fully assigned in Table 1.

TABLE 1 preparation of the above-mentioned compounds¹H-NMR and¹³C-NMR spectroscopic data (acetone-d 6),δH 2.04,δC 29.8ppm)

Example 2

The difference from example 1 is: (1) sun drying 8kg fresh banana flower, extracting with ethanol for 3 times, each time for 1 day, mixing extractive solutions, filtering, and concentrating under reduced pressure at 40 deg.C and-0.1 MPa to obtain ethanol extract; (2) dispersing the ethanol extract into water, sequentially extracting with petroleum ether and ethyl acetate for 3 times respectively to obtain 976g of ethyl acetate extract; the remaining steps and parameters were the same as in example 1 to obtain 26mg of the desired product as a white powder. The obtained white powder was subjected to mass spectrometry and carbon spectrometry to determine the structure of the compound obtained in example 1 as described above.

Example 3

The difference from example 2 is: sun-drying 6kg fresh banana flower, adding ethanol at a material-liquid ratio of 1:40(g/ml), homogenizing under 15MPa for 20min, performing dynamic ultrahigh pressure microjet treatment at 120MPa and 50 deg.C for 80min, continuously leaching for 3 times, each for 1 day, mixing extractive solutions, filtering, and concentrating under reduced pressure at 40 deg.C and-0.1 MPa to obtain ethanol extract; the remaining steps and parameters were the same as in example 1 to give 16mg of the desired product as a white powder. The obtained white powder was subjected to mass spectrometry and carbon spectrometry to determine the structure of the compound obtained in example 1 as described above.

Example 4

The difference from example 2 is: sun drying 10kg fresh banana flower, adding ethanol at a material-liquid ratio of 1:50(g/ml), homogenizing under 20MPa for 15min, performing dynamic ultrahigh pressure microjet treatment at 125MPa and 60 deg.C for 50min, continuously leaching for 3 times, each for 7 days, mixing extractive solutions, filtering, and concentrating under reduced pressure at 40 deg.C and-0.1 MPa to obtain ethanol extract; the remaining steps and parameters were the same as in example 1 to give 29mg of the desired product as a white powder. The obtained white powder was subjected to mass spectrometry and carbon spectrometry to determine the structure of the compound obtained in example 1 as described above.

Comparative example 1

The difference from example 3 is: the dynamic ultrahigh pressure microjet treatment pressure is 115MPa, and the rest steps and parameters are the same as those in example 3, so that 8mg of white powdery target product is obtained. The obtained white powder was subjected to mass spectrometry and carbon spectrometry to determine the structure of the compound obtained in example 1 as described above.

Comparative example 2

The difference from example 3 is: the dynamic ultrahigh pressure microjet treatment pressure was 130MPa, and the remaining steps and parameters were the same as in example 3, to obtain 9mg of a white powdery target product. The obtained white powder was subjected to mass spectrometry and carbon spectrometry to determine the structure of the compound obtained in example 1 as described above.

From examples 2-3 and comparative examples 1-5, it can be seen that the yield of alkaloid compounds can be significantly improved after the leaching raw materials are treated by dynamic ultrahigh pressure micro-jet treatment, and the yield is influenced by too large or too small dynamic ultrahigh pressure micro-jet treatment pressure.

Test example: platelet aggregation inhibitory Activity test of alkaloid Compound of the present invention

1. Experimental Material

SD rat: male, SPF grade, weight 300. + -.20 g, purchased from animal center, Guangxi university of medical.

Reagents and instrumentation: the compound is obtained by separation in example 1; adenosine Diphosphate (ADP), Arachidonic Acid (AA) (beijing solibao technologies ltd); aspirin (Shanghai Aladdin Biotechnology Ltd.); clopidogrel (lep pharmaceuticals, inc); AggRAM^TMType platelet aggregation apparatus (Helena, USA).

2. Experimental methods

(1) Effect of compound on Adenosine Diphosphate (ADP) induced platelet aggregation

Taking blood from SD rat via abdominal aorta, anticoagulating with 129mmol/L sodium citrate, centrifuging at 800r/min for 10min to prepare Platelet Rich Plasma (PRP); after the extraction of the PRP from the supernatant,centrifuging at 3500r/min for 10min to prepare Platelet Poor Plasma (PPP), and adjusting PRP to 200 × 10 platelet count with PPP⁹And (4) one/L is reserved. The PRP is randomly grouped as follows: solvent control group (0.9% sodium chloride solution), positive control group (clopidogrel, 1.6mM), example 1 low dose group (0.05mM), example 1 medium dose group (0.1mM), example 1 high dose group (0.2 mM). In a quartz cup for detecting a platelet aggregation instrument (37 ℃), adding corresponding drugs into each group of PRP with adjusted concentration respectively, incubating for 5min, adding ADP with final concentration of 10 mu M to induce platelet aggregation, recording a 3min platelet aggregation curve, measuring the maximum aggregation rate of each group of platelets, and calculating the inhibition rate of a test product on platelet aggregation according to the following formula:

the inhibition ratio is [ (maximum platelet aggregation rate of the solvent control group-maximum platelet aggregation rate of the test sample)/maximum platelet aggregation rate of the solvent control group ] × 100%.

(2) Effect of Compounds on Arachidonic Acid (AA) induced platelet aggregation

PRP and PPP were prepared as described above, and PRP was adjusted to a platelet count of 500X 10⁹and/L, randomly grouping PRPs, namely: solvent control group (0.9% sodium chloride solution), positive control group (aspirin, 0.4mM), example 1 low dose group (0.05mM), example 1 medium dose group (0.1mM), example 1 high dose group (0.2 mM). And adding AA with the final concentration of 0.5mM after the PRP is incubated by the medicament to induce platelet aggregation for 3min, recording the maximum aggregation rate of the platelets, and calculating the inhibition rate of the medicament on the platelet aggregation.

(3) Effect of Compounds on Normal platelets

Preparation of PRP and PPP, adjustment of PRP to a platelet count of 200X 10⁹PRP was randomly divided into 4 groups, solvent control (physiological saline) group, low concentration group (0.25g/L) in example 1, medium concentration group (0.5g/L) in example 1, and high concentration group (1g/L) in example 1. After the PRP is incubated with the drug, the platelet aggregation curve and the maximum platelet aggregation rate are directly recorded, and the effect of the compound on normal platelets is observed.

3. Results of the experiment

TABLE 2 Effect of Compounds on ADP-induced platelet aggregation

From table 2, the compound of the present invention has significant inhibitory effect on ADP-induced platelet aggregation and is in dosage effect, the dosage is high, the aggregation inhibition rate is high, and the inhibitory effect on ADP-induced platelet aggregation is higher than that of the positive control clopidogrel group when the concentration of the compound reaches 0.2 mM.

TABLE 3 Effect of Compounds on AA-induced platelet aggregation

As can be seen from Table 3, the compound of the present invention has a significant inhibitory effect on platelet aggregation induced by AA, and is in a dosage effect, the dosage is high, the aggregation inhibition rate is high, when the concentration of the compound reaches 0.5g/L, the inhibitory effect on platelet aggregation induced by AA is greater than that of the positive control aspirin group, and when the concentration of the compound reaches 1g/L, the inhibitory effect on platelet aggregation induced by AA is greater than that of the positive control aspirin group.

The results in tables 2 and 3 show that the alkaloid compound of the invention has the activity of inhibiting platelet aggregation and is expected to be used for preparing the medicament for inhibiting platelet aggregation.

The above description is intended to describe in detail the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the claims of the present invention, and all equivalent changes and modifications made within the technical spirit of the present invention should fall within the scope of the claims of the present invention.