阅读说明：本技术 一种中药血盆草中的吲哚类生物碱及其提取纯化方法和应用 (Indole alkaloid in traditional Chinese medicine herba Rubi Corchorifolii, and extraction and purification method and application thereof ) 是由 杨武德 郭正红 陶小艳 於祥 隋怡 于 2020-06-30 设计创作，主要内容包括：本发明涉及一种中药血盆草中的吲哚类生物碱及其提取纯化方法和应用,对血盆草进行止血活性初探、止血活性部位化学成分的分离和止血物质基础的活性筛选等研究,通过这些研究,初步掌握了血盆草的止血药效,分析贵州产血盆草止血活性部位的化学成分组成以及确定血盆草的止血物质基础,在此基础上,能科学评价血盆草的民间用药依据,同时为止血新药的研发、血盆草质量评价标准提供实践基础；为血盆草资源的开发及利用提供科学依据。(The invention relates to indole alkaloid in a traditional Chinese medicine of the blood basin grass and an extraction and purification method and application thereof, wherein the blood basin grass is researched for the initial detection of hemostatic activity, the separation of chemical components of hemostatic active parts, the activity screening of hemostatic material bases and the like, and through the researches, the hemostatic efficacy of the blood basin grass is preliminarily mastered, the chemical component composition of the hemostatic active parts of the blood basin grass in Guizhou province is analyzed, and the hemostatic material bases of the blood basin grass are determined; provides scientific basis for the development and utilization of the blood basin grass resource.)

1. Indole alkaloid in traditional Chinese medicine herba Rubi Corchorifolii is characterized in that the indole alkaloid is indole-3-ethyl formate and 3-carboxyl indole.

2. The indole alkaloid according to claim 1, wherein the indole-3-carboxylic acid ethyl ester compound is characterized by:

structural formula (xvi):

the molecular formula is as follows: c₁₁H₁₁NO₂。

API-ESm/z：212[M+Na]⁺

The state is as follows: white powder

Silica gel thin layer identification: developing with petroleum ether and ethyl acetate at a ratio of 8:1, allowing fluorescence at 254nm, developing and baking with 10% concentrated sulfuric acid-ethanol to obtain orange speckle, and further baking to obtain mauve speckle;

¹H-NMR(600MHz，CDCl₃)、¹³C-NMR(150MHz，CDCl₃) The data are as follows:

3. the indole alkaloid according to claim 1, characterized in that the 3-carboxyindole compound is characterized by:

structural formula (xvi):

the molecular formula is as follows: c₉H₇NO₂

EI-MSm/z：161[M]⁺

Melting point (mp): 232-234 ℃.

The state is as follows: light yellow needle crystal

Silica gel thin layer identification: developing dichloromethane-methanol at a ratio of 35:1, then having fluorescence at 254nm, and developing and baking with 10% concentrated sulfuric acid-ethanol to obtain orange spots;

¹H-NMR(600MHz，CDCl₃)、¹³C-NMR(150MHz，CDCl₃) The data are as follows:

4. the method for extracting and purifying indole alkaloids according to claim 1, wherein the method for extracting and purifying indole alkaloids comprises: crushing 7-12 kg of dried whole herb of rubus bidens into coarse powder, performing reflux extraction for 2-5 times by using 80-98% industrial ethanol, performing concentration under reduced pressure for 1-3 hours each time, and recovering a solvent to obtain 1300-2300 g of ethanol extract; the method comprises the following steps of 1:1, suspending in water, and sequentially performing 1:1, extracting for 4-8 times by using each polar solvent, respectively combining each extraction liquid, and recovering the solvent by using a rotary evaporator to respectively obtain 210-360 g of petroleum ether part extract, 180-320 g of chloroform part extract, 140-240 g of ethyl acetate part extract, 150-265 g of n-butanol part extract and water part;

dissolving the petroleum ether part extract by adding a solvent, mixing the extract with silica gel, applying the mixture to a silica gel column by a dry method, performing gradient elution by the silica gel column with petroleum ether-ethyl acetate in a chromatography of 20: 1-0: 1, tracking by TLC, combining the same parts, and performing coarse segmentation to obtain 12 components Fr.1-12;

fr.6 is subjected to pigment removal by a gel chromatographic column by dichloromethane-methanol with the ratio of 1:1 to obtain 6 components Fr.6.1-Fr.6.6; eluting 36-62 mg of Fr.6.6 by a silica gel column with chromatographic petroleum ether-dichloromethane according to the ratio of 3:1 to obtain 3 components Fr.6.6.1-Fr.6.6.3, and scraping Fr.6.6.2 by a scraper to obtain 4.5-7.8 mg of indole-3-ethyl formate;

removing pigment from 7-13 g of Fr.9 by using a gel chromatographic column with dichloromethane-methanol in a ratio of 1:1, tracking by TLC to obtain 5 components Fr.9.1-Fr.9.5, carrying out wet sample loading on 160-275 mg of Fr.9.5, carrying out column chromatography twice on a dichloromethane-methanol system in a ratio of 40:1 and 35:1, and scraping to obtain 3-carboxyindole 3.1-5.2 mg.

5. The extraction and purification method according to claim 4, wherein the extraction and purification method specifically comprises: crushing 8-11 kg of dried whole herb of the rubus bidens into coarse powder, performing reflux extraction for 2-4 times by using 85-97% industrial ethanol, performing concentration under reduced pressure for 1-3 hours each time, and recovering a solvent to obtain 1500-2100 g of ethanol extract; the method comprises the following steps of 1:1, suspending in water, and sequentially performing 1:1, extracting for 5-7 times by using each polar solvent, respectively combining each extraction liquid, and recovering the solvent by using a rotary evaporator to respectively obtain 230-340 g of petroleum ether extract, 200-300 g of chloroform extract, 160-220 g of ethyl acetate extract, 170-245 g of n-butanol extract and water;

dissolving the petroleum ether part extract by adding a solvent, mixing the extract with silica gel, applying the mixture to a silica gel column by a dry method, performing gradient elution by the silica gel column with petroleum ether-ethyl acetate in a chromatography of 20: 1-0: 1, tracking by TLC, combining the same parts, and performing coarse segmentation to obtain 12 components Fr.1-12;

fr.6 is subjected to pigment removal by a gel chromatographic column by dichloromethane-methanol with the ratio of 1:1 to obtain 6 components Fr.6.1-Fr.6.6; eluting 36-62 mg of Fr.6.6 by a silica gel column with chromatographic petroleum ether-dichloromethane according to the ratio of 3:1 to obtain 3 components Fr.6.6.1-Fr.6.6.3, and scraping Fr.6.6.2 by a scraper to obtain 5-7 mg of indole-3-ethyl formate;

removing pigment from 8-12 g of Fr.9 by using a gel chromatographic column through dichloromethane-methanol according to the ratio of 1:1, tracking by TLC to obtain 5 components Fr.9.1-Fr.9.5, carrying out wet sample loading on 180-255 mg of Fr.9.5, carrying out column chromatography twice through a dichloromethane-methanol system according to the ratio of 40:1 and 35:1, and scraping to obtain 3-carboxyindole 3.5-4.8 mg.

6. The method for extracting and purifying indole alkaloids according to claim 5, wherein the method for extracting and purifying indole alkaloids comprises: pulverizing 9.3kg dried whole plant of herba Rubi Corchorifolii into coarse powder, extracting with 95% industrial ethanol under reflux for 3 times, each for 2 hr, concentrating under reduced pressure, and recovering solvent to obtain alcohol extract 1756 g; the method comprises the following steps of 1:1, suspending in water, and sequentially performing 1:1, extracting for 6 times by each polar solvent, respectively combining each extraction liquid, and recovering the solvent by a rotary evaporator to respectively obtain 286.6g of petroleum ether extract, 244.9g of chloroform extract, 188.6g of ethyl acetate extract, 205.1g of n-butanol extract and water;

dissolving the petroleum ether part extract by adding a solvent, mixing the extract with silica gel, applying the mixture to a silica gel column by a dry method, performing gradient elution by the silica gel column with petroleum ether-ethyl acetate in a chromatography of 20: 1-0: 1, tracking by TLC, combining the same parts, and performing coarse segmentation to obtain 12 components Fr.1-12;

fr.6 is subjected to pigment removal by a gel chromatographic column by dichloromethane-methanol with the ratio of 1:1 to obtain 6 components Fr.6.1-Fr.6.6; eluting 48mg of Fr.6.6 with chromatographic petroleum ether-dichloromethane at a ratio of 3:1 through a silica gel column to obtain 3 components Fr.6.6.1-Fr.6.6.3, and scraping Fr.6.6.2 with a scraper to obtain 6mg of indole-3-ethyl formate;

10g of Fr.9 is depigmented by a gel chromatographic column with dichloromethane-methanol in a ratio of 1:1, TLC tracking is carried out to obtain 5 components Fr.9.1-Fr.9.5, 212mg of Fr.9.5 is subjected to wet sample loading, and a dichloromethane-methanol system in a ratio of 40:1 and 35:1 is subjected to column chromatography twice and then is scraped to obtain 4mg of the compound 3-carboxyl indole.

7. The use of the indole alkaloid of claim 1, wherein said indole-3-carboxylic acid ethyl ester or 3-carboxyindole is used in hemostatic pharmaceuticals.

8. The use of indole alkaloids according to claim 7, wherein said indole-3-carboxylic acid ethyl ester, 3-carboxyindole is used in hemostatic pharmaceutical formulations.

9. The use of the indole alkaloid of claim 8, wherein the preparation is prepared by adding pharmaceutically acceptable adjuvants and conventional process into pharmaceutically acceptable preparations, and the pharmaceutically acceptable preparations are solid preparations or liquid preparations.

10. The use of indole alkaloid according to claim 9, wherein the solid preparation is granule, capsule, tablet, pill, powder, lyophilized powder for injection; the liquid preparation is injection preparation and oral liquid.

Technical Field

The invention relates to the field of pharmacy, in particular to indole alkaloid in traditional Chinese medicine rubus bidens and an extraction and purification method and application thereof.

Background

The hemostatic is a medicine for accelerating blood coagulation, reducing permeability of capillary vessels or promoting contraction of broken ends of blood vessels to stop bleeding; can be used for treating hemorrhage or hemorrhagic diseases caused by various reasons. The hemostatic has wide clinical application and wide application in life, and has important clinical significance and research value.

According to relevant statistics, the dosage of the hemostatic in the Chinese hospital basically keeps a relatively stable rising trend of medication level; if the western medicine is used for treatment, dangerous consequences caused by high coagulation can often occur, and the western medicine is forbidden or cautiously used under the condition of some inapplicable symptoms or postoperative sequelae, such as renal insufficiency or postoperative hematuria; the traditional hemostatic medicines can bring more adverse reactions, and the problems of hemostasis, thrombosis promotion, immunological rejection and the like cannot be solved. The combination of Chinese and Western medicine has become a new idea for researching and developing new hemostatics, and according to the four basic principles of safety, effectiveness, economy and appropriateness of reasonable medicine application, the traditional Chinese medicine dialectically treats and stops bleeding, and can act on three major systems of physiological hemostasis, blood coagulation and fibrinolysis. According to the intervention of the clinical scheme of dialectical treatment of the traditional Chinese medicine, the traditional Chinese medicine is supplemented with western medicines, and the application of the Chinese herbal medicines can also reduce the medical expense and lighten the economic burden of a patient, thereby embodying the superiority of the traditional Chinese medicine in treating diseases.

To date, some hemostatic active ingredients have been isolated and identified from plants while studying the hemostatic effects of traditional Chinese medicines. Reportedly, the quinone compound eriosema chinense element separated from the pyrola whole grass has the anti-bleeding effect and the retinoid K-like activity; isoverbascoside, a phenylpropanoid glycoside compound, is proved to have extremely strong effect of inhibiting oxidation hemolysis; wedelolactone and demethylwedelolactone separated from Hypericum plant also have good hemostatic activity; caffeic acid, one of the main hemostatic components of common cephalanoplos herb, can contract local blood vessels and inhibit fibrinolysis, thereby achieving the hemostatic effect. The action mechanism of other active ingredients is not clear, so the research on the hemostasis mechanism of the traditional Chinese medicine, the national medicine and the monomeric compound for hemostasis is increased, and new hemostatic medicines with good hemostasis effect, clear mechanism and wide clinical application prospect are searched; the monomer compound with hemostatic activity is formulated as the index of quality control of the hemostatic traditional Chinese medicine, and guarantees are provided for reasonable, safe and effective medication in clinic.

The Chinese herbaceous plants (with the scientific name of Salvia cavaliei var. simplicifolia Stib.) are also called as reversed (turning) red-back, and Guizhou copper kernel and most of areas are called Chinese herbaceous plants, are distributed in Guizhou, Hubei, Sichuan, Guangxi and other areas, and have abundant resources in the Guizhou areas; the Chinese basine herb is used as a medicine in folk, is mainly used for treating symptoms such as hematemesis, hemoptysis and the like, and mainly has the effects of cooling blood, detoxifying, dissipating blood stasis, stopping bleeding and the like; the hemostatic drug has wide treatment range and definite curative effect, and is a common hemostatic folk drug for the Guizhou Miao nationality, the Dong nationality and other minority nationalities. The method has abundant resources in each seedling region of Guizhou (including Guiyang city urban areas), and has sufficient resource foundation for reasonably developing and utilizing the sanguinea potted plant. In 1934-2015, reports of Wu Y-B and Pentron and the like are organized and systematically reviewed, and during this period, monomer components separated from the plants are nearly 900 or more, and the main types are diterpenes, triterpenes, phenolic acids, glycosides, alkaloids, flavonoids, lignans, fatty acids and other types of compounds. Literature research shows that the chemical composition research and other related research on the blood basin grass report a few and only how to group the blood basin grass^[12]For the determination of the content of the total flavonoids in the blood basin grass, Zhao Li Min and the like are separated from an n-butanol part of a water extract of the blood basin grass to obtain two trace phenolic glycosides, namely isoquercitrin and milk vetch, Zhang Meng Ke separates 45 compounds from the water extract, Chen Qiong separates 17 compounds from the water extract, and Wang He Ying and the like separate 13 compounds from the water extract, but relevant research reports on the hemostasis aspect are not found.

With the increasingly deep modern research on traditional Chinese medicine and the continuous deepening of understanding on national traditional Chinese medicine, in order to more effectively control the quality of the medicinal materials and more effectively evaluate the medicinal materials, more reasonably utilize medicinal material resources and explore medicinal material resources and ensure more accurate, safe and effective clinical medication, the combination of modern clinical practice and the traditional Chinese medicine theory must be further strengthened, the hemostasis action mechanism and the hemostasis chemical components of the traditional Chinese medicine are researched and summarized by utilizing modern scientific means, and the enrichment and the perfection of the hemostasis components and the hemostasis theory of the national traditional Chinese medicine are realized, so that the theoretical basis of the traditional Chinese medicine can be broken through the theoretical experience of the national traditional Chinese medicine, the better inheritance and the development of the national traditional Chinese medicine is laid, and therefore, the hemostasis components, the medication mechanism and the medication performance of the national hemostatic medicinal herb of the Guizhou national hemostatic herb Xueyi Cai are researched, and the reasonable medicine is developed for the national herb Xueyi Cai, The utilization and resource evaluation have positive significance.

The climate belongs to the mountain type of subtropical plateau in the watershed zone at the east of the cloud plateau and the upstream of the two rivers of the Yangtze river and the Zhujiang river in Guizhou. Due to the special ecological environment and the complex and various natural conditions of Guizhou, the rare and special medicine resources are inoculated, and the difference of the chemical components of the medicine and the same plants is large due to the difference of the growth environment; the basic mechanism of the haemostasis by the sanguinaria japonica in the Guizhou national region is not clear, and in view of multiple purposes of the sanguinaria japonica in the Guizhou national region, the basic research report of the haemostasis is very few, and the effective components are not clear (haemostasis), so that the sanguinaria japonica is required to be subjected to deep systematic research, which is favorable for further mastering and evaluating the scientificity of the application of the sanguinaria japonica of the Guizhou national region.

In order to carry out relevant research on the hemostatic effect of the Guizhou folk medicinal herb, namely the sanguinea japonica, the invention carries out researches such as primary detection on hemostatic activity, separation of chemical components of hemostatic active parts, activity screening of hemostatic material bases and the like on the sanguinea japonica, preliminarily grasps the hemostatic efficacy of the sanguinea japonica, analyzes the chemical component composition of the hemostatic active parts of the Guizhou sanguinea japonica and determines the hemostatic material bases of the sanguinea japonica through the researches, can scientifically evaluate the folk medication basis of the sanguinea japonica on the basis, and simultaneously provides practical bases for the research and development of new hemostatic medicines and the quality evaluation standards of the sanguinea japonica; provides scientific basis for the development and utilization of the blood basin grass resource.

Disclosure of Invention

The invention aims to provide indole alkaloid in traditional Chinese medicine, namely, the hematuria bidentis;

the other purpose of the invention is to provide a method for extracting and purifying indole alkaloid in the traditional Chinese medicine, namely the hematuria bidentis;

the invention also aims to provide the application of indole alkaloid in the traditional Chinese medicine of the hematuria in the aspect of hemostatic medicines.

The indole alkaloid is indole-3-ethyl formate and 3-carboxyl indole.

The indole-3-carboxylic acid ethyl ester compound is characterized in that:

structural formula (xvi):

the molecular formula is as follows: c₁₁H₁₁NO₂。

API-ES m/z：212[M+Na]⁺

The state is as follows: white powder (dichloromethane)

Silica gel thin layer identification: petroleum ether and ethyl acetate (8:1) are developed, fluorescence is generated under 254nm, orange spots are formed after coloration and baking are carried out by 10% concentrated sulfuric acid-ethanol, and the spots are continuously baked to be purple red;

¹H-NMR(600 MHz，CDCl₃)、¹³C-NMR(150 MHz，CDCl₃) The data are shown in figure 1 and figure 2

The 3-carboxyindole compounds of the present invention are characterized as:

structural formula (xvi):

the molecular formula is as follows: c₉H₇NO₂

EI-MS m/z：161[M]⁺

Melting point (mp): 232-234 ℃.

The state is as follows: pale yellow needle crystal (methanol)

Silica gel thin layer identification: the dichloromethane-methanol with the ratio of 35:1 is developed to have fluorescence under 254nm, and the orange spots are shown after the dichloromethane-methanol is developed and baked by 10 percent concentrated sulfuric acid-ethanol.

¹H-NMR(600MHz，CDCl₃)、¹³C-NMR(150MHz，CDCl₃) The data are shown in FIG. 3 and FIG. 4

The method for extracting and purifying the indole alkaloid specifically comprises the following steps: crushing 7-12 kg of dried whole herb of rubus bidens into coarse powder, performing reflux extraction for 2-5 times by using 80-98% industrial ethanol, performing concentration under reduced pressure for 1-3 hours each time, and recovering a solvent to obtain 1300-2300 g of ethanol extract; the method comprises the following steps of 1:1, suspending in water, and sequentially performing 1:1, extracting for 4-8 times by using each polar solvent, respectively combining each extraction liquid, and recovering the solvent by using a rotary evaporator to respectively obtain 210-360 g of petroleum ether part extract, 180-320 g of chloroform part extract, 140-240 g of ethyl acetate part extract, 150-265 g of n-butanol part extract and water part;

dissolving the petroleum ether part extract by adding a solvent, mixing the extract with silica gel, applying the mixture to a silica gel column by a dry method, performing gradient elution by the silica gel column with petroleum ether-ethyl acetate in a chromatography of 20: 1-0: 1, tracking by TLC, combining the same parts, and performing coarse segmentation to obtain 12 components Fr.1-12;

fr.6 is subjected to pigment removal by a gel chromatographic column by dichloromethane-methanol with the ratio of 1:1 to obtain 6 components Fr.6.1-Fr.6.6; eluting 36-62 mg of Fr.6.6 by a silica gel column with chromatographic petroleum ether-dichloromethane according to the ratio of 3:1 to obtain 3 components Fr.6.6.1-Fr.6.6.3, and scraping Fr.6.6.2 by a scraper to obtain 4.5-7.8 mg of indole-3-ethyl formate;

removing pigment from 7-13 g of Fr.9 by using a gel chromatographic column with dichloromethane-methanol in a ratio of 1:1, tracking by TLC to obtain 5 components Fr.9.1-Fr.9.5, carrying out wet sample loading on 160-275 mg of Fr.9.5, carrying out column chromatography twice on a dichloromethane-methanol system in a ratio of 40:1 and 35:1, and scraping to obtain 3-carboxyindole 3.1-5.2 mg.

Preferably, the method for extracting and purifying indole alkaloids provided by the invention specifically comprises the following steps: crushing 8-11 kg of dried whole herb of the rubus bidens into coarse powder, performing reflux extraction for 2-4 times by using 85-97% industrial ethanol, performing concentration under reduced pressure for 1-3 hours each time, and recovering a solvent to obtain 1500-2100 g of ethanol extract; the method comprises the following steps of 1:1, suspending in water, and sequentially performing 1:1, extracting for 5-7 times by using each polar solvent, respectively combining each extraction liquid, and recovering the solvent by using a rotary evaporator to respectively obtain 230-340 g of petroleum ether extract, 200-300 g of chloroform extract, 160-220 g of ethyl acetate extract, 170-245 g of n-butanol extract and water;

dissolving the petroleum ether part extract by adding a solvent, mixing the extract with silica gel, applying the mixture to a silica gel column by a dry method, performing gradient elution by the silica gel column with petroleum ether-ethyl acetate in a chromatography of 20: 1-0: 1, tracking by TLC, combining the same parts, and performing coarse segmentation to obtain 12 components Fr.1-12;

fr.6 is subjected to pigment removal by a gel chromatographic column by dichloromethane-methanol with the ratio of 1:1 to obtain 6 components Fr.6.1-Fr.6.6; eluting 36-62 mg of Fr.6.6 by a silica gel column with chromatographic petroleum ether-dichloromethane according to the ratio of 3:1 to obtain 3 components Fr.6.6.1-Fr.6.6.3, and scraping Fr.6.6.2 by a scraper to obtain 5-7 mg of indole-3-ethyl formate;

removing pigment from 8-12 g of Fr.9 by using a gel chromatographic column through dichloromethane-methanol according to the ratio of 1:1, tracking by TLC to obtain 5 components Fr.9.1-Fr.9.5, carrying out wet sample loading on 180-255 mg of Fr.9.5, carrying out column chromatography twice through a dichloromethane-methanol system according to the ratio of 40:1 and 35:1, and scraping to obtain 3-carboxyindole 3.5-4.8 mg.

Further preferably, the method for extracting and purifying indole alkaloids provided by the invention specifically comprises the following steps: pulverizing 9.3kg dried whole plant of herba Rubi Corchorifolii into coarse powder, extracting with 95% industrial ethanol under reflux for 3 times, each for 2 hr, concentrating under reduced pressure, and recovering solvent to obtain alcohol extract 1756 g; the method comprises the following steps of 1:1, suspending in water, and sequentially performing 1:1, extracting for 6 times by each polar solvent, respectively combining each extraction liquid, and recovering the solvent by a rotary evaporator to respectively obtain 286.6g of petroleum ether extract, 244.9g of chloroform extract, 188.6g of ethyl acetate extract, 205.1g of n-butanol extract and water;

dissolving the petroleum ether part extract by adding a solvent, mixing the extract with silica gel, applying the mixture to a silica gel column by a dry method, performing gradient elution by the silica gel column with petroleum ether-ethyl acetate in a chromatography of 20: 1-0: 1, tracking by TLC, combining the same parts, and performing coarse segmentation to obtain 12 components Fr.1-12;

fr.6 is subjected to pigment removal by a gel chromatographic column by dichloromethane-methanol with the ratio of 1:1 to obtain 6 components Fr.6.1-Fr.6.6; eluting 48mg of Fr.6.6 with chromatographic petroleum ether-dichloromethane at a ratio of 3:1 through a silica gel column to obtain 3 components Fr.6.6.1-Fr.6.6.3, and scraping Fr.6.6.2 with a scraper to obtain 6mg of indole-3-ethyl formate;

10g of Fr.9 is depigmented by a gel chromatographic column with dichloromethane-methanol in a ratio of 1:1, TLC tracking is carried out to obtain 5 components Fr.9.1-Fr.9.5, 212mg of Fr.9.5 is subjected to wet sample loading, and a dichloromethane-methanol system in a ratio of 40:1 and 35:1 is subjected to column chromatography twice and then is scraped to obtain 4mg of the compound 3-carboxyl indole.

The indole-3-ethyl formate and 3-carboxyl indole provided by the invention are applied to hemostatic drugs.

The application of the indole-3-ethyl formate and the 3-carboxyl indole in the aspect of hemostatic pharmaceutical preparations is provided.

The preparation is prepared into a pharmaceutically acceptable preparation by adding pharmaceutically acceptable auxiliary materials according to a conventional process, and the pharmaceutically acceptable preparation is a solid preparation or a liquid preparation.

The solid preparation is granules, capsules, tablets, pills, powder and freeze-dried powder injection; the liquid preparation is injection preparation and oral liquid.

The pharmaceutically acceptable auxiliary materials are not limited, and can be one or more of common auxiliary materials in the field, such as a filling agent, a lubricating agent, a flavoring agent, a disintegrating agent, an antioxidant, a humectant, a surfactant and the like.

The principle of the invention is as follows: 2 indole alkaloids which are indole-3 ethyl formate and 3-carboxyl indole are respectively obtained by separating from a petroleum ether layer of a 95% ethanol extract of the blood basin grass by adopting an animal hemostatic activity guiding method, and the in-vitro plasma recalcification time of the indole alkaloids is determined by adopting a test tube method.

Drawings

FIG. 1 Process for preparing indole-3-carboxylic acid ethyl ester¹³C-NMR chart

FIG. 2 Process for preparing indole-3-carboxylic acid ethyl ester¹H-NMR chart

FIG. 33 Process for preparing carboxyindoles¹³C-NMR chart

FIG. 43 Process for preparing carboxyindoles¹H-NMR chart

Detailed Description

The technical solution of the present invention will be further specifically described below by way of specific examples.