WO2022143513A1 - Oral preparation comprising erigeron breviscapus extract and preparation method therefor - Google Patents

Abstract

The present invention relates to an oral preparation comprising Erigeron breviscapus, and a preparation method therefor and an application thereof, the preparation comprising common oral dosage forms such as capsules, tablets, oral liquids, dripping pills or granules. The preparation process provided in the present invention fully utilizes a caffeic acid ester component of Erigeron breviscapus, reduces the content of other harmful and ineffective substances at the same time, and can form a salt with breviscapine and the caffeic acid ester to improve medicinal efficacy. While providing liver protection, the present invention can also improve weight gain, elevated blood lipids, platelet aggregation and coagulation disorders caused by high-fat diet, providing a more convenient, effective and quality-controllable modern traditional Chinese medicine for clinical practice.

Description

Oral preparation containing breviscapine extract and preparation method thereof technical field

In particular, it relates to a preparation method and application of breviscapine and caffeic acid ester extract or its salt, especially in the preparation and treatment of metabolic-related diseases such as hyperlipidemia, hypertension, diabetes, obesity, non-alcoholic fatty liver disease and heart disease. Application in cerebrovascular disease medicine, and application in preparation of medicine for platelet aggregation and abnormal blood coagulation function caused by metabolism-related disease.

Background technique

In recent years, the spectrum of diseases has undergone significant changes with the acceleration of population aging, deepening urbanization, and gradual changes in lifestyles. Metabolic-related diseases such as hypertension, hyperlipidemia, obesity, and diabetes are widely prevalent, and the incidence of cardiovascular and cerebrovascular diseases is also accelerating.

Dengzhan Asarum (Dengzhanhua) is a dry whole herb of Erigeron breviscapus (Vant.) Hand-Mazz, a plant of the genus Asteraceae. Removing blood stasis, clearing collaterals and relieving pain, used for colds, headaches, rheumatism pain, paralysis caused by cerebrovascular accidents, etc. ("National Chinese Herbal Medicine Compilation"). Dengzhan Xixin can be made into medicine alone, and the currently listed varieties include Dengzhan Xixin Injection, Dengzhan Xixin Capsules, Dengzhan Xixin Soft Capsules, Dengzhan Xixin Dropping Pills, Yimaikang, Dengzhan Xixin Granules, etc. The active ingredients of flavonoids are scutellarin and caffeic acid esters, and the existing products mainly emphasize scutellarin as the main component.

Breviscapine products include breviscapine injection, breviscapine for injection, breviscapine tablets, breviscapine drop pills, etc. Even Dengzhan Asarum capsules, Yimaikang and other Dengzhan Asarum extract products do not regulate the content of caffeic acid esters, let alone enrichment. Dengzhan Xixin injection is the only product that enriches caffeic acid esters from a technological point of view. However, the ratio of the content of caffeic acid ester to flavonoids is only in the range of 1:1.2-1:5.5. Through the research on caffeic acid esters in scutellaria brevis plant, it is found that caffeic acid esters in scutellaria scutellariae are very rich, including monocaffeic acid ester and dicaffeoate. There are many families and genera containing caffeic acid esters in plants, such as Compositae, Honeysuckle, Eucommia, Chuanxiaceae, Rubiaceae, Convolvulaceae and other plants. At present, it is known that Compositae plants are different from other plants containing caffeoyl quinate in that they also contain a class of caffeoyl octulonic acid compounds, including monocaffeoyl, dicaffeoyl and tricaffeoyl octulonic acid, respectively. class substances. The caffeoyl octulonic acid components contained in scutellaria scutellariae are dicaffeoyl octulonic acid substances, including scutellarin B and scutellaria scutellariae. Fenugreek ester B was isolated from the genus Fenugreek of Compositae, while scutellaria scutellariae was only obtained from scutellaria scutellariae. Jianping Zhao, et al. (J. Nat. Prod. 2014, 77, 509-515) reported that six caffeoyl octulonic acids obtained from the genus Syringa spp. have anti-inflammatory and anti-metabolic effects. The content range of these special types of dicaffeoate compounds is listed separately, which can better clearly classify and control the caffeic acid esters obtained from D. The content of the ingredients is easier to control, which also makes the quality of such products more controllable. There is also a class of hepatotoxic substances γ-pyrones in Scutellaria brevis, and the representative compound is pyroconic acid. The method of removing pyroconic acid in the previous patent is to adopt column chromatography, and the properties used are: Pyroconic acid has a high polarity and can be removed by washing with water in column chromatography, but the extraction solvent must be low-concentration alcohol or water extraction, otherwise impurities such as chlorophyll are easy to solidify on the adsorption column, resulting in failure to elute Or lead to difficult column regeneration. In this patent, by adjusting the pH of the acidified supernatant to neutrality, microfiltration is performed first to remove insoluble macromolecular impurities, including most of chlorophyll, protein, tannin, etc., and then concentrated through a nanofiltration membrane. Molecular substances and water are filtered, and pyroconic acid is soluble in water and has a molecular weight of only 112 Da, so 90% of pyroconic acid can be removed by this method. The concentrated solution after removing the pyroconic acid is loaded on the column, so that the column solution can be separated and purified more efficiently. In addition, 50-80% ethanol can be used for extraction in the first step to increase the extraction rate. The extracted and refined scutellarin and caffeic acid ester can be directly dried and used as medicine, or the pH of the scutellarin and caffeic acid ester can be adjusted to neutral, and spray-dried after salt formation to increase their water solubility and even more The druggability of the two components.

SUMMARY OF THE INVENTION

In view of the above-mentioned technical problems, the object of the present invention is to provide an oral preparation containing breviscapine extract, wherein, the oral preparation contains caffeic acid ester or its salt and breviscapine or its salt and pharmaceutically acceptable excipients, coffee The weight ratio of the acid ester or its salt to breviscapine or its salt is 1:1 to 30:1.

The current oral preparation of breviscapine is limited by the extraction method, and usually the preparation is a mixture of caffeic acid ester, breviscapine, and other substances, which cannot be accurately mixed. The invention adopts a unique preparation method, extracts the main components therein, especially prepares and accurately mixes the two main active component salts therein, thereby effectively improving the drug efficacy.

Preferably, the present invention further provides an oral preparation containing scutellaria scutellariae extract, and the caffeic acid ester or its salt includes dicaffeoate or its salt and monocaffeic acid or its salt, wherein dicaffeoate or its salt The weight ratio of its salt to breviscapine or its salt is 1.25:1-6:1, (preferably 1.25:1-5.5:1, more preferably 1.5:1-5:1, most preferably 2:1-4.5: 1), the weight ratio of monocaffeic acid ester or its salt to dicaffeic acid acid ester or its salt is 1:1.2～1:6.2.

Preferably, the present invention further provides an oral preparation containing the extract of Asarum scutellariae, and the dicaffeoate or its salt includes 1,3-O-dicaffeoylquinic acid or its salt, 3,4- O-dicaffeoylquinic acid or its salts, 3,5-O-dicaffeoylquinic acid or its salts, 4,5-O-dicaffeoylquinic acid or its salts, fimbonitrile ethyl or its salts , scutellarin or its salt; monocaffeic acid ester or its salt including 3-O-caffeoylquinic acid or its salt, 4-O-caffeoylquinic acid or its salt, 5-O-caffeoylquinic acid Nitric acid or its salt, scutellarin or its salt.

Preferably, the present invention further provides an oral preparation containing scutellaria scutellariae extract, wherein 1,3-O-dicaffeoylquinic acid or its salt, 3,4- O-dicaffeoylquinic acid or its salt, 3,5-O-dicaffeoylquinic acid or its salt, 4,5-O-dicaffeoylquinic acid or its salt, the above four are the same Isomers; described fiponin ester B or its salts and scutellarin or its salts are isomers, which are dicaffeoyl octulonic acid or its class, wherein simply substituted dicaffeoate or its salts are isomers. The ratio of the salt to dicaffeoyl octanoic acid or its salt is 2:1 to 0.9:1.

The above-mentioned oral preparations are preferably hard capsules, soft capsules, tablets, granules, oral liquids, dripping pills, water pills, powders, and pills.

The scutellarin or its salt of the above-mentioned oral preparation includes scutellarin or its salt, scutellarin or its salt, scutellarin and its salt, etc. scutellarin or its salt-related flavonoid compounds isolated from scutellarin or its salt.

In the above preparation, caffeic acid ester or its salt and breviscapine or its salt account for 80-99% by weight of the preparation, and the balance is auxiliary materials. The oral preparations are preferably capsules, tablets, such as hard capsules.

Further, the dicaffeoate salt and scutellarin salt are both sodium or potassium salts or other medicinal and water-soluble salts; preferably sodium salts, the weight ratio of which is 1:1 to 6:1. That is, other products should also be sodium or potassium salts or other medicinal and water-soluble salts, and similarly, other products should also preferably be sodium salts.

The present invention further provides a preparation method of an oral preparation, the preparation method is as follows:

Take Dengzhan Asarum and add 10-90% concentration of aqueous ethanol to extract, and the extract is concentrated under reduced pressure to form an extract; the extract is dissolved in water, adjusted to pH 7-9, filtered, added acid to adjust pH to 1 to 3, placed overnight, and filtered , collect the filtrate and the precipitation, purify the precipitation with water and ethanol, and dry to obtain scutellarin; after the precipitation is refined, add alkali to adjust the pH value to 7-8, and spray dry to obtain scutellarin salt; the acidified filtrate adjusts the pH value to 7 ~9, use ceramic microfiltration membrane for clarification, and the clarified liquid obtained after clarification is then concentrated with organic nanofiltration membrane, the concentrated liquid is passed through a polyamide chromatography column, eluted with water, eluted with 50-80% ethanol, and collected. The ethanol eluent is concentrated and dried to obtain caffeic acid ester; the ethanol eluent is concentrated and adjusted to pH 7-9, filtered, and the filtrate is spray-dried to obtain caffeic acid ester salt. Dicaffeic acid ester or its salt and scutellaria or its plain salt are 1:1-6:1 (preferably 1.25:1-5.5:1, more preferably 1.5:1-5:1, most preferably 2:1-4.5 :1), adding appropriate auxiliary materials, can be made into tablets, capsules, soft capsules, dropping pills, granules and oral liquid preparations.

In the preparation method of the above oral preparation, the pH value of the alkali adjustment solution is NaOH, Na ₂ CO ₃ , NaHCO ₃ , KOH, K ₂ CO ₃ or KHCO ₃ , or other alkali solutions that can be used to adjust the pH value ; The pH value of the acid-adjusting solution is HCl, H ₂ SO ₄ or H ₃ PO ₄ , or other acid solutions that can be used to adjust the pH value. The pH value of the alkali adjustment solution is preferably NaOH, Na ₂ CO ₃ , NaHCO ₃ , KOH, K ₂ CO ₃ or KHCO ₃ ; the pH value of the acid adjustment solution is preferably HCl, H ₂ SO ₄ Or H ₃ PO ₄ .

The ethanol concentration of the polyamide chromatographic column eluted with ethanol is 50-95%.

The present invention provides the application of the above medicinal composition in the preparation of medicines for treating hypertension, hyperlipidemia, diabetes, obesity, non-alcoholic fatty liver disease and other metabolic related diseases and cardiovascular and cerebrovascular diseases. According to the invention, the pharmaceutical composition obtained by screening the above ratio has the effect of regulating metabolism-related diseases and the resulting platelet aggregation and coagulation disorders, and is more pleased to obtain an unexpected effect, that is, compared with aspirin and clopidogrel, The composition of the invention not only has the anti-platelet aggregation effect equivalent to the positive drug, but also has a better anti-coagulation effect, which is a medical and pharmaceutical advantage that aspirin and clopidogrel do not have, and more importantly, The composition of the present invention does not cause bleeding symptoms when administered beyond the prescribed range, which is very important for preventing atherosclerosis and the cardiovascular and cerebrovascular diseases caused by it.

Currently, the single oral preparation of Asarum brevis on the market does not have the step of removing the hepatotoxic component pyroconic acid, nor the step or process of refining caffeic acid ester or its salts, especially dicaffeoate or its salts. For example, the existing technology of Dengzhan Xixin capsules is to take 2000g of Dengzhan Xixin, and extract it twice with 80% ethanol under reflux for 1.5 hours for the first time and 1 hour for the second time, combine the extracts, add about 640g of activated carbon, boil, filter, and extract the filtrate. Concentrate under reduced pressure to a thick paste, add an appropriate amount of starch, dry under reduced pressure, pulverize, sieve, coat with 3% polyvinyl alcohol solution, add 0.5 g of magnesium stearate, mix well, put into capsules, and make Into 1000 grains, that is.

It has been reported in the literature that caffeoylquinic acid ester compounds have antioxidant, anti-inflammatory, antibacterial, antiviral, hypoglycemic, hypolipidemic, and immunomodulatory effects, and are a class of active ingredients. Therefore, refining such active substances and comparing the two components as quantitative regulations are necessary work to make the products of Asarum brevis serve better clinical services. Both breviscapine and caffeic acid esters have poor water solubility and fat solubility, resulting in low bioavailability and weak druggability. The present invention adopts alkali-soluble acid precipitation in the preparation method, firstly precipitating the flavonoid component breviscapine, and then refining; meanwhile, the acidified supernatant liquid part is retained, and the acidified supernatant liquid part is another type of active ingredient caffeic acid esters, Through membrane separation technology, microfiltration is used to remove macromolecular insoluble substances, and then a nanofiltration membrane is used to concentrate to remove most of the pyroconic acid, and then column chromatography is performed to enrich the dicaffeoate components, and remove the hepatotoxic substances again. Pyroconic acid. After nanofiltration membrane and column chromatography, more than 99% of pyroconic acid can be removed.

This result was confirmed by experiments:

Table 1. Changes in composition of the acidified supernatant of Dengzhan Asarum after membrane filtration after pH adjustment

It can be seen from Table 1 that the acidified supernatant of Asarum scutellariae was adjusted to pH close to neutral and passed through a clarification membrane and a nanofiltration concentration membrane, and the effective substance dicaffeoate was well retained, while the harmful substance pyroconic acid was removed. The rate reached 84%. After column chromatography, the content of dicaffeoate was well retained, and pyroconic acid was further removed.

The microfiltration clarification membrane is preferably 100 nm or 200 nm, and the nanofiltration concentration membrane is preferably 300D-400D.

When the flavonoids and caffeic acid esters are salted, the in vitro dissolution test proves that the dissolution is higher than that of the prototype, so the drugability is better than the prototype. The dissolution profiles of Dengzhanxixin tablets and Dengzhanxixin sodium salt tablets in different dissolution media were evaluated. The results of the study showed that the dissolution rates of the two tablets in 0.1M HCl were both low, reaching saturation in 5 minutes, and the cumulative dissolution percentage was less than 15%; in the dissolution medium of acetate buffer at pH 4.5, the two tablets accumulated The release amount basically reached saturation in 30min, and the cumulative dissolution percentage of Dengzhanxixin sodium salt tablet was higher than that of the prototype tablet, both less than 60%; in the dissolution medium of pH 6.8 phosphate buffer, when the rotation speed was 50rpm, Dengzhanxixin sodium The salt tablets were completely dissolved at 15min, while the cumulative dissolution percentage of Dengzhanxixin tablets was greater than 85% at 45min; the cumulative dissolution percentage of Dengzhanxixin tablets was greater than that of Dengzhanxixin tablets at 30min.

By feeding C57 mice with MCD (Methionine and Choline Deficient L-Amino Acid Diet) methionine and choline deficient diets, a non-alcoholic steatohepatitis (NASH) model was constructed, and the existing Dengzhanxixin capsules and the invented process Dengzhanxixin were explored. The therapeutic effect of capsules on NASH.

1. Test reagents

MCD feed, item number A02082002BR, manufacturer: Research Diets (USA)

2. Test process

(1) Animal grouping and treatment Twenty C57BL/6J male mice were randomly divided into 4 groups after adaptive feeding for 1 week, with 5 mice in each group. The control group is the normal maintenance feed, the model group is fed with the MCD feed, the existing Dengzhanxixin capsule powder group and the invention process Dengzhanxixin capsule powder group (see Example 6 for the prescription), the oral liquid is administered by gavage (100mg/kg body weight). ), once a day. The experimental animals had free access to food and water for 4 weeks.

(2) Determination of mouse body weight and liver weight The body weight and liver wet weight of each group of mice were weighed, and the liver index was calculated.

3. Test results

(1), the effect on mouse body weight, liver weight and liver index

After 4 weeks of feeding, the body weight, liver weight and liver index of mice in the model group, the existing Dengzhan Xixin capsule powder group, and the Dengzhan Xixin capsule powder group with the invention process were significantly reduced compared with the control group, indicating that the test drug could not effectively improve MCD diet-induced reduction in body weight, liver weight. In addition, there was no significant difference in body weight, liver weight and liver index between the two tested drugs and the model group, as shown in Table 2.

Table 2. Effects of Dengzhan Xixin capsule powder on body weight, liver weight and liver index of mice

group	Weight (g)	Liver weight (g)	Liver Index (%)
control group	27.08±1.46	1.43±0.15	5.31±0.36
model group	21.94±1.25	1.91±0.24	8.69±0.75

Invented craft Dengzhan Asarum group	22.12±0.56	1.88±0.11	8.49±0.36
Existing Craft Dengzhan Asarum Group	22.68±0.22	1.97±0.35	8.77±0.22

(2), the effect on the liver function indexes of mice

Compared with the control group, the serum alanine aminotransferase (AST) and aspartate aminotransferase (ALT) levels of the mice in the model group were significantly increased ( ^### P＜0.001). The serum AST and ALT levels of the mice in the Dengzhanxixin capsule powder group with the invention process were higher than those in the control group, but compared with the model group, the Dengzhanxixin capsule powder with the invention process significantly reduced the MCD diet-induced increase in serum AST and ALT in mice. High; the existing process Dengzhanxixin group (such as Yunnan Haobang Pharmaceutical) can significantly reduce the ALT level, and there is a trend of reducing AST, but there is no statistical significance, see Table 3.

Table 3. Effects of Dengzhan Xixin Capsule Powder and Existing Dengzhan Shengmai Capsule Powder on Liver Function Index of NASH Mice

group	Alanine aminotransferase (U/L)	Aspartate aminotransferase (U/L)
control group	37.2±6.57	103.2±22.9
model group	160.8±49.2 ###	181.2±34.1 ###
Invented craft Dengzhan Asarum group	101.6±28.2* #	132.1±22.1*
Existing Craft Dengzhan Asarum Group	100.1±22.4* #	162.2±21.6 ##

^### P<0.005vs control group; ^## P<0.01vs control group; ^# P<0.05vs control group; ^* P<0.05vs model group

4. Analysis and conclusion

(1) Effects on body weight MCD diet-induced weight loss in NASH mice is a disease characterization of this dietary model. This study found that the invented process Dengzhan Xixin capsule powder and the existing technology Dengzhan Xixin capsule powder could not inhibit the weight loss induced by MCD diet, which may be related to the structure of MCD diet.

(2), the effect on liver function MCD diet-fed mice serum ALT and AST levels were significantly increased, indicating that the MCD diet-induced NASH model was successfully established. This study found that both the invented technology Dengzhan Xixin capsule powder and the existing technology Dengzhan Xixin capsule powder can improve the liver function damage induced by MCD diet, while the invented technology Dengzhan Xixin capsule powder can down-regulate both ALT and AST, although Existing literature found that scutellarin (scutellarin, usually by injection) may have a protective effect on the liver, but this experiment shows that compared with the scutellaria scutellariae obtained by the existing process, orally administered scutellarin. The drug (100mg/Kg) has better liver protection.

The existing breviscapine capsule powder is prepared from breviscapine salt and caffeic acid ester salt in the ratio within the scope of the claims.

Description of drawings:

Figure 1 Changes in body weight of golden hamsters;

Fig. 2 Plasma total cholesterol content of golden hamster;

Figure 3. Plasma low-density lipoprotein-cholesterol (LDL-c) content in golden hamsters;

Fig. 4 Plasma triglyceride (TG) content of golden hamster;

Fig. 5 Plasma alanine aminotransferase (ALT) content of golden hamster;

Fig. 6 golden hamster plasma aspartate aminotransferase (AST) content;

Fig. 7 Golden hamster liver index (%);

Figure 8 Fat index (%) of golden hamster;

Fig. 9 liver triglyceride content of golden hamster;

Figure 10 Platelet aggregation rate (%);

Figure 11 Activated partial thromboplastin time;

Figure 12 Prothrombin time;

Figure 13 Plasma thrombin time;

Figure 14. Plasma fibrinogen levels.

The purpose of this experiment is to study the efficacy of different proportions of the main components of calyxarum on metabolic-related diseases caused by high-fat diet, as well as on platelet aggregation and coagulation disorders, so as to provide experimental data support for new clinical uses of the drug.

It is found from the aforementioned experiments that the new technology Dengzhan Xixin Capsule can effectively reduce ALT and AST in the non-alcoholic steatohepatitis model, and then further use the existing technology (Yunnan Haobang Pharmaceutical Co., Ltd.) and this product technology example 6 to carry out. For comparison, at the same time of comparison, the dicaffeoate and scutellarin salts in the examples are calculated, and the exact ratio is carried out, so as to achieve the optimum ratio of dicaffeoate and scutellarin salt. The study used aspirin and clopidogrel as positive control drugs. Since there was no significant difference in the test results of the two drugs, the following experiments only show the test results of aspirin.

2) Experimental method:

2.1) Experimental animal: LVG Syrian golden hamster, male, 6 weeks old, weighing 90-110 g, adapted to the rearing environment for 2 weeks. Animals were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

2.2) Experimental feed: basic feed (Normal chow diet, NC group); high-fat feed (High fat diet model group, HFD group: 1.0% cholesterol, 0.2% sodium cholate, 10.0% lard, 5.0% egg yolk powder and 83.8 % basal feed). Feed was purchased from Beijing Keao Xieli Feed Co., Ltd.

2.3) Mode of administration: Animals were given high-fat feed-based medicated feed for 8 weeks, wherein the feed containing Dengzhan Asarum contained 4g/kg of medicine, which was equivalent to the dosage of 200mg/kg body weight/day in terms of animal food intake; The aspirin-containing feed contains 0.4g/kg of the drug, which is equivalent to the dosage of 20mg/kg body weight/day in the equivalent of animal food intake. In addition, the same batch of golden hamsters were fed with basal feed as the normal control group (NC group).

2.4) Experimental grouping and administration

Table 4. Experimental Grouping and Administration

The control Dengzhanxixin was prepared by the existing process; the Dengzhanxixin groups A-F were prepared by the inventive process.

2.5) Detection indicators and methods:

a. Body weight measurement

During the experiment, animal body weights were recorded once a week.

b. Blood biochemical test

After the experiment, the animals were fasted for 12 h, blood (0.5 ml) was collected from the orbital venous plexus in an anticoagulation tube, centrifuged at 3500 rpm and 4 °C for 10 min, and 100 μL of the supernatant was taken and stored in a -80 °C refrigerator. Then an automatic biochemical analyzer was used to detect the levels of total cholesterol (CHO), low-density lipoprotein-cholesterol (LDL-C), triglyceride (TG), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in plasma . The detection kit was purchased from Zhongsheng Beikong Biotechnology Co., Ltd.

c. Liver wet weight ratio and epididymal fat wet weight ratio

After the experiment, the liver tissue and epididymal fat of the animals were taken and weighed, and the wet weight of the liver and the weight of the epididymal fat of each animal were calculated and their body weight percentage values were calculated respectively.

d. Determination of liver tissue lipid content

After the experiment, each hamster in each group weighed 20 mg of liver tissue, and added 20 times the volume of T-PER (containing 1/100 of protease phosphate) in the ratio of weight (g): volume (ml) = 1:20. Enzyme inhibitor), mechanically homogenize (42HZ, 4-6min) under ice-water bath conditions, then centrifuge at 2500rpm for 10min, take 2.5μL of the upper layer liquid and dilute 5 times to prepare the sample to be tested, and test according to the instructions of the TG detection kit. The remaining liver tissue homogenate was centrifuged at 12000rpm and 4°C for 30min, and the supernatant was aspirated for protein quantification by BCA method. The obtained results were further calculated according to the following formula.

e. Determination of platelet aggregation and coagulation indexes

After the experiment, 5% chloral hydrate (1ml/100g) was anesthetized, and blood was collected from the heart of the syringe. The blood sample was added with anticoagulant in a ratio of 1:9 of sodium citrate (4%) solution and blood, and centrifuged at 200g for 10min. Platelet-rich plasma (PRP) was obtained from the supernatant, and the remaining plasma was centrifuged at 800 g for 10 min to obtain the supernatant to obtain platelet-poor plasma (PPP). In the maximum platelet aggregation rate detection experiment, first use PPP for calibration, pre-warm PRP at 37°C for 180 seconds, put it in the detection tank, and quickly click to start while adding 10ul of adenosine diphosphate (ADP, 10nM), and read the platelets within 300s Maximum Aggregation Ratio (MAR) value. In the four detection experiments of coagulation, PPP was pre-warmed at 37 °C for 180 s and then placed in the detection tank, and the quantitative activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time ( TT) Fibrinogen (FIB) detection reagent quickly click to start, read clotting time and fibrinogen content.

2.6) Data Analysis

All data were entered into Excel files and expressed as Mean±SEM. Statistical analysis of data was performed using Graphpad Prism 8.0 software, one-way or two-way ANOVA comparison method, and *P<0.05 was used as the criterion for significant differences.

3) In vivo test results

3.1) Weight change

The body weight of the animals was measured once a week during the administration period, and the results are shown in Figure 1 . Compared with the normal diet control group (NC), the animals in the high-fat diet model group (HFD) had significantly higher body weight. The antiplatelet drug aspirin did not reduce weight gain caused by a high-fat diet. The control drug (Yunnan Haobang, existing technology) and Dengzhan Asarum (invention process) each proportioning group can reduce the weight gain caused by high-fat diet; Dengzhan Asarum (inventive technology) each proportioning group reduces body weight better than the control Dengzhan Asarum (Yunnan Haobang, existing technology); among them, the two ratios of dicaffeic acid ester salt: breviscapine salt 3:1 and 4.5:1 have the best effect.

3.2) Blood biochemical detection

a. Results for plasma total cholesterol content are shown in FIG. 2 and Table 5.

Table 5. Plasma total cholesterol content

The results showed that the plasma total cholesterol content of high-fat diet model animals was significantly higher than that of the healthy control group (***P<0.001). The experimental Dengzhanxixin (inventive process) has a tendency to reduce plasma total cholesterol, but there is no significant difference compared with the HFD group.

b. Results for plasma low density lipoprotein-cholesterol (LDL-c) levels are shown in FIG. 3 and Table 6. b.

Table 6. Plasma Low Density Lipoprotein-Cholesterol (LDL-c) Content

The results showed that the plasma low-density lipoprotein-cholesterol content of the high-fat diet model animals was significantly higher than that of the healthy control group (***P<0.001); -Cholesterol effect; experimental breviscapine (inventive process) has the effect of lowering plasma LDL-cholesterol, among which the dicaffeoate salt: breviscapine salt 4:1 and 4.5:1 two ratio groups can significantly reduce LDL- c effect (**P<0.01), 3:1 (P=0.085) and 6:1 (P=0.071) groups also had a certain effect on reducing plasma LDL-cholesterol.

c. Results of plasma triglyceride (TG) levels are shown in FIG. 4 and Table 7.

Table 7. Plasma triglyceride (TG) levels

The results showed that the plasma triglyceride (TG) content of the high-fat diet model animals was significantly higher than that of the healthy control group (***P<0.001); aspirin and control Dengzhan Xixin (existing technology) had no effect on reducing plasma triglyceride. ; Experimental breviscapine (inventive process) dicaffeoate salt: breviscapine salt 4.5:1 ratio group can significantly reduce plasma triglyceride (*P<0.05), 3:1 (P=0.0548) ratio The group also had a certain effect of lowering plasma triglyceride.

d. The results of plasma alanine aminotransferase (ALT) levels are shown in Figure 5 and Table 8.

Table 8. Plasma alanine aminotransferase (ALT) levels

The results showed that the plasma alanine aminotransferase (ALT) content of the high-fat diet model animals was significantly higher than that of the healthy control group (*P<0.05); aspirin and the control Dengzhanxixin (existing technology) had no effect on reducing plasma alanine aminotransferase; experimental Dengzhanxixin (inventive process) dicaffeoate salt: scutellarin salt 3:1 ratio group and 4.5:1 ratio group significantly reduced plasma ALT (*P<0.05); 4:1 (P=0.0573) ) ratio group also has a certain effect of reducing plasma alanine aminotransferase.

e. Results for plasma aspartate aminotransferase (AST) levels are shown in Figure 6 and Table 9.

Table 9. Plasma aspartate aminotransferase (AST) levels

The results showed that the plasma aspartate aminotransferase (AST) content of the high-fat diet model animals was significantly higher than that of the healthy control group (*P<0.05); each experimental group had no effect on reducing plasma aspartate aminotransferase.

3.3) Liver and fat index detection

a. The results of the liver index are shown in Figure 7 and Table 10.

Table 10. Liver Index

Liver index refers to the ratio of liver wet weight to body weight. The results showed that compared with the healthy control group, the liver index of the high-fat diet model animals was significantly increased (***P<0.001); aspirin and the control Dengzhanxixin (existing technology) did not have the effect of reducing the liver index; the experimental Dengzhanxixin (the invention). Process) Dicaffeic acid ester salt: Breviscapine salt 4.5:1 ratio group has a certain effect on reducing liver index (P=0.0529), suggesting that it has a certain preventive effect on non-alcoholic fatty liver disease caused by high-fat diet.

b. The results of the fat index are shown in Figure 8 and Table 11

Table 11. Fat Index

Fat index refers to the ratio of epididymal fat wet weight to body weight. The results showed that compared with the healthy control group, the fat index of high-fat diet model animals increased significantly (*P<0.05); aspirin and control Dengzhanxixin (existing technology) had no effect on reducing fat index; experimental Dengzhanxixin (inventive process) Dicaffeic acid ester salt: breviscapine salt 4.5:1 ratio group has a certain effect of reducing fat index.

3.4) Test results of lipid content in liver tissue

The liver lipid content was expressed as liver triglycerides, the results of which are shown in FIG. 9 and Table 12.

Table 12. Liver triglyceride content

The results showed that the TG content in the liver tissue of the high-fat diet model animals was significantly higher than that of the healthy control group (***P<0.001). (Invention process) can significantly reduce TG in liver tissue, among which the two ratio groups of dicaffeoate salt: breviscapine salt 3:1 and 4.5:1 have the most significant effects (**P<0.01), 1.25:1, 2: The 1, 4:1 and 6:1 groups could also significantly reduce the TG of liver tissue (*P<0.05), suggesting that they have a better effect on prevention and treatment of non-alcoholic fatty liver disease.

3.5) Platelet aggregation and coagulation test results

a. Platelet aggregation results are shown in Figure 10 and Table 13.

Table 13. Platelet Aggregation Rate

Increased platelet aggregation rate is easy to form thrombus, block blood vessels, and is also one of the factors of coronary heart disease, which may lead to the formation of atherosclerosis. The results showed that the platelet aggregation rate of the high-fat diet model animals was significantly higher than that of the healthy control group (***P<0.001); aspirin significantly reduced platelet aggregation induced by high-fat diet (***P<0.001); Xin (existing technology) and each ratio group of scutellaria scutellariae (inventive process) in each experimental group can significantly reduce platelet aggregation caused by high-fat diet, wherein dicaffeoate salt: breviscapine salt 3:1 and 4.5:1 The two ratio groups had the most significant effect (**P<0.01), and the 1.25:1, 2:1, 4:1 and 6:1 groups could also significantly reduce platelet aggregation caused by high-fat diet (*P<0.05) and the effect Both are better than the control Dengzhanxixin (existing technology).

b. Activated partial thromboplastin time (APTT) results are shown in Figure 11 and Table 14.

Table 14. Activated partial thromboplastin time

Activated partial thromboplastin time (APTT) is a coagulation function test index that reflects the comprehensive activity of coagulation factors in the endogenous coagulation pathway, especially the first stage. The results showed that the APTT of the high-fat diet model animals was significantly lower than that of the healthy control group (*P<0.05); aspirin and control Dengzhanxixin (existing technology) did not increase APTT; the experimental group Dengzhanxixin (inventive technology) two coffee Acetate salt: Breviscapine salt 4:1 (P=0.052) and 4.5:1 (P=0.089) two ratio groups had a certain effect of increasing APTT, suggesting that it may prevent thrombotic diseases caused by high-fat diet.

c. Prothrombin time (PT) results are shown in Figure 12 and Table 15.

Table 15. Prothrombin Time (PT)

Prothrombin time is an index reflecting the activity of coagulation factors I, II, V, VII and X in plasma. Low prothrombin time is common when the blood is in a hypercoagulable state, such as myocardial infarction or cerebral thrombosis. The results showed that the PT of the high-fat diet model animals was significantly lower than that of the healthy control group (***P<0.001); aspirin and the control Dengzhanxixin (existing technology) did not increase PT; the experimental group Dengzhanxixin (the invention process) The dicaffeoate salt: breviscapine salt 4.5:1 ratio group can significantly increase the PT reduction caused by high-fat diet (*P<0.05), suggesting that it may prevent thrombotic diseases caused by high-fat diet.

d. Thrombin time (TT) results are shown in Figure 13 and Table 16.

Table 16. Thrombin time

Plasma thrombin time refers to the time required for the conversion of plasma fibrinogen into fibrin after adding "standardized" thrombin to the tested plasma. The low thrombin time may be related to the hyperlipidemia in the patient's own blood. May be caused by thromboembolic disease. The results showed that the TT of the high-fat diet model animals was significantly lower than that of the healthy control group (***P<0.001); aspirin and the control Dengzhanxixin (existing technology) did not have the effect of increasing PT; the experimental group Dengzhanxixin (the invention process) The ratio of dicaffeoate salt: breviscapine salt 4.5:1 can significantly increase the TT reduction caused by high-fat diet (*P<0.05), suggesting that it may prevent thrombotic diseases caused by high-fat diet.

d. Fibrinogen (FIB) results are shown in Figure 14 and Table 17.

Table 17. Fibrinogen

Fibrinogen (FIB) is a protein with coagulation function mainly synthesized by hepatocytes and is the most abundant coagulation factor in plasma. Clinical data show that most patients with elevated fibrinogen levels may experience myocardial infarction or sudden death, and the higher the fibrin level, the greater the risk; elevated plasma fibrinogen is an important factor in promoting the pathogenesis of atherosclerosis. Factors; elevated fibrinogen, blood is in a hypercoagulable state, blood flow slows down, blood viscosity increases, and it is easy to produce thrombus; high blood pressure is often accompanied by elevated fibrinogen levels, which induces and aggravates high blood pressure. An important reason; fibrinogen is closely related to fat metabolism; in addition, old age, smoking, obesity, stress and diabetes can promote the level of fibrinogen in the body. The results showed that the plasma fibrinogen of the high-fat diet model animals was significantly higher than that of the healthy control group (***P<0.001); aspirin and control Dengzhanxixin (existing technology) did not reduce the increase in fibrinogen caused by the high-fat diet. High effect; experimental group breviscapine (inventive process) dicaffeoate salt: breviscapine salt 4.5:1 ratio group can significantly reduce the increase of fibrinogen caused by high-fat diet (*P<0.05), dicaffeine Acid salt: Breviscapine 2:1 (P=0.064), 3:1 (P=0.058), 6:1 (P=0.057) groups also had a certain effect on reducing plasma fibrinogen.

4 Conclusion

4.1) High-fat diet for eight weeks can significantly increase the body weight, liver index, fat index, liver lipid, blood lipid and liver enzyme levels of golden hamsters, and the model is successful.

4.2) The anti-platelet drug aspirin has no effect on reducing the weight gain caused by high-fat diet (the other positive drug, clopidogrel, has the same test result as aspirin); the invention technology of Dengzhan Xixin each compounding drug can reduce the weight gain caused by high-fat diet; And the weight-reducing effect is better than that of the control Dengzhanxixin (Haobang Pharmaceutical, the existing technology); among them, the two ratio groups of dicaffeoate salt: breviscapine salt 3:1 and 4.5:1 have the best effect.

4.3) The antiplatelet drug aspirin and the control Dengzhanxixin (Haobang Pharmaceutical, existing technology) have no blood lipid lowering effect, including plasma total cholesterol, low density lipoprotein-cholesterol, triglyceride (another positive drug clopidogrel test results) It is the same as aspirin); the experimental Dengzhanxixin (inventive process) has the effect of lowering blood lipids, and the dicaffeoate salt: breviscapine salt 4.5:1 ratio group is the best.

4.4) Aspirin and control Dengzhanxixin (Haobang Pharmaceutical, existing technology) did not have the effect of lowering plasma liver enzymes (the other positive drug clopidogrel test results were the same as aspirin); experimental Dengzhanxixin (inventive process) had the effect of reducing glutathione Transaminase effect, and the dicaffeoate salt: breviscapine salt 4.5:1 ratio group was the best.

4.5) Aspirin and control Dengzhanxixin (Haobang Pharmaceutical, existing technology) did not reduce the liver index, fat index and lipid accumulation in liver tissue (the other positive drug clopidogrel test results were the same as aspirin); Dengzhanxixin ( The new process) each ratio of the drugs has the effect of adjusting the above indicators, and the dicaffeoate salt: breviscapine salt 4.5:1 ratio group is the best.

4.6) The drugs in the proportions of Dengzhanxixin (inventive process) can significantly improve platelet aggregation and coagulation disorders induced by high-fat diet, including four indicators of platelet aggregation rate and coagulation; its effect is better than that of the control Dengzhanxixin (existing technology). ); among them, the combined effect of dicaffeoate salt: breviscapine salt ratio of 4.5:1 is the best; aspirin can effectively reduce platelet aggregation but has no anticoagulant effect (the other positive drug clopidogrel test results are the same as aspirin ).

7. Based on the above results, the experimental Dengzhan Asarum (inventive process) has a good effect on preventing and treating metabolic-related diseases such as hyperlipidemia, non-alcoholic fatty liver disease, and obesity, and can effectively prevent and treat platelet aggregation and abnormal blood coagulation function caused by metabolic-related diseases. , has the effect of preventing and treating cardiovascular and cerebrovascular diseases; its effect is related to the preparation process and the proportion of main components.

The existing breviscapine capsule powder is prepared from breviscapine salt and caffeic acid ester salt in the ratio within the scope of the claims.

Detailed ways:

Example 1: Preparation of raw material drug extract

Take 2000 g of Asarum radix and add 50% aqueous ethanol to decoct twice, 2 hours each time, filter, combine the filtrates, and concentrate under reduced pressure to form an extract (relative density 1.15-1.25, 75°C); add 5 times the amount of the extract dissolved in water, add 5% sodium hydroxide solution with stirring, adjust pH to 7.5-8.5, filter, add 10% sulfuric acid solution to adjust pH to 1-3, leave overnight, filter, collect filtrate and precipitate, precipitate with ethanol, and then Add 5% sodium hydroxide to adjust the pH value to 7-8, spray dry to obtain powder 1; adjust the pH value of the acidified filtrate to 7-9, use a 100nm ceramic membrane for clarification, and use a 350D organic membrane for the clarified solution obtained after clarification. Concentrated, the concentrate was passed through a 30-60 mesh polyamide chromatography column (diameter to length ratio 1:4), eluted with water for 3 column volumes, eluted with 65% ethanol for 4 column volumes, and collected the ethanol eluent, After concentration, the pH value is adjusted to 7-9 and spray-dried to obtain powder 2; powder 1 and powder 2 are mixed in proportion to obtain a mixture of scutellarin sodium salt powder and caffeate sodium salt powder 1:4 to obtain the medicine of the present invention API composition ingredients of the composition. Among them, powder 1 contains 36.3% of scutellarin sodium salt, a total of 5.4 g. Powder 2 contains 21.0% of dicaffeic acid ester salt, a total of 30.9 g, and 9.1 g of monocaffeic acid ester salt. Among them, the simple substituted dicaffeoate salt was 19.0 g, and the dicaffeoyl octanoate salt was 11.9 g. The obtained total mixed powder of Asarum scutellariae was 162 g.

Example 2: Preparation of raw drug extracts

Take 2000 g of Asarum radix and add 50% aqueous ethanol to decoct twice, 2 hours each time, filter, combine the filtrates, and concentrate under reduced pressure to form an extract (relative density 1.15-1.25, 75°C); add 5 times the amount of the extract Dissolve in water, add 5% sodium hydroxide solution with stirring, adjust pH to 7.5-8.5, filter, add 10% sulfuric acid solution to adjust pH to 1-3, leave overnight, filter, collect filtrate and precipitate, precipitate with ethanol, reduce Press-dried to obtain powder 1; the acidified filtrate was adjusted to pH 7-9, clarified with a 100nm ceramic membrane, the clarified solution obtained after clarification was concentrated with a 350D organic membrane, and the concentrated solution passed through a 30-60 mesh polyamide chromatography column (diameter to length ratio 1:4), after eluting 3 column volumes with water, eluting 4 column volumes with 65% ethanol, collecting the ethanol eluent, concentrating and drying under reduced pressure to obtain powder 2; 2. Mix according to the proportion to obtain a 1:4 mixture of breviscapine powder and caffeic acid ester powder, and obtain the raw material drug composition components of the pharmaceutical composition of the present invention. Among them, powder 1 contains 35.7% of scutellarin 5.6g in total. Powder 2 contained 21.5% of dicaffeic acid ester in a total of 31.4 g and monocaffeic acid ester 9.1 g. Among them, the simple substituted dicaffeoate was 18.5 g, and the dicaffeoyl octanoic acid was 12.9 g. 161.7 g of the total mixed powder of Asarum scutellariae was obtained.

Example 3: Preparation of raw drug extracts

Take 3000 g of Asarum radix and add 60% aqueous ethanol to decoct twice, 2 hours each time, filter, combine the filtrates, and concentrate under reduced pressure to form an extract (relative density 1.15-1.25, 75°C); add 5 times the amount of extract Dissolve in water, add 5% sodium hydroxide solution under stirring, adjust pH 7~9, filter, add 10% hydrochloric acid solution to adjust pH 1~3, leave overnight, filter, collect filtrate and precipitate, precipitate with ethanol, add Adjust the pH value to 7-8 with 5% sodium hydroxide, and spray dry to obtain powder 1;

The acidified filtrate was adjusted to pH 7-9, clarified with a 200nm ceramic membrane, the clarified solution obtained after clarification was concentrated with a 400D organic membrane, and the concentrated solution passed through a 30-60 mesh polyamide chromatography column (diameter to length ratio of 1). : 4), after eluting with 3 column volumes of water, eluting with 3 column volumes of 70% ethanol, collecting the ethanol eluent, adjusting the pH value to 7-9 after concentration and spray drying to obtain powder 2; 1 about 18.7g, powder 2 about 208.3g, mix powder 1 and powder 2 in proportion to obtain a mixture of scutellarin sodium salt powder and caffeic acid ester sodium salt powder 1:3 to obtain the medicinal composition of the present invention. API composition ingredients.

Among them, powder 1 contains 45% of scutellarin sodium salt, a total of 8.4 g. Powder 2 contained 18.6% of dicaffeoate salt in a total of 38.7 g and 7.3 g of monocaffeate salt. Among them, the simple substituted dicaffeoate salt was 18.3 g, and the dicaffeoyl octanoate was 20.4 g.

Example 4: Preparation of raw drug extracts

Take 3000 g of Asarum radix and add 65% aqueous ethanol to decoct twice, 2 hours each time, filter, combine the filtrates, and concentrate under reduced pressure into extract (relative density 1.15-1.25, 75 ° C); add 5 times the amount of extract Dissolve in water, add 5% sodium hydroxide solution under stirring, adjust pH 7~9, filter, add 10% hydrochloric acid solution to adjust pH 1~3, leave overnight, filter, collect filtrate and precipitate, precipitate with ethanol, reduce Pressed powder 1; the acidified filtrate was adjusted to pH 7-9, clarified with a 200nm ceramic membrane, the clarified solution obtained after clarification was concentrated with a 400D organic membrane, and the concentrated solution passed through a 30-60 mesh polyamide chromatography column ( The ratio of diameter to length is 1:4), after eluting with 3 column volumes of water, eluting with 3 column volumes of 70% ethanol, collecting the ethanol eluent, concentrating and adjusting the pH value to 7-9 for spray drying to obtain Powder 2; obtain about 25.3 g of powder 1 and about 223 g of powder 2, mix powder 1 and powder 2 in proportion to obtain a mixture of breviscapine and dicaffeoate sodium salt powder 1:3.5 to obtain the medicinal combination of the present invention ingredient of the drug substance.

Among them, powder 1 contains 32% scutellarin 8.1g in total. Powder 2 contained 12.7% of dicaffeoate salt in a total of 28.4 g and 7.3 g of monocaffeate salt. Among them, the simple substituted dicaffeoate salt was 13.4 g, and the dicaffeoyl octanoate was 15 g.

Example 5: Preparation of API Extract

Take 2500 g of Asarum sinensis and add 75% concentration of water ethanol to decoct twice, 2 hours each time, filter, combine the filtrates, and concentrate under reduced pressure into extract (relative density 1.15-1.25, 75 ° C); add 5 times the amount of extract Dissolved in water, add 5% sodium hydroxide solution with stirring, adjust pH 7.5~9.0, filter, add 10% sulfuric acid solution to adjust pH 1~3, leave overnight, filter, collect filtrate and precipitate, precipitate with ethanol, add Adjust the pH value to 7-8 with 5% sodium hydroxide, and spray dry to obtain powder 1;

The acidified filtrate was adjusted to pH 7-9, clarified with a 100nm ceramic membrane, and the clarified solution obtained after clarification was then concentrated with a 300D organic membrane, and the concentrated solution passed through a 30-60 mesh polyamide chromatography column (diameter to length ratio 1: 4), after eluting with 3.5 column volumes of water, eluting with 3 column volumes of 75% ethanol, collecting the ethanol eluent, concentrating and adjusting the pH value to 7-9 by spray drying to obtain powder 2; obtain 13.5g of Powder 1 and 164 g of powder 2, the obtained scutellarin sodium salt powder and caffeic acid ester sodium salt powder are mixed according to 1:2 to obtain the raw material drug composition components of the pharmaceutical composition of the present invention. Among them, powder 1 contains 51% of scutellarin sodium salt, totaling 6.9 g. Powder 2 contained 19.5% of dicaffeic acid ester salt, a total of 32.0 g, and 9.7 g of monocaffeic acid ester salt. Among them, the simple substituted dicaffeoate salt was 19.6 g, and the dicaffeoyl octanoate was 12.4 g.

The inventor also used 10%, 30%, 40%, 45%, 50%, 55%, 60%, 75%, 90% of different concentrations of aqueous ethanol to extract Asarum sinensis, and other steps were the same as in Example 1. , After being acidified and concentrated through the membrane, spray dry powder is obtained. Experiments have shown that 10-90% ethanol extraction can obtain lampshade extract, but from the yield, membrane passing rate, impurity content, powder bulk density and uniformity, industrial Measured by indicators such as production cost, the extract obtained with more than 40% ethanol is relatively better than the ethanol extract with less than 40%, and the ethanol extract with 40-75% is obviously better and more suitable for the process of the present invention, and 50% is the most preferred. % ethanol to extract Asarum sinensis.

Embodiment 6: the difference of active ingredient content in the inventive process and the existing process

Inventive process: take 2000 g of Asarum radix and add 50% hydrous ethanol to decoct twice, 2 hours each time, filter, combine the filtrates, and concentrate under reduced pressure to form an extract (relative density 1.15-1.25, 75° C.); Dissolve 5 times the amount of water, add 5% sodium hydroxide solution under stirring, adjust pH to 7.5-8.5, filter, add 10% sulfuric acid solution to adjust pH 1-3, leave overnight, filter, collect filtrate and precipitate, and precipitate ethanol After refining, add 5% sodium hydroxide to adjust the pH value to 7-8, spray-dry to obtain powder 1; adjust the pH value of the acidified filtrate to 7-9, use 100nm ceramic membrane for clarification, and use 350D for the clarified liquid obtained after clarification. The organic membrane is concentrated, and the concentrate is passed through a 30-60 mesh polyamide chromatography column (diameter to length ratio of 1:4), eluted with water for 3 column volumes, and eluted with 65% ethanol for 4 column volumes, and the ethanol wash is collected. After deliquoring, the pH value was adjusted to 7-9 after concentration and spray-dried to obtain powder 2; 15 g of powder 1 and 147 g of powder 2 were mixed to obtain breviscapine sodium salt powder and caffeate sodium salt powder.

Existing technology: take 2000g of Asarum radix, extract twice with 80% ethanol under reflux, the first time is 1.5 hours, the second time is 1 hour, the extracts are combined, about 640g of activated carbon is added, boiled, filtered, and the filtrate is concentrated under reduced pressure to Thick paste, add appropriate amount of starch, dry under reduced pressure, pulverize and sieve. Obtain Asarum radix extract powder.

Taking the weight of the total powder as the denominator and the weight of each type of ingredients as the numerator to calculate the content Table 18

Table 18. Comparison of components between the existing process and the inventive process

Breviscapine is the extract specified in the pharmacopoeia, and the main components are scutellarin and other substances, such as scutellarin, apigenin, scutellarin, etc., but the content is expressed as scutellarin.

In view of the high content of the above-mentioned effective substances, the proportion of the experimental group is based on the test results, using powder 1 and powder 2 to prepare.

Example 5: Preparation of capsules

Take 162 g of the extraction mixture prepared in Example 1, add 18 g of starch, mix well, and fill the capsules to obtain the final product.

Example 6: Preparation of capsules

Take 151 g of the extraction mixture prepared in Example 2, add 29 g of starch, mix well, and fill the capsules to obtain the final product.

Example 7: Preparation of capsules

Take 142 g of the extraction mixture prepared in Example 1, add 34 g of starch, and 4 g of magnesium stearate, mix well, and fill the capsules to obtain the final product.

Example 8: Preparation of Tablets

Take 151 g of the extraction mixture prepared in Example 2, 100 g of starch, and 10 g of dextrin, pass through a 14-mesh sieve for granulation, ventilate and dry at 60-70° C., and add 3 g of magnesium stearate. Compressed into tablets and coated.

Example 9: Preparation of dripping pills

Take 15 g of the extraction mixture prepared in Example 3, put in 45 g of polyethylene glycol 4000, mix evenly, melt, drop into low-temperature liquid paraffin, select pellets, and remove the liquid paraffin.

Embodiment 10: the preparation of oral liquid

Take 20 g of the extraction mixture prepared in Example 1, mix it with 300 g of honey, 50 g of sucrose, 2 g of sodium benzoate and 300 ml of distilled water, heat to dissolve, and filter at a temperature to obtain it.

Example 11: Preparation of Granules

Take 9 g of the extraction mixture prepared in Example 3, mix it with 40 g of microcrystalline cellulose, add 3% povidone ethanol solution to make soft material, pass through an 18-mesh sieve to make granules, dry at 600 ° C for 30-45 minutes, granulate, add 4g talcum powder, mix well, granulate, bag, and that’s it.

Example 12: Preparation of Soft Capsules

Take 120 g of the extraction mixture prepared in Example 3, add 5% glycerol, 1% glycine, add 400 to 400 g of polyethylene glycol, mix well, and press into 1000 soft capsules.

Claims (10)

1. An oral preparation containing breviscapine extract, characterized in that the oral preparation contains caffeic acid ester or its salt and breviscapine or its salt and pharmaceutically acceptable excipients, caffeic acid ester or its salt and breviscapine The weight ratio of its salt is 1:1 to 30:1.
2. The oral preparation according to claim 1, wherein the caffeic acid ester or its salt comprises dicaffeoate or its salt and monocaffeic acid or its salt, wherein the dicaffeoate or its salt is a The weight ratio of anthocyanin or its salt is 1:1 to 6:1, and the weight ratio of monocaffeic acid ester or its salt to dicaffeoate or its salt is 1:1.2 to 1:6.2.
3. The oral preparation according to claim 2, wherein the dicaffeoyl ester or its salt comprises 1,3-O-dicaffeoylquinic acid or its salt, 3,4-O-dicaffeoyl Quinic acid or its salts, 3,5-O-dicaffeoylquinic acid or its salts, 4,5-O-dicaffeoylquinic acid or its salts, Fembol ethyl or its salts, scutellarin or its salt; monocaffeic acid ester or its salt including 3-O-caffeoylquinic acid or its salt, 4-O-caffeoylquinic acid or its salt, 5-O-caffeoylquinic acid or its salt , scutellarin or its salt.
4. The oral preparation according to claim 3, wherein in the dicaffeoate or its salt, 1,3-O-dicaffeoylquinic acid or its salt, 3,4-O-dicaffeoyl Quinic acid or its salt, 3,5-O-dicaffeoylquinic acid or its salt, 4,5-O-dicaffeoylquinic acid or its salt are collectively referred to as simple substituted dicaffeoate or its salt; Said fiponin ester B or its salt and scutellarin ester or its salt are isomers, called dicaffeoyl caprylic acid or its salts, wherein simply substituted dicaffeoate or its salt and dicaffeic acid ester or its salt are isomers. The ratio of caffeoyl caprylic acid or its salt is 2:1 to 0.9:1.
5. The oral preparation according to any one of claims 1-4, wherein the oral preparation is a hard capsule, a soft capsule, a tablet, a granule, an oral liquid, a dripping pill, a water pill, a powder, or a pill.
6. The oral preparation of claim 5, wherein the weight ratio of the caffeic acid ester or its salt to breviscapine or its salt is 1:1 to 30:1, and the caffeic acid ester salt and the breviscapine salt are Sodium or potassium salts or other medicinal and water-soluble salts.
7. The preparation method of the oral preparation according to claim 1, characterized in that: taking Dengzhan Asarum and adding 10-90% concentration of aqueous ethanol to extract, the extract is concentrated under reduced pressure to form an extract; the extract is dissolved in water, and the pH is adjusted to 7-9 , filter, add acid to adjust pH to 1~3, leave overnight, filter, collect filtrate and precipitate, precipitate with water and ethanol, and get breviscapine after drying; add alkali to adjust pH to 7~8 after precipitation and purification, spray Drying to obtain scutellarin salt; the acidified filtrate is adjusted to pH 7-9, clarified with a ceramic microfiltration membrane, the clarified solution obtained after clarification is concentrated with an organic nanofiltration membrane, and the concentrated solution is passed through a polyamide chromatography column , eluted with water, eluted with 50-80% ethanol, collected the ethanol eluate, concentrated and dried to obtain caffeic acid ester; the ethanol eluate was concentrated and adjusted to pH 7-9, filtered, and the filtrate was spray-dried to obtain caffeic acid Ester salt; caffeic acid ester or its salt and breviscapine or its salt are mixed in the ratio of 1:1 to 30:1 to obtain the oral preparation of breviscapine, adding appropriate auxiliary materials, can be made into tablets, capsules, soft Capsules, drop pills, granules and oral liquid preparations.
8. The preparation method according to claim 7, characterized in that: the pH value of the alkali-adjusting solution is NaOH, Na ₂ CO ₃ , NaHCO ₃ , KOH, K ₂ CO ₃ or KHCO ₃ , or others that can be used to adjust The pH value of the alkaline solution; the pH value of the acid adjustment solution is HCl, H ₂ SO ₄ or H ₃ PO ₄ , or other acid solutions that can be used to adjust the pH value.
9. The application of the oral preparation of claim 1 in the preparation of medicines for treating hyperlipidemia, hypertension, diabetes, obesity, non-alcoholic fatty liver disease and other metabolic related diseases and cardiovascular and cerebrovascular diseases.
10. The application of the oral preparation of claim 1 in the preparation of a drug for inhibiting or treating platelet aggregation and coagulation disorders caused by metabolic-related diseases.

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