CN1795893A - Medication composition for treating hepatitis, prepartion method and application - Google Patents

Abstract

The invention relates to a pharmaceutical composition for treating hepatitis, which is a medicament prepared from salvia miltiorrhiza and notoginseng as raw materials. The medicament is an oral and injection preparation. The drug dosage is reduced, impurities are removed at the same time, the dosage is more accurate, and the curative effect for treating hepatitis is good, safe, controllable and stable, and provides a new choice of drugs for treating hepatitis in clinic.

Description

A kind of pharmaceutical composition for treating hepatitis and its preparation method and application

technical field

The invention relates to a pharmaceutical composition for treating hepatitis, in particular to a pharmaceutical composition for treating hepatitis prepared from traditional Chinese medicines, and belongs to the field of traditional Chinese medicines.

Background technique

The liver is one of the important viscera of the human body. The theory of traditional Chinese medicine believes that the main physiological function of the liver is to regulate the flow of blood and store blood. The liver governs catharsis, which means that the liver has the functions of relaxing, developing, modulating, dispersing, and circulating to maintain the smooth flow of Qi in the whole body. The normal performance of this function can regulate the spirit and emotions, make people happy and feel comfortable; it can also promote digestion and absorption, coordinate the rise and fall of spleen and stomach qi; it can also maintain the operation of qi and blood and regulate water metabolism. The liver stores blood, which means that the liver also has the function of storing blood and regulating blood volume. The blood stored in the liver is the material basis of its dredging function, and dredging is the functional manifestation of the stored blood. Normal dredging depends entirely on the nourishment of yin and blood; smooth circulation of blood is inseparable from the regulation of liver qi. Once the liver fails to ventilate and qi stagnates, blood stasis in the liver will occur, the spleen will fail to move well, and phlegm will stop inside, which will lead to pathological changes such as hepatic enlargement, spasm and pain under the hypochondriac, and abnormal liver function.

At present, there are three main categories of "anti-hepatitis drugs" clinically used in Western medicine: antiviral drugs, immunomodulatory drugs, and liver-protecting drugs (vitamin drugs, drugs with detoxification functions, drugs that promote energy metabolism, and drugs that promote protein synthesis). drugs). Antiviral drugs mainly include interferon antiviral drugs and nucleotide antiviral drugs, and clinical applications mainly include interferon and lamivudine; immunomodulatory drugs mainly include thymosin, lentinan, etc.; The clinical application of drugs mainly includes diammonium glycyrrhizinate (Ganlixin), matrine and so on. However, since there has been no breakthrough in the etiological treatment of viral hepatitis so far, symptomatic treatment is usually performed for symptoms of abnormal liver function in clinical practice.

In the traditional Chinese medicine treatment of abnormal liver function caused by hepatitis, etc., the main therapeutic principles are clearing away heat and removing dampness, promoting blood circulation and removing blood stasis, strengthening the body and strengthening the root. Among them, traditional Chinese medicines that mainly promote blood circulation and remove blood stasis may contain medicinal materials such as salvia miltiorrhiza and notoginseng, which have the following therapeutic characteristics for hepatitis: ① recovery of liver function is better and faster, and it has obvious effects of lowering enzymes, turbidity, and bilirubin Function; ②Retract liver and spleen, improve stasis; ③Anti-hepatic fibrosis; ④Promote regeneration of liver cells. At present, the existing Chinese patent medicine products are basically oral crude preparations, and there is no effective part injection dosage form. Because the liver is the detoxification organ of the human body, many drugs need to be metabolized by the liver. In the treatment of hepatitis, taking a large amount of drugs often increases the burden on the liver. It needs to be refined and purified to reduce the dosage of drugs. More than 90% of the crude preparations are non-active ingredients, which are absorbed Entering the body will greatly increase the burden on the liver; at the same time, clinically, most patients are admitted to the hospital for obvious liver damage due to the acute attack of hepatitis. At this time, the condition is critical, the change is rapid, and the mortality rate is high. Therefore, for patients with acute abnormal liver function, it is urgent to develop oral preparations and injection preparations made of active parts or active ingredients, so as to increase efficiency, reduce toxicity and quickly exert therapeutic effects.

Today's pharmaceutical preparations containing Danshen and Panax notoginseng raw materials, including oral preparations and injections, such as Danshen injection, compound Danshen dripping pills, etc., have not been strictly extracted and purified, and the content of effective parts is low, and the reported functions and indications And indications for the treatment of cardiovascular and cerebrovascular. At present, there have been reports on the treatment of hepatitis B by injection of Danshen and Panax notoginseng or oral or injection containing two raw materials (Wu Shanming et al., Preliminary observations on the treatment of blood stasis type chronic hepatitis and its blood rheology with Shenshen Sanqi injection, Shanghai Journal of Traditional Chinese Medicine 1983; 8, 14; Tong Shaohong, Observation on the short-term curative effect of Danshen acupoint injection on 39 cases of chronic hepatitis, Fujian Medical Journal 1980; 1, 50; Research and Practice of Traditional Chinese Medicine, No. 03, 1996; Xu Liming, "Huangqi Jiedu Huoxue Decoction" in the treatment of 107 cases of chronic hepatitis B, Jiangsu Traditional Chinese Medicine, No. 10, 2000; Chaiping Qinggan Decoction in the treatment of 146 cases of hepatitis A, Chinese Journal of Practical Traditional Chinese and Western Medicine, 2002 Issue 01), but due to the complex composition of traditional Chinese medicine, the efficacy of medicinal materials in the preparation is different, and the mixed use of single medicine and extract in traditional Chinese medicine injection may produce unpredictable efficacy and toxicity. Therefore, there have been no reports on the combined injection of Danshen and Sanqi to treat hepatitis B, and there is no report on the combined use of raw materials Danshen and Sanqi to treat hepatitis.

Contents of the invention

The technical solution of the present invention is to provide a pharmaceutical composition for treating hepatitis, specifically, a pharmaceutical composition for treating hepatitis prepared from Chinese medicinal materials Salvia miltiorrhiza and Panax notoginseng as raw materials, and the present invention also provides the composition of the pharmaceutical composition Preparation method and use.

The invention provides a pharmaceutical composition for treating hepatitis, which is a medicament prepared from the following raw materials in weight ratio:

3-10 parts of Danshen, 1-3 parts of Panax notoginseng.

Among them, Salvia miltiorrhiza is the dried root and rhizome of Salvia miltiorrhiza Bge. of Labiatae; Sanqi is the dried root of Panax notoginseng (Burk.) F.H.Chen of Araliaceae.

Furthermore, it is a medicament prepared from the following raw materials in weight ratio:

6 parts of Danshen, 1 part of Panax notoginseng.

The pharmaceutical composition is a medicament prepared from the water extract or alcohol extract of medicinal materials Danshen and Panax notoginseng as active components, plus pharmaceutically acceptable auxiliary materials or auxiliary components.

The medicament is oral preparation or injection.

Further, the oral preparations are capsules, tablets, pills, granules, and powders; the injections are injections, intravenous drops, and powder injections.

Wherein, the injection contains 3.24-3.36 mg of water-soluble total phenols of Danshen, 1.69-1.76 mg of total saponins, 0.83-0.91 mg of Danshensu, 0.23-0.25 mg of protocatechualdehyde, and 0.63-0.69 mg of ginsenoside Rg1 per milliliter.

Wherein, the HPLC fingerprint spectrum of this pharmaceutical composition is shown in Figure 1, is made up of 4 characteristic peaks,

The chromatographic conditions are as follows: octadecylsilane bonded silica gel is used as filler; methanol-acetonitrile-1.67% formic acid solution (volume ratio 23:7:70) is used as mobile phase; the detection wavelength is 286nm.

Further, the relative standard deviation RSDs of the relative retention times of the four common peaks are all less than 10.0%, wherein the average relative retention time of No. 1 peak is RT0.134%, No. 2 peak RT0.184, and No. 3 peak RT0.724 , No. 4 peak 1.000.

The present invention also provides a preparation method of the pharmaceutical composition, comprising the following steps:

a, weigh raw materials according to each weight ratio: 3-10 parts of salvia miltiorrhiza, 1-3 parts of notoginseng;

b. Preparation of Salvia miltiorrhiza extract: take medicinal material Salvia miltiorrhiza, add water to decoct and extract, filter, concentrate the filtrate, dry, use 50-95% ethanol as solvent for the dry extract powder, extract by percolation method, obtain percolation liquid, add activated carbon, Extract by heating under reflux, filter, recover ethanol, add water to dissolve, pass the aqueous solution through D ₁₀₁ macroporous adsorption resin, and elute with 30-85% ethanol, recover ethanol from the ethanol eluent to obtain the salvia miltiorrhiza extract;

c. Preparation of Panax notoginseng extract: take the medicinal material Panax notoginseng, use 35-95% ethanol as solvent, extract by percolation method, obtain percolation liquid, add activated carbon to the percolation liquid, reflux extraction, filter, and depressurize the filtrate Recover ethanol to obtain alcoholic extract, dissolve the alcoholic extract with water, filter, pass the aqueous solution through a D ₁₀₁ macroporous adsorption resin column, and then elute the effective part with 30-90% ethanol to obtain ethanol eluent, recover ethanol under reduced pressure , to obtain the extract of Panax notoginseng;

d. Mix the Danshen extract prepared in step b and the Panax notoginseng extract prepared in step c, add water to dissolve, add pharmaceutically acceptable excipients or auxiliary components, and prepare a commonly used pharmaceutical preparation.

Wherein, step d is to mix the salvia miltiorrhiza extract and the notoginseng extract prepared in step b and step c, add water to dissolve, and add common excipients commonly used in pharmaceutical oral preparations to obtain the oral preparation of the pharmaceutical composition of the present invention; or

Step d is to mix the salvia miltiorrhiza extract and Panax notoginseng extract prepared in step b and step c, add water to dissolve, add commonly used additives for injections commonly used in pharmacy, adjust the pH value to 5.5-6.5, and obtain the pharmaceutical composition injection of the present invention .

The invention also provides the use of raw materials in various weight ratios in the preparation of medicines for treating acute and chronic hepatitis.

Further, the use of raw materials in various weight ratios in the preparation of oral medicines for treating acute and chronic hepatitis.

Further, the use of raw materials in various weight ratios in the preparation of injections for the treatment of acute and chronic hepatitis.

Furthermore, the use of raw materials in various weight ratios in the preparation of intravenous injection of medicines for treating acute and chronic hepatitis.

The raw material formula of the pharmaceutical composition of the present invention is based on the guidance of traditional Chinese medicine theory, and the selection of medicines is reasonable and precise. It is suitable for long-term use, and is not easy to cause adverse reactions. Modern research shows that Salvia miltiorrhiza not only has various pharmacological effects on cardiovascular and cerebrovascular, microcirculation, etc., but also has relatively Good blood lipid-lowering effect, for the liver, can protect the liver damage caused by various reasons, and has anti-hepatotoxic effect; it has preventive and therapeutic effects on experimental liver cirrhosis, can also promote cell regeneration, inhibit liver fibrosis, etc., so the prescription It is used as a monarch drug in traditional Chinese medicine. Notoginseng, sweet in nature and flavor, slightly bitter and damp. Mainly belongs to the liver meridian, and also has the function of invigorating qi and nourishing blood (Higher Education Press, facing the 21st century course textbook "Chinese Materia Medica"). Modern research shows that Panax notoginseng can lower blood lipids, promote blood cell production, relieve pain; resist liver damage , improve liver blood circulation, improve chronic indicators of liver function, and enhance immune function and strength. Radix Notoginseng is used to assist the blood circulation of Salvia miltiorrhiza to remove blood stasis, eliminate symptoms and relieve pain, and has the effect of strengthening Qi and replenishing qi and blood, so that the whole prescription can be eliminated without injury, so it is used as an auxiliary drug. The two raw materials are used in proportion to play a synergistic effect.

Because there are too many unknown ingredients in a single Chinese medicine, and the properties of some active ingredients are also unstable, the injections prepared by mixing Chinese medicines are prone to oxidative polymerization of chemical components, reducing the efficacy of the medicine, and even producing precipitation. die. Through the method of extracting and refining the raw materials of the pharmaceutical composition of the present invention, the pharmaceutical composition of the present invention is prepared to treat hepatitis, extract and purify effective parts, use less raw materials, and be convenient to take. The injection solution does not produce precipitation, and the quality is stable; the intravenous injection has no side effects, is safe, and can overcome the defects of the water injection. Moreover, the pharmaceutical composition of the present invention uses fingerprints to control the quality of the injection, so that the pharmaceutical composition of the present invention is controllable and stable.

In summary, the medicine of the present invention uses Danshen and Panax notoginseng as raw materials to prepare the preparation. The mixture of the extracts of Salvia miltiorrhiza in the medicinal materials, the water-soluble total phenols of Danshen and the total saponins of Panax notoginseng, not only reduces the dosage, but also removes impurities, and the dosage is more accurate. The curative effect of treating hepatitis is good, safe, controllable and stable, and provides a new choice of drugs for treating hepatitis in clinic.

Apparently, according to the above content of the present invention, according to common technical knowledge and conventional means in this field, without departing from the above basic technical idea of the present invention, other various forms of modification, replacement or change can also be made.

The above-mentioned content of the present invention will be further described in detail below through specific implementation in the form of examples. However, this should not be construed as limiting the scope of the above-mentioned subject matter of the present invention to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Description of drawings

Fig. 1 HPLC fingerprint of the pharmaceutical composition of the present invention, wherein, No. 1 peak retention time RT is: 3.950min, No. 2 peak retention time RT5.425min, No. 3 peak retention time RT21.892min, and No. 4 peak retention time 30.533min.

Detailed ways

Embodiment 1 The preparation of the pharmaceutical composition of the present invention intravenous drip dosage form

a. Take 1000g of Salvia miltiorrhiza, cut into thick slices, add 8 times the amount of water to decoct and extract 4 times, decoct for 1.5 hours each time, combine the decoction, filter, and concentrate the filtrate to a clear liquid with a relative density of about 1.05 (60°C thermal measurement). paste, spray-dried to obtain extract powder. Get the extract powder and mix with 3 times the amount of diatomaceous earth, according to the percolation method (Appendix 10 of "Chinese Pharmacopoeia" 2000 edition) under the extract and extract item, use 92% ethanol as solvent, and soak for 4 hours Finally, slowly percolate at a speed of 3ml per minute, collect 15000ml of percolation liquid, add 50g of activated carbon, heat and reflux for 60 minutes, let it cool, filter, and recover ethanol from the filtrate under reduced pressure to dryness, add 2500ml of water to dissolve, and let it stand for 24 hour, filter, and the filtrate passes through the D ₁₀₁ macroporous adsorption resin column that has been pretreated, add water 5000ml to elute, discard the water washing solution, continue to use 50% ethanol 20000ml to elute, collect 50% ethanol eluate, reduce pressure The ethanol is recovered to obtain the salvia miltiorrhiza extract, which is the water-soluble total phenols of the salvia miltiorrhiza.

Among them, the transfer rate of water-soluble total phenols, danshensu, and protocatechuic aldehyde of salvia miltiorrhiza in water decoction extraction process is all above 95%, the dry paste yield is about 63%, and the total phenolic acid content of salvia miltiorrhiza in dry paste is about 5.5%; ethanol percolation The transfer rate of salvia miltiorrhiza root water-soluble total phenols, danshensu, and protocatechuic aldehyde in the pre-removal process is all over 85%, the yield of dry paste is reduced to about 10%, and the content of water-soluble total phenolics in salvia miltiorrhiza in dry paste is about 35%; macropores The yield of the extract obtained after purification by adsorption resin is about 3.0%, the content of water-soluble total phenols in the extract is about 85%, the content of Danshensu is about 25%, and the content of protocatechualdehyde is about 8%, which has reached the level of intravenous injection New drug requirements.

B, notoginseng extraction and purification: get notoginseng coarse powder 1000g, according to the percolation method ("Chinese Pharmacopoeia" 2000 edition an appendix I Q) under the liquid extract and the extract item, make solvent with 9 times of amount 65% ethanol, Immerse overnight, percolate at a rate of 5ml per minute, collect the percolation liquid, add 30g of activated carbon, heat and reflux for 60 minutes, let it cool, filter, and recover ethanol from the filtrate under reduced pressure, add 2500ml of water to dissolve, filter, and pass the filtrate through the pre-prepared The treated D ₁₀₁ macroporous adsorption resin column was washed with water until the washing solution was nearly colorless (about 1200ml), discarded the washing solution, and then eluted with 50% ethanol with 11 times the amount of the medicinal materials, collected the eluent, and recovered under reduced pressure ethanol to obtain the Panax notoginseng extract, which is the total saponins of Panax notoginseng.

Among them, the ethanol percolation extraction process of the total saponins of Panax notoginseng and the transfer rate of ginsenoside Rg1 are all above 90%, the yield of the extract is about 20%, and the content of the total saponins of Panax notoginseng in the extract is about 45%; obtained after purification of the macroporous adsorption resin The yield of the notoginseng extract is about 8.5%, and the total saponin content of the notoginseng is more than 90%, which has reached the requirement of intravenous injection.

c. Take 3.3g of Danshen extract and 1.7g of Sanqi extract, add 400ml of water for injection to dissolve, and set aside; take another 9g of sodium chloride for injection, 0.2g of sodium bisulfite, add 100ml of water for injection to dissolve, heat to boil , add 0.2 g of activated carbon, stir well, continue to boil for 15 minutes, filter, mix the filtrate with the above-mentioned medicinal solution, add water for injection to 1000 ml, adjust the pH value to 6.0 with 10% sodium hydroxide solution, and ultra-filter until clarity, Divide into 100ml infusion bottles, crimp the caps, sterilize, check the clarity, pack, and get ready.

Example 2 The preparation method of the drug powder injection of the present invention

a. Take 1000g of Salvia miltiorrhiza, cut into thick slices, add 8 times the amount of water to decoct and extract 4 times, decoct for 1.5 hours each time, combine the decoction, filter, and concentrate the filtrate to a clear liquid with a relative density of about 1.05 (60°C thermal measurement). paste, spray-dried to obtain extract powder. Get the extract powder and mix with 3 times the amount of diatomaceous earth, according to the percolation method (Appendix 10 of "Chinese Pharmacopoeia" 2000 edition) under the extract and extract item, use 92% ethanol as solvent, and soak for 4 hours Finally, slowly percolate at a speed of 3ml per minute, collect 15000ml of percolation liquid, add 50g of activated carbon, heat and reflux for 60 minutes, let it cool, filter, and recover ethanol from the filtrate under reduced pressure to dryness, add 2500ml of water to dissolve, and let it stand for 24 hour, filter, and the filtrate passes through the D ₁₀₁ macroporous adsorption resin column that has been pretreated, add water 5000ml to elute, discard the water washing solution, continue to use 50% ethanol 20000ml to elute, collect 50% ethanol eluate, reduce pressure The ethanol is recovered to obtain the salvia miltiorrhiza extract, which is the water-soluble total phenols of the salvia miltiorrhiza.

Among them, the transfer rate of water-soluble total phenols, danshensu, and protocatechuic aldehyde of salvia miltiorrhiza in water decoction extraction process is all above 95%, the dry paste yield is about 63%, and the total phenolic acid content of salvia miltiorrhiza in dry paste is about 5.5%; ethanol percolation The transfer rate of salvia miltiorrhiza root water-soluble total phenols, danshensu, and protocatechuic aldehyde in the pre-removal process is all over 85%, the yield of dry paste is reduced to about 10%, and the content of water-soluble total phenolics in salvia miltiorrhiza in dry paste is about 35%; macropores The yield of the extract obtained after purification by adsorption resin is about 3.0%, the content of water-soluble total phenols in the extract is about 85%, the content of Danshensu is about 25%, and the content of protocatechualdehyde is about 8%, which has reached the new drug of powder injection requirements.

B, Radix Notoginseng extraction and purification: get Notoginseng coarse powder 1000g, according to the percolation method (Chinese Pharmacopoeia 2000 edition an appendix I Q) under the liquid extract and the extract item, make solvent with 9 times of amount 65% ethanol, soak overnight , percolate at a rate of 5ml per minute, collect the percolation liquid, add 30g of activated carbon, heat and reflux for 60 minutes, let cool, filter, and recover ethanol from the filtrate under reduced pressure, add 2500ml of water to dissolve, filter, and the filtrate has been pretreated D ₁₀₁ macroporous adsorption resin column, washed with water until the washing liquid is nearly colorless (about 1200ml), discarding the washing liquid, followed by elution with 50% ethanol of 11 times the amount of medicinal materials, collecting the eluent, and recovering ethanol under reduced pressure, The notoginseng extract is obtained, and the notoginseng extract is total saponins of notoginseng.

Among them, the ethanol percolation extraction process of the total saponins of Panax notoginseng and the transfer rate of ginsenoside Rg1 are all above 90%, the yield of the extract is about 20%, and the content of the total saponins of Panax notoginseng in the extract is about 45%; obtained after purification of the macroporous adsorption resin The yield of Panax notoginseng extract is about 8.5%, and the total saponin content of Panax notoginseng is more than 90%, which has reached the requirement of new medicine for powder injection.

c. Take 3.3g of Salvia miltiorrhiza extract and 1.7g of Sanqi extract, add 400ml of water for injection to dissolve, and set aside; put 100-200g of excipients for injection into a sterile container, add 450ml of water for injection, stir to dissolve, and mix with the above drugs Mix the solution, add water for injection to make up to 1000ml, stir gently until it is completely dissolved, filter the solution with two microporous filters connected in series under positive pressure, and spray dry it under sterile conditions to make Danqi Aseptic powder, divided into bottles, stoppered, pressed aluminum caps, sealed, sterilized, packaged, ready to be obtained.

Embodiment 3 The preparation of oral drug preparation of the present invention

a. Take 1000g of Salvia miltiorrhiza, cut into thick slices, add 8 times the amount of water to decoct and extract 4 times, decoct for 1.5 hours each time, combine the decoction, filter, and concentrate the filtrate to a clear liquid with a relative density of about 1.05 (60°C thermal measurement). paste, spray-dried to obtain extract powder. Get extract powder and mix with 3 times of amount diatomite, according to the percolation method (Chinese Pharmacopoeia 2000 edition one appendix 10) under flow extract and extract item, make solvent with 92% ethanol, after soaking for 4 hours, Slowly percolate at a rate of 3ml per minute, collect 15000ml of percolation liquid, add 50g of activated carbon, heat and reflux for 60 minutes, let cool, filter, recover ethanol from the filtrate under reduced pressure until dry, add 2500ml of water to dissolve, and let it stand for 24 hours. Filtrate, pass the filtrate through the D ₁₀₁ macroporous adsorption resin column that has been pretreated, add 5000ml of water to elute, discard the water wash, continue to elute with 20000ml of 50% ethanol, collect the 50% ethanol eluate, and recover ethanol under reduced pressure , that is, the extract of salvia miltiorrhiza.

Among them, the transfer rate of water-soluble total phenols, danshensu, and protocatechuic aldehyde of salvia miltiorrhiza in water decoction extraction process is all above 95%, the dry paste yield is about 63%, and the total phenolic acid content of salvia miltiorrhiza in dry paste is about 5.5%; ethanol percolation The transfer rate of salvia miltiorrhiza root water-soluble total phenols, danshensu, and protocatechuic aldehyde in the pre-removal process is all over 85%, the yield of dry paste is reduced to about 10%, and the content of water-soluble total phenolics in salvia miltiorrhiza in dry paste is about 35%; macropores The yield of the extract obtained after purification by adsorption resin is about 3.0%, the content of water-soluble total phenols in the extract is about 85%, the content of Danshensu is about 25%, and the content of protocatechualdehyde is about 8%, which has reached the new drug of powder injection requirements.

B, Radix Notoginseng extraction and purification: get Notoginseng coarse powder 1000g, according to the percolation method (Chinese Pharmacopoeia 2000 edition an appendix I Q) under the liquid extract and the extract item, make solvent with 9 times of amount 65% ethanol, soak overnight , percolate at a rate of 5ml per minute, collect the percolation liquid, add 30g of activated carbon, heat and reflux for 60 minutes, let cool, filter, and recover ethanol from the filtrate under reduced pressure, add 2500ml of water to dissolve, filter, and the filtrate has been pretreated D ₁₀₁ macroporous adsorption resin column, washed with water until the washing liquid is nearly colorless (about 1200ml), discarding the washing liquid, followed by elution with 50% ethanol of 11 times the amount of medicinal materials, collecting the eluent, and recovering ethanol under reduced pressure, That is the extract of Panax notoginseng.

Among them, the ethanol percolation extraction process of the total saponins of Panax notoginseng and the transfer rate of ginsenoside Rg1 are all above 90%, the yield of the extract is about 20%, and the content of the total saponins of Panax notoginseng in the extract is about 45%; obtained after purification of the macroporous adsorption resin The yield of Panax notoginseng extract is about 8.5%, and the total saponin content of Panax notoginseng is more than 90%, which has reached the requirements of new medicine for powder injection;

c. Take 3.3g of Salvia miltiorrhiza extract and 1.7g of Panax notoginseng extract, add 300ml of water to dissolve, and set aside; another 100-200g of auxiliary materials for oral preparations are placed in a sterile container, add 400ml of water, stir to dissolve, and mix with the above liquid medicine , add water to make up to 1000ml, stir gently until completely dissolved, concentrate, granulate, and dry. Made into Danqi tablets, granules or capsules, subpackaged and sterilized. Pack and serve.

Embodiment 4 preparation of injection of the present invention

According to the method of Example 1, 4000 g of medicinal materials Danshen and 400 g of Radix Notoginseng were taken, and 13 g of Danshen extract and 1 g of Radix Notoginseng extract were prepared to prepare aqueous injections.

Embodiment 5 Preparation of injection of the present invention

According to the method of Example 1, 3000 g of medicinal materials Danshen and 1000 g of Radix Notoginseng were taken, and 9.8 g of Danshen extract and 1.8 g of Radix Notoginseng extract were prepared to prepare aqueous injections.

Embodiment 6 Preparation of injection of the present invention

According to the method of Example 1, 5000 g of medicinal materials Danshen and 2000 g of Radix Notoginseng were taken, and 16.6 g of Danshen extract and 3.5 g of Radix Notoginseng extract were prepared to prepare aqueous injections.

Embodiment 7 The HPLC quantitative control method of medicine of the present invention

Get the pharmaceutical composition of the present invention prepared in Examples 1 to 3, measure according to high performance liquid chromatography, use octadecylsilane bonded silica gel as filler; methanol-acetonitrile-1.67% formic acid solution (volume ratio 23:7: 70) is the mobile phase; the detection wavelength is 286nm. Its standard fingerprint spectrum is shown in accompanying drawing 1.

The relative retention time of its standard fingerprint is 0.134(1), 0.184(2), 0.724(3), 1.000(S) successively, and its relative deviation shall not exceed 10%.

Embodiment 8 Drug quantitative control method of the present invention

①Water-soluble total phenols

◆Instruments and reagents: Shimadzu UV-2501 ultraviolet spectrophotometer; reagents and reagents are of analytical grade; Danshensu sodium reference substance was purchased from China Institute for the Control of Pharmaceutical and Biological Products (batch number 0855-200102).

◆Analysis method: Dilute the sample of Danqi injection with water, develop the color with the mixed solution of ferric chloride and potassium ferricyanide, measure the absorbance at 770nm wavelength, and use the standard curve method to calculate the content with Danshensu as the control.

◆Methodological investigation: The feasibility of the method was confirmed by the precision, reproducibility, stability and sample recovery tests.

◆Measuring results: See Table 1.

Table 1 Determination results of water-soluble total phenols in Danqi injection batch number 020401 020415 020416 average value Content (mg/ml) 3.24 3.34 3.36 3.31

②Total saponins

◆Instruments and reagents: Shimadzu UV-2501 ultraviolet spectrophotometer; reagents and reagents are of analytical grade; ginsenoside Rg1 reference substance is provided by China Institute for the Control of Pharmaceutical and Biological Products (batch number 0703-200015).

◆Analysis method: Danqi injection is extracted with n-butanol, dissolved and diluted in ethanol, developed with vanillin-perchloric acid, and the absorbance is measured at a wavelength of 551nm. With ginsenoside Rg1 as a control, the content is calculated by the standard curve method.

◆Methodological investigation: The feasibility of the method was confirmed by the precision, reproducibility, stability and sample recovery tests.

◆Measuring results: See Table 2.

Table 2 Determination Results of Total Saponins in Danqi Injection batch number 020401 020415 020416 average value Content (mg/ml) 1.76 1.69 1.73 1.72

③ Danshensu and protocatechualdehyde

◆Instruments and reagents: Shimadzu LC-10A high performance liquid chromatography, SPD-10AV ultraviolet detector. Methanol in the mobile phase is chromatographically pure, and other reagents are analytically pure. Danshensu sodium and protocatechualdehyde reference substances were purchased from China Institute for the Control of Pharmaceutical and Biological Products (batch numbers are 0855-200102 and 0810-200004, respectively).

◆Analysis method: Danqi injection was diluted with water as the test solution, and determined by HPLC. The chromatographic column is Shimpack CLC-ODS column; the mobile phase is methanol-0.5% glacial acetic acid solution (11:89); the detection wavelength is 280nm; the content is calculated by the external standard method.

◆Methodological investigation: The feasibility of the method was confirmed by the precision, reproducibility, stability and sample recovery tests.

◆Measuring results: See Table 3.

Table 3 Determination Results of Danshensu and Protocatechualdehyde in Danqi Injection batch number 020401 020415 020416 average value Danshensu (mg/ml) Protocatechualdehyde (mg/ml) 0.890.24 0.910.25 0.840.24 0.830.24 0.860.23 0.840.25 0.860.24

④Ginsenoside Rg1

◆Instruments and reagents: Shimadzu LC-10A high performance liquid chromatography, SPD-10AV ultraviolet detector. The reference substance of ginsenoside Rg1 was provided by China Institute for the Control of Pharmaceutical and Biological Products (batch number 0703-200015). Its high-performance liquid chromatography has a single main peak, and its purity measured by normalization method is 99.8%.

◆Analysis method: Danqi injection is directly used as the test solution, and determined by HPLC. The chromatographic column is an octylsilane bonded silica gel column; the mobile phase is acetonitrile-0.05% phosphoric acid solution (19:81); the detection wavelength is 203nm; the content is calculated by the external standard method.

◆Methodological investigation: The feasibility of the method was confirmed by the precision, reproducibility, stability and sample recovery tests.

Measurement results: see Table 4.

Table 4 Determination results of ginsenoside Rg1 in Danqi injection Sample lot number 020401 020415 020416 Ginsenoside Rg1(mg/ml) 0.69 0.64 0.63

Danshen notoginseng is an active ingredient in raw medicines. Therefore, the quality control of the ingredients in the two raw medicines is highly controllable, and the quality is more stable and controllable.

Prove beneficial effect of the present invention by pharmacodynamic test below.

Experimental Example 1 Anti-acute liver injury model test:

To the rat acute liver injury model caused by CCL4, the drug injection of the present invention 17.0mg/kg can significantly raise the value of ALT (alanine aminotransferase) and the value of AST (aspartate aminotransferase). Very significant reducing effect (P<0.001, P<0.01); 8.5mg/kg can very significantly reduce the ALT value (P<0.01). 4.2, 8.5, and 17.0 mg/kg of the drug injection of the present invention have a certain recovery effect on damaged liver cells, but there is no significant effect (P>0.05). Drug oral preparation of the present invention (prepared in embodiment 3) 21.0mg/kg can have extremely significant and AST value raising very significantly reducing effect to ALT value raising (P＜0.001, P＜0.01); 10.5mg/kg can Very significantly lowered the ALT value (P<0.01). 5.2, 10.5, and 21.0 mg/kg of the pharmaceutical oral preparations of the present invention have a certain recovery effect on damaged liver cells, but there is no significant effect (P>0.05).

Table 5 The influence of drug injection of the present invention on rat acute liver injury model caused by CCL4 group Dose (mg/kg) Number of rats (only) Blood and liver function of rats 3h after administration (x±SD) ALT(u/ml) AST(u/ml) normal control group equal volume 10 48.6±10.8 122.3±26.8 model group equal volume 10 292.2±129.7▲▲▲ 355.9±148.4▲▲▲ High-dose group of drug injection of the present invention 17 10 82.6±26.3 ^*** 185.4±43.4 ^*** Middle dose group of drug injection of the present invention 8.5 10 123.7±75.7 ^** 249.4±142.5 Low dose group of drug injection of the present invention 4.2 10 233.3±108.2 454.7±153.9

Note: Comparing each administration group with the model group ^* P<0.05 ^** P<0.01 ^*** P<0.001

Comparison between the model group and the normal control group ▲▲▲P<0.001 (two-sided t-test)

As can be seen from the above table: to the rat acute liver injury model caused by CCL4, the drug injection 17.0mg/kg of the present invention can have a very significant reduction effect on the ALT value increase and the AST value increase (P＜0.001, P<0.01); 8.5mg/kg can significantly reduce the ALT value (P<0.01). 4.2, 8.5, and 17.0 mg/kg of the drug injection of the present invention have a certain recovery effect on damaged liver cells, but there is no significant effect (P>0.05).

To the acute liver injury model of mice caused by CCL4, 17.0mg/kg of the drug injection of the present invention can significantly reduce ALT and AST values (P<0.001, P<0.001); 8.5mg/kg can very significantly reduce ALT, AST value (P<0.01); 4.2mg/kg can significantly reduce ALP (alkaline phosphatase) value (P<0.05). 4.2, 8.5, 17.0mg/kg have a certain recovery effect on the damaged liver cells, but there is no significant significance (P>0.05). 21.0mg/kg of the pharmaceutical oral preparation of the present invention can very significantly reduce ALT, AST value (P＜0.001, P＜0.001); 10.5mg/kg can very significantly reduce ALT, AST value (P＜0.01); 5.2mg/kg kg can significantly reduce the ALP value (P<0.05). 5.2, 10.5, 21.0mg/kg have a certain recovery effect on damaged liver cells, but there is no significant significance (P>0.05).

Table 6 The effect of the drug injection of the present invention on the mouse acute liver injury model caused by CCL4 group Dose (mg/kg) Number of rats (only) Serum liver function test of mice 3h after administration (x±SD) ALT(u/ml) AST(u/ml) ALP(u/ml) normal control group equal volume 10 39.8±18.5 113.9±26.1 154±32 model group equal volume 10 592.0±196.3▲▲▲ 519.3±173.0▲▲▲ 174±11 High-dose group of drug injection of the present invention 17.0 10 118.5±59.5 ^*** 191.5±72.4 ^*** 171±40 Middle dose group of drug injection of the present invention 8.5 10 292.2±221.9 ^** 278.0±133.8 ^** 175±24 Low dose group of drug injection of the present invention 4.2 10 445.8±215.1 471.8±158.2 148±37 ^*

Note: Comparing each administration group with the model group ^* P<0.05 ^** P<0.01 ^*** P<0.001

Comparison between the model group and the normal control group ▲▲▲P<0.001 (two-sided t-test)

To the acute liver injury model of mice caused by CCL4, 17.0mg/kg of the drug injection of the present invention can significantly reduce ALT and AST values (P<0.001, P<0.001); 8.5mg/kg can very significantly reduce ALT, AST value (P<0.01); 4.2mg/kg can significantly reduce ALP value (P<0.05). 4.2, 8.5, 17.0mg/kg have a certain recovery effect on the damaged liver cells, but there is no significant significance (P>0.05).

From the above dose-effect relationship test, it can be seen that the effect of the high and middle dose groups is more obvious than that of the low dose group.

To alcohol-induced acute liver injury model in mice, drug injection 17.0mg/kg of the present invention has very significant and very significant effect of reducing ALT and TB value (P＜0.01, P＜0.001); 8.5mg/kg has significant Decrease the effect of ALT value (P<0.05). 21.0mg/kg of the pharmaceutical oral preparation of the present invention has a very significant and very significant effect of reducing ALT and TB value (P<0.01, P<0.001); 10.5mg/kg has a significant effect of reducing ALT value (P<0.05).

Table 7 Anti-alcohol-induced mouse acute liver injury model test of drug injection of the present invention group Dose (mg/kg) Number of rats (only) Serum liver function test of mice 2.5h after administration (x±SD) ALT(u/ml) AST(u/ml) TB (umol/ml) normal control group -- 10 49.7±8.8 181.2±18.4 3.32±1.17 model group -- 10 127.7±26.2▲▲▲ 428.8±128.7▲▲▲ 9.54±1.21▲▲▲ High-dose group of drug injection of the present invention 17.0 10 95.6±18.8 ^** 365.0±82.7 5.96±21.8 ^*** Middle dose group of drug injection of the present invention 8.5 10 106.2±14.1 ^* 427.5±200.7 8.49±2.82 Low dose group of drug injection of the present invention 4.2 10 115.0±26.8 455.4±151.9 8.62±1.2

Note: Comparing each administration group with the model group ^* P<0.05 ^** P<0.01 ^*** P<0.001

Comparison between the model group and the normal control group ▲▲▲P<0.001 (two-sided t-test)

To alcohol-induced acute liver injury model in mice, drug injection 17.0mg/kg of the present invention has very significant and very significant effect of reducing ALT and TB value (P＜0.01, P＜0.001); 8.5mg/kg has significant Decrease the effect of ALT value (P<0.05).

To TAA (thioacetamide) induced acute liver injury model in rats, drug injection of the present invention 17.0mg/kg can significantly reduce TBil value (P＜0.05); 8.5mg/kg can significantly reduce ALT value (P＜0.05) ); 4.2mg/kg had a significant reduction effect on ALT, AST and TBil (P<0.05). 4.2, 8.5, 17.0 mg/kg all have a certain recovery effect on the damaged liver cells, but there is no significant significance (P>0.05). Drug oral preparation of the present invention 21.0mg/kg can significantly reduce Tbil (total bilirubin) value (P＜0.05); 10.5mg/kg can significantly reduce ALT value (P＜0.05); 5.2mg/kg can significantly reduce ALT, AST and TBil has a significant reduction effect (P <0.05). 5.2, 10.5, 21.0 mg/kg all have a certain recovery effect on damaged liver cells, but there is no significant significance (P>0.05).

To D-GLAN (D-galactosamine)-induced acute liver injury model in rats, the drug injection of the present invention 17.0mg/kg can significantly reduce the ALT value (P<0.01), significantly reduce the AST value (P<0.01) 0.025); 8.5mg/kg can significantly reduce the ALT value (P<0.05, P<0.025). 8.5mg/kg has a significant recovery effect on damaged liver cells (P<0.05); 4.2, 17.0mg/kg has a certain recovery effect on damaged liver cells, but there is no significant effect (P>0.05). 21.0mg/kg of the pharmaceutical oral preparation of the present invention can significantly reduce the ALT value (P<0.01), significantly reduce the AST value (P<0.025); 10.5mg/kg has the effect of significantly reducing the ALT value (P<0.05, P<0.025) ). 10.5mg/kg has a significant recovery effect on damaged liver cells (P<0.05); 5.2, 21.0mg/kg has a certain recovery effect on damaged liver cells, but there is no significant effect (P>0.05).

To D-GLAN induced mouse acute liver injury model, drug injection 17.0mg/kg of the present invention has significantly and very significantly reduced ALT, ALP value effect (P＜0.025, P＜0.001); 8.5mg/kg has significant and Very significant effect on reducing AST, ALT, ALP value (P<0.01, P<0.05, P<0.01); 4.2mg/kg has significant and very significant effect on reducing ALT, ALP (P<0.025, P<0.001). 4.2, 8.5, 17.0mg/kg have a certain recovery effect on the damaged liver cells, but there is no significant significance (P>0.05). 21.0mg/kg of the pharmaceutical oral preparation of the present invention has significant and very significant effect of reducing ALT, ALP value (P＜0.025, P＜0.001); 10.5mg/kg has significant and very significant effect of reducing AST, ALT, ALP value (P <0.01, P<0.05, P<0.01); 5.2 mg/kg had a significant and extremely significant effect on reducing ALT and ALP (P<0.025, P<0.001). 5.2, 10.5, 21.0mg/kg have a certain recovery effect on damaged liver cells, but there is no significant significance (P>0.05).

Experimental example 2 Anti-immune liver injury model test

For the mouse immune liver injury model caused by bacterial endotoxin, 17.0mg/kg of the drug injection of the present invention has a very significant effect of reducing ALT (P<0.01); 8.5mg/kg has a significant effect of reducing ALT and AST (P<0.05 ). 17.0mg/kg can significantly reduce liver cell necrosis (P<0.05); can significantly increase liver cell proliferation (P<0.05); 8.5mg/kg can significantly improve liver cell degeneration (P<0.001); 4.2 mg/kg can significantly improve liver cell degeneration and necrosis (P<0.001); can significantly improve liver cell inflammatory lesions (P<0.05); can significantly increase liver cell proliferation (P<0.001). 21.0 mg/kg of the pharmaceutical oral preparation of the present invention has a very significant effect of reducing ALT (P<0.01); 10.5 mg/kg has a significant effect of reducing ALT and AST (P<0.05). 21.0mg/kg can significantly reduce liver cell necrosis (P<0.05); can significantly increase liver cell proliferation (P<0.05); 10.5mg/kg can significantly improve liver cell degeneration (P<0.001); 5.2 mg/kg can significantly improve liver cell degeneration and necrosis (P<0.001); can significantly improve liver cell inflammatory lesions (P<0.05); can significantly increase liver cell proliferation (P<0.001).

Experimental example 3 yellowing test

In the rat obstructive jaundice model caused by p-naphthalene isothiocyanate, 17.0mg/kg of the drug injection of the present invention has the effect of significantly reducing DBil, ALT and ALP values (P＜0.05); 8.5mg/kg can significantly reduce ALT and ALP value (P<0.05); 4.2mg/kg can significantly reduce ALT (P<0.001), and very significantly reduce ALP (P<0.01). 21.0mg/kg of the pharmaceutical oral preparation of the present invention has significantly reducing DBil, ALT and ALP value effect (P＜0.05); 10.5mg/kg can significantly reduce ALT and ALP value (P＜0.05); 5.2mg/kg can significantly reduce ALT (P<0.001) and ALP were significantly reduced (P<0.01).

Experimental Example 4 Immune Function Test

Drug injection of the present invention 17.0mg/kg can be very significant and 4.2mg/kg can significantly enhance the effect of mouse monocyte phagocytosis foreign matter (P＜0.01, P＜0.05), 17.0,8.5,4.2mg/kg have a significant effect on D - The acute liver injury model mice caused by GALN have extremely significant, very significant and significantly enhanced monocyte phagocytosis of foreign bodies (P<0.001, P<0.01, P<0.05), and have an immune-enhancing effect. 17.0mg/kg can significantly reduce the ALT value (P<0.025).

21.0mg/kg of the pharmaceutical oral preparation of the present invention can be very significant and 5.2mg/kg can significantly enhance the effect of mouse monocyte phagocytosis of foreign matter (P＜0.01, P＜0.05), 21.0, 10.5, 5.2mg/kg to D - The acute liver injury model mice caused by GALN have extremely significant, very significant and significantly enhanced monocyte phagocytosis of foreign bodies (P<0.001, P<0.01, P<0.05), and have an immune-enhancing effect. 21.0mg/kg can significantly reduce the ALT value (P<0.025).

Experimental example 5 Anti-stress ability test

8.5, 17.0, and 34.0 mg/kg of the drug injection of the present invention can improve the survival time of normal mice under normal pressure and hypoxia, but there is no significant effect (P>0.05). 34.0, 17.0 mg/kg can significantly increase the survival time of mice in ice water (P<0.01).

10.5, 21.0, and 42.0 mg/kg of the pharmaceutical oral preparations of the present invention can improve the survival time of normal mice under normal pressure and hypoxia, but there is no significant significance (P>0.05). 42.0, 21.0 mg/kg can significantly increase the survival time of mice in ice water (P<0.01).

Experimental Example 6 Improving hemorheology test

For rat liver qi stagnation and cold stagnation qi stagnation model caused by adrenaline, the drug injection 50.0mg/kg of the present invention has a very significant effect of reducing whole blood viscosity (P<0.001); to whole blood reduced viscosity and whole blood relative index There is a very significant, very significant increase (P<0.001, P<0.01) 25.0, 12.5mg/kg have a very significant, very significant reduction in whole blood viscosity (P<0.01, P<0.001); The reduced viscosity and relative index of whole blood had a very significant and significant increase (P<0.01, P<0.05). 25.0mg/kg can extremely significantly and 12.5mg/kg can significantly reduce whole blood fibrin (P<0.001, P<0.05). The medicine oral preparation 60.0mg/kg of the present invention has extremely significant reduction whole blood viscosity effect (P＜0.001); Whole blood reduced viscosity and whole blood relative index have extremely significant, very significant raising effect (P＜0.001, P＜ 0.01) 30.0 and 15.0mg/kg have very significant and very significant effect of reducing the viscosity of whole blood (P<0.01, P<0.001); they have a very significant and significant effect of increasing the reduced viscosity of whole blood and the relative index of whole blood ( P<0.01, P<0.05). 30.0mg/kg can extremely significantly and 15.0mg/kg can significantly reduce whole blood fibrin (P<0.001, P<0.05).

Experimental example 7 Improve microcirculation test

Both 50.0 and 25.0 mg/kg of the drug injection of the present invention can significantly expand the diameter of microvessels (P<0.05); both can significantly resist the reduction of the diameter of microvessels caused by adrenaline (P<0.05). 12.5, 25.0, 50.0 mg/kg can significantly, very significantly and extremely significantly counteract the effect of adrenaline on blood flow, and improve microcirculation (P<0.05, P<0.01, P<0.001).

Both 60.0 and 30.0 mg/kg of the pharmaceutical oral preparations of the present invention can significantly expand the diameter of microvessels (P<0.05); both can significantly resist the reduction of the diameter of microvessels caused by adrenaline (P<0.05). 15.0, 30.0, 60.0 mg/kg can significantly, very significantly and extremely significantly counteract the effect of adrenaline on blood flow, and have the effect of improving microcirculation (P<0.05, P<0.01, P<0.001).

Experimental Example 8 Repair damaged liver cells and promote liver cell regeneration

To D-GLAN induced acute liver injury model in rats, 8.5 mg/kg of drug injection of the present invention has significant recovery effect on damaged liver cells (P<0.05); to mouse immune liver injury model caused by bacterial endotoxin 17.0mg/kg of the drug injection of the present invention can significantly reduce liver cell necrosis (P<0.05), 8.5mg/kg can significantly improve liver cell degeneration (P<0.001), and 4.2mg/kg can significantly improve liver function. Cell degeneration and necrosis (P<0.001) can also significantly improve the inflammatory lesion effect of liver cells (P<0.05). At the same time, 17.0 mg/kg and 4.2 mg/kg of the drug injection of the present invention can significantly and extremely significantly promote the proliferation of liver cells (P<0.05, P<0.001). 10.5mg/kg of the drug oral preparation of the present invention has a significant recovery effect on damaged liver cells (P<0.05); the mouse immune liver injury model caused by bacterial endotoxin, the drug injection of the present invention 21.0mg/kg has significantly reduced Liver cell necrosis (P<0.05), 10.5mg/kg can significantly improve liver cell degeneration (P<0.001), 5.2mg/kg can significantly improve liver cell degeneration and necrosis (P<0.001), and can also Significantly improved the inflammatory lesion effect of liver cells (P<0.05). At the same time, 21.0 mg/kg and 5.2 mg/kg of the pharmaceutical oral preparations of the present invention can significantly and extremely significantly promote the proliferation of liver cells (P<0.05, P<0.001).

In addition, the experimental results of the acute liver injury model in mice caused by CCL4 and TAA, and the acute liver injury model in mice caused by CCL4 and D-GLAN (D-galactosamine) all show that the drug injection of the present invention is 4.2-17.0 Both mg/kg and 5.2-21.0 mg/kg of the oral preparation of the present invention have a certain recovery effect on damaged liver cells, but there is no significant effect (P>0.05).

The above-mentioned pharmacodynamic study proves that both the drug injection and the oral preparation of the present invention can significantly reduce the activity of ALT, AST, ALP, and TB caused by various causes of acute liver injury and immune liver injury; Hepatocyte degeneration, necrosis, and inflammation in damaged histomorphology can be improved to a certain extent and hepatic cell proliferation can be improved; the jaundice caused by biliary obstruction can be reduced to a certain extent; it can improve the regulation of cellular immunity in normal animals and animals with liver damage Function; it has better effects on improving blood rheology and microcirculation, and has a certain anti-stress effect.

The toxicity and allergies of traditional Chinese medicine injections have always been difficult problems to solve, mainly because Chinese medicinal materials contain a large number of unclear ingredients. In order to evaluate its pharmacodynamic effect and toxicological response, the main pharmacodynamics, general pharmacology, acute toxicity, long-term toxicity (long-term toxicity of Beagle dogs, long-term toxicity of rats) and special safety related to the main functions and indications of the preparation were carried out. The experimental research is described as follows:

Experimental Example 9 Animal Acute Toxicity Test

Calculating and obtaining the LD50 and 95% credible limit of the drug injection of the present invention to the ICR kind mice by intravenous injection with Biliss method is 2684.81 (2554.52～2821.75) mg/Kg; The credible limit of % is 2750.85 (2538.73-2980.70) mg/Kg. Calculated by Biliss method, the LD50 and its 95% credible limit of oral administration of the pharmaceutical preparation of the present invention to ICR mice are 2879.81 (2684.57-3078.63) mg/Kg.

Calculate and obtain the LD50 and 95% credible limit of drug injection of the present invention to SD rat intravenous injection administration with Biliss method be 2189.35 (2073.62～2311.53) mg/Kg; The credible limit of 2682.58 (2493.14 ~ 2886.42) mg/Kg. The LD50 and 95% credible limit of the oral drug preparation of the present invention to SD rats are calculated by Biliss method and are 2714.35 (2531.84-2916.45) mg/Kg.

After intravenous injection, the animals showed symptoms such as dyspnea and decreased activity. Most of the dead animals died within 30 minutes after the injection. The surviving animals returned to normal after 24 hours, with no significant reduction in food intake and body weight. After the animals were sacrificed, no abnormalities were found in the vital organs (heart, liver, spleen, lung, kidney, etc.) observed with naked eyes.

After the intraperitoneal injection, the animals showed symptoms such as dyspnea and decreased activity. Most of the mice died within 8 hours after the injection; most of the rats died within 12 hours after the injection, and the number and time of death decreased with the dose. Prolonged, anatomical observation, no visible lesions. The surviving animals returned to normal after 48 hours, and there was no significant reduction in food intake and body weight. After the animals were sacrificed, no abnormalities were found in the vital organs (heart, liver, spleen, lung, kidney, etc.) with the naked eye.

Symptoms such as dyspnea and decreased activity occurred in the animals after intragastric administration, and returned to normal after 48 hours, with no significant reduction in food intake and body weight. No abnormalities were found.

Conclusion: the above test shows that the safety dose range of the drug injection and the oral preparation of the present invention is relatively large.

Experimental Example 10 Long-term Toxicity Test in Animals

1. Beagle dog long-term toxicity test

The medicine injection of the present invention presses three doses of 250, 125 and 62.5 mg/kg, which are respectively equivalent to 30, 15 and 7.5 times of its clinical intravenous infusion dose, and the Beagle dog is continuously administered intravenously for 45 days and withdrawn for 15 days, The result is as follows:

(1) Observation of growth and general conditions: Observation during administration and withdrawal of the drug, animal mental behavior, activities, fur, respiration, circulation, appetite, defecation, etc. were not obviously abnormal. There were no significant effects on body weight gain and development, and no delayed toxicity.

(2) Routine urine examination: no abnormalities were found.

(3) Effect on blood picture: No abnormality was found.

(4) Effects on liver function: No abnormality was found.

(5) Effects on renal function: No abnormality was found.

(6) Effects on blood sugar and serum cholesterol: no abnormalities were found.

(7) Effects on electrocardiogram: No abnormality was found.

(8) Bone marrow examination of living animals: no abnormalities were found.

(9) Systematic autopsy and organ index: no abnormalities were found.

(10) Histopathological examination: after administration for 45 days, submucosal lymphoid nodules formed in 1 case of male animals in the low-dose group; increased lymphoid follicles appeared in the medulla in 1 case of female animals in the low-dose group. There was no statistical difference between the administration group and the control group (P>0.05). After stopping the drug for 15 days, no abnormalities were found in the organ examinations of each group.

Conclusion: continuous intravenous infusion of 250 mg/Kg of the drug injection of the present invention to Beagle dogs for 45 days has no toxic reaction. The dosage of 250mg/Kg and below has no obvious abnormal effects on various indicators of dogs, and there is no obvious delayed toxic reaction, which belongs to the safe dosage range.

2. Summary of long-term toxicity test data in rats

The medicine injection of the present invention presses three doses of 250, 125 and 62.5 mg/kg, and the medicine oral preparation of the present invention presses three doses of 630, 157.5 and 78 mg/kg, which are respectively equivalent to 30, 15 and 7.5 times of the clinical daily dose, Respectively continuous intraperitoneal injection administration to rats for 45 days and drug withdrawal for 14 days, observed the main adverse reactions of drug injection of the present invention and oral preparations to animals:

(1) General condition and body weight: The behavior of the rats in each group was normal after 45 days of administration and 14 days after withdrawal of the drug, the animal hair was smooth, and there was no abnormal secretion. Animal body weight and food intake increased normally over time, and no animal died in each group.

(2) routine blood test: drug injection of the present invention is continuously administered to rats by intraperitoneal injection for 45 days of blood routine inspection. Compared with the matched group, RBC of the high and low dose female groups of drug injection of the present invention significantly increases respectively (P＜0.05 ), the RBC in the middle-dose female group was significantly increased (P<0.01); the PLT in the high-dose, middle-dose, and low-dose male groups was significantly increased (P<0.001); Very significantly and extremely significantly increased (P<0.05, P<0.01 and P<0.001); MCHC in the high-dose and middle-dose female groups was significantly reduced (P<0.01), and the rest of the indicators were normal (P>0.05) , the indicators of each group are within the normal range, the practical significance is not obvious. Drug withdrawal 14 days inspection compared with the matched group, the RBC of the high-dose female group of drug injection of the present invention significantly reduces (P＜0.05), the PDW of the middle-dose female group significantly reduces (P＜0.01), and each group index is all within the normal range , the actual meaning is not obvious.

The drug oral preparation group of the present invention is continuously administered to rats for 45 days by intraperitoneal injection for blood routine inspection. Compared with the matched group, the high and low dose female group RBC of the drug oral preparation group of the present invention significantly increases respectively (P＜0.05), and the middle dose RBC in the female group was significantly increased (P<0.01); PLT in the high, medium and low dose male groups was significantly increased (P<0.001); RC in the high, medium and low dose female and male groups was significantly, very significantly and extremely Significantly increased (P<0.05, P<0.01 and P<0.001); MCHC in the high-dose and middle-dose female groups was significantly reduced (P<0.01), and no abnormalities were found in the other indicators (P>0.05). All within the normal range, the actual significance is not obvious. Drug withdrawal 14 days inspection is compared with matched group, and the RBC of high-dose female group of drug oral preparation group of the present invention significantly reduces (P＜0.05), and the PDW of middle-dose female group very significantly reduces (P＜0.01), and each group index is all in normal range , the actual meaning is not obvious.

(3) Liver function: the drug injection of the present invention was administered continuously for 45 days and checked. Compared with the matched group, the TP of the female group in the middle dose of the drug injection of the present invention had a very significant increase (P＜0.01), and the liver of all the other groups of animals The functional indicators were all within the normal range. There was no significant difference between the 14-day drug withdrawal examination and the control group (P>0.05). The liver function indexes of the animals in each group were within the normal range.

The medicine oral preparation group of the present invention is administered continuously for 45 days and checks. Compared with the matched group, the dose female group TP in the medicine oral preparation group of the present invention has a very significant increase (P＜0.01), and the liver function indexes of all the other groups of animals are all within within the normal range. There was no significant difference between the 14-day drug withdrawal examination and the control group (P>0.05). The liver function indexes of the animals in each group were within the normal range.

(4) renal function: drug injection of the present invention, oral preparation group is administered continuously 45 days and drug withdrawal 14 days renal function checks, compares with matched group, does not see significant difference (P＞0.05), each group animal renal function index are within the normal range.

(5) Other biochemical indexes: drug injection of the present invention, oral preparation group were administered continuously for 45 days, compared with the matched group, CHOL of the high and low dose female groups of drug injection of the present invention, oral preparation group significantly decreased respectively (P＜0.05 ). Checked 14 days after drug withdrawal, compared with the control group, no significant difference was found (P>0.05).

(6) Systemic autopsy and visceral index: the results showed that there was no significant difference (P>0.05) between each dosage group of the drug injection of the present invention, the oral preparation group and the control group after administration for 45 days and drug withdrawal for 14 days.

(7) Histopathological examination: 45 days of administration and 14 days of drug withdrawal, each dosage group of the drug injection of the present invention and the rat brain (cerebellum), pituitary gland, optic nerve, heart, liver, spleen, lung, kidney, Histomorphological examination of the adrenal gland, thyroid, pancreas, thymus, bladder, stomach, duodenum, ileum, colon, ovary, uterus, testis, epididymis, prostate, lymph nodes, spinal cord, sternum, nasal septum, and retina found that: After 45 days of taking the drug injection of the present invention, there was 1 case of male animal in the high-dose group, and 1 case of female animal in the middle-dose group. The blood vessels in the lower part of the stomach were dilated with neutrophil infiltration. In the high-dose group, there were 2 cases of male animals, 1 case of female animals, 1 case of middle-dose female animals, 1 case of male animals, and 1 case of male animals in the control group. Colonic submucosal lymphoid nodules formed. One male animal in the high-dose group had obvious hemorrhage in part of the muscular layer of the bladder with mononuclear and neutrophil infiltration. One case in the high-dose group had a little neutrophil infiltration in the endometrium and muscle layer. No obvious pathomorphological damage was found in the indexes of other groups. There was no statistical difference between the administration group and the control group (P>0.05). After 14 days of drug withdrawal, no obvious pathomorphological damage was found in each group.

Histopathological examination: 45 days of administration and 14 days of drug withdrawal, each dosage group of the drug oral preparation group of the present invention and rat brain (cerebellum), pituitary gland, optic nerve, heart, liver, spleen, lung, kidney, adrenal gland, Histomorphological examination of the thyroid, pancreas, thymus, bladder, stomach, duodenum, ileum, colon, ovary, uterus, testis, epididymis, prostate, lymph nodes, spinal cord, sternum, nasal septum, and retina. In the oral preparation group of the present invention, there were 2 male animals in the high-dose group, and 1 female animal in the middle-dose group with vasodilation and neutrophil infiltration in the lower part of the stomach. In the high-dose group, 1 male animal, 1 female animal, 1 middle-dose female animal, 1 male animal, and 1 male animal in the control group formed colonic submucosal lymphoid nodules. One male animal in the high-dose group had obvious hemorrhage in part of the muscular layer of the bladder with mononuclear and neutrophil infiltration. One case in the high-dose group had a little neutrophil infiltration in the endometrium and muscle layer. No obvious pathomorphological damage was found in the indexes of other groups. There was no statistical difference between the administration group and the control group (P>0.05). After 14 days of drug withdrawal, no obvious pathomorphological damage was found in each group.

Conclusion: Long-term high-dose intraperitoneal injection of the drug injection of the present invention and the oral preparation of the drug of the present invention did not cause obvious toxic reaction and pathological damage to the tested animals.

3. Summary of special safety test research data

3.1 Hemolysis test

The drug injection of the present invention is mixed with 2% rabbit erythrocyte suspension and incubated for 4 hours, and no hemolysis and coagulation appear as a result.

3.2 Vascular stimulation test

Medicinal injection of the present invention drips 50mg/kg to rabbit's ear edge vein, continuously for 3 days, no stimulation reactions such as hyperemia, edema, degeneration, necrosis are observed by naked eyes, and each structure of pathological histomorphological examination is normal, shows that this medicine has no effect on the injection site. Vascular non-irritating effect.

3.3 Muscle stimulation test

The medicinal injection of the present invention injects 1ml (5 mg) into the quadriceps femoris of rabbits, and after 48 hours, the stimulation reaction situation is observed, and the results have no stimulation reactions such as hyperemia, edema, degeneration, etc., showing that the medicine has no stimulating effect on the muscles of the injection site.

3.4 Systemic active allergy test

The drug injection of the present invention is sensitized to guinea pigs by intraperitoneal injection of 0.5ml every other day, for a total of 3 times. On the 14th day and the 21st day after the first injection of the drug injection of the present invention, the animals are challenged by intravenous injection of 2ml of the drug injection of the present invention. , the results showed no symptoms of systemic active allergic reaction, and all the animals in the ovalbumin-positive control group had positive reactions, indicating that the drug did not cause allergic reactions.

3.5 Antihypertensive substance inspection

The drug injection of the present invention injects 8.3 mg/kg into the femoral vein of anesthetized cats, and the antihypertensive experiment is carried out with histamine as a control. As a result, the drug injection of the present invention does not cause a hypotensive reaction, indicating that the drug has no antihypertensive effect.

Summary: The drug injection of the present invention does not produce hemolysis and coagulation reactions, is not irritating to muscles and blood vessels, does not cause systemic active allergic reactions, has no hypotensive effect, and meets the general safety requirements of injections.

Systematic pharmacology and toxicology studies have proved that the drug injection of the present invention can significantly reduce the activity of ALT, AST, ALP, and TB caused by various causes of acute liver injury and immune liver injury; Histomorphological liver cell degeneration, necrosis, and inflammation have a certain effect on improving and repairing liver cells; it has a certain disappearance of jaundice caused by biliary obstruction; it can improve the cellular immunity of normal animals and liver-injured animals It has a good effect on improving blood rheology and microcirculation, and has a good anti-stress effect on normal animals and liver-injured animals, which provides a pharmacological basis for clinical application; in addition, acute Toxicity test, Beagle dog, rat long-term toxicity test and general pharmacology test results show that the preparation has low toxicity and high safety, which provides a toxicological basis for its clinical safety drug use, and has no effect on various systems of normal animals. No adverse reactions. The special safety test research shows that it meets the general safety requirements for injections.

The compatibility of the drug raw materials of the present invention is determined by pharmacodynamic screening and disassembly research, and the drug injection preparation of the present invention made from the compatibility of raw materials has been proved to have a strong effect on liver damage caused by various reasons through drug efficacy and toxicology studies. Pharmacological activity, low toxicity and side effects. Although it has no pathogenic therapeutic effect, it is characterized by reducing enzymes and promoting liver cell regeneration. It has a wide range of applications and can quickly improve clinical symptoms. The use of oral preparations and injections can enhance the means of emergency treatment of traditional Chinese medicine.

Claims (14)

1. A pharmaceutical composition for treating hepatitis, characterized in that: it is a medicament prepared from the raw materials in the following weight ratio: 3-10 parts of Danshen, 1-3 parts of Panax notoginseng. 2. The pharmaceutical composition for treating hepatitis according to claim 1, characterized in that it is a medicament prepared from the following raw materials in weight ratio: 6 parts of Danshen, 1 part of Panax notoginseng. 3. The pharmaceutical composition for treating hepatitis according to claim 1 or 2, characterized in that it is composed of water extracts or alcohol extracts of medicinal materials Danshen and Panax notoginseng as active ingredients, plus pharmaceutically acceptable Medicaments prepared from excipients or auxiliary ingredients. 4. The pharmaceutical composition for treating hepatitis according to claim 3, characterized in that: said medicament is an oral preparation or an injection. 5. The pharmaceutical composition for treating hepatitis according to claim 4, characterized in that: said oral preparations are capsules, tablets, pills, granules and powders. 6. The pharmaceutical composition for treating hepatitis according to claim 4, characterized in that: said injection is water injection, intravenous drop, or powder injection. 7. The pharmaceutical composition for treating hepatitis according to claim 6, characterized in that each milliliter of injection water contains 3.24-3.36 mg of water-soluble total phenols of Danshen, 1.69-1.76 mg of total saponins, and 0.83-0.91 mg of Danshensu , Protocatechualdehyde 0.23~0.25mg, Ginsenoside Rg1 0.63~0.69mg. 8. The pharmaceutical composition for treating hepatitis according to claim 6 or 7, characterized in that: the HPLC fingerprint of the pharmaceutical composition is shown in Figure 1, consisting of 4 characteristic peaks, The chromatographic conditions are as follows: octadecylsilane bonded silica gel as filler; methanol-acetonitrile-1.67% formic acid solution, mobile phase whose volume ratio is 23:7:70; detection wavelength is 286nm. 9. The pharmaceutical composition for treating hepatitis according to claim 8, characterized in that: the relative standard deviation RSDs of the relative retention times of the four characteristic peaks are all less than 10.0%, wherein the average relative retention time of No. 1 peak is RT 0.134, peak 2 RT 0.184, peak 3 RT 0.724, peak 4 RT 1.000. 10. A method for preparing the pharmaceutical composition for treating hepatitis according to any claim above, which comprises the following steps: a, weigh raw materials according to each weight ratio: 3-10 parts of salvia miltiorrhiza, 1-3 parts of notoginseng; b. Preparation of Salvia miltiorrhiza extract: take medicinal material Salvia miltiorrhiza, add water to decoct and extract, filter, concentrate the filtrate, dry, use 50-95% ethanol as solvent for the dry extract powder, extract by percolation method, obtain percolation liquid, add activated carbon, Extract by heating under reflux, filter, recover ethanol, add water to dissolve, pass the aqueous solution through the macroporous adsorption resin, elute with 30-85% ethanol, ethanol eluate, recover ethanol to obtain the salvia miltiorrhiza extract; c. Preparation of Panax notoginseng extract: take the medicinal material Panax notoginseng, use 35-95% ethanol as solvent, extract by percolation method, obtain percolation liquid, add activated carbon to the percolation liquid, reflux extraction, filter, and depressurize the filtrate Recover ethanol to obtain alcoholic extract, dissolve the alcoholic extract in water, filter, pass the aqueous solution through a macroporous adsorption resin column, and then elute with 30-90% ethanol to obtain ethanol eluate, recover ethanol, and obtain Panax notoginseng extract thing; d. Mix the Danshen extract prepared in step b and the Panax notoginseng extract prepared in step c, add water to dissolve, add pharmaceutically acceptable excipients or auxiliary components, and prepare a commonly used pharmaceutical preparation. 11. The use of the raw materials according to claim 1 in the following weight proportions in the preparation of medicines for treating acute and chronic hepatitis: 3-10 parts of Danshen and 1-3 parts of Panax notoginseng. 12. The use according to claim 11, characterized in that the drug is an oral drug for treating acute and chronic hepatitis. 13. The use according to claim 11, characterized in that the medicine is injection medicine for treating acute and chronic hepatitis. 14. The use according to claim 13, characterized in that: the medicine is intravenously injected to treat acute and chronic hepatitis.

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